Tzehoval E, Dagan S, Eisenbach L, Atsmon J, Feldman M
Department of Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Eur J Immunol. 1989 Jan;19(1):89-96. doi: 10.1002/eji.1830190115.
Two clones, E2-7.7 and E2-10.50, derived from two macrophage(M phi)hybridomas, E2-7 and E2-10, have been studied. The first clone, E2-7.7, is Ia+ and Fc receptor (FcR) negative and manifests a strong antigen-presenting capacity. When we pulsed its cells in vitro with keyhole limpet hemocyanin (KLH) antigen and injected them into syngeneic animals, we found that as small a dose as 10(3) cells initiated an immune response in vivo. On the other hand, antigen-pulsed cells of the E2-10.50 clone, which are Ia- and FcR+, were almost incapable of triggering immunity, even when injected at a dose of 10(5) cells. Thus, the two clones differ in their immunogenic capacity (both cellular and humoral immunity). In experiments aimed at testing the stimulation in vitro of primed lymph node (LN) cells by antigen-pulsed cells of these two hybridoma clones, we observed that E2-7.7 stimulated the unfractionated population of LN cells and the LN-derived population of T cells. The E2-10.50 cells stimulated only the unfractionated population of LN cells, but not the T cell population. Subsequent tests indicated that the E2-10.50 cells require an intermediate Ia+ accessory cell to present the antigen to the T lymphocytes. Analyzing the molecular structure of the M phi hybridomas, we discovered that major histocompatibility complex (MHC) genes of the myeloma haplotype (H-2d), and of the splenic M phi used for fusion (H-2k), which were not expressed in the parental myeloma or in the E2-10.50, were expressed in the E2-7.7. Thus, somatic cell fusion of M phi resulted in the activation of suppressed genes of the myeloma partner. It appears that these antigens participate in controlling the immunogenic properties of the E2-7.7 clone. Testing the effects of interferons on the M phi hybridomas, we observed that interferon-gamma activated, at both the mRNA and the cell surface-antigen levels, the expression of H-2Dk, H-2Kd and H-2Dd in the E2-10.50 cells, but not in the E2-7.7. Consequently, interferon-gamma augmented significantly antigen presentation by E2-10.50 but not by E2-7.7 cells. These two hybridoma clones might represent two distinct subsets of normal M phi, manifesting two different sets of functional properties.(ABSTRACT TRUNCATED AT 400 WORDS)
对源自两个巨噬细胞（M phi）杂交瘤E2 - 7和E2 - 10的两个克隆E2 - 7.7和E2 - 10.50进行了研究。第一个克隆E2 - 7.7，Ia阳性且Fc受体（FcR）阴性，具有很强的抗原呈递能力。当我们在体外用钥孔戚血蓝蛋白（KLH）抗原脉冲其细胞并将它们注射到同基因动物体内时，我们发现低至10³个细胞的剂量就能在体内引发免疫反应。另一方面，E2 - 10.50克隆的抗原脉冲细胞，Ia阴性且FcR阳性，即使以10⁵个细胞的剂量注射，也几乎无法触发免疫反应。因此，这两个克隆在其免疫原性能力（细胞免疫和体液免疫）方面存在差异。在旨在测试这两个杂交瘤克隆的抗原脉冲细胞对致敏淋巴结（LN）细胞的体外刺激作用的实验中，我们观察到E2 - 7.7刺激了未分级的LN细胞群体和LN来源的T细胞群体。E2 - 10.50细胞仅刺激了未分级的LN细胞群体，而未刺激T细胞群体。后续测试表明，E2 - 10.50细胞需要一个中间的Ia阳性辅助细胞将抗原呈递给T淋巴细胞。通过分析M phi杂交瘤的分子结构，我们发现骨髓瘤单倍型（H - 2d）以及用于融合的脾M phi（H - 2k）的主要组织相容性复合体（MHC）基因，在亲本骨髓瘤或E2 - 10.50中未表达，但在E2 - 7.7中表达。因此，M phi的体细胞融合导致了骨髓瘤伙伴中受抑制基因的激活。似乎这些抗原参与控制E2 - 7.7克隆的免疫原性特性。测试干扰素对M phi杂交瘤的影响时，我们观察到干扰素 - γ在mRNA和细胞表面抗原水平上激活了E2 - 10.50细胞中H - 2Dk、H - 2Kd和H - 2Dd的表达，但未激活E2 - 7.7细胞中的表达。因此，干扰素 - γ显著增强了E2 - 10.50细胞的抗原呈递能力，但未增强E2 - 7.7细胞的抗原呈递能力。这两个杂交瘤克隆可能代表正常M phi的两个不同亚群，表现出两组不同的功能特性。（摘要截于400字）
