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Abstracts / Human Immunology 73 (2012) 49–167
DONOR SPECIFIC HLA-DQA ANTIBODY IN RENAL POST-TRANSPLANT PATIENTS. Xuebin Yang, Justine L. Gaspari, Meghan P. Riley, Ronald E. Domen, Dennis F. Habig, Hiroko Shike. Pathology, Penn State Hershey Medical Center, Hershey, PA, USA. Aim: To determine the frequency of donor specific HLA antibody (DSA) to DQ alpha in renal post-transplant recipients. Donor DQA1 typing is not a routine procedure under UNOS. Frequency of post-transplant DQ alpha DSA and possible importance in post-transplant monitoring is not well-studied. Methods: Retrospective review of post-transplant HLA antibody test results. Reviewed are all HLA antibody test results between January 2010 and April 2012 for the patients who received a deceased or living donor kidney at our center. HLA typing of donors and recipients was tested by serology or SSO/SSP. HLA antibody was tested by solid phase methods (LABScreen PRA and LABScreen Single Antigen beads, One Lambda). Current antibody monitoring schedule for low risk patients (no pre-formed or de novo DSA and stable creatinine) is 9 times in the first year, every 6 months until the 3rd year, then annually. Additional tests are performed if higher risk or suspected rejections. If DQ alpha antibody is detected, donor DQA1 was typed retrospectively using frozen cells. Results: Post-transplant HLA antibody was tested on 158 recipients: 27 pediatrics and 131 adults (average 46.3 years old). Results represent data points from two months to 26 years post-transplant (average 5.0 years). In all patients, pre-transplant crossmatch was either compatible by CDC method, or compatible/ weakly incompatible by flow cytometry method. Of the 158 patients, DSA (either de novo or preformed) was detected in 29 patients (18.4%). Detected were class I DSA only in 3 patients, class II DSA only in 20 patients, and both class I/II in 6 patients. Specificities of class II DSAs were DQB1 in 22 patients (85%), DQA1 in 10 patients (39%), DRB1/3/4/5 in 7 patients (27%), and DPB1 in 1 patient (4%). Conclusions: HLA-DQ alpha DSA was detected in a substantial percent of DSA positive renal transplant patients, thus DQA1 donor typing is important in renal post-transplant monitoring.
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SYNERGISTIC EFFECT OF COMPLEMENT FIXING AND NON-COMPLEMENT FIXING ANTIBODIES TO MAJOR HISTOCOMPATIBILITY CLASS I ANTIGENS IN DEVELOPMENT OF OBLITERATIVE AIRWAY DISEASE (CHRONIC LUNG ALLOGRAFT REJECTION). Vijay Subramanian 1, Masashi Takenaka 1, Venkataswarup Tiriveedhi 1, Andrew Gelman 1, Alexander Patterson 1, T. Mohanakumar 2. 1 Department of Surgery, Washington University School of Medicine, St. Louis, MO, USA; 2 Departments of Surgery and Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA. Aim: Our previous results demonstrated that 200 lg complement fixing antibodies (Abs) to MHC class I (cfAbs) or non-complement fixing Abs (ncAbs) result in obliterative airway disease (OAD) in mouse model of intrabronchial anti-MHC administration. In human lung transplantation, recipients may develop both complement fixing and non-complement fixing DSA that may augment chronic rejection. The aim of this study was to test if low levels of cfAbs and ncAbs can synergize leading to OAD. Methods: Low dose anti-MHC class I ncAbs mouse IgG1 (100 lg) or cfAbs IgG2a (25 lg) were given alone or in combination intrabronchially in F1 C57Bl/6 B.10 mice on day 0,1,2,6 and weekly after. Isotype IgG were given in control. Lungs analyzed by histopathology. Serum tested for Abs to Kalpha1Tubulin (Ka1T) and collagenV (ColV) by ELISA. IFNc, IL17 and IL10 cells specific to Ka1T and ColV enumerated by ELISPOT. Cytokines and growth factors in lung determined by RTPCR. Results: Low dose of ncAbs (100 lg) or cfAbs (25 lg) alone did not lead to OAD including isotype control. Combination of ncAbs and cfAbs at same low dose resulted in OAD with cellular infiltration, fibrosis and luminal occlusion of vessels and bronchioles. Mice given both ncAbs and cfAbs developed serum Abs to Ka1T (667 ± 63 lg/mL, p = 0.065) and ColV (323 ± 41, p = 0.0034) compared to ncAbs alone, cfAbs alone or isotype mice. This was associated with increased IFNc (p = 0.002), IL17 (p = 0.004) specific to Ka1T and ColV and suppression of IL10 compared to other groups. Further, there was upregulation of TGFb, MCP-1, IL-6, IL-1b and VEGF. Conclusions: High dose ncAbs and cfAbs to MHC lead to OAD, but low dose of ncAbs or cfAbs given alone did not induce OAD. However, low levels of ncAbs and cfAbs in combination, can synergize and potentiate each other resulting in activation of inflammatory and fibrotic cascades leading to OAD. Hence, in human lung transplant recipients low titers of de novo DSA which are cfAbs and ncAbs may synergize augmenting chronic lung allograft rejection.
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Abstracts / Human Immunology 73 (2012) 49–167
DONOR SPECIFIC HLA-DQA ANTIBODY IN RENAL POST-TRANSPLANT PATIENTS. Xuebin Yang, Justine L. Gaspari, Meghan P. Riley, Ronald E. Domen, Dennis F. Habig, Hiroko Shike. Pathology, Penn State Hershey Medical Center, Hershey, PA, USA. Aim: To determine the frequency of donor specific HLA antibody (DSA) to DQ alpha in renal post-transplant recipients. Donor DQA1 typing is not a routine procedure under UNOS. Frequency of post-transplant DQ alpha DSA and possible importance in post-transplant monitoring is not well-studied. Methods: Retrospective review of post-transplant HLA antibody test results. Reviewed are all HLA antibody test results between January 2010 and April 2012 for the patients who received a deceased or living donor kidney at our center. HLA typing of donors and recipients was tested by serology or SSO/SSP. HLA antibody was tested by solid phase methods (LABScreen PRA and LABScreen Single Antigen beads, One Lambda). Current antibody monitoring schedule for low risk patients (no pre-formed or de novo DSA and stable creatinine) is 9 times in the first year, every 6 months until the 3rd year, then annually. Additional tests are performed if higher risk or suspected rejections. If DQ alpha antibody is detected, donor DQA1 was typed retrospectively using frozen cells. Results: Post-transplant HLA antibody was tested on 158 recipients: 27 pediatrics and 131 adults (average 46.3 years old). Results represent data points from two months to 26 years post-transplant (average 5.0 years). In all patients, pre-transplant crossmatch was either compatible by CDC method, or compatible/ weakly incompatible by flow cytometry method. Of the 158 patients, DSA (either de novo or preformed) was detected in 29 patients (18.4%). Detected were class I DSA only in 3 patients, class II DSA only in 20 patients, and both class I/II in 6 patients. Specificities of class II DSAs were DQB1 in 22 patients (85%), DQA1 in 10 patients (39%), DRB1/3/4/5 in 7 patients (27%), and DPB1 in 1 patient (4%). Conclusions: HLA-DQ alpha DSA was detected in a substantial percent of DSA positive renal transplant patients, thus DQA1 donor typing is important in renal post-transplant monitoring.
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SYNERGISTIC EFFECT OF COMPLEMENT FIXING AND NON-COMPLEMENT FIXING ANTIBODIES TO MAJOR HISTOCOMPATIBILITY CLASS I ANTIGENS IN DEVELOPMENT OF OBLITERATIVE AIRWAY DISEASE (CHRONIC LUNG ALLOGRAFT REJECTION). Vijay Subramanian 1, Masashi Takenaka 1, Venkataswarup Tiriveedhi 1, Andrew Gelman 1, Alexander Patterson 1, T. Mohanakumar 2. 1 Department of Surgery, Washington University School of Medicine, St. Louis, MO, USA; 2 Departments of Surgery and Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA. Aim: Our previous results demonstrated that 200 lg complement fixing antibodies (Abs) to MHC class I (cfAbs) or non-complement fixing Abs (ncAbs) result in obliterative airway disease (OAD) in mouse model of intrabronchial anti-MHC administration. In human lung transplantation, recipients may develop both complement fixing and non-complement fixing DSA that may augment chronic rejection. The aim of this study was to test if low levels of cfAbs and ncAbs can synergize leading to OAD. Methods: Low dose anti-MHC class I ncAbs mouse IgG1 (100 lg) or cfAbs IgG2a (25 lg) were given alone or in combination intrabronchially in F1 C57Bl/6 B.10 mice on day 0,1,2,6 and weekly after. Isotype IgG were given in control. Lungs analyzed by histopathology. Serum tested for Abs to Kalpha1Tubulin (Ka1T) and collagenV (ColV) by ELISA. IFNc, IL17 and IL10 cells specific to Ka1T and ColV enumerated by ELISPOT. Cytokines and growth factors in lung determined by RTPCR. Results: Low dose of ncAbs (100 lg) or cfAbs (25 lg) alone did not lead to OAD including isotype control. Combination of ncAbs and cfAbs at same low dose resulted in OAD with cellular infiltration, fibrosis and luminal occlusion of vessels and bronchioles. Mice given both ncAbs and cfAbs developed serum Abs to Ka1T (667 ± 63 lg/mL, p = 0.065) and ColV (323 ± 41, p = 0.0034) compared to ncAbs alone, cfAbs alone or isotype mice. This was associated with increased IFNc (p = 0.002), IL17 (p = 0.004) specific to Ka1T and ColV and suppression of IL10 compared to other groups. Further, there was upregulation of TGFb, MCP-1, IL-6, IL-1b and VEGF. Conclusions: High dose ncAbs and cfAbs to MHC lead to OAD, but low dose of ncAbs or cfAbs given alone did not induce OAD. However, low levels of ncAbs and cfAbs in combination, can synergize and potentiate each other resulting in activation of inflammatory and fibrotic cascades leading to OAD. Hence, in human lung transplant recipients low titers of de novo DSA which are cfAbs and ncAbs may synergize augmenting chronic lung allograft rejection.