PMC:17827 / 52450-58107 JSONTXT

{"project":"2_test","denotations":[{"id":"11178129-9008595-4386913","span":{"begin":373,"end":375},"obj":"9008595"},{"id":"11178129-8834014-4386914","span":{"begin":376,"end":378},"obj":"8834014"},{"id":"11178129-8752676-4386915","span":{"begin":443,"end":444},"obj":"8752676"},{"id":"11178129-7673404-4386916","span":{"begin":445,"end":447},"obj":"7673404"},{"id":"11178129-7673404-4386917","span":{"begin":559,"end":561},"obj":"7673404"},{"id":"11178129-8912877-4386918","span":{"begin":577,"end":579},"obj":"8912877"},{"id":"11178129-7648435-4386919","span":{"begin":580,"end":582},"obj":"7648435"},{"id":"11178129-8613470-4386920","span":{"begin":583,"end":585},"obj":"8613470"},{"id":"11178129-7561064-4386921","span":{"begin":628,"end":630},"obj":"7561064"},{"id":"11178129-8082888-4386922","span":{"begin":631,"end":633},"obj":"8082888"},{"id":"11178129-1381726-4386923","span":{"begin":634,"end":636},"obj":"1381726"},{"id":"11178129-7704909-4386924","span":{"begin":637,"end":639},"obj":"7704909"},{"id":"11178129-8102558-4386925","span":{"begin":640,"end":642},"obj":"8102558"},{"id":"11178129-3259597-4386926","span":{"begin":643,"end":645},"obj":"3259597"},{"id":"11178129-6352937-4386927","span":{"begin":687,"end":689},"obj":"6352937"},{"id":"11178129-2694400-4386928","span":{"begin":828,"end":830},"obj":"2694400"},{"id":"11178129-6366990-4386929","span":{"begin":831,"end":833},"obj":"6366990"},{"id":"11178129-2442194-4386930","span":{"begin":834,"end":836},"obj":"2442194"},{"id":"11178129-1657009-4386931","span":{"begin":837,"end":839},"obj":"1657009"},{"id":"11178129-9082943-4386932","span":{"begin":870,"end":872},"obj":"9082943"},{"id":"11178129-7927499-4386933","span":{"begin":980,"end":982},"obj":"7927499"},{"id":"11178129-1657009-4386934","span":{"begin":1585,"end":1587},"obj":"1657009"},{"id":"11178129-7632096-4386935","span":{"begin":1929,"end":1930},"obj":"7632096"},{"id":"11178129-7510485-4386936","span":{"begin":1934,"end":1936},"obj":"7510485"},{"id":"11178129-2542542-4386937","span":{"begin":2112,"end":2114},"obj":"2542542"},{"id":"11178129-9082943-4386938","span":{"begin":2245,"end":2247},"obj":"9082943"},{"id":"11178129-2042649-4386939","span":{"begin":2282,"end":2284},"obj":"2042649"},{"id":"11178129-2042650-4386940","span":{"begin":2285,"end":2287},"obj":"2042650"},{"id":"11178129-8912499-4386941","span":{"begin":2609,"end":2610},"obj":"8912499"},{"id":"11178129-8003048-4386942","span":{"begin":2611,"end":2613},"obj":"8003048"},{"id":"11178129-7531793-4386943","span":{"begin":2614,"end":2616},"obj":"7531793"},{"id":"11178129-1380043-4386944","span":{"begin":2617,"end":2619},"obj":"1380043"},{"id":"11178129-1411083-4386945","span":{"begin":2657,"end":2659},"obj":"1411083"},{"id":"11178129-8660103-4386946","span":{"begin":2660,"end":2662},"obj":"8660103"},{"id":"11178129-1870583-4386947","span":{"begin":2663,"end":2665},"obj":"1870583"},{"id":"11178129-7531793-4386948","span":{"begin":3143,"end":3145},"obj":"7531793"},{"id":"11178129-1380043-4386949","span":{"begin":3146,"end":3148},"obj":"1380043"},{"id":"11178129-8912499-4386950","span":{"begin":3259,"end":3260},"obj":"8912499"},{"id":"11178129-8003048-4386951","span":{"begin":3261,"end":3263},"obj":"8003048"},{"id":"11178129-10233853-4386952","span":{"begin":3264,"end":3266},"obj":"10233853"},{"id":"11178129-8912499-4386953","span":{"begin":3281,"end":3282},"obj":"8912499"},{"id":"11178129-8003048-4386954","span":{"begin":3283,"end":3285},"obj":"8003048"},{"id":"11178129-10233853-4386955","span":{"begin":3568,"end":3570},"obj":"10233853"},{"id":"11178129-10820282-4386956","span":{"begin":3571,"end":3573},"obj":"10820282"},{"id":"11178129-1411083-4386957","span":{"begin":3724,"end":3726},"obj":"1411083"},{"id":"11178129-8660103-4386958","span":{"begin":3727,"end":3729},"obj":"8660103"},{"id":"11178129-1870583-4386959","span":{"begin":4189,"end":4191},"obj":"1870583"},{"id":"11178129-1411083-4386960","span":{"begin":5161,"end":5163},"obj":"1411083"},{"id":"11178129-2694400-4386961","span":{"begin":5651,"end":5653},"obj":"2694400"}],"text":"Similarities/discrepancies between the in situ and in vitro phenotype of RA-SFB\n\nSimilarities\nThe cells obtained by negative isolation generally showed features similar to those reported in in situ analyses. The expression of Thy-1, CD13 (aminopeptidaseN), and vimentin exemplify this. The expression of these molecules, particularly their local upregulation by cytokines [54,55] and/or heterogeneous expression at different anatomical sites [9,49], may be pathogenetically relevant, either in terms of defining functionally heterogeneous SFB subpopulations [49] (reviewed in [50,51,52]), by providing pro-inflammatory enzymes [17,47,56,57,58,59], or as targets of autoimmune reactions [60].\nMHC-II. The constitutive MHC-II expression in isolated RA-SFB and OA-SFB, confirming previous in situ observations in the RA and OA SM [13,21,22,28] and in systemic scleroderma [61], is probably related to the inflammatory microenvironment, since normal skin-FB (Table 4) and normal SFB [31] hardly express MHC-II. This is further supported by the a strong decrease of the percentage of MHC-II+ cells upon repeated passage (both conventional and following negative isolation), possibly due to lack of or a progressive decrease of external stimulation with pro-inflammatory mediators (Fig. 7 and Table 5). Notably, there was a significant, negative correlation between the percentage of MHC-II-positive SFB and treatment of the RA patients with Methotrexate (ρ=-0.866, P = 0.01, n = 7), indicating that effective antirheumatic therapy may be reflected by decreased MHC-II expression on RA-SFB [28].\nProcollagen III. The significant increase of the MFI for intracellular procollagen III in RA-SFB and OA-SFB (Table 4) suggests that the pattern of SFB activation may be functional (among other things) to the increase of collagen metabolism; this is supported by the increased expression of collagen α2I and α1III mRNA in the RASM in situ [2,23,24], in comparison with normal or non-RA synovial tissue. Also, in as much as procollagen III is the fetal form of collagen used in wound healing and tissue repair (reviewed in [62,63]), ongoing fibrosis may be a considerable component of the disease process in RA (and OA), in analogy to systemic scleroderma [61] or interstitial kidney fibrosis [11,26]. A striking increase of procollagen I and III expression in RA-SFB from primary culture to fourth passage (increase of positive cells by ≥ 38%; Fig. 7 and Table 5), on the other hand, indicates that primary-culture cells do not exploit their full potential of matrix production.\n\nDiscrepancies\nThe expression of VCAM-1 [1,15,64,65] and Jun and Fos proto-oncogenes [23,25,66,67] showed some in situ / in vitro discrepancies.\nVCAM-1. Surprisingly, only a moderate and variable percentage of RA-SFB (whether conventionally passaged or negatively isolated from primary culture) expressed the adhesion molecule VCAM-1 (Tables 4 and 5). There were also no significant differences between RA-SFB and OA-SFB or normal skin-FB (Table 4). This is in apparent contrast to the enhanced expression of VCAM-1 reported in the lining layer of RA and OA synovial tissue [64,65]; however, it is well compatible with the large variability in VCAM-1 expression observed in vitro (Table 4) [1,15,68] and in situ [1,15]. The significant, positive correlation between VCAM-1 expression in isolated primary-culture RA-SFB and the erythrocyte sedimentation rate in RA patients (ρ=1.00, P = 0.000, n = 4) indicates that the variability may depend on disease activity, as also suggested in recent reports [68,69,70].\nProto-oncogene expression. Negatively isolated RA-SFB expressed c-Fos, c-Jun, and Jun-D (Table 4), in analogy to the features of SFB in RASM [23,25,66]. The expression of c-Jun and Jun-D in the present study could be unequivocally demonstrated by FACS analysis (Table 4), while previous immunohistochemical studies had failed to detect these molecules in situ [23]. Notably, however, the degree of proto-oncogene expression (both percentage of positive cells and MFI) did not significantly differ from that of normal skin-FB (Fig. 6 and Table 4) or, as previously reported, of SFB from traumatic joint injury [67]. This finding supports in situ observations that, at a single-cell level, the degree of proto-oncogene expression in RA, OA, and joint trauma is similar [23], thereby questioning whether proto-oncogene expression in RA reflects per se cell transformation and/or severe metabolic abnormalities.\nNotably, the expression of the proto-oncogenes c-Fos and Jun-D was strikingly increased in conventional fourth-passage RA-SFB as compared with isolated primary-culture RA-SFB (increase of positive cells ≥ 38%; Fig. 7 and Table 5). This increase was limited to RA, since the percentage of cells positive for these molecules was significantly decreased in OA-SFB and numerically decreased in normal skin-FB upon passaging. As a consequence of these reciprocal changes, the percentage of positive cells and/or MFI for c-Fos and Jun-D in conventional fourth-passage RA-SFB was significantly higher than in OA-SFB, under these circumstances confirming previously published data [25].\nFinally, a significant, positive correlation between the percentage of c-fos+ and procollagen I and III+ isolated primary-culture SFB in both RA and OA patients (RA: c-fos/procollagen I, ρ=1.000, P = 0.00, n = 7; c-fos/procollagen III, ρ=0.964, P = 0.00, n = 7) suggests a link between c-fos expression and augmented matrix production by SFB in rheumatic disorders, in line with the known regulation of collagen expression by the activator protein-1 transcription factor (reviewed in [13]).\n"}

{"project":"Colil","denotations":[{"id":"T123","span":{"begin":3281,"end":3282},"obj":"8912499"},{"id":"T124","span":{"begin":3283,"end":3285},"obj":"8003048"},{"id":"T125","span":{"begin":3264,"end":3266},"obj":"10233853"},{"id":"T126","span":{"begin":980,"end":982},"obj":"7927499"},{"id":"T127","span":{"begin":2285,"end":2287},"obj":"2042650"},{"id":"T128","span":{"begin":2282,"end":2284},"obj":"2042649"},{"id":"T129","span":{"begin":373,"end":375},"obj":"9008595"},{"id":"T130","span":{"begin":376,"end":378},"obj":"8834014"},{"id":"T131","span":{"begin":5651,"end":5653},"obj":"2694400"},{"id":"T132","span":{"begin":640,"end":642},"obj":"8102558"},{"id":"T133","span":{"begin":643,"end":645},"obj":"3259597"},{"id":"T134","span":{"begin":577,"end":579},"obj":"8912877"},{"id":"T135","span":{"begin":3261,"end":3263},"obj":"8003048"},{"id":"T136","span":{"begin":1934,"end":1936},"obj":"7510485"},{"id":"T137","span":{"begin":2609,"end":2610},"obj":"8912499"},{"id":"T138","span":{"begin":2611,"end":2613},"obj":"8003048"},{"id":"T139","span":{"begin":2614,"end":2616},"obj":"7531793"},{"id":"T140","span":{"begin":2617,"end":2619},"obj":"1380043"},{"id":"T141","span":{"begin":2657,"end":2659},"obj":"1411083"},{"id":"T142","span":{"begin":2660,"end":2662},"obj":"8660103"},{"id":"T143","span":{"begin":2663,"end":2665},"obj":"1870583"},{"id":"T144","span":{"begin":445,"end":447},"obj":"7673404"},{"id":"T145","span":{"begin":3724,"end":3726},"obj":"1411083"},{"id":"T146","span":{"begin":4189,"end":4191},"obj":"1870583"},{"id":"T147","span":{"begin":2112,"end":2114},"obj":"2542542"},{"id":"T148","span":{"begin":3727,"end":3729},"obj":"8660103"},{"id":"T149","span":{"begin":559,"end":561},"obj":"7673404"},{"id":"T150","span":{"begin":1929,"end":1930},"obj":"7632096"},{"id":"T151","span":{"begin":443,"end":444},"obj":"8752676"},{"id":"T152","span":{"begin":628,"end":630},"obj":"7561064"},{"id":"T153","span":{"begin":631,"end":633},"obj":"8082888"},{"id":"T154","span":{"begin":634,"end":636},"obj":"1381726"},{"id":"T155","span":{"begin":637,"end":639},"obj":"7704909"},{"id":"T156","span":{"begin":3259,"end":3260},"obj":"8912499"},{"id":"T157","span":{"begin":5161,"end":5163},"obj":"1411083"},{"id":"T158","span":{"begin":828,"end":830},"obj":"2694400"},{"id":"T159","span":{"begin":831,"end":833},"obj":"6366990"},{"id":"T160","span":{"begin":834,"end":836},"obj":"2442194"},{"id":"T161","span":{"begin":837,"end":839},"obj":"1657009"},{"id":"T162","span":{"begin":870,"end":872},"obj":"9082943"},{"id":"T163","span":{"begin":580,"end":582},"obj":"7648435"},{"id":"T164","span":{"begin":583,"end":585},"obj":"8613470"},{"id":"T165","span":{"begin":1585,"end":1587},"obj":"1657009"},{"id":"T166","span":{"begin":3568,"end":3570},"obj":"10233853"},{"id":"T167","span":{"begin":3571,"end":3573},"obj":"10820282"},{"id":"T168","span":{"begin":687,"end":689},"obj":"6352937"},{"id":"T169","span":{"begin":3143,"end":3145},"obj":"7531793"},{"id":"T170","span":{"begin":3146,"end":3148},"obj":"1380043"},{"id":"T171","span":{"begin":2245,"end":2247},"obj":"9082943"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/docs/sourcedb/PubMed/sourceid/"}],"text":"Similarities/discrepancies between the in situ and in vitro phenotype of RA-SFB\n\nSimilarities\nThe cells obtained by negative isolation generally showed features similar to those reported in in situ analyses. The expression of Thy-1, CD13 (aminopeptidaseN), and vimentin exemplify this. The expression of these molecules, particularly their local upregulation by cytokines [54,55] and/or heterogeneous expression at different anatomical sites [9,49], may be pathogenetically relevant, either in terms of defining functionally heterogeneous SFB subpopulations [49] (reviewed in [50,51,52]), by providing pro-inflammatory enzymes [17,47,56,57,58,59], or as targets of autoimmune reactions [60].\nMHC-II. The constitutive MHC-II expression in isolated RA-SFB and OA-SFB, confirming previous in situ observations in the RA and OA SM [13,21,22,28] and in systemic scleroderma [61], is probably related to the inflammatory microenvironment, since normal skin-FB (Table 4) and normal SFB [31] hardly express MHC-II. This is further supported by the a strong decrease of the percentage of MHC-II+ cells upon repeated passage (both conventional and following negative isolation), possibly due to lack of or a progressive decrease of external stimulation with pro-inflammatory mediators (Fig. 7 and Table 5). Notably, there was a significant, negative correlation between the percentage of MHC-II-positive SFB and treatment of the RA patients with Methotrexate (ρ=-0.866, P = 0.01, n = 7), indicating that effective antirheumatic therapy may be reflected by decreased MHC-II expression on RA-SFB [28].\nProcollagen III. The significant increase of the MFI for intracellular procollagen III in RA-SFB and OA-SFB (Table 4) suggests that the pattern of SFB activation may be functional (among other things) to the increase of collagen metabolism; this is supported by the increased expression of collagen α2I and α1III mRNA in the RASM in situ [2,23,24], in comparison with normal or non-RA synovial tissue. Also, in as much as procollagen III is the fetal form of collagen used in wound healing and tissue repair (reviewed in [62,63]), ongoing fibrosis may be a considerable component of the disease process in RA (and OA), in analogy to systemic scleroderma [61] or interstitial kidney fibrosis [11,26]. A striking increase of procollagen I and III expression in RA-SFB from primary culture to fourth passage (increase of positive cells by ≥ 38%; Fig. 7 and Table 5), on the other hand, indicates that primary-culture cells do not exploit their full potential of matrix production.\n\nDiscrepancies\nThe expression of VCAM-1 [1,15,64,65] and Jun and Fos proto-oncogenes [23,25,66,67] showed some in situ / in vitro discrepancies.\nVCAM-1. Surprisingly, only a moderate and variable percentage of RA-SFB (whether conventionally passaged or negatively isolated from primary culture) expressed the adhesion molecule VCAM-1 (Tables 4 and 5). There were also no significant differences between RA-SFB and OA-SFB or normal skin-FB (Table 4). This is in apparent contrast to the enhanced expression of VCAM-1 reported in the lining layer of RA and OA synovial tissue [64,65]; however, it is well compatible with the large variability in VCAM-1 expression observed in vitro (Table 4) [1,15,68] and in situ [1,15]. The significant, positive correlation between VCAM-1 expression in isolated primary-culture RA-SFB and the erythrocyte sedimentation rate in RA patients (ρ=1.00, P = 0.000, n = 4) indicates that the variability may depend on disease activity, as also suggested in recent reports [68,69,70].\nProto-oncogene expression. Negatively isolated RA-SFB expressed c-Fos, c-Jun, and Jun-D (Table 4), in analogy to the features of SFB in RASM [23,25,66]. The expression of c-Jun and Jun-D in the present study could be unequivocally demonstrated by FACS analysis (Table 4), while previous immunohistochemical studies had failed to detect these molecules in situ [23]. Notably, however, the degree of proto-oncogene expression (both percentage of positive cells and MFI) did not significantly differ from that of normal skin-FB (Fig. 6 and Table 4) or, as previously reported, of SFB from traumatic joint injury [67]. This finding supports in situ observations that, at a single-cell level, the degree of proto-oncogene expression in RA, OA, and joint trauma is similar [23], thereby questioning whether proto-oncogene expression in RA reflects per se cell transformation and/or severe metabolic abnormalities.\nNotably, the expression of the proto-oncogenes c-Fos and Jun-D was strikingly increased in conventional fourth-passage RA-SFB as compared with isolated primary-culture RA-SFB (increase of positive cells ≥ 38%; Fig. 7 and Table 5). This increase was limited to RA, since the percentage of cells positive for these molecules was significantly decreased in OA-SFB and numerically decreased in normal skin-FB upon passaging. As a consequence of these reciprocal changes, the percentage of positive cells and/or MFI for c-Fos and Jun-D in conventional fourth-passage RA-SFB was significantly higher than in OA-SFB, under these circumstances confirming previously published data [25].\nFinally, a significant, positive correlation between the percentage of c-fos+ and procollagen I and III+ isolated primary-culture SFB in both RA and OA patients (RA: c-fos/procollagen I, ρ=1.000, P = 0.00, n = 7; c-fos/procollagen III, ρ=0.964, P = 0.00, n = 7) suggests a link between c-fos expression and augmented matrix production by SFB in rheumatic disorders, in line with the known regulation of collagen expression by the activator protein-1 transcription factor (reviewed in [13]).\n"}