Marquardt P, Müller-Hermelink H K
Institute of Pathology, University of Würzburg, Federal Republic of Germany.
Environ Health Perspect. 1990 Aug;88:233-5. doi: 10.1289/ehp.9088233.
Using a recently described technique for direct expansion of human T-lymphocytes isolated from small intestine biopsies, we have investigated the local cellular immune response in six patients with B-cell lymphomas of various subtypes. T-cell lines (TCL) were established by seeding tumor-infiltrating T-lymphocytes (T1TL) at limiting dilution in the presence of irradiated feeder cells in culture medium containing rIl-2 and phytohemagglutinin (PHA). About 1/50 T-cells gave rise to a TCL; they all were CD3+. The CD4/CD8 ratio was 3.8:1 before and after cloning. Of 45 TCLs analysed so far from one patient with B-cell lymphoma of the lung, 4 were cytotoxic as shown by their ability to exert lectin-dependent cytotoxicity against allogeneic target cells. Of these, 3 demonstrated specificity for the autologous malignant B-cells. Five TCLs lysed the NK-sensitive K562 cell line in a HLA-unrestricted manner. When tested for antigen-specific proliferative activity, 4 TCLS only responded to the autologous lymphoma cells, but 5 TCLs reacted to the autoantigenic ganglioside GM1. Southern Blot analyses did not show a clonal pattern of T-cell receptor gene rearrangement within all TITL populations. The peripheral T-lymphocytes of the lymphoma patients showed a drastically reduced response to the mitogens PHA. Concanavalin A, and pokeweed mitogen. The present report demonstrates that it is possible to analyze TITL at clonal level. This technique may be the only means of investigating the specificity of the TITL and may help us to identify the relevant tumor-associated autoantigens if tumor-induced autoimmunization is indeed one of the mechanisms that control the growth of tumors and metastases.
我们运用一种最近描述的从小肠活检组织中直接扩增人T淋巴细胞的技术，对6例不同亚型B细胞淋巴瘤患者的局部细胞免疫反应进行了研究。通过在含有重组白细胞介素-2（rIL-2）和植物血凝素（PHA）的培养基中，以有限稀释度接种肿瘤浸润性T淋巴细胞（TIL），并与经辐照的饲养细胞共同培养，建立了T细胞系（TCL）。约1/50的T细胞可产生TCL，这些TCL均为CD3阳性。克隆前后CD4/CD8比值均为3.8:1。到目前为止，对1例肺B细胞淋巴瘤患者的45个TCL进行分析，其中4个具有细胞毒性，表现为它们能够对同种异体靶细胞发挥凝集素依赖性细胞毒性。其中3个对自体恶性B细胞具有特异性。5个TCL以HLA非限制性方式裂解NK敏感的K562细胞系。当检测抗原特异性增殖活性时，4个TCL仅对自体淋巴瘤细胞有反应，但5个TCL对自身抗原性神经节苷脂GM1有反应。Southern印迹分析未显示所有TIL群体中T细胞受体基因重排的克隆模式。淋巴瘤患者的外周T淋巴细胞对丝裂原PHA、刀豆球蛋白A和商陆有丝分裂原的反应显著降低。本报告表明，有可能在克隆水平上分析TIL。如果肿瘤诱导的自身免疫确实是控制肿瘤和转移生长的机制之一，那么这项技术可能是研究TIL特异性的唯一方法，并且可能有助于我们识别相关的肿瘤相关自身抗原。
