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CORD-19:bcef0894cf382a83c2aa4d9c803978780c2809db JSONTXT 8 Projects

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Id Subject Object Predicate Lexical cue
TextSentencer_T1 0-106 Sentence denotes The MHC class-II HLA-DR receptor mediates bat influenza A-like H17N10 virus entry 1 into mammalian cells 2
TextSentencer_T1 0-106 Sentence denotes The MHC class-II HLA-DR receptor mediates bat influenza A-like H17N10 virus entry 1 into mammalian cells 2
TextSentencer_T2 108-116 Sentence denotes Abstract
TextSentencer_T2 108-116 Sentence denotes Abstract
TextSentencer_T3 117-288 Sentence denotes Bats are notorious reservoirs of diverse, potentially zoonotic viruses, exemplified by the 20 evolutionarily distinct, influenza A-like viruses H17N10 and H18N11 (BatIVs).
TextSentencer_T3 117-288 Sentence denotes Bats are notorious reservoirs of diverse, potentially zoonotic viruses, exemplified by the 20 evolutionarily distinct, influenza A-like viruses H17N10 and H18N11 (BatIVs).
TextSentencer_T4 289-565 Sentence denotes The surface 21 glycoproteins [haemagglutinin (H) and neuraminidase (N)] of BatIVs neither bind nor cleave 22 sialic acid receptors, which suggests that these viruses employ cell attachment and entry 23 mechanisms that differ from those of classical influenza A viruses (IAVs).
TextSentencer_T4 289-565 Sentence denotes The surface 21 glycoproteins [haemagglutinin (H) and neuraminidase (N)] of BatIVs neither bind nor cleave 22 sialic acid receptors, which suggests that these viruses employ cell attachment and entry 23 mechanisms that differ from those of classical influenza A viruses (IAVs).
TextSentencer_T5 566-708 Sentence denotes Identifying the 24 cellular factors that mediate entry and determine susceptibility to infection will help assess 25 the host range of BatIVs.
TextSentencer_T5 566-708 Sentence denotes Identifying the 24 cellular factors that mediate entry and determine susceptibility to infection will help assess 25 the host range of BatIVs.
TextSentencer_T6 709-797 Sentence denotes Here, we investigated a range of cell lines from different species for 26 author/funder.
TextSentencer_T6 709-797 Sentence denotes Here, we investigated a range of cell lines from different species for 26 author/funder.
TextSentencer_T7 798-818 Sentence denotes All rights reserved.
TextSentencer_T7 798-818 Sentence denotes All rights reserved.
TextSentencer_T8 819-855 Sentence denotes No reuse allowed without permission.
TextSentencer_T8 819-855 Sentence denotes No reuse allowed without permission.
TextSentencer_T9 856-1102 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint their susceptibility to infection by pseudotyped viruses (PV) bearing bat H17 and/or N10 27 envelope glycoproteins.
TextSentencer_T9 856-1102 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint their susceptibility to infection by pseudotyped viruses (PV) bearing bat H17 and/or N10 27 envelope glycoproteins.
TextSentencer_T10 1103-1270 Sentence denotes We show that a number of human haematopoietic cancer cell lines 28 and the canine kidney MDCK II (but not MDCK I) cells are susceptible to H17-pseudotypes 29 (H17-PV).
TextSentencer_T10 1103-1270 Sentence denotes We show that a number of human haematopoietic cancer cell lines 28 and the canine kidney MDCK II (but not MDCK I) cells are susceptible to H17-pseudotypes 29 (H17-PV).
TextSentencer_T11 1271-1513 Sentence denotes We observed with microarrays and qRT-PCR that the dog leukocyte antigen DLA-30 DRA mRNA is over expressed in late passaged parental MDCK and commercial MDCK II cells, 31 compared to early passaged parental MDCK and MDCK I cells, respectively.
TextSentencer_T11 1271-1513 Sentence denotes We observed with microarrays and qRT-PCR that the dog leukocyte antigen DLA-30 DRA mRNA is over expressed in late passaged parental MDCK and commercial MDCK II cells, 31 compared to early passaged parental MDCK and MDCK I cells, respectively.
TextSentencer_T12 1514-1630 Sentence denotes The human 32 orthologue HLA-DRA encodes the alpha subunit of the MHC class II HLA-DR antigen-binding 33 heterodimer.
TextSentencer_T12 1514-1630 Sentence denotes The human 32 orthologue HLA-DRA encodes the alpha subunit of the MHC class II HLA-DR antigen-binding 33 heterodimer.
TextSentencer_T13 1631-1765 Sentence denotes Small interfering RNA-or neutralizing antibody-targeting HLA-DRA, drastically 34 reduced the susceptibility of Raji B cells to H17-PV.
TextSentencer_T13 1631-1765 Sentence denotes Small interfering RNA-or neutralizing antibody-targeting HLA-DRA, drastically 34 reduced the susceptibility of Raji B cells to H17-PV.
TextSentencer_T14 1766-1926 Sentence denotes Conversely, over expression of HLA-DRA 35 and its paralogue HLA-DRB1 on the surface of the unsusceptible HEK293T/17 cells conferred 36 susceptibility to H17-PV.
TextSentencer_T14 1766-1926 Sentence denotes Conversely, over expression of HLA-DRA 35 and its paralogue HLA-DRB1 on the surface of the unsusceptible HEK293T/17 cells conferred 36 susceptibility to H17-PV.
TextSentencer_T15 1927-2105 Sentence denotes The identification of HLA-DR as an H17N10 entry mediator will 37 contribute to a better understanding of the tropism of the virus and will elucidate its zoonotic 38 transmission.
TextSentencer_T15 1927-2105 Sentence denotes The identification of HLA-DR as an H17N10 entry mediator will 37 contribute to a better understanding of the tropism of the virus and will elucidate its zoonotic 38 transmission.
TextSentencer_T16 2106-2111 Sentence denotes 39 40
TextSentencer_T16 2106-2111 Sentence denotes 39 40
TextSentencer_T17 2113-2142 Sentence denotes to accommodate SA 2, 15, 17 .
TextSentencer_T17 2113-2142 Sentence denotes to accommodate SA 2, 15, 17 .
TextSentencer_T18 2143-2295 Sentence denotes The cell receptors for the BatIVs are as yet unidentified, but they 79 are clearly not SA moieties, a conclusion reached by several studies 15, 18, 19 .
TextSentencer_T18 2143-2295 Sentence denotes The cell receptors for the BatIVs are as yet unidentified, but they 79 are clearly not SA moieties, a conclusion reached by several studies 15, 18, 19 .
TextSentencer_T19 2296-2500 Sentence denotes Furthermore, bat 80 N10 and N11 are structurally similar to classical NAs but lack conserved amino acids for SA 81 binding and cleavage 2, 16, 17 and do not exhibit typical neuraminidase activity 18, 20 .
TextSentencer_T19 2296-2500 Sentence denotes Furthermore, bat 80 N10 and N11 are structurally similar to classical NAs but lack conserved amino acids for SA 81 binding and cleavage 2, 16, 17 and do not exhibit typical neuraminidase activity 18, 20 .
TextSentencer_T20 2501-2612 Sentence denotes Initial 82 efforts to isolate infectious BatIVs directly from bats have failed, mainly because the receptors 83
TextSentencer_T20 2501-2612 Sentence denotes Initial 82 efforts to isolate infectious BatIVs directly from bats have failed, mainly because the receptors 83
TextSentencer_T21 2613-2682 Sentence denotes were unknown and susceptible cell lines were unavailable 21, 22, 23 .
TextSentencer_T21 2613-2682 Sentence denotes were unknown and susceptible cell lines were unavailable 21, 22, 23 .
TextSentencer_T22 2683-3024 Sentence denotes Attempts to circumvent 84 these limitations have included H17-or H18-pseudotyped vesicular stomatitis virus (VSV 19, 85 DLA-DRA is a well-conserved orthologue of the human leukocyte antigen class II DR α-chain 157 (HLA-DRA) (∼90% amino acid identity between canine, human and Desmodus rotundus bat 158 ectodomains; Supplementary material 3).
TextSentencer_T22 2683-3024 Sentence denotes Attempts to circumvent 84 these limitations have included H17-or H18-pseudotyped vesicular stomatitis virus (VSV 19, 85 DLA-DRA is a well-conserved orthologue of the human leukocyte antigen class II DR α-chain 157 (HLA-DRA) (∼90% amino acid identity between canine, human and Desmodus rotundus bat 158 ectodomains; Supplementary material 3).
TextSentencer_T23 3025-3298 Sentence denotes In humans, MHC-II molecules occur as three highly 159 polymorphic isotypes (HLA-DR, HLA-DP and HLA-DQ) which are selectively expressed 160 under normal conditions on the surface of antigen presenting cells (APCs), including B, 161 dendritic and mononuclear phagocyte cells.
TextSentencer_T23 3025-3298 Sentence denotes In humans, MHC-II molecules occur as three highly 159 polymorphic isotypes (HLA-DR, HLA-DP and HLA-DQ) which are selectively expressed 160 under normal conditions on the surface of antigen presenting cells (APCs), including B, 161 dendritic and mononuclear phagocyte cells.
TextSentencer_T24 3299-3498 Sentence denotes These molecules are non-covalently associated 162 heterodimers of two glycosylated, transmembrane polypeptide chains, the monomorphic 163 35-kDa α-chain and the highly polymorphic 28-kDa β-chain 36 .
TextSentencer_T24 3299-3498 Sentence denotes These molecules are non-covalently associated 162 heterodimers of two glycosylated, transmembrane polypeptide chains, the monomorphic 163 35-kDa α-chain and the highly polymorphic 28-kDa β-chain 36 .
TextSentencer_T25 3499-3695 Sentence denotes Both chains have an 164 extracellular portion composed of two domains (α1 and α2, or β1 and β2) that is anchored 165 on the cell membrane by short transmembrane and cytoplasmic domains (Fig. 2c ).
TextSentencer_T25 3499-3695 Sentence denotes Both chains have an 164 extracellular portion composed of two domains (α1 and α2, or β1 and β2) that is anchored 165 on the cell membrane by short transmembrane and cytoplasmic domains (Fig. 2c ).
TextSentencer_T26 3696-3916 Sentence denotes In the 166 classical scenario, the protease-derived foreign antigen peptides bind to MHC class II proteins 167 in the cleft formed by the α1 and β1 domains, and the complex is transported to the cell 168 surface 36, 37 .
TextSentencer_T26 3696-3916 Sentence denotes In the 166 classical scenario, the protease-derived foreign antigen peptides bind to MHC class II proteins 167 in the cleft formed by the α1 and β1 domains, and the complex is transported to the cell 168 surface 36, 37 .
TextSentencer_T27 3917-4106 Sentence denotes When antigenic peptides are not available, endogenous peptides such as the class 169 II associated invariant peptide (CLIP) substitute them and restore MHC class II dimer 170 stability 37 .
TextSentencer_T27 3917-4106 Sentence denotes When antigenic peptides are not available, endogenous peptides such as the class 169 II associated invariant peptide (CLIP) substitute them and restore MHC class II dimer 170 stability 37 .
TextSentencer_T28 4107-4368 Sentence denotes The complex of HLA-DR and endocytosed peptides (usually 9-30 amino acids in 171 length), constitutes a ligand for the T-cell receptor (TCR) and plays a key role in the 172 presentation of foreign antigens to CD4 + T helper cells and immune surveillance 38, 39 .
TextSentencer_T28 4107-4368 Sentence denotes The complex of HLA-DR and endocytosed peptides (usually 9-30 amino acids in 171 length), constitutes a ligand for the T-cell receptor (TCR) and plays a key role in the 172 presentation of foreign antigens to CD4 + T helper cells and immune surveillance 38, 39 .
TextSentencer_T29 4369-4372 Sentence denotes 173
TextSentencer_T29 4369-4372 Sentence denotes 173
TextSentencer_T30 4373-4555 Sentence denotes Since the ultimate goal of our studies is to obtain insight on the zoonotic potential of H17N10, 175 we focused on the possible influence of HLA-DR on cellular susceptibility to H17.
TextSentencer_T30 4373-4555 Sentence denotes Since the ultimate goal of our studies is to obtain insight on the zoonotic potential of H17N10, 175 we focused on the possible influence of HLA-DR on cellular susceptibility to H17.
TextSentencer_T31 4556-4775 Sentence denotes Hence, 176 taking advantage of the high expression of HLA-DR on certain human hematopoietic 177 carcinomas 40 , we further explored H17-PV tropism using a panel of human leukaemia and 178 lymphoma cell lines (Fig. 2d) .
TextSentencer_T31 4556-4775 Sentence denotes Hence, 176 taking advantage of the high expression of HLA-DR on certain human hematopoietic 177 carcinomas 40 , we further explored H17-PV tropism using a panel of human leukaemia and 178 lymphoma cell lines (Fig. 2d) .
TextSentencer_T32 4776-4960 Sentence denotes We found that the Burkitt's lymphoma-derived Raji, Ramos and 179 BJAB B-lymphocytes and the B lymphoblastoid cells (B-LCL) show decreasing susceptibility, 180 in that order, to H17-PV.
TextSentencer_T32 4776-4960 Sentence denotes We found that the Burkitt's lymphoma-derived Raji, Ramos and 179 BJAB B-lymphocytes and the B lymphoblastoid cells (B-LCL) show decreasing susceptibility, 180 in that order, to H17-PV.
TextSentencer_T33 4961-5243 Sentence denotes Kasumi-1 leukaemic cells showed marginal susceptibility in terms of 181 luciferase activity, while Molt-4 and HL-60 leukaemic cells, Jurkat T-cells, pro-monocytic THP-182 1 and U-937 cells, and primary B cells showed no susceptibility to the pseudotypes (Fig. 2d, 183 left Y axis).
TextSentencer_T33 4961-5243 Sentence denotes Kasumi-1 leukaemic cells showed marginal susceptibility in terms of 181 luciferase activity, while Molt-4 and HL-60 leukaemic cells, Jurkat T-cells, pro-monocytic THP-182 1 and U-937 cells, and primary B cells showed no susceptibility to the pseudotypes (Fig. 2d, 183 left Y axis).
TextSentencer_T34 5244-5511 Sentence denotes We hypothesised that the different susceptibility of the various cell types by H17-184 PV were due to disparate expression of HLA-DR, confirmed by qRT-PCR analysis for a non-185 polymorphic region of the α chain of HLA-DR on the same samples (Fig. 2d , right Y-axis).
TextSentencer_T34 5244-5511 Sentence denotes We hypothesised that the different susceptibility of the various cell types by H17-184 PV were due to disparate expression of HLA-DR, confirmed by qRT-PCR analysis for a non-185 polymorphic region of the α chain of HLA-DR on the same samples (Fig. 2d , right Y-axis).
TextSentencer_T35 5512-5519 Sentence denotes The 186
TextSentencer_T35 5512-5519 Sentence denotes The 186
TextSentencer_T36 5520-5850 Sentence denotes presence of the HLA-DR heterodimer was also confirmed by flow cytometry with a FITC-187 conjugated monoclonal antibody (clone Tü36), which specifically binds to a monomorphic 188 epitope on the HLA-DR α/β complex and not the isolated α or β chains (Fig. 2e) cells correspondingly reduced the infection of H17-PV by 50% (Fig. 2f) .
TextSentencer_T36 5520-5850 Sentence denotes presence of the HLA-DR heterodimer was also confirmed by flow cytometry with a FITC-187 conjugated monoclonal antibody (clone Tü36), which specifically binds to a monomorphic 188 epitope on the HLA-DR α/β complex and not the isolated α or β chains (Fig. 2e) cells correspondingly reduced the infection of H17-PV by 50% (Fig. 2f) .
TextSentencer_T37 5851-5854 Sentence denotes 203
TextSentencer_T37 5851-5854 Sentence denotes 203
TextSentencer_T38 5855-5951 Sentence denotes To determine if blocking attachment of virus to the HLA-DR ectodomain can prevent its entry, 205
TextSentencer_T38 5855-5951 Sentence denotes To determine if blocking attachment of virus to the HLA-DR ectodomain can prevent its entry, 205
TextSentencer_T39 5952-6148 Sentence denotes Raji cells were incubated with increasing concentrations of a monoclonal antibody (mAb 206 Clone 302CT2.3.2) targeting a monomorphic, extracellular region of the HLA-DRA antigen 207 author/funder.
TextSentencer_T39 5952-6148 Sentence denotes Raji cells were incubated with increasing concentrations of a monoclonal antibody (mAb 206 Clone 302CT2.3.2) targeting a monomorphic, extracellular region of the HLA-DRA antigen 207 author/funder.
TextSentencer_T40 6149-6169 Sentence denotes All rights reserved.
TextSentencer_T40 6149-6169 Sentence denotes All rights reserved.
TextSentencer_T41 6170-6206 Sentence denotes No reuse allowed without permission.
TextSentencer_T41 6170-6206 Sentence denotes No reuse allowed without permission.
TextSentencer_T42 6207-6375 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint (HLA-DRA epitope: amino acids 48-75).
TextSentencer_T42 6207-6375 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint (HLA-DRA epitope: amino acids 48-75).
TextSentencer_T43 6376-6510 Sentence denotes The presence of the antibody significantly reduced, in 208 a dose-dependent manner, infection with H17-PV but not VSV-G-PV (Fig. 2g) .
TextSentencer_T43 6376-6510 Sentence denotes The presence of the antibody significantly reduced, in 208 a dose-dependent manner, infection with H17-PV but not VSV-G-PV (Fig. 2g) .
TextSentencer_T44 6511-6514 Sentence denotes 209
TextSentencer_T44 6511-6514 Sentence denotes 209
TextSentencer_T45 6515-6675 Sentence denotes We next sought to ascertain whether ectopic expression of HLA-DR was sufficient to increase 211 the susceptibility of non-APC, HEK293T/17, cells to the H17-PVs.
TextSentencer_T45 6515-6675 Sentence denotes We next sought to ascertain whether ectopic expression of HLA-DR was sufficient to increase 211 the susceptibility of non-APC, HEK293T/17, cells to the H17-PVs.
TextSentencer_T46 6676-6792 Sentence denotes HEK293T/17 cells were 212 transiently transfected with the DRA expression vector, alone or in combination with DRB1.
TextSentencer_T46 6676-6792 Sentence denotes HEK293T/17 cells were 212 transiently transfected with the DRA expression vector, alone or in combination with DRB1.
TextSentencer_T47 6793-6955 Sentence denotes Taken together, HLA-DR is shown to function as a bona fide entry mediator for H17 but may 226 function with unknown factors that facilitate virus internalisation.
TextSentencer_T47 6793-6955 Sentence denotes Taken together, HLA-DR is shown to function as a bona fide entry mediator for H17 but may 226 function with unknown factors that facilitate virus internalisation.
TextSentencer_T48 6956-7196 Sentence denotes Interaction between HLA-227 DR and H17 may trigger viral entry through canonical receptor-mediated endocytosis, but 228 could also trigger entry through an activation of cell signalling pathways that the virus 229 subverts to its advantage.
TextSentencer_T48 6956-7196 Sentence denotes Interaction between HLA-227 DR and H17 may trigger viral entry through canonical receptor-mediated endocytosis, but 228 could also trigger entry through an activation of cell signalling pathways that the virus 229 subverts to its advantage.
TextSentencer_T49 7197-7385 Sentence denotes Our finding therefore raises questions on the utility and possible 230 evolutionary advantage(s) that an APC-associated receptor would confer to H17N10 231 infectivity and broader fitness.
TextSentencer_T49 7197-7385 Sentence denotes Our finding therefore raises questions on the utility and possible 230 evolutionary advantage(s) that an APC-associated receptor would confer to H17N10 231 infectivity and broader fitness.
TextSentencer_T50 7386-7553 Sentence denotes Some viruses exploit cells of the immune system, such as 232 macrophages, B and dendritic cells, either as viral reservoirs or as "Trojan horses" to 233 author/funder.
TextSentencer_T50 7386-7553 Sentence denotes Some viruses exploit cells of the immune system, such as 232 macrophages, B and dendritic cells, either as viral reservoirs or as "Trojan horses" to 233 author/funder.
TextSentencer_T51 7554-7574 Sentence denotes All rights reserved.
TextSentencer_T51 7554-7574 Sentence denotes All rights reserved.
TextSentencer_T52 7575-7611 Sentence denotes No reuse allowed without permission.
TextSentencer_T52 7575-7611 Sentence denotes No reuse allowed without permission.
TextSentencer_T53 7612-7785 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint penetrate the epithelial barriers 44, 45 .
TextSentencer_T53 7612-7785 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint penetrate the epithelial barriers 44, 45 .
TextSentencer_T54 7786-8044 Sentence denotes The measles virus for example, exploits macrophages or 234 dendritic cells, which traffic the virus to bronchus-associated lymphoid tissue or regional 235 lymph nodes, resulting in local amplification and subsequent systemic dissemination by 236 viremia 46 .
TextSentencer_T54 7786-8044 Sentence denotes The measles virus for example, exploits macrophages or 234 dendritic cells, which traffic the virus to bronchus-associated lymphoid tissue or regional 235 lymph nodes, resulting in local amplification and subsequent systemic dissemination by 236 viremia 46 .
TextSentencer_T55 8045-8567 Sentence denotes A similar strategy employed by H17N10 could provide an explanation on why viral 237 RNA was detected in different organs and tissues in carrier S. lilium bats (i.e. lung, kidney, 238 liver, intestine) and why the virus fails to grow in vitro in cell lines developed from the same 239 In this study we did not establish the stoichiometry of the HLA-DR: H17 engagement, or 250 clarify how the virus moves to sub-membranous regions and might hijack the receptor-251 mediated signalling pathway to promote its internalization.
TextSentencer_T55 8045-8567 Sentence denotes A similar strategy employed by H17N10 could provide an explanation on why viral 237 RNA was detected in different organs and tissues in carrier S. lilium bats (i.e. lung, kidney, 238 liver, intestine) and why the virus fails to grow in vitro in cell lines developed from the same 239 In this study we did not establish the stoichiometry of the HLA-DR: H17 engagement, or 250 clarify how the virus moves to sub-membranous regions and might hijack the receptor-251 mediated signalling pathway to promote its internalization.
TextSentencer_T56 8568-8651 Sentence denotes It is likely that the determinants 252 of viral entry in vivo are more complicated.
TextSentencer_T56 8568-8651 Sentence denotes It is likely that the determinants 252 of viral entry in vivo are more complicated.
TextSentencer_T57 8652-8806 Sentence denotes We cannot rule out the use by H17N10 (as by 253 other bat-borne viruses, e.g. SARS CoV 51, 52 ) of more than one molecular species as (co-) 254 receptors.
TextSentencer_T57 8652-8806 Sentence denotes We cannot rule out the use by H17N10 (as by 253 other bat-borne viruses, e.g. SARS CoV 51, 52 ) of more than one molecular species as (co-) 254 receptors.
TextSentencer_T58 8807-9009 Sentence denotes Nevertheless, the implication of this study is that H17N10 has the capacity to 255 enter human HLA-DR + cells and our work provides substantial evidence that the H17N10 256 virus has zoonotic potential.
TextSentencer_T58 8807-9009 Sentence denotes Nevertheless, the implication of this study is that H17N10 has the capacity to 255 enter human HLA-DR + cells and our work provides substantial evidence that the H17N10 256 virus has zoonotic potential.
TextSentencer_T59 9010-9182 Sentence denotes The current finding not only sheds light on the understanding 257 of BatIV host range, but also provides additional information on the evolution of influenza 258 A viruses.
TextSentencer_T59 9010-9182 Sentence denotes The current finding not only sheds light on the understanding 257 of BatIV host range, but also provides additional information on the evolution of influenza 258 A viruses.
TextSentencer_T60 9183-9201 Sentence denotes 259 author/funder.
TextSentencer_T60 9183-9201 Sentence denotes 259 author/funder.
TextSentencer_T61 9202-9222 Sentence denotes All rights reserved.
TextSentencer_T61 9202-9222 Sentence denotes All rights reserved.
TextSentencer_T62 9223-9259 Sentence denotes No reuse allowed without permission.
TextSentencer_T62 9223-9259 Sentence denotes No reuse allowed without permission.
TextSentencer_T63 9260-9390 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint
TextSentencer_T63 9260-9390 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint
TextSentencer_T64 9391-9623 Sentence denotes Cell lines (complete description in Supplementary material 1) were kindly provided as supplemented with heat-inactivated 15% fetal bovine serum (Life Technologies), penicillin 283 (100 U/ml) and streptomycin (100 μg/ml; Invitrogen).
TextSentencer_T64 9391-9623 Sentence denotes Cell lines (complete description in Supplementary material 1) were kindly provided as supplemented with heat-inactivated 15% fetal bovine serum (Life Technologies), penicillin 283 (100 U/ml) and streptomycin (100 μg/ml; Invitrogen).
TextSentencer_T65 9624-9673 Sentence denotes All cells were maintained in a 284 author/funder.
TextSentencer_T65 9624-9673 Sentence denotes All cells were maintained in a 284 author/funder.
TextSentencer_T66 9674-9694 Sentence denotes All rights reserved.
TextSentencer_T66 9674-9694 Sentence denotes All rights reserved.
TextSentencer_T67 9695-9731 Sentence denotes No reuse allowed without permission.
TextSentencer_T67 9695-9731 Sentence denotes No reuse allowed without permission.
TextSentencer_T68 9732-10043 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint humidified incubator at 37°C and 5% CO2 and were found free of mycoplasma contamination 285 on repeated testing with the MycoFluor Mycoplasma Detection Kit (Life Technologies, UK).
TextSentencer_T68 9732-10043 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint humidified incubator at 37°C and 5% CO2 and were found free of mycoplasma contamination 285 on repeated testing with the MycoFluor Mycoplasma Detection Kit (Life Technologies, UK).
TextSentencer_T69 10044-10047 Sentence denotes 286
TextSentencer_T69 10044-10047 Sentence denotes 286
TextSentencer_T70 10048-10543 Sentence denotes were either treated with the endosomal acidification reagent ammonium chloride (1, 10 or 288 100 mM), or pre-treated with neuraminidase from Clostridium perfrigens for 2 h (1, 10 or 100 289 mM) or pronase (a mixture of endo-and exoproteases from Streptomyces griseus at a final 290 concentration of 5, 10 or 50 µg/ml; Calbiochem, UK) for 30 min, an N-glycosylation inhibitor 291 (tunicamycin from Streptomyces sp. at a final concentration of 0.01, 0.1 or 1 µg/ml; Sigma-292 Aldrich, UK) for 5 h.
TextSentencer_T70 10048-10543 Sentence denotes were either treated with the endosomal acidification reagent ammonium chloride (1, 10 or 288 100 mM), or pre-treated with neuraminidase from Clostridium perfrigens for 2 h (1, 10 or 100 289 mM) or pronase (a mixture of endo-and exoproteases from Streptomyces griseus at a final 290 concentration of 5, 10 or 50 µg/ml; Calbiochem, UK) for 30 min, an N-glycosylation inhibitor 291 (tunicamycin from Streptomyces sp. at a final concentration of 0.01, 0.1 or 1 µg/ml; Sigma-292 Aldrich, UK) for 5 h.
TextSentencer_T71 10544-10686 Sentence denotes Pre-treated cells were washed with phosphate buffer saline (PBS) 3 293 times, and then incubated/infected as before with PVs for another 24 h.
TextSentencer_T71 10544-10686 Sentence denotes Pre-treated cells were washed with phosphate buffer saline (PBS) 3 293 times, and then incubated/infected as before with PVs for another 24 h.
TextSentencer_T72 10687-10754 Sentence denotes Cell viabilities were 294 assessed by a trypan blue exclusion test.
TextSentencer_T72 10687-10754 Sentence denotes Cell viabilities were 294 assessed by a trypan blue exclusion test.
TextSentencer_T73 10755-10762 Sentence denotes 295 296
TextSentencer_T73 10755-10762 Sentence denotes 295 296
TextSentencer_T74 10763-10850 Sentence denotes Pseudotypes expressing H17 and N10 genes were produced as described previously 32, 53 .
TextSentencer_T74 10763-10850 Sentence denotes Pseudotypes expressing H17 and N10 genes were produced as described previously 32, 53 .
TextSentencer_T75 10851-10854 Sentence denotes 298
TextSentencer_T75 10851-10854 Sentence denotes 298
TextSentencer_T76 10855-11140 Sentence denotes Briefly, the lentiviral packaging plasmid p8.91 54 , the pCSFLW firefly luciferase lentiviral 299 vector 55 or the GFP expressing vector pCSGW, the expression plasmids for H17 and/or N10 300 [vector pI.18 56 and the protease encoding plasmid pCAGGS-HAT (a kind gift by Eva Böttcher-301
TextSentencer_T76 10855-11140 Sentence denotes Briefly, the lentiviral packaging plasmid p8.91 54 , the pCSFLW firefly luciferase lentiviral 299 vector 55 or the GFP expressing vector pCSGW, the expression plasmids for H17 and/or N10 300 [vector pI.18 56 and the protease encoding plasmid pCAGGS-HAT (a kind gift by Eva Böttcher-301
TextSentencer_T77 11141-11379 Sentence denotes Friebertshäuser, Philipps University of Marburg, Germany) were co-transfected using 302 polyethylenimine transfection reagent (Sigma Aldrich, UK) into HEK293T/17 cells, plated on 303 6-well Nunclon © plates (Thermo Fisher Scientific, UK).
TextSentencer_T77 11141-11379 Sentence denotes Friebertshäuser, Philipps University of Marburg, Germany) were co-transfected using 302 polyethylenimine transfection reagent (Sigma Aldrich, UK) into HEK293T/17 cells, plated on 303 6-well Nunclon © plates (Thermo Fisher Scientific, UK).
TextSentencer_T78 11380-11492 Sentence denotes Supernatants were collected 48-72 h 304 post transfection and filtered through a 0.45 µm filter (Millipore, UK).
TextSentencer_T78 11380-11492 Sentence denotes Supernatants were collected 48-72 h 304 post transfection and filtered through a 0.45 µm filter (Millipore, UK).
TextSentencer_T79 11493-11625 Sentence denotes To remove viral titer 305 bias between different PV stocks, pseudotypes were concentrated and (re-) titrated by serial 306 dilution.
TextSentencer_T79 11493-11625 Sentence denotes To remove viral titer 305 bias between different PV stocks, pseudotypes were concentrated and (re-) titrated by serial 306 dilution.
TextSentencer_T80 11626-11766 Sentence denotes Concentration was carried out by ultra-centrifugation for 2 h at 25,000 rpm, 4°C in 307 the SW32 rotor of a L2-65B Beckman ultra-centrifuge.
TextSentencer_T80 11626-11766 Sentence denotes Concentration was carried out by ultra-centrifugation for 2 h at 25,000 rpm, 4°C in 307 the SW32 rotor of a L2-65B Beckman ultra-centrifuge.
TextSentencer_T81 11767-11770 Sentence denotes 308
TextSentencer_T81 11767-11770 Sentence denotes 308
TextSentencer_T82 11771-11940 Sentence denotes Two-fold serial dilutions of PV-containing supernatant were performed as previously 309 described 32 using white 96-well Nunclon © plates (Thermo Fisher Scientific, UK).
TextSentencer_T82 11771-11940 Sentence denotes Two-fold serial dilutions of PV-containing supernatant were performed as previously 309 described 32 using white 96-well Nunclon © plates (Thermo Fisher Scientific, UK).
TextSentencer_T83 11941-11959 Sentence denotes 310 author/funder.
TextSentencer_T83 11941-11959 Sentence denotes 310 author/funder.
TextSentencer_T84 11960-11980 Sentence denotes All rights reserved.
TextSentencer_T84 11960-11980 Sentence denotes All rights reserved.
TextSentencer_T85 11981-12017 Sentence denotes No reuse allowed without permission.
TextSentencer_T85 11981-12017 Sentence denotes No reuse allowed without permission.
TextSentencer_T86 12018-12283 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint Subsequently, approximately 1x10 4 (for adherent) and 3 x10 4 cells (for suspension) cells were 311 added in 50 µl of medium per well.
TextSentencer_T86 12018-12283 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint Subsequently, approximately 1x10 4 (for adherent) and 3 x10 4 cells (for suspension) cells were 311 added in 50 µl of medium per well.
TextSentencer_T87 12284-12340 Sentence denotes Plates were incubated for 48 h, after which 50 µl of 312
TextSentencer_T87 12284-12340 Sentence denotes Plates were incubated for 48 h, after which 50 µl of 312
TextSentencer_T88 12341-12387 Sentence denotes Bright-Glo™ substrate (Promega, UK) was added.
TextSentencer_T88 12341-12387 Sentence denotes Bright-Glo™ substrate (Promega, UK) was added.
TextSentencer_T89 12388-12585 Sentence denotes Luciferase readings were conducted with a 313 luminometer (FLUOstar OPTIMA, BMG Labtech) after a 5-minute incubation period and 314 luciferase reading recorded in relative luminescence units (RLU).
TextSentencer_T89 12388-12585 Sentence denotes Luciferase readings were conducted with a 313 luminometer (FLUOstar OPTIMA, BMG Labtech) after a 5-minute incubation period and 314 luciferase reading recorded in relative luminescence units (RLU).
TextSentencer_T90 12586-12674 Sentence denotes Data were normalized using 315 Δ-env and cell-only measurements and expressed as RLU/ml.
TextSentencer_T90 12586-12674 Sentence denotes Data were normalized using 315 Δ-env and cell-only measurements and expressed as RLU/ml.
TextSentencer_T91 12675-12678 Sentence denotes 316
TextSentencer_T91 12675-12678 Sentence denotes 316
TextSentencer_T92 12679-12760 Sentence denotes Mammalian expression plasmids (pcDNA3.1 + /C -(K)DYK) for HLA-DRA (NM_019111) and
TextSentencer_T92 12679-12760 Sentence denotes Mammalian expression plasmids (pcDNA3.1 + /C -(K)DYK) for HLA-DRA (NM_019111) and
TextSentencer_T93 12762-13025 Sentence denotes Total RNA was isolated from biological triplicates of early and late passage MDCK from T25 331 flasks that had been seeded with 8 x 10 5 cells and allowed to become confluent and polarize 332 over 4 days in culture cells using a Ribopure kit (Ambion, Austin, TX).
TextSentencer_T93 12762-13025 Sentence denotes Total RNA was isolated from biological triplicates of early and late passage MDCK from T25 331 flasks that had been seeded with 8 x 10 5 cells and allowed to become confluent and polarize 332 over 4 days in culture cells using a Ribopure kit (Ambion, Austin, TX).
TextSentencer_T94 13026-13221 Sentence denotes Acquired RNA was 333 precipitated with EtOH and subsequently purified employing columns, procedures and 334 reagents from an RNEasy kit (Qiagen, Germantown, MD) and resuspended in RNAse-free H2O.
TextSentencer_T94 13026-13221 Sentence denotes Acquired RNA was 333 precipitated with EtOH and subsequently purified employing columns, procedures and 334 reagents from an RNEasy kit (Qiagen, Germantown, MD) and resuspended in RNAse-free H2O.
TextSentencer_T95 13222-13225 Sentence denotes 335
TextSentencer_T95 13222-13225 Sentence denotes 335
TextSentencer_T96 13226-13330 Sentence denotes Complementary DNA and RNA synthesis were performed according to Affymetrix Expression 336 author/funder.
TextSentencer_T96 13226-13330 Sentence denotes Complementary DNA and RNA synthesis were performed according to Affymetrix Expression 336 author/funder.
TextSentencer_T97 13331-13351 Sentence denotes All rights reserved.
TextSentencer_T97 13331-13351 Sentence denotes All rights reserved.
TextSentencer_T98 13352-13388 Sentence denotes No reuse allowed without permission.
TextSentencer_T98 13352-13388 Sentence denotes No reuse allowed without permission.
TextSentencer_T99 13389-13564 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint Analysis protocols (see www.affymetrix.com).
TextSentencer_T99 13389-13564 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint Analysis protocols (see www.affymetrix.com).
TextSentencer_T100 13565-13708 Sentence denotes Briefly, double-stranded cDNA was synthesized 337 from 5 μg of total RNA using the Superscript double-stranded cDNA synthesis kit (Invitrogen).
TextSentencer_T100 13565-13708 Sentence denotes Briefly, double-stranded cDNA was synthesized 337 from 5 μg of total RNA using the Superscript double-stranded cDNA synthesis kit (Invitrogen).
TextSentencer_T101 13709-13712 Sentence denotes 338
TextSentencer_T101 13709-13712 Sentence denotes 338
TextSentencer_T102 13713-14084 Sentence denotes Following phenol/chloroform extraction and ethanol precipitation, a biotin-labeled in-vitro 339 transcription reaction was carried out using the cDNA template (Enzo Life Sciences, 340 In order to evaluate the interaction of HLA-DR with H17 we used the HLA DRA mAb 361 (302CT2.3.2), which is generated from mice immunized with a KLH conjugated synthetic 362 author/funder.
TextSentencer_T102 13713-14084 Sentence denotes Following phenol/chloroform extraction and ethanol precipitation, a biotin-labeled in-vitro 339 transcription reaction was carried out using the cDNA template (Enzo Life Sciences, 340 In order to evaluate the interaction of HLA-DR with H17 we used the HLA DRA mAb 361 (302CT2.3.2), which is generated from mice immunized with a KLH conjugated synthetic 362 author/funder.
TextSentencer_T103 14085-14105 Sentence denotes All rights reserved.
TextSentencer_T103 14085-14105 Sentence denotes All rights reserved.
TextSentencer_T104 14106-14142 Sentence denotes No reuse allowed without permission.
TextSentencer_T104 14106-14142 Sentence denotes No reuse allowed without permission.
TextSentencer_T105 14143-14327 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint peptide between 48-75 amino acids from human HLA-DRA.
TextSentencer_T105 14143-14327 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint peptide between 48-75 amino acids from human HLA-DRA.
TextSentencer_T106 14328-14499 Sentence denotes After a 1 h pre-incubation with Bio-Techne Ltd) targeting a monomorphic general framework determinant of HLA-DR Class II 386 antigen, diluted in 5% BSA/PBS for 1 hr at RT.
TextSentencer_T106 14328-14499 Sentence denotes After a 1 h pre-incubation with Bio-Techne Ltd) targeting a monomorphic general framework determinant of HLA-DR Class II 386 antigen, diluted in 5% BSA/PBS for 1 hr at RT.
TextSentencer_T107 14500-14651 Sentence denotes The cover slips were then washed 3X with 387 0.02% Tween 20 and 1% BSA in PBS, followed by incubation with Alexafluor 488 conjugated 388 author/funder.
TextSentencer_T107 14500-14651 Sentence denotes The cover slips were then washed 3X with 387 0.02% Tween 20 and 1% BSA in PBS, followed by incubation with Alexafluor 488 conjugated 388 author/funder.
TextSentencer_T108 14652-14672 Sentence denotes All rights reserved.
TextSentencer_T108 14652-14672 Sentence denotes All rights reserved.
TextSentencer_T109 14673-14709 Sentence denotes No reuse allowed without permission.
TextSentencer_T109 14673-14709 Sentence denotes No reuse allowed without permission.
TextSentencer_T110 14710-14904 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint anti-mouse (Thermo Fisher Scientific, UK) for 30 minutes at RT.
TextSentencer_T110 14710-14904 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint anti-mouse (Thermo Fisher Scientific, UK) for 30 minutes at RT.
TextSentencer_T111 14905-14936 Sentence denotes After washing 3X with 0.02% 389
TextSentencer_T111 14905-14936 Sentence denotes After washing 3X with 0.02% 389
TextSentencer_T112 14937-15046 Sentence denotes Tween 20 and 1% BSA in PBS, the cover slips were mounted using Prolong Gold containing 390 DAPI (Invitrogen).
TextSentencer_T112 14937-15046 Sentence denotes Tween 20 and 1% BSA in PBS, the cover slips were mounted using Prolong Gold containing 390 DAPI (Invitrogen).
TextSentencer_T113 15047-15150 Sentence denotes Images were acquired on EVOS fluorescent microscope (EVOS FL imaging 391 system; Life Technology, USA).
TextSentencer_T113 15047-15150 Sentence denotes Images were acquired on EVOS fluorescent microscope (EVOS FL imaging 391 system; Life Technology, USA).
TextSentencer_T114 15151-15186 Sentence denotes Experiments were carried out twice.
TextSentencer_T114 15151-15186 Sentence denotes Experiments were carried out twice.
TextSentencer_T115 15187-15195 Sentence denotes Reverse:
TextSentencer_T115 15187-15195 Sentence denotes Reverse:
TextSentencer_T116 15196-15256 Sentence denotes 5'-TCTGGAGGTACATTGGTGTTCG-3'), for canine DLA-DRB1 (Forward:
TextSentencer_T116 15196-15256 Sentence denotes 5'-TCTGGAGGTACATTGGTGTTCG-3'), for canine DLA-DRB1 (Forward:
TextSentencer_T117 15257-15278 Sentence denotes 5'-414 author/funder.
TextSentencer_T117 15257-15278 Sentence denotes 5'-414 author/funder.
TextSentencer_T118 15279-15299 Sentence denotes All rights reserved.
TextSentencer_T118 15279-15299 Sentence denotes All rights reserved.
TextSentencer_T119 15300-15336 Sentence denotes No reuse allowed without permission.
TextSentencer_T119 15300-15336 Sentence denotes No reuse allowed without permission.
TextSentencer_T120 15337-15500 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint AGCACCAAGTTTGACAAGC-3', Reverse:
TextSentencer_T120 15337-15500 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint AGCACCAAGTTTGACAAGC-3', Reverse:
TextSentencer_T121 15501-15571 Sentence denotes 5-AAGAGCAGACCCAGGACAAAG-3'). and RQ Manager v1.2 (Applied Biosystems).
TextSentencer_T121 15501-15571 Sentence denotes 5-AAGAGCAGACCCAGGACAAAG-3'). and RQ Manager v1.2 (Applied Biosystems).
TextSentencer_T122 15572-15767 Sentence denotes Gene expression data were normalized against 419 the housekeeping gene GAPDH, and compared with the mock controls using the comparative 420 CT method (also referred to as the 2 -ΔΔCT method 58 ).
TextSentencer_T122 15572-15767 Sentence denotes Gene expression data were normalized against 419 the housekeeping gene GAPDH, and compared with the mock controls using the comparative 420 CT method (also referred to as the 2 -ΔΔCT method 58 ).
TextSentencer_T123 15768-15934 Sentence denotes Absolute copy numbers of HLA-DRA in The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint
TextSentencer_T123 15768-15934 Sentence denotes Absolute copy numbers of HLA-DRA in The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint
TextSentencer_T124 15935-16037 Sentence denotes The buffy coat residues for the isolation of CD19 + primary B cells were purchased from the UK 440 20.
TextSentencer_T124 15935-16037 Sentence denotes The buffy coat residues for the isolation of CD19 + primary B cells were purchased from the UK 440 20.
TextSentencer_T125 16038-16054 Sentence denotes Garcia-Sastre A.
TextSentencer_T125 16038-16054 Sentence denotes Garcia-Sastre A.
TextSentencer_T126 16055-16121 Sentence denotes The neuraminidase of bat influenza viruses is not a neuraminidase.
TextSentencer_T126 16055-16121 Sentence denotes The neuraminidase of bat influenza viruses is not a neuraminidase.
TextSentencer_T127 16122-16125 Sentence denotes 520
TextSentencer_T127 16122-16125 Sentence denotes 520
TextSentencer_T128 16126-16176 Sentence denotes Proc Natl Acad Sci U S A 109, 18635-18636 (2012) .
TextSentencer_T128 16126-16176 Sentence denotes Proc Natl Acad Sci U S A 109, 18635-18636 (2012) .
TextSentencer_T129 16177-16195 Sentence denotes 521 author/funder.
TextSentencer_T129 16177-16195 Sentence denotes 521 author/funder.
TextSentencer_T130 16196-16216 Sentence denotes All rights reserved.
TextSentencer_T130 16196-16216 Sentence denotes All rights reserved.
TextSentencer_T131 16217-16253 Sentence denotes No reuse allowed without permission.
TextSentencer_T131 16217-16253 Sentence denotes No reuse allowed without permission.
TextSentencer_T132 16254-16384 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint
TextSentencer_T132 16254-16384 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint
TextSentencer_T133 16385-16435 Sentence denotes Ciminski K, Thamamongood T, Zimmer G, Schwemmle M.
TextSentencer_T133 16385-16435 Sentence denotes Ciminski K, Thamamongood T, Zimmer G, Schwemmle M.
TextSentencer_T134 16436-16484 Sentence denotes Novel insights into bat 523 influenza A viruses.
TextSentencer_T134 16436-16484 Sentence denotes Novel insights into bat 523 influenza A viruses.
TextSentencer_T135 16485-16647 Sentence denotes J Gen Virol 98, 2393-2400 (2017 The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint
TextSentencer_T135 16485-16647 Sentence denotes J Gen Virol 98, 2393-2400 (2017 The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/507467 doi: bioRxiv preprint