A novel Raji-Burkitt's lymphoma model for preclinical and mechanistic evaluation of CD52-targeted immunotherapeutic agents
A novel Raji-Burkitt's lymphoma model for preclinical and mechanistic evaluation of CD52-targeted immunotherapeutic agents
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DOI：
10.1158/1078-0432.ccr-07-1006
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发表时间：
2008-01-15
影响因子：
11.5
通讯作者：
Byrd, John C.
中科院分区：
文献类型：
--
作者：
Lapalombella, Rosa;Zhao, Xiaobin;Byrd, John C.
目的：迄今为止，由于缺乏稳定表达CD 52的转化B细胞系和动物模型，研究CD 52靶向治疗（如Alemtuzumab）的努力受到限制。我们描述了产生和利用的细胞系，稳定表达CD 52在体外和在vivo.Experimental Design：通过有限稀释，我们已经建立了几个克隆的Raji-Burkitt的淋巴瘤细胞系，表达表面CD 52。对这些克隆进行免疫表型和细胞遗传学表征。在体内有用的CD 52（高）细胞系，以评估CD 52-导向抗体的治疗效果进行了研究，使用SCID小鼠异种移植model.Results：CD 52的稳定表达，证实在体外培养的细胞连续生长长达52周。使用Alemtuzumab显示了表达的CD 52分子的功能完整性，Alemtuzumab在体外诱导了CD 52（高）而非CD 52（低）克隆的细胞毒性效应。与对照抗体相比，CD 52（高）接种小鼠中的阿仑单抗治疗导致中位生存期显着增加。在CD 52 high克隆和原发性B-慢性淋巴细胞白血病细胞中，CD 52靶向的直接细胞毒性、补体依赖性细胞毒性和抗体依赖性细胞毒性以及抗CD 52免疫脂质体介导的合成寡脱氧核糖核苷酸的递送水平相当，这暗示了该模型用于评价CD 52靶向抗体和包封治疗剂的免疫脂质体的体内应用的潜力。这些结果显示了稳定表达高水平CD 52的克隆的Raji细胞系的体外效用。本文所述的使用这些稳定细胞系的播散性白血病-淋巴瘤小鼠模型可用作针对CD 52分子的现有和新型抗体的体内治疗和机制评价的优异系统。
Purpose: To date, efforts to study CD52-targeted therapies, such as alemtuzumab, have been limited due to the lack of stable CD52 expressing transformed B-cell lines and animal models. We describe generation and utilization of cell lines that stably express CD52 both in vitro and in vivo.Experimental Design: By limiting dilution, we have established several clones of Raji-Burkitt's lymphoma cell line that express surface CD52. Immunophenotype and cytogenetic characterization of these clones was done. In vivo usefulness of the CD52(high) cell line to evaluate the therapeutic efficacy of CD52-directed antibody was investigated using a SCID mouse xenograft model.Results: Stable expression of CD52 was confirmed in cells cultured in vitro up to 52 weeks of continuous growth. The functional integrity of the expressed CD52 molecule was shown using alemtuzumab, which induced cytotoxic effects in vitro in the CD52(high) but not the CD52(low) clone. Compared with control antibody, alemtuzumab treatment in CD52(high) inoculated mice resulted in significantly increased median survival. Comparable levels of CD52-targeted direct cytotoxicity, complement-dependent cytotoxicity, and antibody-dependent cytotoxicity and anti-CD52 immunoliposome-mediated delivery of synthetic oligodeoxyribo nucleotides in CD52high clone and primary B-chronic lymphocytic leukemia cells implicated potential in vivo application of this model for evaluation of CD52-targeted antibody and immunoliposomes encapsulating therapeutic agents.Conclusions: These results show the in vitro utility of the cloned Raji cell lines that stably express high levels CD52. The disseminated leukemia-lymphoma mouse model described herein using these stable cell lines can serve as an excellent system for in vivo therapeutic and mechanistic evaluation of existing and novel antibodies directed against CD52 molecule.