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S antigen specific effector T cell activation detected by cytokine flow cytometry

¹ The Larry Donoso Research Laboratory, Division of Ophthalmology and Visual Sciences, Eye Ear Nose and Throat Centre, Queen's Medical Centre, University of Nottingham, Nottingham NG7 2UH, UK
² Department of Immunology, University of Nottingham, UK
³ The Larry Donoso Research Laboratory, Division of Ophthalmology and Visual Sciences, University of Nottingham, UK
⁴ Departments of Ophthalmology and Immunology, University of Nottingham, UK

Correspondence to:
Correspondence to:
James P Morgan;
James.Morgan{at}nottingham.ac.uk

ABSTRACT
Background/aims: Effector T cell activation is particularly important in the initiation of autoimmune uveitis. This pilot study seeks to demonstrate activation of human peripheral effector T cells in response to the uveitis candidate autoantigen, retinal S antigen (SAg), using cytokine flow cytometry (CFC).

Methods: Peripheral blood mononuclear cell (PBMC) suspensions from uveitis patients and controls were stimulated with bovine SAg. Activation responses were detected by CFC.

Results: Electronic gating enabled analysis of CD69+, IFN-+ CD4+ lymphocytes. An SAg specific response was detectable in four of 13 patients and four of eight controls.

Conclusion: SAg specific, peripheral, effector T cell activation can be detected by CFC. Similar levels of responsiveness were seen in patient and control groups. More detailed cytokine profiling may demonstrate functional differences between the groups.

Keywords: uveoretinitis; cytokine; flow cytometry; S antigen

Abbreviations: CFC, cytokine flow cytometry; EAU, experimental autoimmune uveoretinitis; PBMC, peripheral blood mononuclear cell; PCR, polymerase chain reaction; PMA, phorbol myristate acetate; SAg, retinal S antigen