Quantification of IFNγ- and IL17-producing cells after stimulation with citrullinated proteins in healthy subjects and RA patients

Steendam, Katleen Van; Ceuleneer, Marlies De; Tilleman, Kelly; Elewaut, Dirk; Keyser, Filip De; Deforce, Dieter

doi:10.1007/s00296-012-2470-9

Cited by 9 publications

(5 citation statements)

References 8 publications

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“…In line with this, ACPAs are of the IgG subtype, which indicates that ACPA-producing B cells have undergone T cell-dependent class switching [ 23 ]. Moreover, citrullinated epitope specific T cells have been identified in ACPA + patients [ 24 – 26 ].…”

Section: Introductionmentioning

confidence: 99%

CCR6+ Th cell populations distinguish ACPA positive from ACPA negative rheumatoid arthritis

Paulissen¹,

Hamburg²,

Davelaar³

et al. 2015

Arthritis Res Ther

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IntroductionPatients with rheumatoid arthritis (RA) can be separated into two major subpopulations based on the absence or presence of serum anti-citrullinated protein antibodies (ACPAs). The more severe disease course in ACPA+ RA and differences in treatment outcome between these subpopulations suggest that ACPA+ and ACPA− RA are different disease subsets. The identification of T-helper (Th) cells specifically recognizing citrullinated peptides, combined with the strong association between HLA-DRB1 and ACPA positivity, point toward a pathogenic role of Th cells in ACPA+ RA. In this context we recently identified a potential pathogenic role for CCR6+ Th cells in RA. Therefore, we examined whether Th cell population distributions differ by ACPA status.MethodsWe performed a nested matched case–control study including 27 ACPA+ and 27 ACPA− treatment-naive early RA patients matched for disease activity score in 44 joints, presence of rheumatoid factor, sex, age, duration of complaints and presence of erosions. CD4+CD45RO+ (memory) Th cell distribution profiles from these patients were generated based on differential chemokine receptor expression and related with disease duration.ResultsACPA status was not related to differences in total CD4+ T cell or memory Th cell proportions. However, ACPA+ patients had significantly higher proportions of Th cells expressing the chemokine receptors CCR6 and CXCR3. Similar proportions of CCR4+ and CCR10+ Th cells were found. Within the CCR6+ cell population, four Th subpopulations were distinguished based on differential chemokine receptor expression: Th17 (CCR4+CCR10−), Th17.1 (CXCR3+), Th22 (CCR4+CCR10+) and CCR4/CXCR3 double-positive (DP) cells. In particular, higher proportions of Th22 (p = 0.02), Th17.1 (p = 0.03) and CCR4/CXCR3 DP (p = 0.01) cells were present in ACPA+ patients. In contrast, ACPA status was not associated with differences in Th1 (CCR6−CXCR3+; p = 0.90), Th2 (CCR6−CCR4+; p = 0.27) and T-regulatory (CD25hiFOXP3+; p = 0.06) cell proportions. Interestingly, CCR6+ Th cells were inversely correlated with disease duration in ACPA− patients (R2 = −0.35; p < 0.01) but not in ACPA+ (R2 < 0.01; p = 0.94) patients.ConclusionsThese findings demonstrate that increased peripheral blood CCR6+ Th cells proportions distinguish ACPA+ RA from ACPA− RA. This suggests that CCR6+ Th cells are involved in the differences in disease severity and treatment outcome between ACPA+ and ACPA− RA.

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Section: Introductionmentioning

confidence: 99%

CCR6+ Th cell populations distinguish ACPA positive from ACPA negative rheumatoid arthritis

Paulissen¹,

Hamburg²,

Davelaar³

et al. 2015

Arthritis Res Ther

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“…One type of these modifications is citrullination, the modification of peptidylarginine to citrulline. Citrullination occurs under physiological conditions, but insufficient regulation can induce the formation of neoepitopes that may promote autoimmune reactions to modified Ags (16) and possibly also to the native protein (17). Notably, ApoE (18) as well as other apolipoproteins (19) have previously been identified as part of the "citrullinome" in RA patient SF.…”

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confidence: 99%

Apolipoprotein E Triggers Complement Activation in Joint Synovial Fluid of Rheumatoid Arthritis Patients by Binding C1q

Vogt

Kwasniewicz

Talens

et al. 2020

The Journal of Immunology

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We identified apolipoprotein E (ApoE) as one of the proteins that are found in complex with complement component C4d in pooled synovial fluid of rheumatoid arthritis (RA) patients. Immobilized human ApoE activated both the classical and the alternative complement pathways. In contrast, ApoE in solution demonstrated an isoform-dependent inhibition of hemolysis and complement deposition at the level of sC5b-9. Using electron microscopy imaging, we confirmed that ApoE interacts differently with C1q depending on its context; surface-bound ApoE predominantly bound C1q globular heads, whereas ApoE in a solution favored the hinge/stalk region of C1q. As a model for the lipidated state of ApoE in lipoprotein particles, we incorporated ApoE into phosphatidylcholine/phosphatidylethanolamine liposomes and found that the presence of ApoE on liposomes increased deposition of C1q and C4b from serum when analyzed using flow cytometry. In addition, posttranslational modifications associated with RA, such as citrullination and oxidation, reduced C4b deposition, whereas carbamylation enhanced C4b deposition on immobilized ApoE. Posttranslational modification of ApoE did not alter C1q interaction but affected binding of complement inhibitors factor H and C4b-binding protein. This suggests that changed ability of C4b to deposit on modified ApoE may play an important role. Our data show that posttranslational modifications of ApoE alter its interactions with complement. Moreover, ApoE may play different roles in the body depending on its solubility, and in diseased states such as RA, deposited ApoE may induce local complement activation rather than exert its typical role of inhibition.

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“…In addition, as staining reagents, they can facilitate in-depth, exploratory single cell analyses, such as TCR sequencing, transcriptomics, assessment of oligoclonal expansion, and cloning ( 6 , 23 ). Furthermore, because they can be applied to flow cytometry or cell sorting, multimers are much more sensitive and versatile than ex vivo analyses such as ELISPOT, which struggle to detect low level cytokine responses above background produced by autoantigen-specific memory T cells ( 24 – 26 ).…”

Section: Discussionmentioning

confidence: 99%

Flow Cytometric Clinical Immunomonitoring Using Peptide–MHC Class II Tetramers: Optimization of Methods and Protocol Development

et al. 2018

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With the advent of novel strategies to induce tolerance in autoimmune and autoimmune-like conditions, clinical trials of antigen-specific tolerizing immunotherapy have become a reality. Besides safety, it will be essential to gather mechanistic data on responding CD4+ T cells to assess the effects of various immunomodulatory approaches in early-phase trials. Peptide–MHC class II (pMHCII) multimers are an ideal tool for monitoring antigen-specific CD4+ T cell responses in unmanipulated cells directly ex vivo. Various protocols have been published but there are reagent and assay limitations across laboratories that could hinder their global application to immune monitoring. In this methodological analysis, we compare protocols and test available reagents to identify sources of variability and to determine the limitations of the tetramer binding assay. We describe a robust pMHCII flow cytometry-based assay to quantify and phenotype antigen-specific CD4+ T cells directly ex vivo from frozen peripheral blood mononuclear cell samples, which we suggest should be tested across various laboratories to standardize immune-monitoring results.

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Quantification of IFNγ- and IL17-producing cells after stimulation with citrullinated proteins in healthy subjects and RA patients

Cited by 9 publications

References 8 publications

CCR6+ Th cell populations distinguish ACPA positive from ACPA negative rheumatoid arthritis

CCR6+ Th cell populations distinguish ACPA positive from ACPA negative rheumatoid arthritis

Apolipoprotein E Triggers Complement Activation in Joint Synovial Fluid of Rheumatoid Arthritis Patients by Binding C1q

Flow Cytometric Clinical Immunomonitoring Using Peptide–MHC Class II Tetramers: Optimization of Methods and Protocol Development

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