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

PLoS One. 2022 Oct 13;17(10):e0275564. doi: 10.1371/journal.pone.0275564. eCollection 2022.

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.

MeSH terms

  • Animals
  • B-Cell Activating Factor
  • Cell Proliferation
  • Graft Rejection
  • Immunoglobulin G
  • Isoantibodies
  • Isoantigens
  • Kidney Transplantation*
  • Rats
  • Rats, Inbred Lew
  • Rodentia
  • Tumor Necrosis Factor Ligand Superfamily Member 13

Substances

  • B-Cell Activating Factor
  • Immunoglobulin G
  • Isoantibodies
  • Isoantigens
  • TNFSF13 protein, human
  • Tumor Necrosis Factor Ligand Superfamily Member 13

Grants and funding

RR- KL2 career development award (4KL2TR000428-10), https://ictr.wisc.edu/career-development-awards-2/; American Society of Transplant Surgeons Faculty Development Grant (MSN183242), https://asts.org/asts-foundation/grants-and-eligibility; American College of Surgeons Franklin Martin, MD, FACS Faculty Research Fellowship (MSN192116), https://www.facs.org/member-services/scholarships/research/acsfaculty. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.