HLA分型技术 PPT课件.pptVIP

Tempo of rejection reaction type of rejection time taken cause chronic months-years unclear causes: cross reactive Ab, immune complexes, slow cellular reaction, tolerance breakdown, disease recurrence accelerated days reactivation of sensitized T cells (secondary response) acute days-weeks primary activation of T cells hyperacute minutes- hours preformed anti-donor antibodies and complement A, B DR matching and graft survival Removal of T cells from marrow graft Magnet Magnetic antibodies HLA and disease association Disease Associated alleles Frequency in Relative risk patients control Ankylsoing spondylitis Reiter’s disease Acute anterior uveitis Psoriasis vulgaris Dermatitis herpetiformis B27 B27 B27 CW6 DR3 9 79 52 87 85 9 9 9 33 26 87.4 37.0 10.4 13.3 15.4 HLA抗原检测法 微量细胞毒试验 原理 　　　当特异性抗体与淋巴细胞表面HLA分子结合，激活补体，使细胞溶解。在显微镜下可见细胞被活性染料着色。 　　　Ｔ淋巴细胞只表达HLA I分子，如欲测定HLA II类分子，需用Ｂ细胞。 微量细胞毒试验 HLA I类分型法 方法 加板：Terasaki板中加入特异性抗HLA I类分子抗体。 分离PBMC。制成浓度为1?106/ml的细胞悬液。 每孔内加入1?l细胞悬液。 室温培育30min。 1 ?l兔补体。室温培育60min。 每孔内加入5?l 5%伊红水溶液。室温5min。 每孔内加37%甲醛固定。 倒置相差显微镜下观察结果。 HLA分型技术 Major Histopatibility Complex(MHC) What is MHC? HLA H-2 Minor histopatibility antigens Significance of the MHC role in immune response role in organ transplantation role in predisposition to disease Progress in organ transplantation Genetic barriers to transplantation autologous: in the same individual (autograft) isologous: between genetically Identical individuals (isograft), i.e., identical twins (inbred animals) homologous: between individuals of the same species (allograft) heterologous: between individuals different species (xenograft) Principles of transplantation Minor histopatibility antigensand graft survival minor histopatibility antigens also cause rejection The rejection time is variable but longer than that for major histocompatibility antigen They have additive effects Graft versus host (GVH) disease GVH disease in humans The human MHC genes The mouse MHC genes Polymorphism of MHC antige