Moonsom S, Khunkeawla P, Kasinrerk W
Department of Clinical Immunology, Faculty of Associated Medical Sciences, Chiang Mai University, 50200, Chiang Mai, Thailand.
Immunol Lett. 2001 Feb 1;76(1):25-30. doi: 10.1016/s0165-2478(00)00321-7.
DNA immunization, in theory, is of great interest as a source of specific antibodies against different antigens. In an attempt to produce polyclonal and monoclonal antibodies against cell surface molecules by using the DNA immunization strategy, intramuscular and intrasplenic routes of DNA injection were compared. Two to five, but not a single, intramuscular DNA immunizations induced anti-CD54 and anti-CD147 antibody production. In contrast, a single intrasplenic immunization of CD54-encoding DNA could induce anti-CD54 antibody production. To produce monoclonal antibody (mAb), spleen cells obtained from an intrasplenic CD54-encoding DNA immunized mouse were fused with myeloma cells using the standard hybridoma technique. A hybridoma secreting specific mAb to CD54 was established. The generated mAb reacted to CD54 protein expressed on transfected COS cells and various cell types, the same as using standard CD54 mAb MEM-111. Our results demonstrated that direct immunization of antigen-encoding DNA into spleen is an effective route for production of both polyclonal and monoclonal antibodies to cell surface molecules. This finding is very useful for the production of antibodies to cell surface molecules where the protein antigen is not available or difficult to prepare, but cDNA encoding the corresponding protein is available.
理论上，DNA免疫作为一种针对不同抗原产生特异性抗体的来源具有很大的吸引力。为了通过DNA免疫策略产生针对细胞表面分子的多克隆和单克隆抗体，比较了肌肉内和脾内DNA注射途径。两到五次而非单次肌肉内DNA免疫可诱导抗CD54和抗CD147抗体产生。相比之下，单次脾内注射编码CD54的DNA可诱导抗CD54抗体产生。为了产生单克隆抗体（mAb），使用标准杂交瘤技术将从脾内注射编码CD54的DNA免疫的小鼠获得的脾细胞与骨髓瘤细胞融合。建立了分泌针对CD54的特异性mAb的杂交瘤。所产生的mAb与转染的COS细胞和各种细胞类型上表达的CD54蛋白发生反应，与使用标准CD54 mAb MEM-111的情况相同。我们的结果表明，将编码抗原的DNA直接免疫到脾脏中是产生针对细胞表面分子的多克隆和单克隆抗体的有效途径。这一发现对于在蛋白质抗原不可用或难以制备但编码相应蛋白质的cDNA可用的情况下产生针对细胞表面分子的抗体非常有用。
