ACR Meeting Abstracts

ACR Meeting Abstracts

Abstract Number: 1949

Generation and Immunophenotyping of Citrullin-Specific T Cell Lines from Peripheral Blood and Synovial Fluid of ACPA+ RA Patients

Muhammad Sohel Mia1, Christina Gerstner1, Daniel Ramsköld1, Jessica Herrath2, Anatoly Dubnovitsky1,3, Marline Marline4, Annika van Vollenhoven1, Qiaolin Deng4, Adnane Achour3 and Vivianne Malmström5, 1Department of Medicine, Rheumatology Unit, Karolinska University Hospital, Solna, Stockholm, Sweden, 2Department of Medicine,, Rheumatology Unit, Department of Medicine, Solna, Karolinska Institutet, Stockholm, Sweden, 3Department of Medicine, Science for Life Laboraories, Karolinska Institute, Stockholm, Sweden, 4Cell and Molecular Biology Department, Ludwig Institute for Cancer Research, Karolinska Institute, Solna, Stockholm, Sweden, 5Rheumatology Unit, Department of Medicine, Karolinska Institutet, Stockholm, Sweden

Meeting: 2015 ACR/ARHP Annual Meeting

Date of first publication: September 29, 2015

Keywords:

Session Information

Date: Monday, November 9, 2015

Title: T cell Biology and Targets in Autoimmune Disease Poster I

Session Type: ACR Poster Session B

Session Time: 9:00AM-11:00AM

Background/Purpose: Citrulline autoimmunity is a characteristic of ACPA+ Rheumatoid Arthritis (RA). Many autoantigens have been implicated even within patients of the same HLA such as HLA-DR*0401. By HLA-tetramer technology, autoreactive T cells can be captured and studied, but ACPA status cannot predict which T cell specificity will be most abundant. The present study aimed to evaluate the functional and transcriptional character of citrullin-specific T cells from RA synovial fluid and peripheral blood.  

Methods: Peripheral blood mononuclear (PBMC) and synovial fluid mononuclear cells (SFMC) were isolated HLA-DR*04:01+ subjects. A pool of HLA DRB1*0401 class II tetramers (Tmr) were used to isolate autoreactive CD4+ T cells recognizing validated T cell epitopes from citrullinated alpha-enolase and vimentin T cell lines were generated following polyclonal stimuli and expansion and their specificity determined by stimulating with the same set of peptides used in the tetramers. Proliferation and cytokine profiling was measured by thymidine incorporation and cytokine bead array respectively. Smart-seq-2 RNA seq methods were used to perform transcriptomical analyses. 

Results: By the tetramer pool technology we can reproducibly capture and grow autoreactive T cells even from synovial fluid and from modest cell numbers. Approximately half of the Tmr positive cell lines from both SFMC (42%) and PBMC (47%) were positive upon rechallenge. Citrulline reactivity was confirmed by both proliferation and cytokine output. We also demonstrate proof of principle of single cells transcriptomics (n=24) from RA synovial fluid with identification of approx 3000 genes per cell.

Conclusion: We demonstrated that citrulline specific T cell lines could be generated from both peripheral blood and synovial fluid. By this method we can capture autoreactive T cells even from modest number of cells without knowing the dominant T cell subset. This will enable identification of autoreactive T cell receptors (TCR) and down stream studies of e.g. T cell – APC interaction.

Disclosure: M. S. Mia, None; C. Gerstner, None; D. Ramsköld, None; J. Herrath, None; A. Dubnovitsky, None; M. Marline, None; A. van Vollenhoven, None; Q. Deng, None; A. Achour, None; V. Malmström, No commercial Interest, 2.

To cite this abstract in AMA style:

Mia MS, Gerstner C, Ramsköld D, Herrath J, Dubnovitsky A, Marline M, van Vollenhoven A, Deng Q, Achour A, Malmström V. Generation and Immunophenotyping of Citrullin-Specific T Cell Lines from Peripheral Blood and Synovial Fluid of ACPA+ RA Patients [abstract]. Arthritis Rheumatol. 2015; 67 (suppl 10). https://acrabstracts.org/abstract/generation-and-immunophenotyping-of-citrullin-specific-t-cell-lines-from-peripheral-blood-and-synovial-fluid-of-acpa-ra-patients/. Accessed .

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