Dessain Scott K, Adekar Sharad P, Stevens Jennifer B, Carpenter Katherine A, Skorski Maria L, Barnoski Barry L, Goldsby Richard A, Weinberg Robert A
The Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, MA 02142, USA.
J Immunol Methods. 2004 Aug;291(1-2):109-22. doi: 10.1016/j.jim.2004.05.005.
The native human antibody repertoire holds unexplored potential for the development of novel monoclonal antibody therapeutics. Current techniques that fuse immortal cells and primary B-lymphocytes are sub-optimal for the routine production of hybridomas that secrete human monoclonal antibodies. We have found that a murine cell line that ectopically expresses murine interleukin-6 (mIL-6) and human telomerase (hTERT) efficiently forms stable human antibody-secreting heterohybridomas through cell fusion with primary human B-lymphocytes. The hybrid cells maintain secretion of human antibodies derived from the primary B-lymphocytes through multiple rounds of cloning. Using splenic B-lymphocytes from a patient immunized with a Streptococcus pneumoniae capsular polysaccharide vaccine, we have succeeded in creating hybridomas that secrete human monoclonal antibodies specific for S. pneumoniae antigens. Using peripheral blood lymphocytes, we have similarly cloned a human antibody that binds a viral antigen. These experiments establish that SP2/0-derived cell lines ectopically expressing mIL-6 and hTERT will enable the rapid cloning of native human monoclonal antibodies.
天然人类抗体库在新型单克隆抗体治疗药物的开发方面具有尚未被探索的潜力。目前将永生细胞与原代B淋巴细胞融合的技术在常规生产分泌人单克隆抗体的杂交瘤方面并不理想。我们发现，一种异位表达小鼠白细胞介素-6（mIL-6）和人端粒酶（hTERT）的小鼠细胞系，通过与原代人B淋巴细胞进行细胞融合，能够高效地形成稳定的分泌人抗体的异种杂交瘤。这些杂交细胞通过多轮克隆维持源自原代B淋巴细胞的人抗体的分泌。利用来自接种肺炎链球菌荚膜多糖疫苗患者的脾B淋巴细胞，我们成功创建了分泌针对肺炎链球菌抗原的人单克隆抗体的杂交瘤。利用外周血淋巴细胞，我们同样克隆出了一种结合病毒抗原的人抗体。这些实验表明，异位表达mIL-6和hTERT的源自SP2/0的细胞系将能够快速克隆天然人单克隆抗体。
