Production of non-pathogenic human monoclonal antibodies to desmoglein 3 from pemphigus vulgaris patient
Production of non-pathogenic human monoclonal antibodies to desmoglein 3 from pemphigus vulgaris patient
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DOI：
10.1080/08916930290016655
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发表时间：
2002-03-01
期刊：
影响因子：
3.5
通讯作者：
Ahmed, AR
中科院分区：
文献类型：
--
作者：
Bhol, KC;Ahmed, AR
寻常型天疱疮是一种潜在的致命性自身免疫性黏膜皮肤病，与桥粒蛋白3自身抗体的产生有关。桥粒蛋白3是一种130 kDa的表皮蛋白。为了进一步鉴定寻常型天疱疮抗原的表位(S)，我们用人和小鼠的杂交瘤细胞与外周血单核细胞融合，建立了两株人-人杂交瘤。免疫荧光、免疫印迹和ELISA法分析表明，这两株杂交瘤细胞均能与细胞间粘连物质和皮肤中存在的130 kDa蛋白结合，并与重组桥粒蛋白3结合，但不与桥粒蛋白1蛋白结合。免疫球蛋白亚类分布研究表明，这两种抗体均为IgG1亚类，均为非致病性抗体，在体外均不能在器官培养的正常人皮肤中产生棘层松解，也不能在BALB/c新生小鼠体内致病。讨论了这些单抗在寻常型天疱疮发病机制中的相关性和价值。
Pemphigus vulgaris is a potentially fatal autoimmune mucocutaneous disease associated with production of IgG autoantibodies to desmoglein 3, a 130 kDa epidermal protein. To further characterize the epitope(s) of pemphigus vulgaris antigen we established two human-human hybridoma by fusion of the peripheral blood mononuclear cells with a human and mouse heterohybridoma. These hybridomas designated as MAb Dsg-3: 06 and MAb Dsg-3: 10 and stable in culture and demonstrated yield of monoclonal antibodies specific for pemphigus vulgaris.Immunofluorescence, immunoblot, ELISA assays demonstrated that both the monoclonal antibodies bind to the intercellular cement substance and to 130 kDa protein present in the skin and specifically binds to recombinant desmoglein 3 protein, but not to desmoglein 1 protein. The IgG subclass distribution study demonstrated that both the antibodies are of IgG1 subclass in nature, Both the antibodies were non-pathogenic as demonstrated in vitro by their inability to produce acantholysis in normal human skin in organ culture or in vivo by the induction of disease in neonatal BALB/c mice. The relevance and value of these monoclonal antibodies in the pathogenesis of pemphigus vulgaris is discussed.