Yang W C, Esquenazi V, Carreno M, Vallone T, Fuller L, Roth D, Nery J, Burke G, Miller J
Department of Surgery, University of Miami School of Medicine, Florida.
Transplantation. 1994 Jul 27;58(2):233-40.
An IgG1 monoclonal antibody (mAb), designated as 15F1.5, was generated against surface determinants of a dog peripheral blood-derived PHA-induced IL-2-dependent T cell line. It reacted with 65-80% of peripheral blood mononuclear cells (PBMCs), 90-95% of polymorphonuclear cells (PMNs), 65-70% of thymocytes, 85-95% of Thy-1 positive cells and 85-95% of IL-2-dependent T lymphoid cells in flow cytometry. It was nonreactive with peripheral blood red cells and platelets. It immunoprecipitated 95 and 150 Kd proteins derived from detergent solubilized lymphocyte membranes. Indirect immunofluorescent and immunoperoxidase staining of frozen tissue sections demonstrated positive reactivity to cells in lymphoid but not nonlymphoid tissues. The 15F1.5 antibody was not directly mitogenic for PBMC's. It caused significant decrease (P < or = 0.05) in the lymphoproliferative response to T-dependent B cell mitogens, such as pokeweed mitogen (PWM) and staphage lysate (SPL), without significant effects on responses to the T cell mitogens, phytohemagglutinin (PHA), and concanavalin A (Con A). The mixed lymphocyte culture (MLC) response and both the proliferative and effector arms of the cell-mediated cytotoxicity reactions (CMC) were inhibited in a dose-dependent manner. The mAb also inhibited the auto- and allolymphoproliferative reactivity of mixed lymphocyte kidney or islet cell cultures (MLKC and MLIC), and the adhesion of T lymphoblasts and PMA-treated PMNs to endothelial cells. In vivo administration of the 15F1.5 (20 mg/day for 5 days) caused an immediate and prolonged reduction in MLC responses, associated with cell binding of the mAb to PBMC and epitope modulation during the course of treatment, as indicated by flow cytometry. These results suggest that 15F1.5 is an immunomodulating antibody reacting with canine LFA-1. Thus, this mAb would be useful in studying the role of LFA-1/ICAM-1 in graft rejection as well as other inflammatory responses. It would also allow the use of an animal model to investigate the immunoregulatory effects of in vivo administration of anti-CD11/CD18 antibodies in organ/tissue transplants.
一种名为15F1.5的IgG1单克隆抗体（mAb）是针对犬外周血来源的PHA诱导的IL-2依赖性T细胞系的表面决定簇产生的。在流式细胞术中，它与65 - 80%的外周血单核细胞（PBMC）、90 - 95%的多形核细胞（PMN）、65 - 70%的胸腺细胞、85 - 95%的Thy-1阳性细胞以及85 - 95%的IL-2依赖性T淋巴细胞发生反应。它与外周血红细胞和血小板无反应。它免疫沉淀了来自去污剂溶解的淋巴细胞膜的95和150 Kd蛋白质。冷冻组织切片的间接免疫荧光和免疫过氧化物酶染色显示，对淋巴组织而非非淋巴组织中的细胞有阳性反应。15F1.5抗体对PBMC不具有直接促有丝分裂作用。它导致对T细胞依赖性B细胞有丝分裂原（如商陆有丝分裂原（PWM）和葡萄球菌溶素（SPL））的淋巴细胞增殖反应显著降低（P≤0.05），而对T细胞有丝分裂原植物血凝素（PHA）和刀豆球蛋白A（Con A）的反应无显著影响。混合淋巴细胞培养（MLC）反应以及细胞介导的细胞毒性反应（CMC）的增殖和效应臂均呈剂量依赖性抑制。该mAb还抑制混合淋巴细胞肾或胰岛细胞培养（MLKC和MLIC）的自身和同种淋巴细胞增殖反应，以及T淋巴母细胞和PMA处理的PMN与内皮细胞的黏附。体内给予15F1.5（20 mg/天，共5天）导致MLC反应立即且持续降低，流式细胞术显示在治疗过程中mAb与PBMC发生细胞结合以及表位调节。这些结果表明15F1.5是一种与犬LFA-1反应的免疫调节抗体。因此，这种mAb在研究LFA-1/ICAM-1在移植排斥以及其他炎症反应中的作用方面将是有用的。它还将允许使用动物模型来研究体内给予抗CD11/CD18抗体在器官/组织移植中的免疫调节作用。
