In vitro immunization of human lymphocytes. I. Production of human monoclonal antibodies against bombesin and tetanus toxoid.
In vitro immunization of human lymphocytes. I. Production of human monoclonal antibodies against bombesin and tetanus toxoid.
复制标题
人淋巴细胞的体外免疫。
作者：
M. Ho;N. Rand;J. Murray;K. Kato;H. Rabin
介绍了一种体外增敏人淋巴细胞对破伤风类毒素(BTT)结合蛙皮素的方法。蛙皮素是一种与小细胞肺癌相关的十四肽。我们发现，用尼龙分离的人脾淋巴细胞与脂多糖、植物血凝素激活的淋巴细胞上清液、人AB血清以及与破伤风类毒素结合的蛙皮素在体外培养6天，可以产生针对蛙皮素和破伤风类毒素的抗体反应。用这种方法致敏的细胞与小鼠骨髓瘤细胞NS-1融合。通过酶联免疫分析和竞争抑制实验分析得到的杂交瘤细胞的特异性。获得分泌抗蛙皮素(IgM)或抗破伤风类毒素(IgM或Ig G)的杂交瘤细胞。用琼脂糖珠偶联抗原可以提高破伤风类毒素抗体的免疫球蛋白和免疫球蛋白的比例。本文所述的致敏方法为体外研究人B淋巴细胞的抗原刺激和制备具有所需特异性的人的单抗提供了一种系统。
A procedure for in vitro sensitization of human lymphocytes against bombesin conjugated to tetanus toxoid (BTT) is described. Bombesin is a tetradecapeptide associated with small cell lung carcinoma. We found that antibody responses against bombesin as well as tetanus toxoid could be generated in vitro by culturing nylon-separated human splenic lymphocytes for 6 days with lipopolysaccharide, phytohemagglutinin-activated lymphocyte supernatants, human AB serum, and bombesin conjugated to tetanus toxoid. Cells sensitized by this procedure were fused to murine myeloma cells, NS-1. The specificities of resulting hybrids were analyzed by enzyme-linked immunoassays and competitive inhibition experiments. Hybrids secreting anti-bombesin (IgM) or anti-tetanus toxoid (IgM or IgG) were obtained. The ratio of IgG to IgM antibodies against tetanus toxoid could be increased by using antigen coupled to Sepharose beads. The sensitization procedure described here offers a system for the study of antigenic stimulation of human B lymphocytes in vitro and for the production of human monoclonal antibodies with the desired specificities.