人生的成长经历必须失去一些东西才.ppt

EPITOPE SPECIFICITIES OF ANTIBODIES REACTIVE WITH SINGLE HLA ANTIGENS IN LUMINEX ASSAYS Aurora D, Balazs I, Davies A, Ebner D, Hariharan J, Jiang N, Mostecki J, Ray B, Zaslavskaia L TEPNEL LIFECODES CORPORATION, STAMFORD, CT, USA Objective Identify the sensitizing HLA Class I and Class II epitopes in transplant cases. The pattern of reactivity of antibody eluted from each bead was compared with the reactivities predicted by the epitopes listed in HLAMatchmaker. Lifecodes Class 2 Single Antigen Panel (LSA2) Example of HLA Class I eplets Transplant # 1 Patient: DR1, 15; DR51; DQB5, 6 Donor: DR4, 15; DR51, 53; DQB6, 8 Post transplant HLA Ab: DRB1*0401, 0701, 0901, 1001, 1401 DRB4*0101; DQB*0301, 0302, 0303 Specificity of eluted Class II Ab Patient: DR11, 13; DR52; DQ2, 9 Donor: DR7, 15; DR51, 53; DQ2, 6 Post transplant HLA Ab: DRB1*0901, 1001, 1401, 1501 DRB4*0101; DRB5*0202; DQB*0503, 0601 Specificity of eluted Class II Ab Conclusions Eplets, as defined in HLAMatchmaker, may be used to define the sensitizing epitopes of antibodies developed in transplant patients. These type of studies could be used: to define the most common epitopes involved in humoral rejection. to understand the properties of Ab/Ag interaction for different epitopes and their effect in antidoy assays (e.g. relative sensitivity of individual Ag). to determine the effect of competition among Ab, in a polyclonal Ab, for the same Ag. How does this affect relative signal? * Procedure outline Sera samples used for this study were derived from transplanted patients positive for anti-HLA Class I and/or II Ab using LSA (Tepnel Lifecodes). Individual HLA single antigens were bound to latex beads. Based on the Ag of the donor and the anti-HLA reactivity(ies), serum was incubated with different antigen-beads. Following incubation, anti-HLA bound to each antigen-bead was eluted and tested with the Lifecodes Single Antigen assay. DRB5*0101 ? ? ? ? DPA1*0202 DPB1*2801 DRB4*0101 ? ? DPA1*0301 DPB1*1901