天疱疮抗独特型单克隆抗体的研制和鉴定

PreparationandCharacterizationofAnti-idiotypeMonoclonalAntibodySpecifictoPemphigus

用纯化的天疱疮抗体ＩｇＧ作为抗原免疫ＢＡＬＢ／ｃ小鼠，取免疫鼠的脾细胞与ＳＰ２／０骨髓瘤细胞融合，经过反复筛选、克隆化获得两株（ＰＩ１，ＰＩ２）分泌抗天疱疮独特型的单克隆抗体的杂交瘤细胞系。对其中之一ＰＩ１作了深入鉴定。ＰＩ１的抗体亚类属于ＩｇＧ２ｂ。核型分析结果染色体数为８６条。通过免疫荧光竞争抑制试验和ＥＬＩＳＡ中和抑制试验对ＰＩ１进行鉴定，发现ＰＩ１既能干扰抗原与天疱疮抗体的结合，又能被不同来源的另一天疱疮抗体所中和。无论竞争性抑制还是中和抑制，都呈现剂量依赖关系。表明ＰＩ１杂交瘤细胞所分泌的单克隆抗独特型抗体为结合部位相关型抗体，并对ＰＩ１的可能应用意义进行了讨论。

WepurifiedIgGintheserumofapatientwithpemphigusvulgaris.AfterimmunizingBALB/cmicewithpurifiedpemphigusautoantibody(Pab),spleencelsofthemicewerefusedwithamyelomacellineSP2/0,andweselectedtwohybridomacellines(PI1,PI2)whichproducedanti-idio-typeantibodyspecifictothepemphigusantibodyIgG.Theanti-idiotypeantibodyproducedbyPI1hy-bridomacellinewascharacterized,itwasoftheIgG2bsubclass.PI1couldinhibitthebindingofPabtopemphigusantigen.ThebindingofPI1toPabcouldbeinhibitedbytheIgGantibodyintheseraofotherpatientswithpemphigus.Bothcompetitiveinhibitionandneutralizationinhibitionweredose-depen-dent.ThissuggeststhatidiotypeantibodyrecognisedbyPI1iscloselyasociatedwiththeparatopyofPab.ThepotentialusefulnessofPI1inthestudyoftheclinicaldiagnosis,treatmentandimmuneregu-lationofpatientswithpemphigusisdiscussed.