Recent data from clinical transplantations suggest a critical role for alloantibodies in acute and chronic rejection. A role has also been hypothesized for alloreactive B cells as antigen-presenting cells, stimulating alloreactive T cells via the indirect pathway. However, the diversity of the B cell repertoire makes it difficult to study the behavior of alloreactive B cells in vivo. We hypothesize that a mouse with a monoclonal population of MHC Class I-specific B cells allow in-depth studies into the role of alloreactive B cells in allograft rejection. In accordance with the stated intent of an R03 award of developing new reagents and model systems, the goal of this application is to generate an alloreactive BCR-knock-in (anti-BALB/c) mouse on a C57BL/6 background. We have identified an H-2Kd- and H-2Kk-specific HB-24 hybridoma, and have cloned and sequenced the genes encoding the light and heavy chains. BCR-transgenic (Tg) mice expressing both the light and heavy chain transgenes have been generated in C57BL/6 mice using standard transgenesis techniques. Alloreactive BCR-transgenic (Tg) B cells are enriched in HB-24 BCR-Tg C57BL/6 mice, and respond to immunization with Kd- or Kk-expressing cells by secreting antibodies of HB-24 specificity. We now propose to generate HB-24 BCR-knock-in mice. The specific experiments are organized into two Specific Aims.
Specific Aim I : Generate anti-H2-KdDk BCR-knock-in mice.
Specific Aim 2 : Perform phenotypic and functional characterizations of the B cells in the anti-H2-KdDk BCR-knock-in mice. The successful generation of alloreactive BCR-knock-in mice will facilitate the investigation of several central questions in transplant immunology, including the role of B cells in rejection and tolerance. Recent data from clinical transplantation suggest a critical role for allo-reactive B cells and antibodies in acute and chronic rejection. We propose to generate a mouse expressing a high frequency of alloreactive B cells to study the fate of these B cells after allograft transplantation. Such studies with this new mouse model may lead to the identification of new therapies for controlling alloreactive B cell responses and facilitating the acquisition of long-term allograft tolerance.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Small Research Grants (R03)
Project #
5R03AI069284-02
Application #
7685360
Study Section
Cellular and Molecular Immunology - A Study Section (CMIA)
Program Officer
Rice, Jeffrey S
Project Start
2008-09-12
Project End
2011-08-31
Budget Start
2009-09-01
Budget End
2011-08-31
Support Year
2
Fiscal Year
2009
Total Cost
$77,000
Indirect Cost
Name
University of Chicago
Department
Surgery
Type
Schools of Medicine
DUNS #
005421136
City
Chicago
State
IL
Country
United States
Zip Code
60637
Chen, J; Yin, H; Xu, J et al. (2013) Reversing endogenous alloreactive B cell GC responses with anti-CD154 or CTLA-4Ig. Am J Transplant 13:2280-92
Chang, Anthony; Moore, Jocelyn M; Cowan, Michelle L et al. (2012) Plasma cell densities and glomerular filtration rates predict renal allograft outcomes following acute rejection. Transpl Int 25:1050-8
Cowan, Michelle L; Sciammas, Roger; Chong, Anita S (2012) Experimental models of B cell tolerance in transplantation. Semin Immunol 24:77-85
Ma, Lianli; Xiang, Zhidan; Sherrill, Taylor P et al. (2008) Bioluminescence imaging visualizes activation of nuclear factor-kappaB in mouse cardiac transplantation. Transplantation 85:903-10
Alegre, Maria-Luisa; Goldstein, Daniel R; Chong, Anita S (2008) Toll-like receptor signaling in transplantation. Curr Opin Organ Transplant 13:358-65
Lee, D D; Grossman, E; Chong, A S (2008) Cellular therapies for type 1 diabetes. Horm Metab Res 40:147-54
Wang, Tongmin; Chen, Luqiu; Ahmed, Emily et al. (2008) Prevention of allograft tolerance by bacterial infection with Listeria monocytogenes. J Immunol 180:5991-9
Li, Yijin; Ma, Lianli; Yin, Dengping et al. (2008) Long-term control of alloreactive B cell responses by the suppression of T cell help. J Immunol 180:6077-84
Xiang, Zhidan; Ma, Lian-Li; Manicassamy, Santhakumar et al. (2008) CD4+ T cells are sufficient to elicit allograft rejection and major histocompatibility complex class I molecule is required to induce recurrent autoimmune diabetes after pancreas transplantation in mice. Transplantation 85:1205-11