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CTLA4-Ig (abatacept) therapy modulates T cell effector functions in autoantibody-positive rheumatoid arthritis patients.

Pieper J, Herrath J, Raghavan S, Muhammad K, Vollenhoven Rv, Malmström V - BMC Immunol. (2013)

Bottom Line: Additionally within the ACPA-positive group significant down-regulation of all key T cell effector subsets including Th1, Th2, and Th17 was observed by analyzing cytokines by intracellular flow cytometry and in cell culture supernatants.RA synovial fluid samples were cultured in vitro in the presence or absence of CTLA4-Ig (abatacept).Our immunological study of T cell functionality in RA patients, both ACPA-positive and ACPA-negative starting biological therapy with the co-stimulation blockade abatacept (CTLA4-Ig) supports the recently published registry study implicating ACPA seropositivity as an independent predictive factor to treatment response as we observed the most striking effect on T cell subset modulation in ACPA-positive patients.These data further support the notion of RA as a disease with several sub-entities, where the ACPA-positive fraction represents a classical HLA-associated autoimmune disorder while ACPA-negative patients may have other driving forces apart from classical adaptive immune responses.

View Article: PubMed Central - HTML - PubMed

Affiliation: Rheumatology Unit, Department of Medicine at Karolinska University Hospital, Karolinska Institute, Solna, Stockholm, Sweden.

ABSTRACT

Background: Rheumatoid arthritis is a chronic inflammatory disease with a strong MHC class II component and where many patients develop characteristic autoantibodies towards the noncoding amino acid citrulline. Such anti-citrullinated protein antibodies (ACPA) have recently been put forward as an independent predictive factor for treatment response by co-stimulation blockade by CTLA4-Ig (abatacept). We have performed a mechanism of action study to dissect T cell functionality in RA patients with long-standing disease undergoing abatacept treatment and the influence of ACPA status.

Results: Peripheral blood samples were collected from RA patients as they started CTLA4-Ig treatment and 3 and 6 months later. A general decrease of regulatory T cell subsets was observed in the cohort. Additionally within the ACPA-positive group significant down-regulation of all key T cell effector subsets including Th1, Th2, and Th17 was observed by analyzing cytokines by intracellular flow cytometry and in cell culture supernatants.RA synovial fluid samples were cultured in vitro in the presence or absence of CTLA4-Ig (abatacept). T cell cytokine production was diminished, but without increasing the functional capacity of CD4+CD25hi regulatory T cells as previously demonstrated in the context of TNF-blockade and anti-IL6R therapy.

Conclusions: Our immunological study of T cell functionality in RA patients, both ACPA-positive and ACPA-negative starting biological therapy with the co-stimulation blockade abatacept (CTLA4-Ig) supports the recently published registry study implicating ACPA seropositivity as an independent predictive factor to treatment response as we observed the most striking effect on T cell subset modulation in ACPA-positive patients. These data further support the notion of RA as a disease with several sub-entities, where the ACPA-positive fraction represents a classical HLA-associated autoimmune disorder while ACPA-negative patients may have other driving forces apart from classical adaptive immune responses.

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Flow cytometry staining for intracellular cytokines TNF, IFN-γ, IL-17A. PBMCs were stimulated in vitro with α-CD3 for 6 h or 5 days and intracellular cytokine staining was performed for IFN-γ, TNF and IL-17A. The plotted cells are gated as CD14-CD3+CD4+CD28+ T cells. (A) A representative FACS staining. Left row: Unstimulated cells. Subsequent rows: α-CD3 stimulation at baseline and 6 months following treatment. (B-E) Intracellular cytokine stainings. (B) ACPA-positive patients, 6h stimulation (n=12). (C) ACPA-negative patients, 6h stimulation (n=7). (D) ACPA-positive patients, 5days stimulation (n=12). (E) ACPA-negative patients, 5days stimulation (n=6).
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Figure 2: Flow cytometry staining for intracellular cytokines TNF, IFN-γ, IL-17A. PBMCs were stimulated in vitro with α-CD3 for 6 h or 5 days and intracellular cytokine staining was performed for IFN-γ, TNF and IL-17A. The plotted cells are gated as CD14-CD3+CD4+CD28+ T cells. (A) A representative FACS staining. Left row: Unstimulated cells. Subsequent rows: α-CD3 stimulation at baseline and 6 months following treatment. (B-E) Intracellular cytokine stainings. (B) ACPA-positive patients, 6h stimulation (n=12). (C) ACPA-negative patients, 6h stimulation (n=7). (D) ACPA-positive patients, 5days stimulation (n=12). (E) ACPA-negative patients, 5days stimulation (n=6).

Mentions: At baseline and six months following abatacept therapy, PBMCs from 19 patients were utilized for polyclonal T cell stimulation for both 6 hrs and 5 days, and subsequently intracellular cytokine stainings were performed. By this approach, each individual is its own control. Representative flow cytometry stainings are depicted in Figure 2A. While unstimulated cells did not produce cytokines, robust production was observed following polyclonal stimulation (Figure 2A).


CTLA4-Ig (abatacept) therapy modulates T cell effector functions in autoantibody-positive rheumatoid arthritis patients.

Pieper J, Herrath J, Raghavan S, Muhammad K, Vollenhoven Rv, Malmström V - BMC Immunol. (2013)

Flow cytometry staining for intracellular cytokines TNF, IFN-γ, IL-17A. PBMCs were stimulated in vitro with α-CD3 for 6 h or 5 days and intracellular cytokine staining was performed for IFN-γ, TNF and IL-17A. The plotted cells are gated as CD14-CD3+CD4+CD28+ T cells. (A) A representative FACS staining. Left row: Unstimulated cells. Subsequent rows: α-CD3 stimulation at baseline and 6 months following treatment. (B-E) Intracellular cytokine stainings. (B) ACPA-positive patients, 6h stimulation (n=12). (C) ACPA-negative patients, 6h stimulation (n=7). (D) ACPA-positive patients, 5days stimulation (n=12). (E) ACPA-negative patients, 5days stimulation (n=6).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750242&req=5

Figure 2: Flow cytometry staining for intracellular cytokines TNF, IFN-γ, IL-17A. PBMCs were stimulated in vitro with α-CD3 for 6 h or 5 days and intracellular cytokine staining was performed for IFN-γ, TNF and IL-17A. The plotted cells are gated as CD14-CD3+CD4+CD28+ T cells. (A) A representative FACS staining. Left row: Unstimulated cells. Subsequent rows: α-CD3 stimulation at baseline and 6 months following treatment. (B-E) Intracellular cytokine stainings. (B) ACPA-positive patients, 6h stimulation (n=12). (C) ACPA-negative patients, 6h stimulation (n=7). (D) ACPA-positive patients, 5days stimulation (n=12). (E) ACPA-negative patients, 5days stimulation (n=6).
Mentions: At baseline and six months following abatacept therapy, PBMCs from 19 patients were utilized for polyclonal T cell stimulation for both 6 hrs and 5 days, and subsequently intracellular cytokine stainings were performed. By this approach, each individual is its own control. Representative flow cytometry stainings are depicted in Figure 2A. While unstimulated cells did not produce cytokines, robust production was observed following polyclonal stimulation (Figure 2A).

Bottom Line: Additionally within the ACPA-positive group significant down-regulation of all key T cell effector subsets including Th1, Th2, and Th17 was observed by analyzing cytokines by intracellular flow cytometry and in cell culture supernatants.RA synovial fluid samples were cultured in vitro in the presence or absence of CTLA4-Ig (abatacept).Our immunological study of T cell functionality in RA patients, both ACPA-positive and ACPA-negative starting biological therapy with the co-stimulation blockade abatacept (CTLA4-Ig) supports the recently published registry study implicating ACPA seropositivity as an independent predictive factor to treatment response as we observed the most striking effect on T cell subset modulation in ACPA-positive patients.These data further support the notion of RA as a disease with several sub-entities, where the ACPA-positive fraction represents a classical HLA-associated autoimmune disorder while ACPA-negative patients may have other driving forces apart from classical adaptive immune responses.

View Article: PubMed Central - HTML - PubMed

Affiliation: Rheumatology Unit, Department of Medicine at Karolinska University Hospital, Karolinska Institute, Solna, Stockholm, Sweden.

ABSTRACT

Background: Rheumatoid arthritis is a chronic inflammatory disease with a strong MHC class II component and where many patients develop characteristic autoantibodies towards the noncoding amino acid citrulline. Such anti-citrullinated protein antibodies (ACPA) have recently been put forward as an independent predictive factor for treatment response by co-stimulation blockade by CTLA4-Ig (abatacept). We have performed a mechanism of action study to dissect T cell functionality in RA patients with long-standing disease undergoing abatacept treatment and the influence of ACPA status.

Results: Peripheral blood samples were collected from RA patients as they started CTLA4-Ig treatment and 3 and 6 months later. A general decrease of regulatory T cell subsets was observed in the cohort. Additionally within the ACPA-positive group significant down-regulation of all key T cell effector subsets including Th1, Th2, and Th17 was observed by analyzing cytokines by intracellular flow cytometry and in cell culture supernatants.RA synovial fluid samples were cultured in vitro in the presence or absence of CTLA4-Ig (abatacept). T cell cytokine production was diminished, but without increasing the functional capacity of CD4+CD25hi regulatory T cells as previously demonstrated in the context of TNF-blockade and anti-IL6R therapy.

Conclusions: Our immunological study of T cell functionality in RA patients, both ACPA-positive and ACPA-negative starting biological therapy with the co-stimulation blockade abatacept (CTLA4-Ig) supports the recently published registry study implicating ACPA seropositivity as an independent predictive factor to treatment response as we observed the most striking effect on T cell subset modulation in ACPA-positive patients. These data further support the notion of RA as a disease with several sub-entities, where the ACPA-positive fraction represents a classical HLA-associated autoimmune disorder while ACPA-negative patients may have other driving forces apart from classical adaptive immune responses.

Show MeSH
Related in: MedlinePlus