HUMAN MONOCLONAL-ANTIBODIES PRODUCED BY PRIMARY INVITRO IMMUNIZATION OF PERIPHERAL-BLOOD LYMPHOCYTES
HUMAN MONOCLONAL-ANTIBODIES PRODUCED BY PRIMARY INVITRO IMMUNIZATION OF PERIPHERAL-BLOOD LYMPHOCYTES
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DOI：
10.1073/pnas.85.11.3995
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发表时间：
1988-06-01
影响因子：
11.1
通讯作者：
MOLLER, SA
中科院分区：
文献类型：
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作者：
BORREBAECK, CAK;DANIELSSON, L;MOLLER, SA
一般程序描述了生产人单克隆抗体从外周血淋巴细胞免疫体外抗t细胞依赖抗原。这些在培养中免疫的淋巴细胞用于产生人-人或人-小鼠杂交瘤，分泌地高梅、血青素、人免疫缺陷病毒（PB1）的gp120包膜糖蛋白重组片段或黑色素瘤相关抗原（p97）的特异性单克隆抗体。富含溶酶体的细胞群，包括大颗粒淋巴细胞、单核细胞、细胞毒性T细胞和cd8阳性T细胞亚群，在细胞体外免疫之前被证明是至关重要的。这种消耗是通过用促溶体剂l -亮氨酸甲酯治疗外周血淋巴细胞完成的。此外，体外免疫必须有白细胞介素2，γ支持。干扰素，以及b细胞生长和分化因子，这些因子来源于辐照的、罂粟花有丝分裂原刺激的人T细胞。从原代抗原特异性激活的外周血淋巴细胞中产生人单克隆抗体可能会避免对志愿者或患者进行免疫。
A general procedure is described for the production of human monoclonal antibodies from peripheral blood lymphocytes immunized in vitro against T-cell-dependent antigens. These lymphocytes immunized in culture were used to produce human-human or human-mouse hybridomas secreting monoclonal antibodies specific for digoxin, hemocyanin, a recombinant fragment of the gp120 envelope glycoprotein of human immunodeficiency virus (PB1), or melanoma-associated antigen (p97). Depletion of a lysosome-rich cell population, containing large granular lymphocytes, monocytes, cytotoxic T cells, and a subset of CD8-positive T cells, was shown to be crucial before the cells could be immunized in vitro. This depletion was accomplished by treating the peripheral blood lymphocytes with the lysosomotropic agent L-leucine methyl ester. In addition, the in vitro immunization had to be supported by interleukin 2, .gamma.-interferon, and B-cell growth and differentiation factors, derived from irradiated, pokeweed-mitogen-stimulated human T cells. The production of human monoclonal antibodies from primary, antigen-specifically activated peripheral lymphocytes might obviate the need to immunize volunteers or patients.