Pope R M, Wallis R S, Sailer D, Buchanan T M, Pahlavani M A
Department of Medicine, Northwestern University, Chicago, Illinois.
Cell Immunol. 1991 Mar;133(1):95-108. doi: 10.1016/0008-8749(91)90182-b.
To define which mycobacterial antigens were responsible for the activation of synovial fluid T lymphocytes, acetone-precipitated Mycobacterium tuberculosis (AP-MT) antigens were separated into five fractions following polyacrylamide gel electrophoresis and added to the mononuclear cell cultures of patients with inflammatory synovitis. Fractions 2 (50 to 70 kDa) and 5 (less than 28 kDa) resulted in significantly more proliferation than that of fractions 1, 3, and 4. The response to a purified mycobacterial 65-kDa heat shock protein (hsp), which migrated in fraction 2, was highly correlated (r = 0.89, P less than 0.001) with the response to the crude AP-MT. The proliferative response to a different hsp. the Escherichia coli DnaK, by synovial fluid lymphocytes was marginal. Analysis of the synovial fluid T cell response to mycobacterial culture filtrates by T cell Western blotting revealed dominant responses to antigen(s) in the range of 31 to 21 kDa in each responding patient, although no other consistent pattern of T cell activation was noted. Three lines of evidence suggested that the response to the low molecular weight fractions was directed against degradation fragments of the 65-kDa protein. These observations suggest that the activation of T lymphocytes obtained from inflammatory synovial fluids by crude mycobacterial antigens was due in large part to recognition of the 65-kDa mycobacterial hsp.
为了确定哪些分枝杆菌抗原可激活滑液T淋巴细胞，将丙酮沉淀的结核分枝杆菌（AP-MT）抗原经聚丙烯酰胺凝胶电泳分离成五个组分，并添加到炎性滑膜炎患者的单核细胞培养物中。第2组分（50至70 kDa）和第5组分（小于28 kDa）导致的增殖明显多于第1、3和4组分。对迁移至第2组分中的纯化的65 kDa分枝杆菌热休克蛋白（hsp）的反应与对粗制AP-MT的反应高度相关（r = 0.89，P < 0.001）。滑液淋巴细胞对另一种hsp即大肠杆菌DnaK的增殖反应很微弱。通过T细胞蛋白质印迹法分析滑液T细胞对分枝杆菌培养滤液的反应，结果显示，尽管未观察到其他一致的T细胞激活模式，但在每个有反应的患者中，对31至21 kDa范围内的抗原均有明显反应。三条证据表明，对低分子量组分的反应针对的是65 kDa蛋白的降解片段。这些观察结果表明，粗制分枝杆菌抗原激活炎性滑液中的T淋巴细胞，很大程度上是由于对65 kDa分枝杆菌hsp的识别。
