APRIL/BLyS deficient rats prevent donor specific antibody (DSA) production and cell proliferation in rodent kidney transplant model

Natalie M. Bath, Department of Surgery, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
Bret M. Verhoven, Department of Surgery, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
Nancy A. Wilson, Department of Surgery, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
Weifeng Zeng, Division of Plastic Surgery, Department of Surgery, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
Weixiong Zhong, Department of Pathology, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
Lauren Coons, Department of Surgery, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
Arjang Djamali, Department of Medicine, Maine Medical Center, Portland, Maine, United States of America.
Robert R. Redfield, Division of Transplant, Department of Surgery, Hospital of the University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.

Abstract

APRIL (A proliferation inducing ligand) and BLyS (B Lymphocyte Stimulator) are two critical survival factors for B lymphocytes and plasma cells, the main source of alloantibody. We sought to characterize the specific effects of these cytokines in a kidney transplant model of antibody mediated rejection (AMR). We engineered APRIL-/- and BLyS-/- Lewis rats using CRISPR/Cas9. APRIL-/- and BLyS-/- rats were sensitized with Brown Norway (BN) blood (complete MHC mismatch). Twenty-one days following sensitization, animals were harvested and collected tissues were analyzed using flow cytometry, ELISPOT, and immunohistochemistry. Flow cross match and a 3 day mixed lymphocyte reaction (MLR) was performed to assess donor specific antibody (DSA) production and T-cell proliferation, respectively. Sensitized dual knock out Lewis rats (APRIL-/-/BLyS-/-) underwent kidney transplantation and were sacrificed on day 7 post-transplant. Sensitized BLyS-/- had significant decreases in DSA and cell proliferation compared to WT and APRIL-/- (p<0.02). Additionally, BLyS-/- rats had a significant reduction in IgG secreting cells in splenic marginal zone B lymphocytes, and in cell proliferation when challenged with alloantigen compared to WT and APRIL-/-. Transplanted APRIL-/-/BLyS-/- rodents had significantly less DSA and antibody secreting cells compared to WT (p<0.05); however, this did not translate into a significant difference in AMR seen between groups. In summary, our studies suggest that APRIL and BLyS play a greater role in DSA generation rather than AMR, highlighting the role of cellular pathways that regulate AMR.