CORD-19:fccc2cff358d6c08f512003b88bab9934e4ffa7a JSONTXT

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CORD-19-PD-HP

Id Subject Object Predicate Lexical cue hp_id
T1 1767-1773 Phenotype denotes cancer http://purl.obolibrary.org/obo/HP_0002664
T2 1800-1806 Phenotype denotes cancer http://purl.obolibrary.org/obo/HP_0002664
T3 2401-2407 Phenotype denotes cancer http://purl.obolibrary.org/obo/HP_0002664
T4 5625-5631 Phenotype denotes cancer http://purl.obolibrary.org/obo/HP_0002664
T5 6206-6214 Phenotype denotes Oncology http://purl.obolibrary.org/obo/HP_0002664
T6 6225-6231 Phenotype denotes Cancer http://purl.obolibrary.org/obo/HP_0002664
T7 23772-23778 Phenotype denotes cancer http://purl.obolibrary.org/obo/HP_0002664
T8 23835-23841 Phenotype denotes cancer http://purl.obolibrary.org/obo/HP_0002664
T9 23962-23968 Phenotype denotes cancer http://purl.obolibrary.org/obo/HP_0002664
T10 24112-24118 Phenotype denotes cancer http://purl.obolibrary.org/obo/HP_0002664
T11 24272-24278 Phenotype denotes cancer http://purl.obolibrary.org/obo/HP_0002664
T12 26891-26907 Phenotype denotes immunodeficiency http://purl.obolibrary.org/obo/HP_0002721
T13 30062-30068 Phenotype denotes cancer http://purl.obolibrary.org/obo/HP_0002664

CORD-19-PD-MONDO

Id Subject Object Predicate Lexical cue mondo_id
T98253 492-501 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T1 492-501 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T21310 660-669 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T2 660-669 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T91212 1746-1755 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T3 1746-1755 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T60790 1758-1773 Disease denotes Cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T4 1758-1773 Disease denotes Cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T71878 1767-1773 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T6 1767-1773 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T74264 1800-1806 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T7 1800-1806 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T93379 1900-1909 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T8 1900-1909 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T70110 2337-2346 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T9 2337-2346 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T76336 2392-2407 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T10 2392-2407 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T89253 2401-2407 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T12 2401-2407 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T14832 3223-3232 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T13 3223-3232 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T5569 3376-3385 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T14 3376-3385 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T35405 3517-3526 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T15 3517-3526 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T39100 3602-3617 Disease denotes virus infection http://purl.obolibrary.org/obo/MONDO_0005108
T16 3602-3617 Disease denotes virus infection http://purl.obolibrary.org/obo/MONDO_0005108
T18224 3608-3617 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T17 3608-3617 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T45000 4134-4148 Disease denotes herpes simplex http://purl.obolibrary.org/obo/MONDO_0004609
T18 4134-4148 Disease denotes herpes simplex http://purl.obolibrary.org/obo/MONDO_0004609
T83465 4167-4172 Disease denotes Ebola http://purl.obolibrary.org/obo/MONDO_0005737
T19 4167-4172 Disease denotes Ebola http://purl.obolibrary.org/obo/MONDO_0005737
T46229 4607-4616 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T20 4607-4616 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T41976 4774-4783 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T21 4774-4783 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T60539 5139-5148 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T22 5139-5148 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T66569 5588-5597 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T23 5588-5597 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T72501 5616-5631 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T24 5616-5631 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T92951 5625-5631 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T26 5625-5631 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T50602 5752-5755 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T27 5752-5755 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T70884 6225-6231 Disease denotes Cancer http://purl.obolibrary.org/obo/MONDO_0004992
T28 6225-6231 Disease denotes Cancer http://purl.obolibrary.org/obo/MONDO_0004992
T9897 11837-11839 Disease denotes VL http://purl.obolibrary.org/obo/MONDO_0024555
T29 11837-11839 Disease denotes VL http://purl.obolibrary.org/obo/MONDO_0024555
T63136 13915-13918 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T30 13915-13918 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T40274 14060-14063 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T31 14060-14063 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T58873 14292-14295 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T32 14292-14295 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T16781 14563-14566 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T33 14563-14566 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T37354 14640-14649 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T34 14640-14649 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T70173 15529-15538 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T35 15529-15538 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T20990 19347-19362 Disease denotes viral infection http://purl.obolibrary.org/obo/MONDO_0005108
T36 19347-19362 Disease denotes viral infection http://purl.obolibrary.org/obo/MONDO_0005108
T13934 19353-19362 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T37 19353-19362 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T70183 19998-20007 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T38 19998-20007 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T51180 23714-23724 Disease denotes infections http://purl.obolibrary.org/obo/MONDO_0005550
T39 23714-23724 Disease denotes infections http://purl.obolibrary.org/obo/MONDO_0005550
T44091 23772-23778 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T40 23772-23778 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T3996 23826-23841 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T41 23826-23841 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T85947 23835-23841 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T43 23835-23841 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T30268 23939-23948 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T44 23939-23948 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T70872 23953-23968 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T45 23953-23968 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T76081 23962-23968 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T47 23962-23968 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T79143 24027-24036 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T48 24027-24036 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T99811 24103-24118 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T49 24103-24118 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T49377 24112-24118 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T51 24112-24118 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T85338 24217-24226 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T52 24217-24226 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T17262 24263-24278 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T53 24263-24278 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T34269 24272-24278 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T55 24272-24278 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T62244 24360-24376 Disease denotes viral infections http://purl.obolibrary.org/obo/MONDO_0005108
T56 24360-24376 Disease denotes viral infections http://purl.obolibrary.org/obo/MONDO_0005108
T30800 24814-24824 Disease denotes infections http://purl.obolibrary.org/obo/MONDO_0005550
T57 24814-24824 Disease denotes infections http://purl.obolibrary.org/obo/MONDO_0005550
T99920 25652-25661 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T58 25652-25661 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T955 26197-26212 Disease denotes virus infection http://purl.obolibrary.org/obo/MONDO_0005108
T59 26197-26212 Disease denotes virus infection http://purl.obolibrary.org/obo/MONDO_0005108
T55247 26203-26212 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T60 26203-26212 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T34962 26826-26835 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T61 26826-26835 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T18803 26871-26880 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T62 26871-26880 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T49967 26891-26907 Disease denotes immunodeficiency http://purl.obolibrary.org/obo/MONDO_0021094
T63 26891-26907 Disease denotes immunodeficiency http://purl.obolibrary.org/obo/MONDO_0021094
T19704 26915-26918 Disease denotes SIV http://purl.obolibrary.org/obo/MONDO_0010029
T64 26915-26918 Disease denotes SIV http://purl.obolibrary.org/obo/MONDO_0010029
T40661 26983-26992 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T65 26983-26992 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T19646 27077-27080 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T66 27077-27080 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T26968 27086-27089 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T67 27086-27089 Disease denotes HSA http://purl.obolibrary.org/obo/MONDO_0011848
T56936 28212-28221 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T68 28212-28221 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T30483 29943-29952 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T69 29943-29952 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T66253 30010-30019 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T70 30010-30019 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T71028 30053-30068 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T71 30053-30068 Disease denotes cervical cancer http://purl.obolibrary.org/obo/MONDO_0005131
T16327 30062-30068 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T73 30062-30068 Disease denotes cancer http://purl.obolibrary.org/obo/MONDO_0004992
T15430 30691-30700 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T74 30691-30700 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T39949 30988-30993 Disease denotes wound http://purl.obolibrary.org/obo/MONDO_0021178
T75 30988-30993 Disease denotes wound http://purl.obolibrary.org/obo/MONDO_0021178
T27644 31114-31123 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T76 31114-31123 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T50345 31515-31524 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T77 31515-31524 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T47962 31936-31945 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T78 31936-31945 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T87688 32651-32660 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T79 32651-32660 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T36128 33170-33179 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T80 33170-33179 Disease denotes infection http://purl.obolibrary.org/obo/MONDO_0005550
T18195 33467-33469 Disease denotes VL http://purl.obolibrary.org/obo/MONDO_0024555
T81 33467-33469 Disease denotes VL http://purl.obolibrary.org/obo/MONDO_0024555

CORD-19-PD-UBERON

Id Subject Object Predicate Lexical cue uberon_id
T1 531-546 Body_part denotes cervical region http://purl.obolibrary.org/obo/UBERON_0005434
T2 1176-1182 Body_part denotes mucosa http://purl.obolibrary.org/obo/UBERON_0000344
T3 1548-1553 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T4 3022-3027 Body_part denotes shell http://purl.obolibrary.org/obo/UBERON_0006612
T5 4652-4658 Body_part denotes mucosa http://purl.obolibrary.org/obo/UBERON_0000344
T6 5307-5313 Body_part denotes mucosa http://purl.obolibrary.org/obo/UBERON_0000344
T7 5471-5476 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T8 5560-5566 Body_part denotes vagina http://purl.obolibrary.org/obo/UBERON_0000996
T9 5737-5742 Body_part denotes serum http://purl.obolibrary.org/obo/UBERON_0001977
T10 5818-5823 Body_part denotes serum http://purl.obolibrary.org/obo/UBERON_0001977
T11 9217-9222 Body_part denotes serum http://purl.obolibrary.org/obo/UBERON_0001977
T12 12161-12166 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T13 12269-12274 Body_part denotes serum http://purl.obolibrary.org/obo/UBERON_0001977
T14 13896-13901 Body_part denotes serum http://purl.obolibrary.org/obo/UBERON_0001977
T15 22277-22282 Body_part denotes Blood http://purl.obolibrary.org/obo/UBERON_0000178
T16 23069-23074 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T17 23162-23168 Body_part denotes vagina http://purl.obolibrary.org/obo/UBERON_0000996
T18 23172-23176 Body_part denotes anus http://purl.obolibrary.org/obo/UBERON_0001245
T19 23203-23208 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T20 23328-23333 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T21 23491-23496 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T22 23669-23674 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T23 28229-28244 Body_part denotes cervical mucosa http://purl.obolibrary.org/obo/UBERON_0012248
T24 28238-28244 Body_part denotes mucosa http://purl.obolibrary.org/obo/UBERON_0000344
T25 28301-28311 Body_part denotes cell layer http://purl.obolibrary.org/obo/UBERON_0000119
T26 28360-28370 Body_part denotes epithelium http://purl.obolibrary.org/obo/UBERON_0000483
T27 28926-28935 Body_part denotes epithelia http://purl.obolibrary.org/obo/UBERON_0000483
T28 29056-29061 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T29 29099-29105 Body_part denotes vagina http://purl.obolibrary.org/obo/UBERON_0000996
T30 29109-29113 Body_part denotes anus http://purl.obolibrary.org/obo/UBERON_0001245
T31 29361-29367 Body_part denotes vagina http://purl.obolibrary.org/obo/UBERON_0000996
T32 29371-29375 Body_part denotes anus http://purl.obolibrary.org/obo/UBERON_0001245
T33 29402-29407 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T34 29519-29524 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T35 29914-29924 Body_part denotes epithelium http://purl.obolibrary.org/obo/UBERON_0000483
T36 30112-30118 Body_part denotes vagina http://purl.obolibrary.org/obo/UBERON_0000996
T37 30471-30489 Body_part denotes mucosal epithelium http://purl.obolibrary.org/obo/UBERON_0003350
T38 30479-30489 Body_part denotes epithelium http://purl.obolibrary.org/obo/UBERON_0000483
T39 30572-30578 Body_part denotes mucosa http://purl.obolibrary.org/obo/UBERON_0000344
T40 30660-30678 Body_part denotes mucosal epithelium http://purl.obolibrary.org/obo/UBERON_0003350
T41 30668-30678 Body_part denotes epithelium http://purl.obolibrary.org/obo/UBERON_0000483
T42 30775-30785 Body_part denotes epithelium http://purl.obolibrary.org/obo/UBERON_0000483
T43 30981-30987 Body_part denotes mucosa http://purl.obolibrary.org/obo/UBERON_0000344
T44 31583-31589 Body_part denotes mucosa http://purl.obolibrary.org/obo/UBERON_0000344
T45 31720-31725 Body_part denotes blood http://purl.obolibrary.org/obo/UBERON_0000178
T46 33191-33197 Body_part denotes mucosa http://purl.obolibrary.org/obo/UBERON_0000344

CORD-19-Sentences

Id Subject Object Predicate Lexical cue
TextSentencer_T1 0-146 Sentence denotes The Underlying Mechanism of 3-Hydroxyphthalic Anhydride- Modified Bovine Beta-Lactoglobulin to Block Human Papillomavirus Entry Into the Host Cell
TextSentencer_T1 0-146 Sentence denotes The Underlying Mechanism of 3-Hydroxyphthalic Anhydride- Modified Bovine Beta-Lactoglobulin to Block Human Papillomavirus Entry Into the Host Cell
TextSentencer_T2 148-156 Sentence denotes Abstract
TextSentencer_T2 148-156 Sentence denotes Abstract
TextSentencer_T3 157-404 Sentence denotes We have previously demonstrated that 3-hydroxyphthalic anhydride (3HP)-modified bovine beta-lactoglobulin (3HP-β-LG) is highly effective in inhibiting entry of pseudovirus (PsV) of high-and low-risk human papillomavirus (HPV) into the target cell.
TextSentencer_T3 157-404 Sentence denotes We have previously demonstrated that 3-hydroxyphthalic anhydride (3HP)-modified bovine beta-lactoglobulin (3HP-β-LG) is highly effective in inhibiting entry of pseudovirus (PsV) of high-and low-risk human papillomavirus (HPV) into the target cell.
TextSentencer_T4 405-547 Sentence denotes Intravaginally applied 3HP-β-LG-containing vaginal gel could significantly inhibit HPV infection and reduce viral load in the cervical region.
TextSentencer_T4 405-547 Sentence denotes Intravaginally applied 3HP-β-LG-containing vaginal gel could significantly inhibit HPV infection and reduce viral load in the cervical region.
TextSentencer_T5 548-670 Sentence denotes However, we still do not understand the underlying molecular mechanism by which 3HP-β-LG is able to inhibit HPV infection.
TextSentencer_T5 548-670 Sentence denotes However, we still do not understand the underlying molecular mechanism by which 3HP-β-LG is able to inhibit HPV infection.
TextSentencer_T6 671-835 Sentence denotes Here, though, we showed that 3HP-β-LG did not inactivate HPV PsV, but rather blocked entry of HPV PsV into the target cell via its interaction with virus, not cell.
TextSentencer_T6 671-835 Sentence denotes Here, though, we showed that 3HP-β-LG did not inactivate HPV PsV, but rather blocked entry of HPV PsV into the target cell via its interaction with virus, not cell.
TextSentencer_T7 836-1183 Sentence denotes It bound to the positively charged region in the HPV L1 protein, suggesting that 3HP-β-LG binds to HPV L1 protein through the interaction between the negatively charged region in 3HP-β-LG and the positively charged region in HPV L1 protein, thus competitively blocking the binding of HPV to the receptor on the basement membrane in vaginal mucosa.
TextSentencer_T7 836-1183 Sentence denotes It bound to the positively charged region in the HPV L1 protein, suggesting that 3HP-β-LG binds to HPV L1 protein through the interaction between the negatively charged region in 3HP-β-LG and the positively charged region in HPV L1 protein, thus competitively blocking the binding of HPV to the receptor on the basement membrane in vaginal mucosa.
TextSentencer_T8 1184-1483 Sentence denotes Although 3HP-modified chicken ovalbumin (3HP-OVA) also carries high net negative charges, it exhibited no anti-HPV activity, suggesting that the interaction between 3HP-modified protein and HPV L1 protein relies on both electrostatic and matchable conformation of the binding sites in both proteins.
TextSentencer_T8 1184-1483 Sentence denotes Although 3HP-modified chicken ovalbumin (3HP-OVA) also carries high net negative charges, it exhibited no anti-HPV activity, suggesting that the interaction between 3HP-modified protein and HPV L1 protein relies on both electrostatic and matchable conformation of the binding sites in both proteins.
TextSentencer_T9 1484-1566 Sentence denotes When topically applied, 3HP-β-LG did not enter the host cell or blood circulation.
TextSentencer_T9 1484-1566 Sentence denotes When topically applied, 3HP-β-LG did not enter the host cell or blood circulation.
TextSentencer_T10 1567-1756 Sentence denotes These findings suggest that 3HP-β-LG targets HPV L1 protein and blocks HPV entry into the host cell, thus being safe and effective for topical application in the treatment of HPV infection.
TextSentencer_T10 1567-1756 Sentence denotes These findings suggest that 3HP-β-LG targets HPV L1 protein and blocks HPV entry into the host cell, thus being safe and effective for topical application in the treatment of HPV infection.
TextSentencer_T11 1758-1849 Sentence denotes Cervical cancer is the second most common cancer among women in China (Chen et al., 2016) .
TextSentencer_T11 1758-1849 Sentence denotes Cervical cancer is the second most common cancer among women in China (Chen et al., 2016) .
TextSentencer_T12 1850-1962 Sentence denotes Almost all these cases result from the persistent infection of human papillomavirus (HPV) (Bosch et al., 2002) .
TextSentencer_T12 1850-1962 Sentence denotes Almost all these cases result from the persistent infection of human papillomavirus (HPV) (Bosch et al., 2002) .
TextSentencer_T13 1963-2057 Sentence denotes HPVs are a large viral family consisting of about 200 different types (Bernard et al., 2010) .
TextSentencer_T13 1963-2057 Sentence denotes HPVs are a large viral family consisting of about 200 different types (Bernard et al., 2010) .
TextSentencer_T14 2058-2190 Sentence denotes Among them, 18 types have high oncogenic properties, and these are regarded as the high-risk types, such as HPV16, HPV18, and HPV58.
TextSentencer_T14 2058-2190 Sentence denotes Among them, 18 types have high oncogenic properties, and these are regarded as the high-risk types, such as HPV16, HPV18, and HPV58.
TextSentencer_T15 2191-2321 Sentence denotes The high-risk types of HPV have relatively low self-clearance rate compared to the low-risk types of HPV (Safaeian et al., 2008) .
TextSentencer_T15 2191-2321 Sentence denotes The high-risk types of HPV have relatively low self-clearance rate compared to the low-risk types of HPV (Safaeian et al., 2008) .
TextSentencer_T16 2322-2593 Sentence denotes To prevent the infection of these highrisk HPVs and the occurrence of cervical cancer, several prophylactic vaccines, like Cervarix, Gardasil, and Gardasil 9 (Deschuyteneer et al., 2010; McCormack, 2014) , have been developed and approved for marketing in many countries.
TextSentencer_T16 2322-2593 Sentence denotes To prevent the infection of these highrisk HPVs and the occurrence of cervical cancer, several prophylactic vaccines, like Cervarix, Gardasil, and Gardasil 9 (Deschuyteneer et al., 2010; McCormack, 2014) , have been developed and approved for marketing in many countries.
TextSentencer_T17 2594-2703 Sentence denotes However, no therapeutic vaccines or drugs have been approved for clinical use to treat HPV-infected patients.
TextSentencer_T17 2594-2703 Sentence denotes However, no therapeutic vaccines or drugs have been approved for clinical use to treat HPV-infected patients.
TextSentencer_T18 2704-2754 Sentence denotes HPV is a non-enveloped, double-stranded DNA virus.
TextSentencer_T18 2704-2754 Sentence denotes HPV is a non-enveloped, double-stranded DNA virus.
TextSentencer_T19 2755-2832 Sentence denotes Its capsid is made up of two proteins, major protein L1 and minor protein L2.
TextSentencer_T19 2755-2832 Sentence denotes Its capsid is made up of two proteins, major protein L1 and minor protein L2.
TextSentencer_T20 2833-2892 Sentence denotes They are both involved in receptor binding and viral entry.
TextSentencer_T20 2833-2892 Sentence denotes They are both involved in receptor binding and viral entry.
TextSentencer_T21 2893-3074 Sentence denotes Analysis of the atomic structure of native T = 7 HPV virus-like particle (VLP) reveals that 72 L1 pentamers form the icosahedral shell of HPV (Baker et al., 1991; Li et al., 2016) .
TextSentencer_T21 2893-3074 Sentence denotes Analysis of the atomic structure of native T = 7 HPV virus-like particle (VLP) reveals that 72 L1 pentamers form the icosahedral shell of HPV (Baker et al., 1991; Li et al., 2016) .
TextSentencer_T22 3075-3255 Sentence denotes The L1 pentamer strongly binds to HPV receptors, such as heparan sulfate proteoglycans (HSPGs), on the basement membrane to mediate viral entry and infection (Kines et al., 2009) .
TextSentencer_T22 3075-3255 Sentence denotes The L1 pentamer strongly binds to HPV receptors, such as heparan sulfate proteoglycans (HSPGs), on the basement membrane to mediate viral entry and infection (Kines et al., 2009) .
TextSentencer_T23 3256-3405 Sentence denotes The highly potent neutralizing antibodies elicited by HPV vaccines were found to bind to the L1 protein and prevent HPV infection (Li et al., 2018) .
TextSentencer_T23 3256-3405 Sentence denotes The highly potent neutralizing antibodies elicited by HPV vaccines were found to bind to the L1 protein and prevent HPV infection (Li et al., 2018) .
TextSentencer_T24 3406-3527 Sentence denotes Thus, the L1 pentamer is regarded as the most important target for development of antiviral agents against HPV infection.
TextSentencer_T24 3406-3527 Sentence denotes Thus, the L1 pentamer is regarded as the most important target for development of antiviral agents against HPV infection.
TextSentencer_T25 3528-3662 Sentence denotes Viral entry inhibitors represent a class of antiviral agents that inhibit virus infection by blocking virus entry into the host cells.
TextSentencer_T25 3528-3662 Sentence denotes Viral entry inhibitors represent a class of antiviral agents that inhibit virus infection by blocking virus entry into the host cells.
TextSentencer_T26 3663-3856 Sentence denotes Jiang et al. reported the first peptide-based HIV entry inhibitor, SJ-2176 , in 1993 (Jiang et al., 1993 and the first HIV entry inhibitor-based anti-HIV drug, enfuvirtide, was approved by U.S.
TextSentencer_T26 3663-3856 Sentence denotes Jiang et al. reported the first peptide-based HIV entry inhibitor, SJ-2176 , in 1993 (Jiang et al., 1993 and the first HIV entry inhibitor-based anti-HIV drug, enfuvirtide, was approved by U.S.
TextSentencer_T27 3857-3911 Sentence denotes FDA for clinical use in 2003 (Dando and Perry, 2003) .
TextSentencer_T27 3857-3911 Sentence denotes FDA for clinical use in 2003 (Dando and Perry, 2003) .
TextSentencer_T28 3912-4205 Sentence denotes Later, Jiang and colleagues have reported that anhydride-modified proteins such as 3-hydroxyphthalic anhydride (3HP)-modified proteins are potent virus entry inhibitors against a number of enveloped viruses, such as HIV , herpes simplex virus (HSV) , and Ebola virus (EBOV) (Li et al., 2017) .
TextSentencer_T28 3912-4205 Sentence denotes Later, Jiang and colleagues have reported that anhydride-modified proteins such as 3-hydroxyphthalic anhydride (3HP)-modified proteins are potent virus entry inhibitors against a number of enveloped viruses, such as HIV , herpes simplex virus (HSV) , and Ebola virus (EBOV) (Li et al., 2017) .
TextSentencer_T29 4206-4457 Sentence denotes In 2012, Jiang's group reported that 3HP-modified bovine beta-lactoglobulin (3HPβ-LG) could also inhibit entry into the target cell of the pseudovirus (PsV) of non-enveloped virus, HPV (high-risk HPV16 and HPV18, and low-risk HPV6) (Lu et al., 2013) .
TextSentencer_T29 4206-4457 Sentence denotes In 2012, Jiang's group reported that 3HP-modified bovine beta-lactoglobulin (3HPβ-LG) could also inhibit entry into the target cell of the pseudovirus (PsV) of non-enveloped virus, HPV (high-risk HPV16 and HPV18, and low-risk HPV6) (Lu et al., 2013) .
TextSentencer_T30 4458-4682 Sentence denotes In the clinical trial, the topical application of vaginal gel containing 3HP-β-LG was proven to be very safe and highly effective in suppressing HPV infection and reducing viral load in vaginal mucosa (Guo et al., 2016a,b) .
TextSentencer_T30 4458-4682 Sentence denotes In the clinical trial, the topical application of vaginal gel containing 3HP-β-LG was proven to be very safe and highly effective in suppressing HPV infection and reducing viral load in vaginal mucosa (Guo et al., 2016a,b) .
TextSentencer_T31 4683-4784 Sentence denotes However, we still do not understand the molecular mechanism by which 3HP-β-LG inhibits HPV infection.
TextSentencer_T31 4683-4784 Sentence denotes However, we still do not understand the molecular mechanism by which 3HP-β-LG inhibits HPV infection.
TextSentencer_T32 4785-4968 Sentence denotes In this study, we found that while 3HP-β-LG could not inactivate HPV PsV, it was effective in inhibiting HPV PsV entry into the host cell via its interaction with virus, not the cell.
TextSentencer_T32 4785-4968 Sentence denotes In this study, we found that while 3HP-β-LG could not inactivate HPV PsV, it was effective in inhibiting HPV PsV entry into the host cell via its interaction with virus, not the cell.
TextSentencer_T33 4969-5386 Sentence denotes Unlike β-LG, 3HP-β-LG could bind with L1 protein and the peptides derived from the positively charged region in the HPV L1 protein, suggesting that 3HP-β-LG inhibits HPV infection by binding, via its negatively charged region, to L1 protein, and thus competitively blocking HPV binding to the receptor on the basement membrane in vaginal mucosa, which then inhibits HPV from entering and replicating in the host cell.
TextSentencer_T33 4969-5386 Sentence denotes Unlike β-LG, 3HP-β-LG could bind with L1 protein and the peptides derived from the positively charged region in the HPV L1 protein, suggesting that 3HP-β-LG inhibits HPV infection by binding, via its negatively charged region, to L1 protein, and thus competitively blocking HPV binding to the receptor on the basement membrane in vaginal mucosa, which then inhibits HPV from entering and replicating in the host cell.
TextSentencer_T34 5387-5489 Sentence denotes We have also shown that topically applied 3HP-β-LG does not enter the host cell and blood circulation.
TextSentencer_T34 5387-5489 Sentence denotes We have also shown that topically applied 3HP-β-LG does not enter the host cell and blood circulation.
TextSentencer_T35 5490-5644 Sentence denotes Therefore, the 3HP-β-LG-containing formulations can be safely used in vagina for treatment of HPV infection and prevention of cervical cancer development.
TextSentencer_T35 5490-5644 Sentence denotes Therefore, the 3HP-β-LG-containing formulations can be safely used in vagina for treatment of HPV infection and prevention of cervical cancer development.
TextSentencer_T36 5645-5864 Sentence denotes Reagents 3-Hydroxyphthalic anhydride (HP), 2,4,6-trinitrobenzenesulfonic acid (TNBS), human serum albumin (HSA), β-lactoglobulin (β-LG), chicken ovalbumin (OVA), and bovine serum albumin (BSA) were purchased from Sigma.
TextSentencer_T36 5645-5864 Sentence denotes Reagents 3-Hydroxyphthalic anhydride (HP), 2,4,6-trinitrobenzenesulfonic acid (TNBS), human serum albumin (HSA), β-lactoglobulin (β-LG), chicken ovalbumin (OVA), and bovine serum albumin (BSA) were purchased from Sigma.
TextSentencer_T37 5865-5975 Sentence denotes Horseradish peroxidase (HRP)-conjugated goat anti-human IgA and IgE antibodies were purchased from Abcam (UK).
TextSentencer_T37 5865-5975 Sentence denotes Horseradish peroxidase (HRP)-conjugated goat anti-human IgA and IgE antibodies were purchased from Abcam (UK).
TextSentencer_T38 5976-6056 Sentence denotes HRP-conjugated rabbit anti-mouse IgG antibody was purchased from Dako (Denmark).
TextSentencer_T38 5976-6056 Sentence denotes HRP-conjugated rabbit anti-mouse IgG antibody was purchased from Dako (Denmark).
TextSentencer_T39 6057-6256 Sentence denotes The HPV16-and HPV-58-L1/L2-expressing plasmids and Luciferase pCLucf plasmid were kindly provided by Dr. John Schiller at the Laboratory of Cellular Oncology, National Cancer Institute, NIH, MD, USA.
TextSentencer_T39 6057-6256 Sentence denotes The HPV16-and HPV-58-L1/L2-expressing plasmids and Luciferase pCLucf plasmid were kindly provided by Dr. John Schiller at the Laboratory of Cellular Oncology, National Cancer Institute, NIH, MD, USA.
TextSentencer_T40 6257-6342 Sentence denotes Anhydride-modified proteins were produced as previously described (Lu et al., 2013) .
TextSentencer_T40 6257-6342 Sentence denotes Anhydride-modified proteins were produced as previously described (Lu et al., 2013) .
TextSentencer_T41 6343-6461 Sentence denotes Briefly, each kind of protein was dissolved with 0.1 M phosphate buffer (pH 8.5) at a final concentration of 20 mg/ml.
TextSentencer_T41 6343-6461 Sentence denotes Briefly, each kind of protein was dissolved with 0.1 M phosphate buffer (pH 8.5) at a final concentration of 20 mg/ml.
TextSentencer_T42 6462-6697 Sentence denotes Afterward, protein solutions were mixed with anhydrides (HP at 1 M in DMSO) to a final concentration of 60 mM by the addition of five equal aliquots at 20-min intervals, while the pH was adjusted to 9.0 with 4 M NaOH after each mixing.
TextSentencer_T42 6462-6697 Sentence denotes Afterward, protein solutions were mixed with anhydrides (HP at 1 M in DMSO) to a final concentration of 60 mM by the addition of five equal aliquots at 20-min intervals, while the pH was adjusted to 9.0 with 4 M NaOH after each mixing.
TextSentencer_T43 6698-6778 Sentence denotes All mixtures were kept at 25°C for two more hours and then dialyzed against PBS.
TextSentencer_T43 6698-6778 Sentence denotes All mixtures were kept at 25°C for two more hours and then dialyzed against PBS.
TextSentencer_T44 6779-6839 Sentence denotes The HPV pseudovirus was produced as previously described 1 .
TextSentencer_T44 6779-6839 Sentence denotes The HPV pseudovirus was produced as previously described 1 .
TextSentencer_T45 6840-7204 Sentence denotes Briefly, 293 T cells, which had been seeded in a 10-cm culture dish (4 × 10 6 cells) at 16 h before transfection, were transfected with a mixture of HPV16-L1/L2-expressing plasmid (p16sheLL) and pCLucf plasmid for HPV16 PsV, or a mixture of HPV58-L1/ L2-expressing plasmid (p58sheLL) and pCLucf plasmid for HPV58 PsV, using VigoFect (Vigorous Biotechnology Corp.).
TextSentencer_T45 6840-7204 Sentence denotes Briefly, 293 T cells, which had been seeded in a 10-cm culture dish (4 × 10 6 cells) at 16 h before transfection, were transfected with a mixture of HPV16-L1/L2-expressing plasmid (p16sheLL) and pCLucf plasmid for HPV16 PsV, or a mixture of HPV58-L1/ L2-expressing plasmid (p58sheLL) and pCLucf plasmid for HPV58 PsV, using VigoFect (Vigorous Biotechnology Corp.).
TextSentencer_T46 7205-7306 Sentence denotes The cells were suspended in 0.5 ml of lysis buffer and incubated for 24 h at 37°C with slow rotation.
TextSentencer_T46 7205-7306 Sentence denotes The cells were suspended in 0.5 ml of lysis buffer and incubated for 24 h at 37°C with slow rotation.
TextSentencer_T47 7307-7459 Sentence denotes The lysate was cooled on ice for 5 min, mixed with 5 M NaCl solutions to adjust the concentration of NaCl to 0.85 M, and further kept on ice for 10 min.
TextSentencer_T47 7307-7459 Sentence denotes The lysate was cooled on ice for 5 min, mixed with 5 M NaCl solutions to adjust the concentration of NaCl to 0.85 M, and further kept on ice for 10 min.
TextSentencer_T48 7460-7593 Sentence denotes Then, the lysate was centrifuged at 5,000 × g for 10 min at 4°C, and quantified for the concentration of L1/L2 capsid protein levels.
TextSentencer_T48 7460-7593 Sentence denotes Then, the lysate was centrifuged at 5,000 × g for 10 min at 4°C, and quantified for the concentration of L1/L2 capsid protein levels.
TextSentencer_T49 7594-7692 Sentence denotes To remove aggregates, the pseudovirion stock was filtered before use for the neutralization assay.
TextSentencer_T49 7594-7692 Sentence denotes To remove aggregates, the pseudovirion stock was filtered before use for the neutralization assay.
TextSentencer_T50 7693-7907 Sentence denotes The Frontiers in Microbiology | www.frontiersin.org inhibitory activity of 3HP-β-LG against HPV PsV entry into HeLa or HaCaT cells was detected as previously described (Surviladze et al., 2012; Kwak et al., 2014) .
TextSentencer_T50 7693-7907 Sentence denotes The Frontiers in Microbiology | www.frontiersin.org inhibitory activity of 3HP-β-LG against HPV PsV entry into HeLa or HaCaT cells was detected as previously described (Surviladze et al., 2012; Kwak et al., 2014) .
TextSentencer_T51 7908-8076 Sentence denotes Briefly, HeLa or HecCatT cells were seeded at 1.5 × 10 4 cells in 100 μl of 10% FBS DMEM (DMEM-10) per well in a 96-well plate, followed by a culture at 37°C overnight.
TextSentencer_T51 7908-8076 Sentence denotes Briefly, HeLa or HecCatT cells were seeded at 1.5 × 10 4 cells in 100 μl of 10% FBS DMEM (DMEM-10) per well in a 96-well plate, followed by a culture at 37°C overnight.
TextSentencer_T52 8077-8180 Sentence denotes 3HP-β-LG or β-LG was serially diluted in DMEM and incubated with 100 μl of HPV PsV (equal to 35 ng L1).
TextSentencer_T52 8077-8180 Sentence denotes 3HP-β-LG or β-LG was serially diluted in DMEM and incubated with 100 μl of HPV PsV (equal to 35 ng L1).
TextSentencer_T53 8181-8255 Sentence denotes Mixtures were added to HeLa or HaCaT cells and incubated at 37°C for 16 h.
TextSentencer_T53 8181-8255 Sentence denotes Mixtures were added to HeLa or HaCaT cells and incubated at 37°C for 16 h.
TextSentencer_T54 8256-8476 Sentence denotes After replacement of the culture supernatants with free medium and culture for additional 72 h, cells were lysed for measurement of luciferase activity, according to the manufacturer's manual (Promega, Madison, WI, USA).
TextSentencer_T54 8256-8476 Sentence denotes After replacement of the culture supernatants with free medium and culture for additional 72 h, cells were lysed for measurement of luciferase activity, according to the manufacturer's manual (Promega, Madison, WI, USA).
TextSentencer_T55 8477-8590 Sentence denotes 3HP-β-LG diluted in PBS was coated onto all wells of a 96-well polystyrene plate (Corning, USA) at 4°C overnight.
TextSentencer_T55 8477-8590 Sentence denotes 3HP-β-LG diluted in PBS was coated onto all wells of a 96-well polystyrene plate (Corning, USA) at 4°C overnight.
TextSentencer_T56 8591-8799 Sentence denotes After blocking with protein-free blocking buffer (Thermo Fisher Scientific, USA), HPV L1 (type16 or 18) protein at 1 μg/ml was added into the wells for 50 μl/well, and the plate was incubated at 37°C for 1 h.
TextSentencer_T56 8591-8799 Sentence denotes After blocking with protein-free blocking buffer (Thermo Fisher Scientific, USA), HPV L1 (type16 or 18) protein at 1 μg/ml was added into the wells for 50 μl/well, and the plate was incubated at 37°C for 1 h.
TextSentencer_T57 8800-8908 Sentence denotes After washing with PBST three times, mouse anti-HPV L1 (type16 or 18) antibody (Abcam, UK) was added at a 1:
TextSentencer_T57 8800-8908 Sentence denotes After washing with PBST three times, mouse anti-HPV L1 (type16 or 18) antibody (Abcam, UK) was added at a 1:
TextSentencer_T58 8909-8924 Sentence denotes 1,000 dilution.
TextSentencer_T58 8909-8924 Sentence denotes 1,000 dilution.
TextSentencer_T59 8925-9056 Sentence denotes After incubation at 37°C for 1 h and washing again, HRP-conjugated rabbit antimouse IgG antibody (Dako, Denmark) was added for 1 h.
TextSentencer_T59 8925-9056 Sentence denotes After incubation at 37°C for 1 h and washing again, HRP-conjugated rabbit antimouse IgG antibody (Dako, Denmark) was added for 1 h.
TextSentencer_T60 9057-9155 Sentence denotes After washing and adding tetramethylbenzidine (Sigma, USA), the absorbance was measured at 450 nm.
TextSentencer_T60 9057-9155 Sentence denotes After washing and adding tetramethylbenzidine (Sigma, USA), the absorbance was measured at 450 nm.
TextSentencer_T61 9156-9481 Sentence denotes Similar protocols were used for detection of 3HP-β-LG in the serum of rhesus monkeys and 3HP-β-LG-specific IgA and IgE in the vaginal swab eluates, which were leftover samples from the clinical trials for evaluating the in vivo safety and efficacy of the intravaginally applied 3HP-β-LG-containing vaginal gels (Registry No.:
TextSentencer_T61 9156-9481 Sentence denotes Similar protocols were used for detection of 3HP-β-LG in the serum of rhesus monkeys and 3HP-β-LG-specific IgA and IgE in the vaginal swab eluates, which were leftover samples from the clinical trials for evaluating the in vivo safety and efficacy of the intravaginally applied 3HP-β-LG-containing vaginal gels (Registry No.:
TextSentencer_T62 9482-9526 Sentence denotes ChiCTR-TRC-12002016) (Guo et al., 2016a,b) .
TextSentencer_T62 9482-9526 Sentence denotes ChiCTR-TRC-12002016) (Guo et al., 2016a,b) .
TextSentencer_T63 9527-9897 Sentence denotes Measurement of the Binding Ability of 3-Hydroxyphthalic Anhydride Modified Bovine Beta-Lactoglobulin and Beta-Lactoglobulin to Peptides Two positively charged peptides of HPV L1 protein, L1-I (residues 474-488: LKAKPKFTLGKRKAT) and L1-II (residues 493-505: STSTTAKRKKRKL), were synthesized by a standard solid-phase Fmoc method as previously reported (He et al., 2007) .
TextSentencer_T63 9527-9897 Sentence denotes Measurement of the Binding Ability of 3-Hydroxyphthalic Anhydride Modified Bovine Beta-Lactoglobulin and Beta-Lactoglobulin to Peptides Two positively charged peptides of HPV L1 protein, L1-I (residues 474-488: LKAKPKFTLGKRKAT) and L1-II (residues 493-505: STSTTAKRKKRKL), were synthesized by a standard solid-phase Fmoc method as previously reported (He et al., 2007) .
TextSentencer_T64 9898-10016 Sentence denotes The peptide solution (1 mM) was prepared in 50 mM phosphate buffer (pH 7.4) and added into the titration sample cells.
TextSentencer_T64 9898-10016 Sentence denotes The peptide solution (1 mM) was prepared in 50 mM phosphate buffer (pH 7.4) and added into the titration sample cells.
TextSentencer_T65 10017-10123 Sentence denotes 3HP-β-LG and β-LG were also dissolved in phosphate buffer to 50 μM and then added to the titration needle.
TextSentencer_T65 10017-10123 Sentence denotes 3HP-β-LG and β-LG were also dissolved in phosphate buffer to 50 μM and then added to the titration needle.
TextSentencer_T66 10124-10257 Sentence denotes Isothermal titration calorimetry (ITC) analysis was carried out at constant temperature of 18°C with stirring speed of 1,000 rev/min.
TextSentencer_T66 10124-10257 Sentence denotes Isothermal titration calorimetry (ITC) analysis was carried out at constant temperature of 18°C with stirring speed of 1,000 rev/min.
TextSentencer_T67 10258-10370 Sentence denotes Each titration volume was 2 μl, and the titration interval was 60 s for the first drop and 120 s for the others.
TextSentencer_T67 10258-10370 Sentence denotes Each titration volume was 2 μl, and the titration interval was 60 s for the first drop and 120 s for the others.
TextSentencer_T68 10371-10446 Sentence denotes HeLa cells were plated in a 96-well plate (for 1.0 × 10 5 /well) overnight.
TextSentencer_T68 10371-10446 Sentence denotes HeLa cells were plated in a 96-well plate (for 1.0 × 10 5 /well) overnight.
TextSentencer_T69 10447-10671 Sentence denotes Then, cells were incubated with 3HP-β-LG at a final concentration of 15 μg/ml at 0.5 h before or 0, 0.5, 1, 2, 4, 8, 12, 24, and 48 h after addition of HPV PsV (equal to 36 ng L1), followed by an incubation at 37°C for 16 h.
TextSentencer_T69 10447-10671 Sentence denotes Then, cells were incubated with 3HP-β-LG at a final concentration of 15 μg/ml at 0.5 h before or 0, 0.5, 1, 2, 4, 8, 12, 24, and 48 h after addition of HPV PsV (equal to 36 ng L1), followed by an incubation at 37°C for 16 h.
TextSentencer_T70 10672-10877 Sentence denotes After replacement of culture supernatant with fresh medium and an incubation for additional 72 h, cells were lysed to determine the entry inhibition ratio, according to the manufacturer's manual (Promega).
TextSentencer_T70 10672-10877 Sentence denotes After replacement of culture supernatant with fresh medium and an incubation for additional 72 h, cells were lysed to determine the entry inhibition ratio, according to the manufacturer's manual (Promega).
TextSentencer_T71 10878-11052 Sentence denotes HeLa cells in 96-well plates were preincubated with 100 μl of 3HP-β-LG (5 μM) or β-LG (5 μM) at 37°C for 1 h and then washed with DMEM three times before addition of HPV PsV.
TextSentencer_T71 10878-11052 Sentence denotes HeLa cells in 96-well plates were preincubated with 100 μl of 3HP-β-LG (5 μM) or β-LG (5 μM) at 37°C for 1 h and then washed with DMEM three times before addition of HPV PsV.
TextSentencer_T72 11053-11150 Sentence denotes For the control, 3HP-β-LG-or β-LG-pretreated HeLa cells were not washed before HPV PsV was added.
TextSentencer_T72 11053-11150 Sentence denotes For the control, 3HP-β-LG-or β-LG-pretreated HeLa cells were not washed before HPV PsV was added.
TextSentencer_T73 11151-11213 Sentence denotes HPV PsV entry into HeLa cells was detected as described above.
TextSentencer_T73 11151-11213 Sentence denotes HPV PsV entry into HeLa cells was detected as described above.
TextSentencer_T74 11214-11296 Sentence denotes The plated HeLa cells and 3HP-β-LG mixture were both prechilled at 4°C for 30 min.
TextSentencer_T74 11214-11296 Sentence denotes The plated HeLa cells and 3HP-β-LG mixture were both prechilled at 4°C for 30 min.
TextSentencer_T75 11297-11370 Sentence denotes Then, the cells were incubated with the 3HP-β-LG mixture at 4°C for 12 h.
TextSentencer_T75 11297-11370 Sentence denotes Then, the cells were incubated with the 3HP-β-LG mixture at 4°C for 12 h.
TextSentencer_T76 11371-11463 Sentence denotes After washing with cold PBS buffer twice, the cells were further incubated at 37°C for 72 h.
TextSentencer_T76 11371-11463 Sentence denotes After washing with cold PBS buffer twice, the cells were further incubated at 37°C for 72 h.
TextSentencer_T77 11464-11587 Sentence denotes Then the cells were lysed to determine luciferase activity, according to the luciferase assay system manual (Promega, USA).
TextSentencer_T77 11464-11587 Sentence denotes Then the cells were lysed to determine luciferase activity, according to the luciferase assay system manual (Promega, USA).
TextSentencer_T78 11588-11850 Sentence denotes The experiment on rhesus macaques was conducted under ethical guidelines and approved by the Ethics Committee of the Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences and Peking Union Medical College (approval number: ILAS-VL-2015-001).
TextSentencer_T78 11588-11850 Sentence denotes The experiment on rhesus macaques was conducted under ethical guidelines and approved by the Ethics Committee of the Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences and Peking Union Medical College (approval number: ILAS-VL-2015-001).
TextSentencer_T79 11851-11942 Sentence denotes In brief, the rhesus macaques were randomly divided into treatment group and control group.
TextSentencer_T79 11851-11942 Sentence denotes In brief, the rhesus macaques were randomly divided into treatment group and control group.
TextSentencer_T80 11943-12093 Sentence denotes In the treatment group, vaginal gel containing 3HP-β-LG (1.8 mg per dose) was dispersed into carbomer gel and administered intravaginally or rectally.
TextSentencer_T80 11943-12093 Sentence denotes In the treatment group, vaginal gel containing 3HP-β-LG (1.8 mg per dose) was dispersed into carbomer gel and administered intravaginally or rectally.
TextSentencer_T81 12094-12146 Sentence denotes In the control group, only carbomer gel was applied.
TextSentencer_T81 12094-12146 Sentence denotes In the control group, only carbomer gel was applied.
TextSentencer_T82 12147-12264 Sentence denotes Then, 2 ml of blood was collected before or after injection at −1 h, 0 h, 30 min, 1 h, 2 h, 3 h, 6 h, 12 h, and 24 h.
TextSentencer_T82 12147-12264 Sentence denotes Then, 2 ml of blood was collected before or after injection at −1 h, 0 h, 30 min, 1 h, 2 h, 3 h, 6 h, 12 h, and 24 h.
TextSentencer_T83 12265-12308 Sentence denotes The serum was separated after condensation.
TextSentencer_T83 12265-12308 Sentence denotes The serum was separated after condensation.
TextSentencer_T84 12309-12441 Sentence denotes To determine whether 3HP-β-LG could enter into HeLa or 293 T cells, cells (1 × 10 5 ) were seeded onto coverslips in a 6-well plate.
TextSentencer_T84 12309-12441 Sentence denotes To determine whether 3HP-β-LG could enter into HeLa or 293 T cells, cells (1 × 10 5 ) were seeded onto coverslips in a 6-well plate.
TextSentencer_T85 12442-12488 Sentence denotes Then, 3HP-β-LG (0.5 μg/ml) was added for 12 h.
TextSentencer_T85 12442-12488 Sentence denotes Then, 3HP-β-LG (0.5 μg/ml) was added for 12 h.
TextSentencer_T86 12489-12552 Sentence denotes Then, the supernatant was replaced with DMEM containing 2% FBS.
TextSentencer_T86 12489-12552 Sentence denotes Then, the supernatant was replaced with DMEM containing 2% FBS.
TextSentencer_T87 12553-12732 Sentence denotes After 24 days, the cells were fixed by 4% paraformaldehyde (PFA; Sigma-Aldrich, St. Louis, MO), perforated by 0.2% Triton X-100, and blocked with 3% BSA (Amresco, LLC, Solon, OH).
TextSentencer_T87 12553-12732 Sentence denotes After 24 days, the cells were fixed by 4% paraformaldehyde (PFA; Sigma-Aldrich, St. Louis, MO), perforated by 0.2% Triton X-100, and blocked with 3% BSA (Amresco, LLC, Solon, OH).
TextSentencer_T88 12733-12830 Sentence denotes Next, cells were incubated overnight with anti-3HP-β-LG mAb or anti-β-actin mAb (1:1,000) at 4°C.
TextSentencer_T88 12733-12830 Sentence denotes Next, cells were incubated overnight with anti-3HP-β-LG mAb or anti-β-actin mAb (1:1,000) at 4°C.
TextSentencer_T89 12831-13012 Sentence denotes After five washes, the cells were incubated with Alexa Fluor 488-labeled donkey anti-mouse IgG (1, 1,000, Thermo Fisher Scientific, Wilmington, DE, USA) at room temperature for 1 h.
TextSentencer_T89 12831-13012 Sentence denotes After five washes, the cells were incubated with Alexa Fluor 488-labeled donkey anti-mouse IgG (1, 1,000, Thermo Fisher Scientific, Wilmington, DE, USA) at room temperature for 1 h.
TextSentencer_T90 13013-13205 Sentence denotes After five washes, the coverslips were sealed with Prolong Gold Antifade reagent with 4,6-diamidino-2phenylindole (Thermo Fisher Scientific) and scanned with the Leica SP8 confocal microscope.
TextSentencer_T90 13013-13205 Sentence denotes After five washes, the coverslips were sealed with Prolong Gold Antifade reagent with 4,6-diamidino-2phenylindole (Thermo Fisher Scientific) and scanned with the Leica SP8 confocal microscope.
TextSentencer_T91 13206-13253 Sentence denotes Frontiers in Microbiology | www.frontiersin.org
TextSentencer_T91 13206-13253 Sentence denotes Frontiers in Microbiology | www.frontiersin.org
TextSentencer_T92 13254-13481 Sentence denotes Our previous studies have shown that 3-hydroxyphthalic anhydride-modified bovine beta-lactoglobulin (3HP-β-LG) is a promising HPV entry inhibitor against entry of PsV of both high-risk and low-risk HPV types (Lu et al., 2013) .
TextSentencer_T92 13254-13481 Sentence denotes Our previous studies have shown that 3-hydroxyphthalic anhydride-modified bovine beta-lactoglobulin (3HP-β-LG) is a promising HPV entry inhibitor against entry of PsV of both high-risk and low-risk HPV types (Lu et al., 2013) .
TextSentencer_T93 13482-13653 Sentence denotes This inhibitory activity was attributed to the interaction of the increased net negative charges on β-LG after 3-hydroxyphthalic anhydride modification (Lu et al., 2013) .
TextSentencer_T93 13482-13653 Sentence denotes This inhibitory activity was attributed to the interaction of the increased net negative charges on β-LG after 3-hydroxyphthalic anhydride modification (Lu et al., 2013) .
TextSentencer_T94 13654-13766 Sentence denotes One may ask whether all proteins with increased net negative charges can inhibit HPV entry into the target cell.
TextSentencer_T94 13654-13766 Sentence denotes One may ask whether all proteins with increased net negative charges can inhibit HPV entry into the target cell.
TextSentencer_T95 13767-14085 Sentence denotes To answer this question, we compared the inhibitory activity of 3HP-β-LG with that of 3-hydroxyphthalic anhydride-modified human serum albumin (3HP-HSA) and 3-hydroxyphthalic anhydride-modified chicken ovalbumin (3HP-OVA) against entry of HPV16 PsV into HeLa cells, using the unmodified β-LG, HSA, and OVA as controls.
TextSentencer_T95 13767-14085 Sentence denotes To answer this question, we compared the inhibitory activity of 3HP-β-LG with that of 3-hydroxyphthalic anhydride-modified human serum albumin (3HP-HSA) and 3-hydroxyphthalic anhydride-modified chicken ovalbumin (3HP-OVA) against entry of HPV16 PsV into HeLa cells, using the unmodified β-LG, HSA, and OVA as controls.
TextSentencer_T96 14086-14304 Sentence denotes As shown in Figure 1A , 3HP-β-LG exhibited potent anti-HPV activity in a dose-dependent manner with an IC 50 (the half maximal inhibitory concentration) of 0.59 μg/ml, which is more potent than that of 3HP-HSA (IC 50 :
TextSentencer_T96 14086-14304 Sentence denotes As shown in Figure 1A , 3HP-β-LG exhibited potent anti-HPV activity in a dose-dependent manner with an IC 50 (the half maximal inhibitory concentration) of 0.59 μg/ml, which is more potent than that of 3HP-HSA (IC 50 :
TextSentencer_T97 14305-14317 Sentence denotes 2.28 μg/ml).
TextSentencer_T97 14305-14317 Sentence denotes 2.28 μg/ml).
TextSentencer_T98 14318-14488 Sentence denotes 3HP-OVA and all three unmodified proteins at the concentration as high as 10 μg/ml showed no significant inhibitory activity against HPV16 PsV entry into the target cell.
TextSentencer_T98 14318-14488 Sentence denotes 3HP-OVA and all three unmodified proteins at the concentration as high as 10 μg/ml showed no significant inhibitory activity against HPV16 PsV entry into the target cell.
TextSentencer_T99 14489-14650 Sentence denotes These results confirm that not all, just some, proteins, such as β-LG and HSA, modified by 3-hydroxyphthalic anhydride gain the ability to inhibit HPV infection.
TextSentencer_T99 14489-14650 Sentence denotes These results confirm that not all, just some, proteins, such as β-LG and HSA, modified by 3-hydroxyphthalic anhydride gain the ability to inhibit HPV infection.
TextSentencer_T100 14651-14836 Sentence denotes In our previous studies, we proved the antiviral activity of 3HP-β-LG against entry of the high-risk HPV types 16 and 18 and low-risk HPV type 6 into the target cell (Lu et al., 2013) .
TextSentencer_T100 14651-14836 Sentence denotes In our previous studies, we proved the antiviral activity of 3HP-β-LG against entry of the high-risk HPV types 16 and 18 and low-risk HPV type 6 into the target cell (Lu et al., 2013) .
TextSentencer_T101 14837-14975 Sentence denotes Aside from these three representative HPV types worldwide, another important high-risk type, HPV58, is found with high incidence in China.
TextSentencer_T101 14837-14975 Sentence denotes Aside from these three representative HPV types worldwide, another important high-risk type, HPV58, is found with high incidence in China.
TextSentencer_T102 14976-15131 Sentence denotes To determine whether 3HP-β-LG is also effective against HPV58, we constructed the HPV58 pseudovirus and tested its sensitivity to 3HP-β-LG (control: β-LG).
TextSentencer_T102 14976-15131 Sentence denotes To determine whether 3HP-β-LG is also effective against HPV58, we constructed the HPV58 pseudovirus and tested its sensitivity to 3HP-β-LG (control: β-LG).
TextSentencer_T103 15132-15333 Sentence denotes As shown in Figure 1B , 3HP-β-LG could also inhibit HPV58 PsV entry into the target cell in a dose-dependent manner (IC 50 of 0.28 μg/ml), confirming that 3HP-β-LG has broad-spectrum anti-HPV activity.
TextSentencer_T103 15132-15333 Sentence denotes As shown in Figure 1B , 3HP-β-LG could also inhibit HPV58 PsV entry into the target cell in a dose-dependent manner (IC 50 of 0.28 μg/ml), confirming that 3HP-β-LG has broad-spectrum anti-HPV activity.
TextSentencer_T104 15334-15645 Sentence denotes Considering that the HaCaT cells express abundant integrin and heparan sulfate proteoglycan (HSPG), the receptors for HPV (Aksoy et al., 2014; Kumar et al., 2014) and act as target cells for HPV infection, we also evaluated the inhibitory activity of 3HP-β-LG against HPV16 and HPV58 PsV entry into HaCaT cells.
TextSentencer_T104 15334-15645 Sentence denotes Considering that the HaCaT cells express abundant integrin and heparan sulfate proteoglycan (HSPG), the receptors for HPV (Aksoy et al., 2014; Kumar et al., 2014) and act as target cells for HPV infection, we also evaluated the inhibitory activity of 3HP-β-LG against HPV16 and HPV58 PsV entry into HaCaT cells.
TextSentencer_T105 15646-15846 Sentence denotes As shown in Figures 1C,D Frontiers in Microbiology | www.frontiersin.org against entry of HPV16 and HPV58 PsV into HaCaT cells, suggesting that 3HP-β-LG acts on the virus, rather than the target cell.
TextSentencer_T105 15646-15846 Sentence denotes As shown in Figures 1C,D Frontiers in Microbiology | www.frontiersin.org against entry of HPV16 and HPV58 PsV into HaCaT cells, suggesting that 3HP-β-LG acts on the virus, rather than the target cell.
TextSentencer_T106 15847-16008 Sentence denotes Bovine Beta-Lactoglobulin Binds to the L1 Protein and the Peptides Derived From the Positively Charged Residue-Enriched Region in Human Papillomavirus L1 Protein
TextSentencer_T106 15847-16008 Sentence denotes Bovine Beta-Lactoglobulin Binds to the L1 Protein and the Peptides Derived From the Positively Charged Residue-Enriched Region in Human Papillomavirus L1 Protein
TextSentencer_T107 16009-16356 Sentence denotes As we proposed before, 3HP-β-LG inhibited HPV PsV entry into the target cell possibly through the binding of the negatively charged residues in 3HP-β-LG with the positively charged residues in a protein on the surface of the viral particle, thereby blocking the interaction between viral protein and receptor on the target cell (Lu et al., 2013) .
TextSentencer_T107 16009-16356 Sentence denotes As we proposed before, 3HP-β-LG inhibited HPV PsV entry into the target cell possibly through the binding of the negatively charged residues in 3HP-β-LG with the positively charged residues in a protein on the surface of the viral particle, thereby blocking the interaction between viral protein and receptor on the target cell (Lu et al., 2013) .
TextSentencer_T108 16357-16457 Sentence denotes However, it is still unclear which protein on the virus surface may be involved in this interaction.
TextSentencer_T108 16357-16457 Sentence denotes However, it is still unclear which protein on the virus surface may be involved in this interaction.
TextSentencer_T109 16458-16518 Sentence denotes The most abundant protein on the HPV particle is L1 protein.
TextSentencer_T109 16458-16518 Sentence denotes The most abundant protein on the HPV particle is L1 protein.
TextSentencer_T110 16519-16597 Sentence denotes Here we detected the interaction between 3HP-β-LG and HPV L1 protein by ELISA.
TextSentencer_T110 16519-16597 Sentence denotes Here we detected the interaction between 3HP-β-LG and HPV L1 protein by ELISA.
TextSentencer_T111 16598-16742 Sentence denotes As shown in Figures 2Aa,Ab, 3HP-β-LG could strongly bind to the L1 protein of both HPV16 and HPV18, while the unmodified β-LG protein could not.
TextSentencer_T111 16598-16742 Sentence denotes As shown in Figures 2Aa,Ab, 3HP-β-LG could strongly bind to the L1 protein of both HPV16 and HPV18, while the unmodified β-LG protein could not.
TextSentencer_T112 16743-16900 Sentence denotes The C-terminal positively charged region of HPV L1 was proved to regulate the binding of HPV to the receptor (Sibbet et al., 2000; Bousarghin et al., 2003) .
TextSentencer_T112 16743-16900 Sentence denotes The C-terminal positively charged region of HPV L1 was proved to regulate the binding of HPV to the receptor (Sibbet et al., 2000; Bousarghin et al., 2003) .
TextSentencer_T113 16901-17028 Sentence denotes This region was exposed on the viral surface, and involved in the binding to cell surface receptors or neutralizing antibodies.
TextSentencer_T113 16901-17028 Sentence denotes This region was exposed on the viral surface, and involved in the binding to cell surface receptors or neutralizing antibodies.
TextSentencer_T114 17029-17170 Sentence denotes Therefore, we synthesized two positively charged peptides in this region of L1 protein, L1-I (residues 474-488) and L1-II (residues 493-505).
TextSentencer_T114 17029-17170 Sentence denotes Therefore, we synthesized two positively charged peptides in this region of L1 protein, L1-I (residues 474-488) and L1-II (residues 493-505).
TextSentencer_T115 17171-17285 Sentence denotes The binding ability of 3HP-β-LG and β-LG to these peptides was measured by isothermal titration calorimetry (ITC).
TextSentencer_T115 17171-17285 Sentence denotes The binding ability of 3HP-β-LG and β-LG to these peptides was measured by isothermal titration calorimetry (ITC).
TextSentencer_T116 17286-17537 Sentence denotes As shown in Figure 2B , β-LG had no obvious binding to L1-I or L1-II peptide, while 3HP-β-LG showed high binding ability against both L1-I (binding constant K α value equals 2.31 × 10 5 /M) and L1-II (binding constant K α value equals 1.04 × 10 5 /M).
TextSentencer_T116 17286-17537 Sentence denotes As shown in Figure 2B , β-LG had no obvious binding to L1-I or L1-II peptide, while 3HP-β-LG showed high binding ability against both L1-I (binding constant K α value equals 2.31 × 10 5 /M) and L1-II (binding constant K α value equals 1.04 × 10 5 /M).
TextSentencer_T117 17538-17745 Sentence denotes These results suggested that 3HP-β-LG may bind to the positively charged sites in the C-terminal region of L1 protein on the HPV surface to block the interaction between the viral particle and cell receptor.
TextSentencer_T117 17538-17745 Sentence denotes These results suggested that 3HP-β-LG may bind to the positively charged sites in the C-terminal region of L1 protein on the HPV surface to block the interaction between the viral particle and cell receptor.
TextSentencer_T118 17746-17917 Sentence denotes To determine whether 3HP-β-LG could inactivate HPV PsV, we incubated HPV PsV with 3HP-β-LG at room temperature for 2 h and then removed 3HP-β-LG by PEG-8000 precipitation.
TextSentencer_T118 17746-17917 Sentence denotes To determine whether 3HP-β-LG could inactivate HPV PsV, we incubated HPV PsV with 3HP-β-LG at room temperature for 2 h and then removed 3HP-β-LG by PEG-8000 precipitation.
TextSentencer_T119 17918-18223 Sentence denotes As shown in Figure 3A , the treated virus could still infect the target host cell with no reduced activity, compared the control virus (DMEM-or β-LG-treated), suggesting that the binding between 3HP-β-LG and HPV PsV may be noncovalent and reversible so that 3HP-β-LG cannot permanently inactive the virus.
TextSentencer_T119 17918-18223 Sentence denotes As shown in Figure 3A , the treated virus could still infect the target host cell with no reduced activity, compared the control virus (DMEM-or β-LG-treated), suggesting that the binding between 3HP-β-LG and HPV PsV may be noncovalent and reversible so that 3HP-β-LG cannot permanently inactive the virus.
TextSentencer_T120 18224-18527 Sentence denotes HPV entry into the host cell takes more than 10 h, a relatively slow process compared to other viruses (Aksoy et al., 2017) , which provides a long window period for an entry inhibitor to bind with the viral protein responsible for interaction with the receptor on the target cell and block viral entry.
TextSentencer_T120 18224-18527 Sentence denotes HPV entry into the host cell takes more than 10 h, a relatively slow process compared to other viruses (Aksoy et al., 2017) , which provides a long window period for an entry inhibitor to bind with the viral protein responsible for interaction with the receptor on the target cell and block viral entry.
TextSentencer_T121 18528-18658 Sentence denotes Here, we performed a time-addition assay to pinpoint the window of functional 3HP-β-LG blocking of HPV entry into the target cell.
TextSentencer_T121 18528-18658 Sentence denotes Here, we performed a time-addition assay to pinpoint the window of functional 3HP-β-LG blocking of HPV entry into the target cell.
TextSentencer_T122 18659-19021 Sentence denotes As shown in Figure 3B , 3HP-β-LG (15 μg/ml) could fully inhibit HPV16 PsV entry when it was added to the cells 0.5 h before and 0.5, 1, 2, 4, and 8 h after addition of HPV16 PsV, respectively, while about 70, 10, and 5% of HPV16 PsV entry were blocked when 3HP-β-LG was added to the cells 12, 24, and 48 h after addition of HPV16 PsV, respectively ( Figure 3B ).
TextSentencer_T122 18659-19021 Sentence denotes As shown in Figure 3B , 3HP-β-LG (15 μg/ml) could fully inhibit HPV16 PsV entry when it was added to the cells 0.5 h before and 0.5, 1, 2, 4, and 8 h after addition of HPV16 PsV, respectively, while about 70, 10, and 5% of HPV16 PsV entry were blocked when 3HP-β-LG was added to the cells 12, 24, and 48 h after addition of HPV16 PsV, respectively ( Figure 3B ).
TextSentencer_T123 19022-19226 Sentence denotes This result suggests that during this relatively long period of entry process (Aksoy et al., 2017) , the entry inhibitor 3HP-β-LG remains sufficiently active to inhibit HPV PsV entry into the target cell.
TextSentencer_T123 19022-19226 Sentence denotes This result suggests that during this relatively long period of entry process (Aksoy et al., 2017) , the entry inhibitor 3HP-β-LG remains sufficiently active to inhibit HPV PsV entry into the target cell.
TextSentencer_T124 19227-19371 Sentence denotes To confirm the functional stage of 3HP-β-LG in blocking viral entry, we used a temperature shift assay to slow down the viral infection process.
TextSentencer_T124 19227-19371 Sentence denotes To confirm the functional stage of 3HP-β-LG in blocking viral entry, we used a temperature shift assay to slow down the viral infection process.
TextSentencer_T125 19372-19572 Sentence denotes At 4°C, HPV PsV could bind to cell receptors, but could not fully enter the cytoplasm because the speeds of membrane fusion and cytoskeletal transformation are largely reduced at such low temperature.
TextSentencer_T125 19372-19572 Sentence denotes At 4°C, HPV PsV could bind to cell receptors, but could not fully enter the cytoplasm because the speeds of membrane fusion and cytoskeletal transformation are largely reduced at such low temperature.
TextSentencer_T126 19573-19668 Sentence denotes We incubated HPV PsV and HeLa cells at 4°C for 12 h so that HPV could bind to the cell surface.
TextSentencer_T126 19573-19668 Sentence denotes We incubated HPV PsV and HeLa cells at 4°C for 12 h so that HPV could bind to the cell surface.
TextSentencer_T127 19669-19770 Sentence denotes Then we washed away the unbound HPV before shifting the cells to 37°C for further 72 h of incubation.
TextSentencer_T127 19669-19770 Sentence denotes Then we washed away the unbound HPV before shifting the cells to 37°C for further 72 h of incubation.
TextSentencer_T128 19771-19893 Sentence denotes When 3HP-β-LG was added into the cells at the beginning and then removed by washing, no viral entry occurred (Figure 3C ).
TextSentencer_T128 19771-19893 Sentence denotes When 3HP-β-LG was added into the cells at the beginning and then removed by washing, no viral entry occurred (Figure 3C ).
TextSentencer_T129 19894-20008 Sentence denotes This proved that 3HP-β-LG blocked the attachment between PsV and cell receptor at the earliest stage of infection.
TextSentencer_T129 19894-20008 Sentence denotes This proved that 3HP-β-LG blocked the attachment between PsV and cell receptor at the earliest stage of infection.
TextSentencer_T130 20009-20188 Sentence denotes To determine whether 3HP-β-LG could also bind to the receptor on the target cell surface to block HPV entry, we incubated HeLa cells with 3HP-β-LG (control: β-LG) at 37°C for 1 h.
TextSentencer_T130 20009-20188 Sentence denotes To determine whether 3HP-β-LG could also bind to the receptor on the target cell surface to block HPV entry, we incubated HeLa cells with 3HP-β-LG (control: β-LG) at 37°C for 1 h.
TextSentencer_T131 20189-20308 Sentence denotes The cells were washed with DMEM to remove the unbound proteins, or not washed (as control), before addition of HPV PsV.
TextSentencer_T131 20189-20308 Sentence denotes The cells were washed with DMEM to remove the unbound proteins, or not washed (as control), before addition of HPV PsV.
TextSentencer_T132 20309-20488 Sentence denotes Under the washing conditions, HPV PsV entry into the HeLa cells was not blocked, while under the non-washing conditions, HPV PsV entry into the HeLa cells was effectively blocked.
TextSentencer_T132 20309-20488 Sentence denotes Under the washing conditions, HPV PsV entry into the HeLa cells was not blocked, while under the non-washing conditions, HPV PsV entry into the HeLa cells was effectively blocked.
TextSentencer_T133 20489-20631 Sentence denotes No inhibition was observed in the β-LG control groups, no matter whether the β-LG-treated HeLa cells were LG was unable to inactivate HPV PsV.
TextSentencer_T133 20489-20631 Sentence denotes No inhibition was observed in the β-LG control groups, no matter whether the β-LG-treated HeLa cells were LG was unable to inactivate HPV PsV.
TextSentencer_T134 20632-20773 Sentence denotes HPV PsV was incubated with 3HP-β-LG (control: β-LG) at room temperature for 2 h and then separated by PEG-8000 to analyze its entry activity.
TextSentencer_T134 20632-20773 Sentence denotes HPV PsV was incubated with 3HP-β-LG (control: β-LG) at room temperature for 2 h and then separated by PEG-8000 to analyze its entry activity.
TextSentencer_T135 20774-20827 Sentence denotes The DMEM-treated virus was taken as negative control.
TextSentencer_T135 20774-20827 Sentence denotes The DMEM-treated virus was taken as negative control.
TextSentencer_T136 20828-20884 Sentence denotes (B) 3HP-β-LG blocked HPV PsV entry into the target cell.
TextSentencer_T136 20828-20884 Sentence denotes (B) 3HP-β-LG blocked HPV PsV entry into the target cell.
TextSentencer_T137 20885-21121 Sentence denotes HeLa cells were incubated with HPV PsV, while 3HP-β-LG (15 μg/ml) was added at different time points (−0.5, 0, 0.5, 1, 2, 4, 8, 12, 24, and 48 h) to analyze the inhibitory activity. (C) 3HP-β-LG blocked HPV PsV entry by targeting virus.
TextSentencer_T137 20885-21121 Sentence denotes HeLa cells were incubated with HPV PsV, while 3HP-β-LG (15 μg/ml) was added at different time points (−0.5, 0, 0.5, 1, 2, 4, 8, 12, 24, and 48 h) to analyze the inhibitory activity. (C) 3HP-β-LG blocked HPV PsV entry by targeting virus.
TextSentencer_T138 21122-21196 Sentence denotes Temperature shift assay for anti-HPV activity of 3HP-β-LG (control: β-LG).
TextSentencer_T138 21122-21196 Sentence denotes Temperature shift assay for anti-HPV activity of 3HP-β-LG (control: β-LG).
TextSentencer_T139 21197-21335 Sentence denotes Diluted 3HP-β-LG or β-LG at 15 μg/ml was mixed with HPV PsV and prechilled at 4°C for 30 min; then they were added to HeLa cells for 12 h.
TextSentencer_T139 21197-21335 Sentence denotes Diluted 3HP-β-LG or β-LG at 15 μg/ml was mixed with HPV PsV and prechilled at 4°C for 30 min; then they were added to HeLa cells for 12 h.
TextSentencer_T140 21336-21540 Sentence denotes After washing twice, the cells were further incubated at 37°C for 72 h and then analyzed for fluorescent integrated density. (D) 3HP-β-LG inhibited HPV PsV entry by targeting the virus, not the host cell.
TextSentencer_T140 21336-21540 Sentence denotes After washing twice, the cells were further incubated at 37°C for 72 h and then analyzed for fluorescent integrated density. (D) 3HP-β-LG inhibited HPV PsV entry by targeting the virus, not the host cell.
TextSentencer_T141 21541-21669 Sentence denotes HeLa cells were incubated with 3HP-β-LG (control: β-LG) at 37°C for 1 h with/without washing by DMEM before addition of HPV PsV.
TextSentencer_T141 21541-21669 Sentence denotes HeLa cells were incubated with 3HP-β-LG (control: β-LG) at 37°C for 1 h with/without washing by DMEM before addition of HPV PsV.
TextSentencer_T142 21670-21746 Sentence denotes Each sample was tested in triplicate, and the experiment was repeated twice.
TextSentencer_T142 21670-21746 Sentence denotes Each sample was tested in triplicate, and the experiment was repeated twice.
TextSentencer_T143 21747-21812 Sentence denotes Data from a representative experiment are presented as mean ± SD.
TextSentencer_T143 21747-21812 Sentence denotes Data from a representative experiment are presented as mean ± SD.
TextSentencer_T144 21813-21871 Sentence denotes Asterisks represent significant differences. ***p < 0.001.
TextSentencer_T144 21813-21871 Sentence denotes Asterisks represent significant differences. ***p < 0.001.
TextSentencer_T145 21872-21955 Sentence denotes Frontiers in Microbiology | www.frontiersin.org washed or not washed ( Figure 3D) .
TextSentencer_T145 21872-21955 Sentence denotes Frontiers in Microbiology | www.frontiersin.org washed or not washed ( Figure 3D) .
TextSentencer_T146 21956-22104 Sentence denotes These results suggest that 3HP-β-LG inhibition of HPV PsV entry into HeLa cells does not result from its binding to HPV's receptor on the host cell.
TextSentencer_T146 21956-22104 Sentence denotes These results suggest that 3HP-β-LG inhibition of HPV PsV entry into HeLa cells does not result from its binding to HPV's receptor on the host cell.
TextSentencer_T147 22105-22294 Sentence denotes Bovine Beta-Lactoglobulin Could Not Penetrate Into the Cell and the Vaginally Applied 3-Hydroxyphthalic Anhydride Modified Bovine Beta-Lactoglobulin Did Not Enter Into the Blood Circulation
TextSentencer_T147 22105-22294 Sentence denotes Bovine Beta-Lactoglobulin Could Not Penetrate Into the Cell and the Vaginally Applied 3-Hydroxyphthalic Anhydride Modified Bovine Beta-Lactoglobulin Did Not Enter Into the Blood Circulation
TextSentencer_T148 22295-22494 Sentence denotes To determine whether intravaginally applied 3HP-β-LG could enter into epithelial cells, we incubated HeLa cells and 293 T cells with 3HP-β-LG for 24 h and then washed the cells three times using PBS.
TextSentencer_T148 22295-22494 Sentence denotes To determine whether intravaginally applied 3HP-β-LG could enter into epithelial cells, we incubated HeLa cells and 293 T cells with 3HP-β-LG for 24 h and then washed the cells three times using PBS.
TextSentencer_T149 22495-22641 Sentence denotes The cells were stained with mouse anti-3HPβ-LG antibody or anti-β-actin antibody, as a control, and Alexa Fluor 488-labeled donkey anti-mouse IgG.
TextSentencer_T149 22495-22641 Sentence denotes The cells were stained with mouse anti-3HPβ-LG antibody or anti-β-actin antibody, as a control, and Alexa Fluor 488-labeled donkey anti-mouse IgG.
TextSentencer_T150 22642-22861 Sentence denotes Different from the anti-β-actin antibody-treated cells that showed strong fluorescence signals, anti-3HP-β-LG antibody-treated cells displayed no significant fluorescence signals inside or outside the cells (Figure 4) .
TextSentencer_T150 22642-22861 Sentence denotes Different from the anti-β-actin antibody-treated cells that showed strong fluorescence signals, anti-3HP-β-LG antibody-treated cells displayed no significant fluorescence signals inside or outside the cells (Figure 4) .
TextSentencer_T151 22862-23003 Sentence denotes These results indicate that 3HP-β-LG could neither enter into the cell nor bind the cell surface proteins, including the receptor(s) for HPV.
TextSentencer_T151 22862-23003 Sentence denotes These results indicate that 3HP-β-LG could neither enter into the cell nor bind the cell surface proteins, including the receptor(s) for HPV.
TextSentencer_T152 23004-23196 Sentence denotes To investigate whether the topically applied 3HP-β-LG enters the blood circulation, a carbomer gel with or without 3HP-β-LG was topically administered in the vagina or anus of rhesus macaques.
TextSentencer_T152 23004-23196 Sentence denotes To investigate whether the topically applied 3HP-β-LG enters the blood circulation, a carbomer gel with or without 3HP-β-LG was topically administered in the vagina or anus of rhesus macaques.
TextSentencer_T153 23197-23290 Sentence denotes Their blood samples were collected at different time points before and after gel application.
TextSentencer_T153 23197-23290 Sentence denotes Their blood samples were collected at different time points before and after gel application.
TextSentencer_T154 23291-23435 Sentence denotes The concentration of 3HP-β-LG in the blood samples and that in the gel (as positive control) were quantified with ELISA using anti-3HP-β-LG mAb.
TextSentencer_T154 23291-23435 Sentence denotes The concentration of 3HP-β-LG in the blood samples and that in the gel (as positive control) were quantified with ELISA using anti-3HP-β-LG mAb.
TextSentencer_T155 23436-23687 Sentence denotes As shown in Figure 5 , no 3HP-β-LG was detected in the blood samples of rhesus macaques vaginally or rectally administered with carbomer gel with or without 3HP-β-LG, confirming that the topically applied 3HP-β-LG does not enter the blood circulation.
TextSentencer_T155 23436-23687 Sentence denotes As shown in Figure 5 , no 3HP-β-LG was detected in the blood samples of rhesus macaques vaginally or rectally administered with carbomer gel with or without 3HP-β-LG, confirming that the topically applied 3HP-β-LG does not enter the blood circulation.
TextSentencer_T156 23688-23842 Sentence denotes About 10-20% of acute HPV infections become persistent, leading to various forms of cancer (Shanmugasundaram and You, 2017) , such as the cervical cancer.
TextSentencer_T156 23688-23842 Sentence denotes About 10-20% of acute HPV infections become persistent, leading to various forms of cancer (Shanmugasundaram and You, 2017) , such as the cervical cancer.
TextSentencer_T157 23843-24119 Sentence denotes Although several multivalent prophylactic HPV vaccines have been used in clinics to prevent HPV infection and cervical cancer development, they have no effect against pre-existing HPV infection in middle-aged and elderly women, the population at high risk for cervical cancer.
TextSentencer_T157 23843-24119 Sentence denotes Although several multivalent prophylactic HPV vaccines have been used in clinics to prevent HPV infection and cervical cancer development, they have no effect against pre-existing HPV infection in middle-aged and elderly women, the population at high risk for cervical cancer.
TextSentencer_T158 24120-24279 Sentence denotes Therefore, it is essential to develop effective and safe therapeutic agents for treatment of HPV infection in order to reduce the morbidity of cervical cancer.
TextSentencer_T158 24120-24279 Sentence denotes Therefore, it is essential to develop effective and safe therapeutic agents for treatment of HPV infection in order to reduce the morbidity of cervical cancer.
TextSentencer_T159 24280-24473 Sentence denotes Viral entry inhibitors have been proven effective and safe for the treatment of viral infections (Jiang et al., 1993; Dando and Perry, 2003; Lu et al., 2016; Li et al., 2017; Su et al., 2019) .
TextSentencer_T159 24280-24473 Sentence denotes Viral entry inhibitors have been proven effective and safe for the treatment of viral infections (Jiang et al., 1993; Dando and Perry, 2003; Lu et al., 2016; Li et al., 2017; Su et al., 2019) .
TextSentencer_T160 24474-24659 Sentence denotes We have previously reported that an HPV entry inhibitor, 3HP-β-LG, is highly effective in blocking the entry of HPV, both high-and low-risk types, into the host cell (Lu et al., 2013) .
TextSentencer_T160 24474-24659 Sentence denotes We have previously reported that an HPV entry inhibitor, 3HP-β-LG, is highly effective in blocking the entry of HPV, both high-and low-risk types, into the host cell (Lu et al., 2013) .
TextSentencer_T161 24660-24848 Sentence denotes In a randomized clinical trial, topical application of the vaginal gel containing 3HP-β-LG has shown excellent efficacy and safety to treat high-risk HPV infections (Guo et al., 2016a,b) .
TextSentencer_T161 24660-24848 Sentence denotes In a randomized clinical trial, topical application of the vaginal gel containing 3HP-β-LG has shown excellent efficacy and safety to treat high-risk HPV infections (Guo et al., 2016a,b) .
TextSentencer_T162 24849-24928 Sentence denotes However, how 3HP-β-LG inhibits HPV entry into the target cell is still unclear.
TextSentencer_T162 24849-24928 Sentence denotes However, how 3HP-β-LG inhibits HPV entry into the target cell is still unclear.
TextSentencer_T163 24929-24986 Sentence denotes FIGURE 4 | 3HP-β-LG could not enter into the target cell.
TextSentencer_T163 24929-24986 Sentence denotes FIGURE 4 | 3HP-β-LG could not enter into the target cell.
TextSentencer_T164 24987-25164 Sentence denotes After incubation with HeLa and 293 T cells, 3HP-β-LG in the cells was detected by immunofluorescence assay using anti-3HP-β-LG mAb (green). β-actin was used as positive control.
TextSentencer_T164 24987-25164 Sentence denotes After incubation with HeLa and 293 T cells, 3HP-β-LG in the cells was detected by immunofluorescence assay using anti-3HP-β-LG mAb (green). β-actin was used as positive control.
TextSentencer_T165 25165-25229 Sentence denotes Cell nuclei were stained by 4,6-diamidino-2-phenylindole (blue).
TextSentencer_T165 25165-25229 Sentence denotes Cell nuclei were stained by 4,6-diamidino-2-phenylindole (blue).
TextSentencer_T166 25230-25662 Sentence denotes In our previous study, we demonstrated that the inhibitory activity of 3HP-β-LG on HPV PsV entry into the target cell is closely correlated with the number of the positively charged lysine and arginine residues in 3HP-β-LG, as modified, i.e., the net negative charges on 3HP-β-LG (Lu et al., 2013) , suggesting that the negatively charged residues on 3HP-β-LG play an important role in 3HP-β-LG-mediated inhibition of HPV infection.
TextSentencer_T166 25230-25662 Sentence denotes In our previous study, we demonstrated that the inhibitory activity of 3HP-β-LG on HPV PsV entry into the target cell is closely correlated with the number of the positively charged lysine and arginine residues in 3HP-β-LG, as modified, i.e., the net negative charges on 3HP-β-LG (Lu et al., 2013) , suggesting that the negatively charged residues on 3HP-β-LG play an important role in 3HP-β-LG-mediated inhibition of HPV infection.
TextSentencer_T167 25663-25880 Sentence denotes As shown in Figure 6A , the unmodified bovine β-LG contains 18 positively charged residues (15 Lys and 3 Arg) and 26 negatively charged residues (16 Asp and 10 Glu) (Qin et al., 1999) , thus having a net charge of −8.
TextSentencer_T167 25663-25880 Sentence denotes As shown in Figure 6A , the unmodified bovine β-LG contains 18 positively charged residues (15 Lys and 3 Arg) and 26 negatively charged residues (16 Asp and 10 Glu) (Qin et al., 1999) , thus having a net charge of −8.
TextSentencer_T168 25881-26043 Sentence denotes In 3HP-β-LG, 14 out of the 15 Lys and all 3 Arg were modified by 3HP at 6 mM (Lu et al., 2013) , resulting in the neutralization of 17 of the 18 positive charges.
TextSentencer_T168 25881-26043 Sentence denotes In 3HP-β-LG, 14 out of the 15 Lys and all 3 Arg were modified by 3HP at 6 mM (Lu et al., 2013) , resulting in the neutralization of 17 of the 18 positive charges.
TextSentencer_T169 26044-26213 Sentence denotes Therefore, the net charge of 3HP-β-LG becomes −25, making 3HP-β-LG be active in binding to a viral protein with high positive net charges and inhibiting virus infection.
TextSentencer_T169 26044-26213 Sentence denotes Therefore, the net charge of 3HP-β-LG becomes −25, making 3HP-β-LG be active in binding to a viral protein with high positive net charges and inhibiting virus infection.
TextSentencer_T170 26214-26372 Sentence denotes In this study, we demonstrated that 3HP-β-LG could strongly bind to the L1 protein on the HPV16 PsV, while the unmodified β-LG protein could not (Figure 2A) .
TextSentencer_T170 26214-26372 Sentence denotes In this study, we demonstrated that 3HP-β-LG could strongly bind to the L1 protein on the HPV16 PsV, while the unmodified β-LG protein could not (Figure 2A) .
TextSentencer_T171 26373-26705 Sentence denotes Using ITC analysis, we showed that unlike β-LG, 3HP-β-LG could strongly interact with the positively charged residue-enriched C-terminal regions (residues 474-488 and 493-505) of L1 protein, which is responsible for the attachment of the HPV particle to the host cell, possibly through its interaction with the cellular receptor(s).
TextSentencer_T171 26373-26705 Sentence denotes Using ITC analysis, we showed that unlike β-LG, 3HP-β-LG could strongly interact with the positively charged residue-enriched C-terminal regions (residues 474-488 and 493-505) of L1 protein, which is responsible for the attachment of the HPV particle to the host cell, possibly through its interaction with the cellular receptor(s).
TextSentencer_T172 26706-26836 Sentence denotes One may argue that 3HP-β-LG may bind to any virus with high net positive charges on its surface protein and inhibit its infection.
TextSentencer_T172 26706-26836 Sentence denotes One may argue that 3HP-β-LG may bind to any virus with high net positive charges on its surface protein and inhibit its infection.
TextSentencer_T173 26837-27033 Sentence denotes Indeed, 3HP-β-LG can also inhibit infection of simian immunodeficiency virus (SIV) (Wyand et al., 1999) and HSV , but it is not effective against infection of MERS-CoV and VSV (unpublished data) .
TextSentencer_T173 26837-27033 Sentence denotes Indeed, 3HP-β-LG can also inhibit infection of simian immunodeficiency virus (SIV) (Wyand et al., 1999) and HSV , but it is not effective against infection of MERS-CoV and VSV (unpublished data) .
TextSentencer_T174 27034-27278 Sentence denotes In this study, we showed that 3HP-modified HSA (3HP-HSA) was also effective, while 3HP-modified OVA (3HP-OVA) was not effective in inhibiting HPV PsV entry (Figure 1) , even though 3HP-OVA also contains high net negative charges on its surface.
TextSentencer_T174 27034-27278 Sentence denotes In this study, we showed that 3HP-modified HSA (3HP-HSA) was also effective, while 3HP-modified OVA (3HP-OVA) was not effective in inhibiting HPV PsV entry (Figure 1) , even though 3HP-OVA also contains high net negative charges on its surface.
TextSentencer_T175 27279-27639 Sentence denotes Thus, aside from having high net negative charges, these results suggest that the active site in 3HP-β-LG for binding to the viral protein must possess a conformation that fits with that in the corresponding site in the viral protein (Figure 6B) , enabling 3HP-β-LG to strongly and closely interact with the viral protein to block HPV entry into the host cell.
TextSentencer_T175 27279-27639 Sentence denotes Thus, aside from having high net negative charges, these results suggest that the active site in 3HP-β-LG for binding to the viral protein must possess a conformation that fits with that in the corresponding site in the viral protein (Figure 6B) , enabling 3HP-β-LG to strongly and closely interact with the viral protein to block HPV entry into the host cell.
TextSentencer_T176 27640-27864 Sentence denotes Indeed, the molecular docking analysis indicated that 3HP-β-LG could strongly bind to the positively charged region of HPV L1 protein, which is the binding site for the negatively charged HSPG receptor for HPV ( Figure 6B ).
TextSentencer_T176 27640-27864 Sentence denotes Indeed, the molecular docking analysis indicated that 3HP-β-LG could strongly bind to the positively charged region of HPV L1 protein, which is the binding site for the negatively charged HSPG receptor for HPV ( Figure 6B ).
TextSentencer_T177 27865-28005 Sentence denotes 3HP-β-LG has many more negative charges on the surface, and the interface between 3HP-β-LG and HPV-L1 is much larger than the HSPG molecule.
TextSentencer_T177 27865-28005 Sentence denotes 3HP-β-LG has many more negative charges on the surface, and the interface between 3HP-β-LG and HPV-L1 is much larger than the HSPG molecule.
TextSentencer_T178 28006-28101 Sentence denotes Therefore, 3HP-β-LG can competitively bind to HPV L1 to block the binding of the HSPG receptor.
TextSentencer_T178 28006-28101 Sentence denotes Therefore, 3HP-β-LG can competitively bind to HPV L1 to block the binding of the HSPG receptor.
TextSentencer_T179 28102-28259 Sentence denotes Based on the results obtained from this study, we proposed a mechanistic model of 3HP-β-LG for inhibiting HPV infection in the cervical mucosa ( Figure 6C) .
TextSentencer_T179 28102-28259 Sentence denotes Based on the results obtained from this study, we proposed a mechanistic model of 3HP-β-LG for inhibiting HPV infection in the cervical mucosa ( Figure 6C) .
TextSentencer_T180 28260-28548 Sentence denotes The newly invaded HPV accesses the basal cell layer through micro lesions or breaks in the cervical epithelium and binds to the HSPG receptor on the basement membrane through interaction between the positively charged region in L1 protein of HPV and the negatively charged region in HSPG.
TextSentencer_T180 28260-28548 Sentence denotes The newly invaded HPV accesses the basal cell layer through micro lesions or breaks in the cervical epithelium and binds to the HSPG receptor on the basement membrane through interaction between the positively charged region in L1 protein of HPV and the negatively charged region in HSPG.
TextSentencer_T181 28549-28799 Sentence denotes This interaction results in a series of conformational changes in the capsid, which leads to protease digestion of L2 and exposure of its N terminus, thus initiating entry of HPV into to the epithelial cells (Kines et al., 2009; Pyeon et al., 2009) .
TextSentencer_T181 28549-28799 Sentence denotes This interaction results in a series of conformational changes in the capsid, which leads to protease digestion of L2 and exposure of its N terminus, thus initiating entry of HPV into to the epithelial cells (Kines et al., 2009; Pyeon et al., 2009) .
TextSentencer_T182 28800-28991 Sentence denotes After HPV infects the epithelial cell, it gradually matures along with the host cell cycles and moves toward the top layer of epithelia, followed by eventual release from the epidermal cells.
TextSentencer_T182 28800-28991 Sentence denotes After HPV infects the epithelial cell, it gradually matures along with the host cell cycles and moves toward the top layer of epithelia, followed by eventual release from the epidermal cells.
TextSentencer_T183 28992-29275 Sentence denotes The newly released A B FIGURE 5 | 3HP-β-LG could not enter into blood circulation when applied through the vagina or anus. (A) Concentration of 3HP-β-LG in sera of monkeys through vaginal administration. (B) Concentration of 3HP-β-LG in sera of monkeys through rectal administration.
TextSentencer_T183 28992-29275 Sentence denotes The newly released A B FIGURE 5 | 3HP-β-LG could not enter into blood circulation when applied through the vagina or anus. (A) Concentration of 3HP-β-LG in sera of monkeys through vaginal administration. (B) Concentration of 3HP-β-LG in sera of monkeys through rectal administration.
TextSentencer_T184 29276-29395 Sentence denotes 3HP-β-LG was formulated in a carbomer gel (1.8 mg/each) and administered through the vagina or anus of rhesus macaques.
TextSentencer_T184 29276-29395 Sentence denotes 3HP-β-LG was formulated in a carbomer gel (1.8 mg/each) and administered through the vagina or anus of rhesus macaques.
TextSentencer_T185 29396-29502 Sentence denotes Their blood samples were collected at different time points before and after the application (−1 to 24 h).
TextSentencer_T185 29396-29502 Sentence denotes Their blood samples were collected at different time points before and after the application (−1 to 24 h).
TextSentencer_T186 29503-29555 Sentence denotes 3HP-β-LG in the blood samples was analyzed by ELISA.
TextSentencer_T186 29503-29555 Sentence denotes 3HP-β-LG in the blood samples was analyzed by ELISA.
TextSentencer_T187 29556-29632 Sentence denotes Each sample was tested in triplicate, and the experiment was repeated twice.
TextSentencer_T187 29556-29632 Sentence denotes Each sample was tested in triplicate, and the experiment was repeated twice.
TextSentencer_T188 29633-29698 Sentence denotes Data from a representative experiment are presented as mean ± SD.
TextSentencer_T188 29633-29698 Sentence denotes Data from a representative experiment are presented as mean ± SD.
TextSentencer_T189 29699-29784 Sentence denotes The asterisks represent significant differences: *p < 0.05; **p < 0.01; ***p < 0.001.
TextSentencer_T189 29699-29784 Sentence denotes The asterisks represent significant differences: *p < 0.05; **p < 0.01; ***p < 0.001.
TextSentencer_T190 29785-30069 Sentence denotes Frontiers in Microbiology | www.frontiersin.org HPV particles access the basement membrane through micro lesions of the cervical epithelium and start another infection and replication cycle, gradually establishing persistent infection and promoting the development of cervical cancer.
TextSentencer_T190 29785-30069 Sentence denotes Frontiers in Microbiology | www.frontiersin.org HPV particles access the basement membrane through micro lesions of the cervical epithelium and start another infection and replication cycle, gradually establishing persistent infection and promoting the development of cervical cancer.
TextSentencer_T191 30070-30323 Sentence denotes 3HP-β-LG that is topically applied in the vagina can bind to the L1 protein of the newly invaded or the newly released HPV and block the attachment of HPV to the basement membrane, thereby inhibiting the entry of HPV into the host cells for replication.
TextSentencer_T191 30070-30323 Sentence denotes 3HP-β-LG that is topically applied in the vagina can bind to the L1 protein of the newly invaded or the newly released HPV and block the attachment of HPV to the basement membrane, thereby inhibiting the entry of HPV into the host cells for replication.
TextSentencer_T192 30324-30701 Sentence denotes After treatment with 3HP-β-LG for a certain period of time (e.g., 2 months), the newly produced HPV-free epithelial cells in the lower part of the mucosal epithelium are expected to move upward and push the HPV-infected cells away from the vaginal mucosa (this movement is accelerated during the menstrual period), making all layers of mucosal epithelium free of HPV infection.
TextSentencer_T192 30324-30701 Sentence denotes After treatment with 3HP-β-LG for a certain period of time (e.g., 2 months), the newly produced HPV-free epithelial cells in the lower part of the mucosal epithelium are expected to move upward and push the HPV-infected cells away from the vaginal mucosa (this movement is accelerated during the menstrual period), making all layers of mucosal epithelium free of HPV infection.
TextSentencer_T193 30702-31124 Sentence denotes Considering that the presence of micro lesions or breaks in the cervical epithelium, possibly caused by sexual activity, is the requirement for HPV to access the HSPG receptor on basement membrane (Bousarghin et al., 2003) , combinational use of some biocompatible materials for mucosa wound healing, such as human collagen proteins (Hua et al., 2019) , may have synergistic or complementary effects against HPV infection.
TextSentencer_T193 30702-31124 Sentence denotes Considering that the presence of micro lesions or breaks in the cervical epithelium, possibly caused by sexual activity, is the requirement for HPV to access the HSPG receptor on basement membrane (Bousarghin et al., 2003) , combinational use of some biocompatible materials for mucosa wound healing, such as human collagen proteins (Hua et al., 2019) , may have synergistic or complementary effects against HPV infection.
TextSentencer_T194 31125-31405 Sentence denotes The results from the time-of-addition assay, cell washout assay, and virus inactivation indicate that 3HP-β-LG could interact with the virions, rather than the cells, to effectively block the entry of HPV PsV into the target cell, but could not inactivate the virions (Figure 3) .
TextSentencer_T194 31125-31405 Sentence denotes The results from the time-of-addition assay, cell washout assay, and virus inactivation indicate that 3HP-β-LG could interact with the virions, rather than the cells, to effectively block the entry of HPV PsV into the target cell, but could not inactivate the virions (Figure 3) .
TextSentencer_T195 31406-31670 Sentence denotes These biological properties of 3HP-β-LG make it an ideal anti-HPV agent for topical application to treat HPV infection because it can interact with HPV particles on the vaginal mucosa and block the virions binding to the receptor, e.g., HSPG, on basement membrane.
TextSentencer_T195 31406-31670 Sentence denotes These biological properties of 3HP-β-LG make it an ideal anti-HPV agent for topical application to treat HPV infection because it can interact with HPV particles on the vaginal mucosa and block the virions binding to the receptor, e.g., HSPG, on basement membrane.
TextSentencer_T196 31671-31841 Sentence denotes Since 3HP-β-LG cannot enter into the cell or the blood circulation, it is not expected to cause systemic toxicity to human or induce harmful 3HP-β-LG-specific antibodies.
TextSentencer_T196 31671-31841 Sentence denotes Since 3HP-β-LG cannot enter into the cell or the blood circulation, it is not expected to cause systemic toxicity to human or induce harmful 3HP-β-LG-specific antibodies.
TextSentencer_T197 31842-31946 Sentence denotes Therefore, the topical formulations containing 3HP-β-LG can be safely used to treat local HPV infection.
TextSentencer_T197 31842-31946 Sentence denotes Therefore, the topical formulations containing 3HP-β-LG can be safely used to treat local HPV infection.
TextSentencer_T198 31947-32068 Sentence denotes The increased net negative charges on β-lactoglobulin after modification of the protein with 3-hydroxyphthalic anhydride.
TextSentencer_T198 31947-32068 Sentence denotes The increased net negative charges on β-lactoglobulin after modification of the protein with 3-hydroxyphthalic anhydride.
TextSentencer_T199 32069-32184 Sentence denotes The structure of 3HP-β-LG was modeled by COOT (Emsley et al., 2010) using the crystal structure of β-LG (PDB entry:
TextSentencer_T199 32069-32184 Sentence denotes The structure of 3HP-β-LG was modeled by COOT (Emsley et al., 2010) using the crystal structure of β-LG (PDB entry:
TextSentencer_T200 32185-32191 Sentence denotes 1BEB).
TextSentencer_T200 32185-32191 Sentence denotes 1BEB).
TextSentencer_T201 32192-32346 Sentence denotes The structures were shown as an electrostatic surface by Pymol (Alexander et al., 2011) . (B) Predicted interactions between HPV L1 pentamer and 3HP-β-LG.
TextSentencer_T201 32192-32346 Sentence denotes The structures were shown as an electrostatic surface by Pymol (Alexander et al., 2011) . (B) Predicted interactions between HPV L1 pentamer and 3HP-β-LG.
TextSentencer_T202 32347-32406 Sentence denotes Crystal structure of HSPG-bound HPV L1 pentamer (PDB entry:
TextSentencer_T202 32347-32406 Sentence denotes Crystal structure of HSPG-bound HPV L1 pentamer (PDB entry:
TextSentencer_T203 32407-32661 Sentence denotes 5W1O) was shown as electrostatic surface (Dasgupta et al., 2011) , and the docking structure between HPV L1 and 3HP-β-LG was generated by AutoDock (Trott and Olson, 2010) . (C) Schematic illustration showing how 3HP-β-LG inhibits HPV entry and infection.
TextSentencer_T203 32407-32661 Sentence denotes 5W1O) was shown as electrostatic surface (Dasgupta et al., 2011) , and the docking structure between HPV L1 and 3HP-β-LG was generated by AutoDock (Trott and Olson, 2010) . (C) Schematic illustration showing how 3HP-β-LG inhibits HPV entry and infection.
TextSentencer_T204 32662-32909 Sentence denotes The newly invaded or newly released HPV particles access the basal layer through the mucosal lesion, or breaks, and bind to the HSPG receptor on the basement membrane, resulting in the entry of HPV into and replication in mucosal epithelial cells.
TextSentencer_T204 32662-32909 Sentence denotes The newly invaded or newly released HPV particles access the basal layer through the mucosal lesion, or breaks, and bind to the HSPG receptor on the basement membrane, resulting in the entry of HPV into and replication in mucosal epithelial cells.
TextSentencer_T205 32910-33198 Sentence denotes 3HP-β-LG can bind with HPV L1 protein, through the electrostatic interaction between the negatively charged region in 3HP-β-LG and the positively charged region in L1 protein, to block the attachment of HPV to the HSPG receptor, resulting in inhibition of HPV infection in vaginal mucosa.
TextSentencer_T205 32910-33198 Sentence denotes 3HP-β-LG can bind with HPV L1 protein, through the electrostatic interaction between the negatively charged region in 3HP-β-LG and the positively charged region in L1 protein, to block the attachment of HPV to the HSPG receptor, resulting in inhibition of HPV infection in vaginal mucosa.
TextSentencer_T206 33199-33288 Sentence denotes All datasets generated for this study are included in the manuscript/supplementary files.
TextSentencer_T206 33199-33288 Sentence denotes All datasets generated for this study are included in the manuscript/supplementary files.
TextSentencer_T207 33289-33480 Sentence denotes The animal study was reviewed and approved by Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences and Peking Union Medical College (approval number: ILAS-VL-2015-001).
TextSentencer_T207 33289-33480 Sentence denotes The animal study was reviewed and approved by Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences and Peking Union Medical College (approval number: ILAS-VL-2015-001).
TextSentencer_T208 33481-33536 Sentence denotes SJ, LSu, and YZ conceived and designed the experiments.
TextSentencer_T208 33481-33536 Sentence denotes SJ, LSu, and YZ conceived and designed the experiments.
TextSentencer_T209 33537-33583 Sentence denotes CH, CW, LSi, and QW conducted the experiments.
TextSentencer_T209 33537-33583 Sentence denotes CH, CW, LSi, and QW conducted the experiments.
TextSentencer_T210 33584-33687 Sentence denotes YZ performed the computer modeling and analysis of the interaction between 3HP-β-LG and HPV L1 protein.
TextSentencer_T210 33584-33687 Sentence denotes YZ performed the computer modeling and analysis of the interaction between 3HP-β-LG and HPV L1 protein.
TextSentencer_T211 33688-33751 Sentence denotes CH, YZ, LSu, and SJ analyzed the data and wrote the manuscript.
TextSentencer_T211 33688-33751 Sentence denotes CH, YZ, LSu, and SJ analyzed the data and wrote the manuscript.
TextSentencer_T212 33752-33808 Sentence denotes All authors have read and approved the final manuscript.
TextSentencer_T212 33752-33808 Sentence denotes All authors have read and approved the final manuscript.
TextSentencer_T213 33809-33929 Sentence denotes This work was supported by an intramural fund of Fudan-Jinbo Functional Protein Joint Research Center, Fudan University.
TextSentencer_T213 33809-33929 Sentence denotes This work was supported by an intramural fund of Fudan-Jinbo Functional Protein Joint Research Center, Fudan University.

CORD-PICO

Id Subject Object Predicate Lexical cue
p2_i0 28-91 Pharmacological denotes 3-Hydroxyphthalic Anhydride- Modified Bovine Beta-Lactoglobulin
p1_all0 28-91 Intervention denotes 3-Hydroxyphthalic Anhydride- Modified Bovine Beta-Lactoglobulin
p2_i1 194-273 Pharmacological denotes 3-hydroxyphthalic anhydride (3HP)-modified bovine beta-lactoglobulin (3HP-β-LG)
p1_all1 194-273 Intervention denotes 3-hydroxyphthalic anhydride (3HP)-modified bovine beta-lactoglobulin (3HP-β-LG)
p2_i2 428-459 Pharmacological denotes 3HP-β-LG-containing vaginal gel
p1_all2 428-459 Intervention denotes 3HP-β-LG-containing vaginal gel
p2_o2 488-501 Physical denotes HPV infection
p1_all3 488-523 Outcome denotes HPV infection and reduce viral load
p1_all4 628-636 Intervention denotes 3HP-β-LG
p1_all5 700-708 Intervention denotes 3HP-β-LG
p2_i3 1193-1233 Pharmacological denotes 3HP-modified chicken ovalbumin (3HP-OVA)
p1_all6 1193-1233 Intervention denotes 3HP-modified chicken ovalbumin (3HP-OVA)
p2_o6 1290-1307 Physical denotes anti-HPV activity
p1_all7 1508-1516 Intervention denotes 3HP-β-LG
p2_o7 1535-1565 Physical denotes host cell or blood circulation