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Logo of annrheumdAnnals of the Rheumatic DiseasesCurrent TOCInstructions for authors
Ann Rheum Dis. May 2005; 64(5): 722–729.
Published online Nov 11, 2004. doi:  10.1136/ard.2004.024455
PMCID: PMC1755485

Analysis of the CD8+ T cell response to the G1 domain of aggrecan in ankylosing spondylitis

Abstract

Background: CD4+ T cell responses to the G1 domain of aggrecan in patients with ankylosing spondylitis (AS) were recently reported. Whether such an immune response can be seen in the CD8+ subpopulation has not yet been determined.

Objective: To determine if HLA-B27 restricted G1-specific CD8+ T cells are present in AS and to analyse immunodominant CD8+ T cell epitopes.

Methods: Peripheral blood mononuclear cells of 45 patients with AS were stimulated with overlapping 18-mer peptides covering the whole G1 protein. Results were compared with those for patients with rheumatoid arthritis (RA) and healthy controls. For epitope analysis, G1-specific interferon gamma positive (IFNγ+) T cells were isolated by magnetic activated cell sorting. After in vitro expansion, CD8+ T cells were restimulated with 14 subpools of G1 peptides. T cells responding to G1 peptide subpools were quantified by flow cytometry according to IFNγ secretion. Predicted peptides were subsequently confirmed by stimulation with single peptides.

Results: G1-specific CD8+ T cell responses were found in 29/45 (64%) patients with AS, 18/35 (51%) patients with RA, but not in healthy controls. Five CD8+ T cell epitopes were identified as immunodominant in five patients. However, the T cell response was not HLA-B27 restricted. Nonamer peptides with an HLA-B27 binding motif did not induce a T cell response.

Conclusion: A G1 peptide-specific CD8+ T cell response is present in AS but also in patients with RA. It does not seem to be HLA-B27 restricted. Whether such a response has a role in the pathogenesis of AS needs clarification.

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Selected References

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Figures and Tables

Figure 1
 Percentage of patients with AS (64%), with RA (51%), or healthy controls (0%) responding to in vitro stimulation with the whole peptide pool of overlapping G1 peptides of the proteoglycan aggrecan. Response was measured by IFNγ production ...
Figure 2
 Example of an antigen-specific response to the whole peptide pool of aggrecan derived G1 peptides compared with stimulation without antigen in a patient with AS. After staining for T cell surface markers and intracellular cytokines a gate for ...
Figure 6
 Example of 18-mer peptides derived from the G1 domain of aggrecan inducing an antigen-specific CD8+ T cell response but not CD4+ T cell response compared with stimulation without antigen (Ag) in patient 1. After staining for T cell surface markers ...
Figure 7
 Antigen-specific CD8+ T cell response to 18-mer single peptides (peptides 16,17) which cannot be blocked by an anti-HLA B27 antibody in patient 1. T cells were taken from PB.
Figure 3
 Example of an antigen-specific IFNγ secretion assay after in vitro stimulation with aggrecan G1 peptides compared with stimulation without antigen in a patient with AS. After stimulation, IFNγ+ T cells were isolated twice by ...
Figure 4
 (A) Example of a patient with AS (patient 1) with an antigen-specific CD8+ T cell response to the subpools of peptides (pools 1, 8, 10, 11) derived from the G1 domain of aggrecan compared with stimulation without antigen. After staining for ...
Figure 5
 Example of a patient with AS with an antigen-specific CD8+ T cell response to 18-mer single peptide 17, but not to a nonamer peptide that is part of this 18-mer peptide, derived from the G1 domain of aggrecan, compared with stimulation without ...

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