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Platelet-Specific CD8+ T Cells in Patients with Immune Thrombocytopenia

J. Vrbensky1, I. Nazy1, J. Kelton1, M. Larché2, R. Clare1, H. Bhakta1, M. Roussakis1, V. Rybenchuk1, D. Arnold1

1Platelet Immunology Laboratory at McMaster University, Hamilton, Ontario, Canada, 2McMaster University, Hamilton, Ontario, Canada

Abstract Number: VPB1223

Meeting: ISTH 2022 Congress

Theme: Platelet Disorders, von Willebrand Disease and Thrombotic Microangiopathies » Acquired Thrombocytopenias

Background: Immune thrombocytopenia (ITP) is a bleeding disorder characterized by a low platelet count. T cells have been implicated in the pathogenesis of ITP, but evidence of platelet-specific CD8+ T cells is lacking.

Aims: Detect platelet-specific CD8+ T cells in ITP patients. Characterize CD8+ T cell regulation and activity in ITP.

Methods: Peripheral blood mononuclear cells from ITP patients (n=11) and healthy individuals (n=10) were labelled with carboxyfluorescein succinimidyl ester (CFSE) to detect platelet-specific T cell proliferation in response to platelet autoantigens. CD8+ T cell proliferation was assessed by flow cytometry after 9 days of stimulation with GP IIbIIIa or Ibα. Anti-GP IIbIIIa and anti-GP IbIX were measured with the direct antigen capture assay. Plasma TGF-β, IL-6, GrA, GrB, and sCD137 were assessed by multiplex (Eve Technologies) in ITP patients, healthy individuals, and non-ITP patients (n=5). P values < 0.05 were considered significant. Informed consent was obtained, and the study was approved by the Hamilton Integrated Research Ethics Board.

Results: After stimulation with GP IIbIIIa, 5 of 11 ITP patients and 6 of 10 healthy individuals showed a CD8+ T cell proliferative response. After stimulation with GP Ibα, 9 of 11 ITP patients and 7 of 10 healthy individuals showed a CD8+ T cell proliferative response (Figure 1). Platelet counts and autoantibody levels were comparable in patients with or without platelet-specific T cells. Plasma TGF-β was significantly lower in ITP patients, while IL-6, GrA, GrB, and sCD137 were significantly elevated in ITP patients compared to healthy individuals and non-ITP patients (Figure 2).

Conclusion(s): Platelet-specific CD8+ T cells can be found in ITP and in healthy individuals, and CD8+ T cell activity is dysregulated in ITP patients. Platelet-specific CD8+ T cells should be isolated are characterized for their ability to promote platelet destruction and underproduction in future investigations.

Image

Figure 1: Platelet-Specific CD8+ T Cells.

Image

Figure 2: Plasma Multiplex Analysis.

To cite this abstract in AMA style:

Vrbensky J, Nazy I, Kelton J, Larché M, Clare R, Bhakta H, Roussakis M, Rybenchuk V, Arnold D. Platelet-Specific CD8+ T Cells in Patients with Immune Thrombocytopenia [abstract]. https://abstracts.isth.org/abstract/platelet-specific-cd8-t-cells-in-patients-with-immune-thrombocytopenia/. Accessed October 1, 2023.

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