Anti-human LFA-1 monoclonal antibodies for prevention of graft failure in HLA-mismatched bone marrow transplantation.

1.抗LFA-1抗体在HLA-单倍型不匹配的骨髓移植（BMT）中预防移植物衰竭的用途。1例范可尼贫血患者因缺乏HLA相合的供者而接受了T细胞耗尽的HLA单倍型不匹配的母体骨髓移植。为了防止移植失败，使用针对人LFA-1的α链的鼠单克隆抗体（mAb）。mAb（25.3）由Immunotech（马赛，法国）提供。检查抗体制备物的无菌性和无热原性。mAb以0.1mg/kg的剂量以6小时静脉输注给药，从-3到+6连续给药，没有任何不良反应的迹象。实现了部分血液学重建，但最终导致移植失败。范可尼贫血的结果与其他机构的经验雅阁。然而，在使用抗LFA-1单克隆抗体时未发现不良反应，这鼓励在所选患者中继续进行进一步的临床研究.针对人LFA-1的mAb的产生。为获得能诱导免疫耐受并可用于临床的抗LFA-1单克隆抗体，将K4细胞免疫的BALB/C小鼠脾细胞与SP2/0细胞融合。经流式细胞仪和生化分析，其中1株产生抗α链抗体（MAY-035），2株产生抗β链抗体（MAY-017和MAY-044）。获得的mAb的同种型均为IgGl。这三种抗体强烈抑制淋巴细胞增殖反应同种抗原和有丝分裂原。

1. Use of the anti-LFA-1 antibody to prevent graft failure in HLA-haplotype mismatched bone marrow transplantation (BMT). A patient with Fanconi anemia underwent T cell- devpleted HLA-haplotype mismatched maternal BMT because of the lack of HLA-identical donor. To prevent graft failure a murine monoclonal antibody (mAb) directed against the alpha chain of human LFA-1 was used. The mAb (25.3) was kindly supplied by Immunotech (Marseille, France). The antibody preparations were checked for sterility and lack of pyrogenicity. The mAb was given in a 6-h iv infusion m dosis of 0.1 mg/kg for consecutive days from-3 to +6 without any signs of adverse reactions. Partial;hematological reconstitution was achieved but finally resulted in graft failure. The result in Fanconi anemia is in accord with experiences in other institutions. However, no adverse reactions in the use of the anti-LFA-1 mAb encourage to continue further clinical studies in patients selected.2. Production of mAb against human LFA-1. To obtain anti-LFA-1 mAb capable of inducing immune tolerance and available for clinical use spleen cells from BALB/C mice immunized with K4 cells were fused with SP2/0 cells. Hybridoma supernatants obtained were screened for the capability of inhibiting the aggregation of K4 cells stimulated by phorbol ester.Three hybridoma clones secreting anti-LFA-1 antibodies were obtained; one clone was determined by flowcytometry and biochemical analysis to produce the antibody directed at a chain (MAY-035) and other two the antibodies directed at 8 chain (MAY-017, and MAY-044). The isotype of the mAbs obtained was all IgGl. The three antibodies strongly inhibited lymphocyte proliferation in response to alloantigens and mitogens.