MONOCLONAL T LYMPHOCYTE FACTOR REGULATION OF MYELOMA

BALB/c或CB 6 F的免疫1 小鼠异源红细胞诱导 多个调节性T细胞，最初可以通过它们的 选择性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在 存在TNP结合的异源红细胞。 的目标 增殖和分化辅助性和抑制性T细胞 似乎是小的，非分泌性淋巴细胞样MOPC-315肿瘤干 细胞 这些实验的目的是产生单克隆细胞系， 绵羊红细胞免疫诱导的各辅助性和抑制性T细胞 关于CB 6 F1 小鼠，并表征每种单克隆调节细胞， 其对增殖和调节因子的触发要求 分泌，细胞表面受体的特异性， 受体表达和调节因子分泌。 在过去的一年里，我们已经成功地产生了三个辅助性T细胞杂交瘤 其组成性分泌轻链独特表位 315- 特异性助手 因子，诱导几乎所有的分泌分化， MOPC-315细胞，即使是密度梯度纯化的非分泌型315细胞， 亚群 这些杂交瘤是通过将SRBC-免疫Ly 1 T融合 细胞粘附IgAlambda2 315- 包被的培养皿， HGPRT- BW 5147 T淋巴瘤细胞，然后通过生长选择杂交体 在HAT媒体 315细胞分化培养阳性有帮助，但 通过限制性克隆那些不增强315-细胞增殖的基因， 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无影响 细胞生长已经产生。 其中一种能特异性结合 α-315一个是特异性的，另一个是SRBC特异性的。 我也 产生了一个克隆的反抑制I-细胞杂交瘤，其特异于 SRBC。 这种杂交瘤使315个细胞对伊加具有抗性， 315- 绑定 不S 细胞分泌的因子，但本身没有辅助活性。 我在 使用流式细胞术和扩增技术对这些T细胞杂交瘤进行表型分型的过程 开始通过亲和层析纯化IgAlambda2 315- 产生的特定监管因素。 我也在确定 IL-2、IL-3和γ-干扰素由这些杂交瘤中的任一种分泌 并开始培育这些类型T细胞的正常克隆系。 （LB）

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)