Allo-Specific Humoral Responses: New Methods for Screening Donor-Specific Antibody and Characterization of HLA-Specific Memory B Cells.

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2021-01

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Abstract

Antibody-mediated allograft rejection (AMR) causes more kidney transplant failure than any other single cause. AMR is mediated by antibodies recognizing antigens expressed by the graft, and antibodies generated against major histocompatibility complex (MHC) mismatches are especially problematic. Most research directed towards the management of clinical AMR has focused on identifying and characterizing circulating donor-specific HLA antibody (DSA) and optimizing therapies that reduce B-cell activation and/or block antibody secretion by inhibiting plasmacyte survival. Here we describe a novel set of reagents and techniques to allow more specific measurements of MHC sensitization across different animal transplant models. Additionally, we have used these approaches to isolate and clone individual HLA-specific B cells from patients sensitized by pregnancy or transplantation. We have identified and characterized the phenotypes of individual HLA-specific B cells, determined the V(D)J rearrangements of their paired H and L chains, and generated recombinant antibodies to determine affinity and specificity. Knowledge of the BCR genes of individual HLA-specific B cells will allow identification of clonally related B cells by high-throughput sequence analysis of peripheral blood mononuclear cells and permit us to re-construct the origins of HLA-specific B cells and follow their somatic evolution by mutation and selection.

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B-Lymphocyte Subsets, Cells, Cultured, Cell Line, Clone Cells, Animals, Mice, Inbred C57BL, Macaca mulatta, Humans, Mice, Immunoglobulin G, beta 2-Microglobulin, RNA, Guide, Isoantibodies, Histocompatibility Antigens, HLA Antigens, Indicators and Reagents, Skin Transplantation, Lymphocyte Activation, Models, Animal, Specific Pathogen-Free Organisms, Antibody Specificity, Species Specificity, Graft Rejection, Gene Rearrangement, B-Lymphocyte, Immunologic Memory, Genes, Reporter, Female, V(D)J Recombination, Clonal Evolution

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https://hdl.handle.net/10161/26169

Published Version (Please cite this version)

10.3389/fimmu.2021.705140

Publication Info

Song, Shengli, Miriam Manook, Jean Kwun, Annette M Jackson, Stuart J Knechtle and Garnett Kelsoe (2021). Allo-Specific Humoral Responses: New Methods for Screening Donor-Specific Antibody and Characterization of HLA-Specific Memory B Cells. Frontiers in immunology, 12. p. 705140. 10.3389/fimmu.2021.705140 Retrieved from https://hdl.handle.net/10161/26169.

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Scholars@Duke

Kwun

Jean Kwun

Associate Professor in Surgery

Dr. Kwun is currently the Director of Transplant Basic Science Research at the Duke Transplant Center.

Research interests include humoral tolerance to organ transplants in animal model and humans, developing a clinically relevant animal model to study the mechanisms of antibody-mediated rejection (AMR), and establishing a conceptual basis that will translate into therapeutic intervention of AMR.

Jackson

Annette M Jackson

Associate Professor in Surgery
Knechtle

Stuart Johnston Knechtle

William R. Kenan, Jr. Distinguished Professor of Surgery

During my career as an academic surgeon, I have had the privilege of leading and/or participating in a diverse portfolio of hypothesis-driven research projects.  These projects have centered on the immunology of surgery and transplantation, including both cellular and antibody-mediated immune responses.  During my training I studied the response of hyper-sensitized recipients to allogeneic liver transplantation, and am currently studying means of reducing immunologic memory that might allow more successful transplantation in sensitized recipients.  This immune response involves pathways of coagulation, antibody-mediated rejection, and cellular rejection and current work in my lab involves these three pathways.  The other major focuses of my work have been co-stimulation blockade and immune cell depletion as approaches to immunologic unresponsiveness or tolerance.  My research group has been involved in translational and clinical research to develop these mechanistic tools for the benefit of human organ transplant recipients.

Recent Publications

Knechtle SJ, Shaw JM, Hering BJ, Kraemer K, Madsen JC. Translational impact of NIH-funded nonhuman primate research in transplantation. Sci Transl Med. 2019 Jul 10;11(500). pii: eaau0143. Reprint | Full Text

Kelsoe

Garnett H. Kelsoe

James B. Duke Distinguished Professor of Immunology
  1. Lymphocyte development and antigen-driven diversification of immunoglobulin and T cell antigen receptor genes.
    2. The germinal center reaction and mechanisms for clonal selection and self - tolerance. The origins of autoimmunity.
    3. Interaction of innate- and adaptive immunity and the role of inflammation in lymphoid organogenesis.
    4. The role of secondary V(D)J gene rearrangment in lymphocyte development and malignancies.
    5. Mathematical modeling of immune responses, DNA motifs, collaborations in bioinformatics.
    6. Humoral immunity to influenza and HIV-1.

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