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IgM+ Antigen Binding B Cells in Peripheral Blood Regulate IFNγ Production in Response to HLA Proteins

L. McLaughlin,1 K. Shiu,1 G. Lombardi,1 J. Spencer,2 R. Vaughan,1 A. Dorling.1

1MRC Centre for Transplantation, King's College London, London, United Kingdom
2Peter Gorer Department of Immunobiology, King's College London, London, United Kingdom.

Meeting: 2015 American Transplant Congress

Abstract number: C15

Keywords: Allorecognition, B cells, Indirect pathway, T cell reactivity

Session Information

Session Name: Poster Session C: Antigen Presenting Cells in Alloimmune Responses/B Cells and Antibody in Alloimmune Responses

Session Type: Poster Session

Date: Monday, May 4, 2015

Session Time: 5:30pm-6:30pm

 Presentation Time: 5:30pm-6:30pm

Location: Exhibit Hall E

Chronic antibody mediated rejection (CAMR) is a major cause of graft loss in renal transplant recipients. Previous work utilising an IFNγ ELISPOT assay revealed two broad patterns of B cell involvement in indirect anti-donor alloreactivity in response to whole donor antigens in CAMR: 1. B cell-dependent anti-donor reactivity, and 2. apparent suppression of anti-donor reactivity by B cells. We hypothesised that these different ELISPOT patterns were due to the presence of distinct phenotypes of alloantigen-specific B cells, and that an understanding of the phenotype of antigen-specific B cells is required if we are ever to achieve our goal of highly tailored treatment of CAMR in each individual.

Pure™ HLA proteins A*01:01 or A*02:01 were used in the indirect IFNγ ELISPOT assay and to detect HLA binding B cells in a cohort of patients with HLA A1 or A2 mismatched grafts respectively. IFNγ production in response to A1 or A2 was seen in 12/34 samples: when B cells were depleted, IFNγ spots reduced in 8/12 (=Bdep); increased in 2/12 (=Bsup) and were unaffected in 2/12 reactive and the remaining 22 non-reactive samples (=Bnon). Analysis of whole B cells revealed no differences in gross phenotype when comparing Bdep, Bsup or Bnon. HLA A1 binding B cells were detected in 4/33 samples from patients with A1 or A2 mismatched grafts Detailed phenotypic analysis was performed on 8/14 samples where the frequency of Ag-binding B cells was >50% above background. Ag-binding B cells were predominantly non transitional and IgM+ in all of these samples. 3/8 Bdep samples had HLA binding B cells that could be phenotyped in depth; IgMhiCD45RB+ (a memory phenotype) cells represented 76±1.5% of Ag-binding cells, whereas IgM+CD45RB- cells accounted for 13±5.4%. 1/2 Bsup and 4/24 Bnon had HLA-binding B cells that could be phenotyped. In these the phenotype was in contrast to the Bdep samples as IgMhiCD45RB+ and IgM+CD45RB- cells made up 45.9±13% and 38.8±8% respectively. These results complement previous data from our lab and indicate that ELISPOT reactivity correlates with the predominance of Ag-binding IgM+ memory cells in patients with CAMR.

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To cite this abstract in AMA style:

McLaughlin L, Shiu K, Lombardi G, Spencer J, Vaughan R, Dorling A. IgM+ Antigen Binding B Cells in Peripheral Blood Regulate IFNγ Production in Response to HLA Proteins [abstract]. Am J Transplant. 2015; 15 (suppl 3). https://atcmeetingabstracts.com/abstract/igm-antigen-binding-b-cells-in-peripheral-blood-regulate-ifn-production-in-response-to-hla-proteins/. Accessed May 30, 2025.

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