PDF(Pubmed)
Abstract：
OBJECTIVE: To investigate whether stimulation with toll-like receptor (TLR) 7 leads to pathways that proceed to tripartite motif-containing protein 21 (TRIM21) or Ro52/SS-A antigen presentation through major histocompatibility complex (MHC) class I in salivary gland epithelial cells (SGECs) from Sjögren\'s syndrome (SS) patients.
METHODS: Cultured SGECs from SS patients were stimulated with TLR7 agonist, loxoribine, and interferon-β. Cell lysates immunoprecipitated by anti-MHC class I antibody were analyzed by Western blotting. The immunofluorescence of salivary gland tissue from SS and non-SS subjects and cultured TLR7-stimulated SGECs was examined.
RESULTS: Significantly increased MHC class I expression was observed in SS patients\' ducts versus non-SS ducts; no significant difference was detected for ubiquitin. Upregulated MHC class I in the cell membrane and cytoplasm and augmented Ro52 expression were observed in SGECs stimulated with TLR7. The formation of peptide-loading complex (PLC), including tapasin, calreticulin, transporter associated with antigen processing 1, and endoplasmic reticulum-resident protein 57 in labial salivary glands (LSGs) from SS patients, was dominantly observed and colocalized with MHC class I, which was confirmed in TLR7-stimulated SGEC samples.
CONCLUSIONS: These findings suggest that the TLR7 stimulation of SS patients\' SGECs advances the process toward the antigen presentation of TRIM21/Ro52-SS-A via MHC class I.
METHODS: Cultured SGECs from SS patients were stimulated with TLR7 agonist, loxoribine, and interferon-β. Cell lysates immunoprecipitated by anti-MHC class I antibody were analyzed by Western blotting. The immunofluorescence of salivary gland tissue from SS and non-SS subjects and cultured TLR7-stimulated SGECs was examined.
RESULTS: Significantly increased MHC class I expression was observed in SS patients\' ducts versus non-SS ducts; no significant difference was detected for ubiquitin. Upregulated MHC class I in the cell membrane and cytoplasm and augmented Ro52 expression were observed in SGECs stimulated with TLR7. The formation of peptide-loading complex (PLC), including tapasin, calreticulin, transporter associated with antigen processing 1, and endoplasmic reticulum-resident protein 57 in labial salivary glands (LSGs) from SS patients, was dominantly observed and colocalized with MHC class I, which was confirmed in TLR7-stimulated SGEC samples.
CONCLUSIONS: These findings suggest that the TLR7 stimulation of SS patients\' SGECs advances the process toward the antigen presentation of TRIM21/Ro52-SS-A via MHC class I.
摘要：
目的：研究Toll样受体（TLR）7的刺激是否导致通过主要组织相容性复合物（MHC）I类在Sjögren综合征（SS）患者的唾液腺上皮细胞（SGECs）中进入含三联基序蛋白21（TRIM21）或Ro52/SS-A抗原呈递的途径。
方法：用TLR7激动剂刺激培养的SS患者SGECs，洛索利滨,和干扰素-β。通过Western印迹分析通过抗MHCI类抗体免疫沉淀的细胞裂解物。检查了SS和非SS受试者的唾液腺组织以及培养的TLR7刺激的SGEC的免疫荧光。
结果：在SS患者导管与非SS导管中观察到MHCI类表达显著增加；泛素没有检测到显著差异。在用TLR7刺激的SGEC中观察到细胞膜和细胞质中I类MHC上调和Ro52表达增强。肽负载复合物（PLC）的形成，包括Tapasin,钙网蛋白,SS患者唇腺（LSGs）中与抗原加工1相关的转运蛋白和内质网驻留蛋白57，主要观察到并与I类MHC共定位，这在TLR7刺激的SGEC样品中得到证实。
结论：这些发现表明，SS患者SGECs的TLR7刺激促进了TRIM21/Ro52-SS-A通过MHCI类的抗原呈递过程。
方法：用TLR7激动剂刺激培养的SS患者SGECs，洛索利滨,和干扰素-β。通过Western印迹分析通过抗MHCI类抗体免疫沉淀的细胞裂解物。检查了SS和非SS受试者的唾液腺组织以及培养的TLR7刺激的SGEC的免疫荧光。
结果：在SS患者导管与非SS导管中观察到MHCI类表达显著增加；泛素没有检测到显著差异。在用TLR7刺激的SGEC中观察到细胞膜和细胞质中I类MHC上调和Ro52表达增强。肽负载复合物（PLC）的形成，包括Tapasin,钙网蛋白,SS患者唇腺（LSGs）中与抗原加工1相关的转运蛋白和内质网驻留蛋白57，主要观察到并与I类MHC共定位，这在TLR7刺激的SGEC样品中得到证实。
结论：这些发现表明，SS患者SGECs的TLR7刺激促进了TRIM21/Ro52-SS-A通过MHCI类的抗原呈递过程。
