BISPECIFIC MONOCLONAL-ANTIBODIES DIRECTED TO CD16 AND TO A TUMOR-ASSOCIATED ANTIGEN INDUCE TARGET-CELL LYSIS BY RESTING NK CELLS AND BY A SUBSET OF NK CLONES
BISPECIFIC MONOCLONAL-ANTIBODIES DIRECTED TO CD16 AND TO A TUMOR-ASSOCIATED ANTIGEN INDUCE TARGET-CELL LYSIS BY RESTING NK CELLS AND BY A SUBSET OF NK CLONES
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DOI：
10.1002/ijc.2910480213
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发表时间：
1991-05-10
影响因子：
6.4
通讯作者：
MORETTA, L
中科院分区：
文献类型：
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作者：
FERRINI, S;PRIGIONE, I;MORETTA, L
CD 16表面抗原代表CD 3 -16 + NK细胞中的活化分子。为了靶向NK细胞对抗相对NK抗性的卵巢癌，我们使用抗CD 16单克隆抗体（MAb）（VD 4），连同抗卵巢癌相关抗原（MOV 19），来构建biMAb。为此，通过将TK缺陷型VD 4杂交瘤突变体与HGPRT缺陷型MOV 19杂交瘤融合来产生杂交瘤。在4小时Cr-51释放测定中，筛选来自已在HAT培养基中选择的杂交杂交瘤的上清液诱导CD 3 -16+ NK克隆以裂解MOV 19+卵巢癌细胞系的能力。NMB.45杂交杂交瘤分泌触发MOV 19+靶细胞裂解但不触发MOV 19-靶细胞裂解的biMAb。用MOV 19 MAb也观察到一定程度的靶细胞溶解（由于ADCC机制），而VD 4 MAb无效。杂交杂交瘤分泌的MAb的HPLC分级分离使得可以鉴定4个不同的峰，其中一个峰似乎含有功能性biMAb分子。HPLC纯化的biMAb（100 ng/ml）诱导静息PBL裂解“NK抗性”IGROVI卵巢癌细胞系。新鲜的MOV 19+肿瘤细胞也被裂解，尽管效率较低。当IL-2激活的淋巴细胞被用作效应源时，biMAb仅引起IL-2诱导的细胞溶解活性的轻微增加。在克隆水平上对biMAb的作用进行进一步分析。在CD 3 -16+ NK细胞克隆中，仅在GLI 83+克隆中观察到明显的增强作用，但在GLI 83-克隆中未观察到。在CD 3+细胞毒性克隆中，可以在4个TCR γ/δ +克隆中的一个中检测到触发效应，但在TCR α/β +克隆中未检测到。
CDI6 surface antigens represent activatory molecules in CD3-16 + NK cells. In order to target NK cells against relatively NK-resistant ovarian carcinomas, we used an anti-CD16 monoclonal antibody (MAb) (VD4), together with an anti-ovarian carcinoma-associated antigen (MOV19), to construct biMAbs. To this end, hybrid hybridomas were generated by fusing a TK-deficient VD4 hybridoma mutant with a HGPRT-deficient MOV19 hybrid. Supernatants from hybrid hybridomas that had been selected in HAT medium were screened for their ability to induce a CD3-16+ NK clone to Iyse an MOV19+ ovarian carcinoma cell line in a 4-hr Cr-51-release assay. The NMB.45 hybrid hybridoma secreted a biMAb which triggered lysis of MOV19+ but not of MOV19-target cells. Some degree of target cell lysis was also observed with MOV19 MAb (due to ADCC mechanisms), while the VD4 MAb was ineffective. HPLC fractionation of MAbs secreted by the hybrid hybridoma made it possible to identify 4 different peaks, one of which appeared to contain functional biMAb molecules. HPLC-purified biMAb (100 ng/ml) induced resting PBL to lyse the "NK-resistant" IGROVI ovarian carcinoma cell line. Fresh MOV19+ tumor cells were also lysed, although with lower efficiency. When IL-2-activated lymphocytes were used as a source of effectors, biMAb caused only minor increases in the IL-2-induced cytolytic activity. Further analyses of the effect of biMAb were performed at the clonal level. Among CD3-16+ NK cell clones a clear enhancing effect could be observed only in GLI83+ but not in GLI83-clones. In CD3+ cytotoxic clones a triggering effect could be detected in one out of 4 TCR gamma/delta+ clones but not in TCR alpha/beta+ clones.