Memory T cells specific to citrullinated α-enolase are enriched in the rheumatic joint

Pieper, Jennifer; Dubnovitsky, Anatoly; Gerstner, Christina; James, Eddie A.; Rieck, Mary; Kozhukh, Genadiy; Tandre, Karolina; Pellegrino, Sara; Gebe, John A.; Rönnblom, Lars; Sandalova, Tatyana; Kwok, William W.; Klareskog, Lars; Buckner, Jane H.; Achour, Adnane; Malmström, Vivianne

doi:10.1016/j.jaut.2018.04.004

Cited by 42 publications

(46 citation statements)

References 385 publications

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“…Table 2 lists the sequence and position of the peptides as well as the tetramer fluorophores. As autoreactive T cells have been shown to be present at low frequencies in the circulation of RA patients and healthy individuals [5,6] we first determined the sensitivity of our multi-tetramer assay by applying it to PBMC from seven HLA-DRB1*04:01-positive healthy control subjects. Here, we unambiguously detected T cells specific for α-enolase, CILP and fibrinogen at frequencies ranging up to 10 per million CD4+ T cells (median frequency 4.7 and 1.7 per million CD4+ T cells, respectively; Figure 1a and b).…”

Section: The Multi-tetramer Approach Allows Stable Detection Of Autormentioning

confidence: 99%

“…HLA class II tetramers allow direct ex vivo detection and phenotypic characterization of antigenspecific T cells [1] and are increasingly used in a range of different settings, like allergy, vaccination and autoimmune diseases [2][3][4][5][6]. This technology requires knowledge of both the antigenic peptide as well as the HLA restriction element, and hereby is utilised in conditions with known antigenic targets and a strong HLA component.…”

Section: Introductionmentioning

confidence: 99%

“…To detect rare antigen-specific CD4+ T cells ex vivo, others and we made use of a protocol first published by Wucherpfennig and colleagues [15] that combines tetramer staining with magnetic bead enrichment. By this means, we previously quantified and phenotypically characterized T cells specific for epitopes from several RA-associated candidate antigens in a variety of different RA samples, including peripheral blood mononuclear cells (PBMC), synovial mononuclear cells (SFMC) and synovial tissue [5,6]. However, a major shortcoming of this method is the need for large samples,…”

Section: Introductionmentioning

confidence: 99%

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Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis

Gerstner

Turcinov

Hensvold

et al. 2020

Preprint

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Background: HLA class II tetramers can be used for ex vivo enumeration and phenotypic characterization of antigen-specific CD4+ T cells. They are increasingly applied in settings like allergy, vaccination and autoimmune diseases. Rheumatoid arthritis (RA) is a chronic autoimmune disorder for which many autoantigens have been described.Results: Using multi-parameter flow cytometry, we developed a multi-HLA class II tetramer approach to simultaneously study several antigen specificities in RA patient samples. We focused on previously described citrullinated HLA-DRB1*04:01-restricted T cell epitopes from α-enolase, fibrinogen-b, vimentin as well as cartilage intermediate layer protein (CILP). First, we examined inter-assay variability and the sensitivity of the assay in peripheral blood from healthy donors (n=7). Next, we confirmed the robustness and sensitivity in a cohort of RA patients with repeat blood draws (n=14). We then applied our method in two different settings. We assessed lymphoid tissue from seropositive arthralgia (n=5) and early RA patients (n=5) and could demonstrate autoreactive T cells in individuals at risk of developing RA. Lastly, we studied peripheral blood from early RA patients (n=10) and found that the group of patients achieving minimum disease activity (DAS28 <2.6) at 6 months follow-up displayed a decrease in the frequency of citrulline-specific T cells. Conclusions: Our study demonstrates the development of a sensitive tetramer panel allowing simultaneous characterization of antigen-specific T cells in ex vivo patient samples including RA ‘at risk’ subjects. This multi-tetramer approach can be useful for longitudinal immune-monitoring in any disease with known HLA-restriction element and several candidate antigens.

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Section: The Multi-tetramer Approach Allows Stable Detection Of Autormentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis

Gerstner

Turcinov

Hensvold

et al. 2020

Preprint

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“…Early RA patients achieving drug-induced remission show a reduction in citrulline-specific T cells a+b) Frequency of (a) influenza-and citrulline-specific T cells in patients achieving remission (DAS28 at 6m <2.6) versus patients not achieving remission (DAS28 at 6m >2. 6) and (b) of T cells reactive to α-enolase, CILP/FGB and vimentin is depicted at baseline (0m) and follow-up visit (6m) for 10 early RA patients.…”

Section: Figurementioning

confidence: 99%

Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis

Gerstner

Turcinov

Hensvold

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

Background: HLA class II tetramers can be used for ex vivo enumeration and phenotypic characterization of antigen-specific CD4+ T cells. They are increasingly applied in settings like allergy, vaccination and autoimmune diseases. Rheumatoid arthritis (RA) is a chronic autoimmune disorder for which many autoantigens have been described. Results: Using multi-parameter flow cytometry, we developed a multi-HLA class II tetramer approach to simultaneously study several antigen specificities in RA patient samples. We focused on previously described citrullinated HLA-DRB1*04:01-restricted T cell epitopes from α-enolase, fibrinogen-b, vimentin as well as cartilage intermediate layer protein (CILP).First, we examined inter-assay variability and the sensitivity of the assay in peripheral blood from healthy donors (n=7). Next, we confirmed the robustness and sensitivity in a cohort of RA patients with repeat blood draws (n=14). We then applied our method in two different settings. We assessed lymphoid tissue from seropositive arthralgia (n=5) and early RA patients (n=5) and could demonstrate autoreactive T cells in individuals at risk of developing RA. Lastly, we studied peripheral blood from early RA patients (n=10) and found that the group of patients achieving minimum disease activity (DAS28 <2.6) at 6 months follow-up displayed a decrease in the frequency of citrulline-specific memory T cells. Conclusions: Our study demonstrates the development of a sensitive tetramer panel allowing simultaneous characterization of antigen-specific T cells in ex vivo patient samples including RA ‘at risk’ subjects. This multi-tetramer approach can be useful for longitudinal immune-monitoring in any disease with known HLA-restriction element and several candidate antigens.

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“…There have then been attempts to find the missing link between anti-citrullinated peptides (Cit-P) production and carriage of certain alleles of the HLA-DRB1 molecule that favours RA. In RA patients, few studies have identified that SE carriage increased T-cell response to certain Cit-P compared with healthy controls 6–8. Binding studies showed that SE would have a higher affinity to Cit-P 9.…”

Section: Introductionmentioning

confidence: 99%

Valine 11 and phenylalanine 13 have a greater impact on the T-cell response to citrullinated peptides than the 70–74 shared epitope of the DRB1 molecule in macaques

Bitoun

Roques

Maillère³

et al. 2019

Ann Rheum Dis

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ObjectivesVarious rheumatoid arthritis (RA) HLA-DRB-1 risk haplotypes have been regrouped under the shared epitope (SE) in position 70–74. The presence of Valine in position 11 (Val11) and phenylalanine in position 13 (Phe13) are also associated with RA, but it is impossible to differentiate their role compared with the SE since they are in strong linkage disequilibrium (LD) in humans. Similar to humans, certain macaques express the SE (H6). We analysed the effect of various DRB1 haplotypes on T-cell response to citrullinated peptides (Cit-P) in macaques.MethodsSix H6 and six non-H6 macaques were immunized with four Cit-P. T-cell response was assessed using Interferon γ enzyme-linked immunospot.ResultsAnimals developed a specific anti-Cit-P T-cell response. Surprisingly, H6 animals had a significantly lower T-cell response than non-H6. In macaques, the 70–74 SE and the Val11 are on separate haplotypes. Presence of Val11 was strongly associated with the anti-Cit-P T-cell response, whatever the 70–74 sequence was. This response was amplified in case of presence of Phe13.ConclusionThe absence of LD between Val11 and SE in macaques allowed us to demonstrate that the most important HLA positions to induce a T-cell response against Cit-P were Val11 and Phe13 and not the 70–74 SE.

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Memory T cells specific to citrullinated α-enolase are enriched in the rheumatic joint

Cited by 42 publications

References 385 publications

Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis

Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis

Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis

Valine 11 and phenylalanine 13 have a greater impact on the T-cell response to citrullinated peptides than the 70–74 shared epitope of the DRB1 molecule in macaques

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