Cleveland W L, Wood I, Cone R E, Iverson G M, Rosenstein R W, Gershon R K, Erlanger B F
Proc Natl Acad Sci U S A. 1981 Dec;78(12):7697-701. doi: 10.1073/pnas.78.12.7697.
A single-cell secretion assay was used to detect cells that secrete products which react with an antiserum that binds T cell antigen-binding polypeptides. The antiserum (R11), which was produced by immunization of rabbits with a murine trinitrophenyl-specific suppressor factor, reacts with T cells and their products and with a suppressor T-cell clone but not with B cells or their products. The secretory cells this antiserum detected were found to be unevenly distributed among various organs (spleen, lymph node, and thymus) and, to different degrees, in spleens of various strains of mice. In unimmunized CBA/J mice, approximately 1-3% of spleen cells secreted macromolecules precipitable by R11. The majority of the secretory cells could be removed by panning with a mixture of Ly 1 and Ly 2 antibodies but not with either antibody alone. This is consistent with the cells having low surface antigen densities as a result of being either "pre-T" cells or mature secretory cells analogous to B-lineage plasma cells. In agreement with the latter possibility was our finding that the secretory activity of cells detected with antisuppressor factor was comparable to that of Ig-secretory cells as detected with an anti-Ig antiserum. However, higher numbers of R11+ secretory cells were seen in the immunoglobulin-negative fraction of spleen cells from nude mice, which could be interpreted to favor the first possibility. In either case this study shows that the single-cell assay technique is well suited for the detection and characterization of molecules released by immunoregulatory T cells.
采用单细胞分泌试验来检测那些分泌能与结合T细胞抗原结合多肽的抗血清发生反应的产物的细胞。该抗血清(R11)是用鼠三硝基苯特异性抑制因子免疫兔子产生的,它能与T细胞及其产物以及一个抑制性T细胞克隆发生反应,但不与B细胞及其产物发生反应。发现这种抗血清检测到的分泌细胞在不同器官(脾脏、淋巴结和胸腺)中分布不均,并且在不同品系小鼠的脾脏中也有不同程度的分布。在未免疫的CBA/J小鼠中,约1% - 3%的脾细胞分泌可被R11沉淀的大分子物质。大多数分泌细胞可用Ly 1和Ly 2抗体的混合物进行淘选去除,但单独使用任何一种抗体都无法去除。这与这些细胞由于是“前T”细胞或类似于B系浆细胞的成熟分泌细胞而具有低表面抗原密度是一致的。与后一种可能性相符的是我们的发现,即抗抑制因子检测到的细胞的分泌活性与抗Ig抗血清检测到的Ig分泌细胞的分泌活性相当。然而,在裸鼠脾细胞的免疫球蛋白阴性部分中观察到更多数量的R11 +分泌细胞,这可以解释为支持第一种可能性。无论哪种情况,这项研究都表明单细胞试验技术非常适合检测和表征免疫调节性T细胞释放的分子。