MONOCLONAL T LYMPHOCYTE FACTOR REGULATION OF MYELOMA

BALB/c或CB6F1小鼠对异种红细胞的免疫诱导 多个调节性T细胞，最初可以通过其 选择性地调节克隆生长或分泌的能力 分泌抗TNP-IgA的BALB/c骨髓瘤MOPC-315细胞的分化 TNP结合的异种红细胞的存在。的目标 增殖和分化的辅助和抑制T细胞 看起来是小的非分泌性淋巴细胞样MOPC-315肿瘤干 细胞。这些实验的目的是为了获得单克隆系 绵羊红细胞免疫诱导的辅助性和抑制性T细胞 并鉴定每一种单抗调节细胞的特性 它对扩散和调控因素的触发要求 分泌，细胞表面受体的特异性，细胞周期的改变 受体的表达和调节因子的分泌。 在过去的一年里，我们成功地培育了三个辅助T细胞杂交瘤 其构成地分泌轻链独特型315特异性辅助分子 诱导几乎所有细胞分泌分化的因子 MOPC-315细胞，甚至密度梯度纯化的非分泌型315细胞 亚群。这些杂交瘤是通过融合SRBC免疫的Ly 1 T而产生的 细胞，贴附在IgAlambda2315包被的培养板上， HGPRT-BW5147 T淋巴瘤细胞及其生长筛选 在HAT媒体上。315细胞分化阳性的培养有帮助，但 那些没有促进315细胞增殖的基因是通过限制 稀释。同样，两个克隆的抑制性T细胞杂交瘤 抑制MOPC-315细胞分泌分化但对315细胞无影响 已经产生了细胞生长。其中一个特别地绑定了 阿尔法-315个独特异体，而另一个是SRBC特有的。我也有过 克隆了一株抗抑制因子I细胞杂交瘤，该杂交瘤是针对 SRBC。该杂交瘤使315细胞对Ig A 315结合产生抗药性 TS细胞分泌的因子，但本身没有辅助活性。我现在在 应用流式细胞术和AM对这些T细胞杂交瘤进行表型鉴定的过程 开始用亲和层析纯化IgAlambda2 产生了315个特定的监管因素。我还在决定是否 IL-2、IL-3和γ-干扰素由这些杂交瘤中的任何一个分泌。 我开始发展这些类型的T细胞的正常克隆株。 (磅)

Immunization of BALB/c or CB6F1 mice to heterologous erythrocytes induces multiple regulatory T cells that can initially be segregated by their ability to selectively modulate either clone growth or secretory differentiation of the anti-TNP IgA-secreting BALB/c myeloma MOPC-315 in the presence of TNP-conjugated heterologous erythrocytes. The target for both the proliferation and differentiation helper and suppressor T cells appears to be the small, nonsecretory lymphocytoid MOPC-315 tumor stem cells. The purpose of the experiments is to produce monoclonal lines of each helper and suppressor T cell induced by sheep erythrocyte immunization of CB6F1 mice and to characterize each monoclonal regulatory cell as to its triggering requirements for proliferation and regulatory factor secretion, specificity of cell surface receptors, cell cycle alterations in receptor expression, and regulatory factor secretion. In past year we have successfully produced three helper T-cell hybridomas which constitutively secrete a light chain idiotope 315-specific helper factor that induces secretory differentiation of virtually all of the MOPC-315 cells, even a density gradient-purified nonsecretory 315-cell subpopulation. These hybridomas were produced by fusing SRBC-immune Ly 1 T cells, which adhered to IgAlambda2 315-coated Petri plates, with HGPRT- BW5147 T lymphoma cells and then selecting for hybrids by growth in HAT media. Cultures positive for 315-cell differentiation help, but those which did not enhance 315-cell proliferation were cloned by limiting dilution. Similarly, two cloned suppressor T-cell hybridomas which suppress MOPC-315 cell secretory differentiation but have no effect on 315 cell growth have been produced. One of these specifically binds alpha-315idiotopes while the other is SRBC specific. I have also produced a cloned contrasuppressor I-cell hybridoma which is specific for SRBC. This hybridoma makes 315 cells resistant to the IgA 315-binding TS cell-secreted factors but has no helper activity itself. I am in the process of phenotyping these T-cell hybridomas using flow cytometry and am beginning to purify, by affinity chromatography, the IgAlambda2 315-specific regulatory factors produced. I am also determining whether IL-2, IL-3, and gamma-interferon are secreted by any of these hybridomas and am beginning to develop normal cloned lines of these types of T cells. (LB)