Clinical study of LAK adoptive immunotherapy for angiosarcoma enhanced by anti-angiosarcoma × anti-CD3 bispecific antibody

LAK过继免疫治疗是治疗血管肉瘤的有效方法。为了加强这种免疫治疗的效果，我们设计了一种双特异性抗体(BSAb)来在肿瘤细胞和LAK细胞之间进行交联。我们以人血管肉瘤细胞系ISO-HAS为免疫原，建立了抗血管肉瘤特异性单抗HEW-3。癌症杂志81：305-308,1999)。HEW-3是一种IgM抗体，可识别免疫原中43kD的分子。根据HEW-3的F(ab‘)_2区分别制备抗血管肉瘤细胞抗体和抗LAK细胞CD3抗体。BSAb以剂量和时间依赖的方式增强LAK细胞对ISO-HAS细胞的杀伤作用。此外，细胞毒活性与体外测定上清液中肿瘤坏死因子α的剂量有关。在我们建立的人血管肉瘤SCID小鼠模型(WB-SCID)中，研究了BSAb在LAK免疫治疗中的体内作用。SCI。27：88-94,2001)。与单纯LAK免疫治疗相比，最初4次皮内注射BSAb免疫治疗对生长的血管肉瘤有明显的抑制作用。基于这些基本结果，我们用这种疗法治疗了三名血管肉瘤患者。2例头皮血管肉瘤患者皮损内注射2.5μg BSAb联合LAK细胞，1例选择性动脉注射15μg BSAb联合LAK细胞治疗1例颅内血管肉瘤。人类抗鼠抗体的增加阻止了持续给药，但我们检测了这种免疫疗法在每个患者中的临床确切但暂时的效果。这种免疫治疗的副作用仅有1例发热，2例无内脏损害的患者C反应蛋白升高。这一前瞻性研究有望在不久的将来推动LAK免疫治疗血管肉瘤的人型BSAb的研制。

LAK adoptive immunotherapy is effective in the treatment of angiosarcoma. To enhance the effect of this immunotherapy, we devised the utility of a bispecific antibody (BSAb) to crosslink between tumor cells and LAK cells. We established an anti-angiosarcoma specific monoclonal antibody HEW-3 using a human angiosarcoma cell line ISO-HAS as an immunogen (Int. J. Cancer 81 : 305-308, 1999). HEW-3 was an IgM antibody and recognizes a 43 Kd molecule in the immunogen. The BSAb was made from the respective F(ab') _2 regions of HEW-3 for angiosarcoma cell and anti-CD3 antibody for LAK cell. The BSAb enhanced LAK cytotoxicity against ISO-HAS cells in a dose- and time-dependent manner. Moreover, the cytotoxic activity was related to a dose of TNFα measured in the supernates in vitro. The in vivo effect of the BSAb in LAK immunotherapy was studied in a human angiosarcoma SCID mouse model (WB-SCID) established by us (J. Dermatol. Sci. 27 : 88-94, 2001). The growing angiosarcoma was suppressed significantly only by initial 4-times intra-lesional injection of the BSAb immunotherapy in comparison with LAK immunotherapy alone. Based on these basic results, we treated three patients with angiosarcoma by this therapy. The 2.5μg BSAb combined with LAK cells was administered to two patients with angiosarcoma of the scalp by intra-lesional injection, and the 15 μg BSAb combined with LAK cells to one patient with intracranial angiosarcoma by selective intra-arterial injection. The increase in the human anti-mouse antibody prevented continuous administration, but we detected the clinical exact but temporal effect of this immunotherapy in each patient. Side effects in this immunotherapy were only fever in one patient and CRP elevation in two patients without visceral damage. This prospective study could promote the making of human-type BSAb for LAK immunotherapy of angiosarcoma in the near future.