Mor F, Cohen I R
Department of Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
J Clin Invest. 1992 Dec;90(6):2447-55. doi: 10.1172/JCI116136.
To characterize the cellular immune response in an autoimmune lesion, we investigated the accumulation of specific T cells in the central nervous system in actively induced experimental autoimmune encephalomyelitis (EAE) in Lewis rats, using a limiting dilution analysis (LDA) assay for T cells that proliferate in response to antigens. Lymphocytes isolated from the spinal cord infiltrate were compared with cells from the popliteal lymph nodes with respect to frequency of cells responding to basic protein (BP), mycobacterium tuberculosis (MT), the 65-kD heat shock protein (hsp65), allogeneic brown norway spleen cells, and concanavalin A. Additionally, we compared the BP frequency in acute EAE of cells from the spinal cord, peripheral blood, spleen and lymph nodes, and the spinal cord and lymph node after recovery from EAE. We found that acute EAE was associated with marked enrichment of BP-reactive T cells in the spinal cord relative to their frequency in the lymphoid organs and peripheral blood. The infiltrate was also enriched for T cells responding to hsp65; alloreactive T cells, in contrast, were not enriched. The frequency of BP reactive T cells in the spinal cord was highest at the peak of paralysis; however, BP-reactive T cells could still be detected at moderate frequencies after clinical recovery. We established BP- and Mycobacteria-reactive T cell lines from the spinal infiltrates that were CD4+ and TcR alpha beta +. Most of the BP lines were found to react to the major encephalitogenic epitope of guinea pig BP for rats (amino acids 71-90); these lines were found to mediate EAE in naive recipients. T cell lines recognizing other epitopes of BP were not encephalitogenic. All of the lines responsive to Mycobacteria recognized hsp65 or hsp70. These results indicating that the immune infiltrate in active EAE is enriched with cells responding to the autoantigen and to hsp65 were confirmed in EAE adoptively transferred by anti-BP T cell clone.
为了表征自身免疫性病变中的细胞免疫反应，我们利用针对因抗原而增殖的T细胞的极限稀释分析（LDA）检测法，研究了Lewis大鼠主动诱导实验性自身免疫性脑脊髓炎（EAE）中中枢神经系统内特异性T细胞的积聚情况。将从脊髓浸润物中分离出的淋巴细胞与腘窝淋巴结中的细胞，就对碱性蛋白（BP）、结核分枝杆菌（MT）、65-kD热休克蛋白（hsp65）、同种异体棕色挪威大鼠脾细胞以及伴刀豆球蛋白A产生反应的细胞频率进行了比较。此外，我们比较了急性EAE时脊髓、外周血、脾脏和淋巴结中的细胞以及EAE恢复后脊髓和淋巴结中的BP反应频率。我们发现，相对于它们在淋巴器官和外周血中的频率，急性EAE与脊髓中BP反应性T细胞的显著富集有关。浸润物中对hsp65产生反应的T细胞也有所富集；相比之下，同种异体反应性T细胞并未富集。脊髓中BP反应性T细胞频率在麻痹高峰期最高；然而，临床恢复后仍能以中等频率检测到BP反应性T细胞。我们从脊髓浸润物中建立了对BP和分枝杆菌产生反应的T细胞系，这些细胞系为CD4+且TcRαβ+。发现大多数BP细胞系对大鼠豚鼠BP的主要致脑炎性表位（氨基酸71 - 90）有反应；这些细胞系被发现可在未接触过抗原的受体中介导EAE。识别BP其他表位的T细胞系不具有致脑炎性。所有对分枝杆菌有反应的细胞系都识别hsp65或hsp70。这些结果表明，在抗BP T细胞克隆过继转移的EAE中证实，活动性EAE中的免疫浸润富含对自身抗原和hsp65产生反应的细胞。
