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Simultaneous analysis of T cell clonality and cytokine production in rheumatoid arthritis using three-colour flow cytometry

Simultaneous analysis of T cell clonality and cytokine production in rheumatoid arthritis using three-colour flow cytometry
Simultaneous analysis of T cell clonality and cytokine production in rheumatoid arthritis using three-colour flow cytometry
In this study we examined the cytokine production by T cells and TCRV? subsets in peripheral blood (PB) and synovial fluid (SF) from six RA patients and PB from 10 normal subjects, using three-colour flow cytometry. In two RA subjects we assessed T cell clonality by RT PCR using TCRBV family-specific primers and analysed the CDR3 (complementarity determining region 3) length by GeneScan analysis. A high percentage of IFN-?- and IL-2- producing cells was observed among the PB T cells in both the RA patients and normal controls and among the SF T cells in RA patients. In contrast, the percentage of T cells producing IL-4 and IL-5 was small among PB T cells in both RA patients and normal controls and among SF T cells in RA patients.
There was no significant difference in the production of IFN-?, IL-2 and IL-5 between the two compartments (PB and SF); however, there were significantly more IL-4-producing cells in SF. Molecular analysis revealed clonal expansions of four TCRBV families in SF of two of the RA patients studied: TCRBV6·7, TCRBV13·1 and TCRBV22 in one and TCRBV6·7, TCRBV21·3 and TCRBV22 in the second. These expansions demonstrated cytokine expression profiles that differed from total CD3+ cells, implying that T cell subsets bearing various TCR-V? families may have the potential to modulate the immune response in RA patients.
cytokine production, rheumatoid arthritis, synovial fluid, t cell receptor clonality
0009-9104
370-378
Bakakos, P.
3570861d-e41b-4ec5-91e2-9fec071e696c
Pickard, C.
e21117b3-6345-4d09-a876-ac9965ec4d6a
Wong, W.M.
6718f842-98bc-4e91-94e2-a2c56dece5d9
Ayre, K.R.
6badcdeb-7793-47d0-bdd0-26c41b5765fa
Madden, J.
0771e352-d432-41ea-8a7e-4704c1efca46
Frew, A.J.
c00e9630-a5f0-44b3-add0-44b68836bbcb
Hodges, E.
85c114a2-4533-48d7-8a6c-7b82a4138251
Cawley, M.I.
2b9c2f70-6486-43ed-a13b-c288d3b53676
Smith, J.L.
f9291742-f9d3-4e27-bfe6-94d41e816fa8
Bakakos, P.
3570861d-e41b-4ec5-91e2-9fec071e696c
Pickard, C.
e21117b3-6345-4d09-a876-ac9965ec4d6a
Wong, W.M.
6718f842-98bc-4e91-94e2-a2c56dece5d9
Ayre, K.R.
6badcdeb-7793-47d0-bdd0-26c41b5765fa
Madden, J.
0771e352-d432-41ea-8a7e-4704c1efca46
Frew, A.J.
c00e9630-a5f0-44b3-add0-44b68836bbcb
Hodges, E.
85c114a2-4533-48d7-8a6c-7b82a4138251
Cawley, M.I.
2b9c2f70-6486-43ed-a13b-c288d3b53676
Smith, J.L.
f9291742-f9d3-4e27-bfe6-94d41e816fa8

Bakakos, P., Pickard, C., Wong, W.M., Ayre, K.R., Madden, J., Frew, A.J., Hodges, E., Cawley, M.I. and Smith, J.L. (2002) Simultaneous analysis of T cell clonality and cytokine production in rheumatoid arthritis using three-colour flow cytometry. Clinical and Experimental Immunology, 129 (2), 370-378. (doi:10.1046/j.1365-2249.2002.01868.x).

Record type: Article

Abstract

In this study we examined the cytokine production by T cells and TCRV? subsets in peripheral blood (PB) and synovial fluid (SF) from six RA patients and PB from 10 normal subjects, using three-colour flow cytometry. In two RA subjects we assessed T cell clonality by RT PCR using TCRBV family-specific primers and analysed the CDR3 (complementarity determining region 3) length by GeneScan analysis. A high percentage of IFN-?- and IL-2- producing cells was observed among the PB T cells in both the RA patients and normal controls and among the SF T cells in RA patients. In contrast, the percentage of T cells producing IL-4 and IL-5 was small among PB T cells in both RA patients and normal controls and among SF T cells in RA patients.
There was no significant difference in the production of IFN-?, IL-2 and IL-5 between the two compartments (PB and SF); however, there were significantly more IL-4-producing cells in SF. Molecular analysis revealed clonal expansions of four TCRBV families in SF of two of the RA patients studied: TCRBV6·7, TCRBV13·1 and TCRBV22 in one and TCRBV6·7, TCRBV21·3 and TCRBV22 in the second. These expansions demonstrated cytokine expression profiles that differed from total CD3+ cells, implying that T cell subsets bearing various TCR-V? families may have the potential to modulate the immune response in RA patients.

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Published date: 2002
Keywords: cytokine production, rheumatoid arthritis, synovial fluid, t cell receptor clonality
Organisations: Dev Origins of Health & Disease

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Local EPrints ID: 26923
URI: http://eprints.soton.ac.uk/id/eprint/26923
ISSN: 0009-9104
PURE UUID: 42f60c52-6277-4807-bc01-fbfbea6e1742

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Date deposited: 25 Apr 2006
Last modified: 15 Jul 2019 19:13

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Contributors

Author: P. Bakakos
Author: C. Pickard
Author: W.M. Wong
Author: K.R. Ayre
Author: J. Madden
Author: A.J. Frew
Author: E. Hodges
Author: M.I. Cawley
Author: J.L. Smith

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