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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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Abatacept reduces T cell proliferation in vitro. (A-C) SFMC from RA patients (n=6) were sorted into CD4+CD25- T effector cells as well as CD4+CD25++ Treg and were co-cultured with CD3-APC for 6 days in the presence of plate-bound α-CD3, either in the presence or absence of 10 μg/ml abatacept. (A) Synovial CD4+CD25- T effector cells were cultured alone and in the presence of different ratios of CD4+CD25++ Treg. The graph depicts the percentage of suppression by CD4+CD25++ Treg in co-culture either in the absence (black line) or the presence of abatacept (grey line). A summary of six experiments is shown and all values are expressed as mean+SD. (B) The graph displays the suppression of proliferation at the 1:1 ratio of T effector cells and Tregs. (C) Proliferation of CD4+CD25- T effector cells alone was measured by thymidine incorporation in the absence or presence of abatacept (n=6). (D-F) SFMC from RA patients were stimulated with α-CD3 (white bars) or influenza vaccine (grey bars) in the presence of 10 μg/ml abatacept or a control compound. Proliferation of SFMC was measured by thymidine incorporation following 72 hours (α-CD3) or 6 days (influenza) in culture. All values are expressed as mean+SD. (D) All patients are displayed, (α-CD3: n=15 patients; Influenza: n=7). (E) Only ACPA-positive patients are displayed, (α-CD3: n=9 patients; Influenza: n=5). (F) Only ACPA-negative patients are displayed, (α-CD3: n=6 patients; Influenza: n=2).
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Figure 5: Abatacept reduces T cell proliferation in vitro. (A-C) SFMC from RA patients (n=6) were sorted into CD4+CD25- T effector cells as well as CD4+CD25++ Treg and were co-cultured with CD3-APC for 6 days in the presence of plate-bound α-CD3, either in the presence or absence of 10 μg/ml abatacept. (A) Synovial CD4+CD25- T effector cells were cultured alone and in the presence of different ratios of CD4+CD25++ Treg. The graph depicts the percentage of suppression by CD4+CD25++ Treg in co-culture either in the absence (black line) or the presence of abatacept (grey line). A summary of six experiments is shown and all values are expressed as mean+SD. (B) The graph displays the suppression of proliferation at the 1:1 ratio of T effector cells and Tregs. (C) Proliferation of CD4+CD25- T effector cells alone was measured by thymidine incorporation in the absence or presence of abatacept (n=6). (D-F) SFMC from RA patients were stimulated with α-CD3 (white bars) or influenza vaccine (grey bars) in the presence of 10 μg/ml abatacept or a control compound. Proliferation of SFMC was measured by thymidine incorporation following 72 hours (α-CD3) or 6 days (influenza) in culture. All values are expressed as mean+SD. (D) All patients are displayed, (α-CD3: n=15 patients; Influenza: n=7). (E) Only ACPA-positive patients are displayed, (α-CD3: n=9 patients; Influenza: n=5). (F) Only ACPA-negative patients are displayed, (α-CD3: n=6 patients; Influenza: n=2).

Mentions: First we studied the effect of abatacept on synovial T cells in vitro by investigating suppression in the presence of abatacept. Treg co-culture experiments were performed but no difference in suppressive capacity was seen in cultures with abatacept compared to control cultures (Figure 5A and B). However, we observed a significant reduction of CD25-negative T effector cell proliferation in the presence of abatacept (Figure 5C).


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)

Abatacept reduces T cell proliferation in vitro. (A-C) SFMC from RA patients (n=6) were sorted into CD4+CD25- T effector cells as well as CD4+CD25++ Treg and were co-cultured with CD3-APC for 6 days in the presence of plate-bound α-CD3, either in the presence or absence of 10 μg/ml abatacept. (A) Synovial CD4+CD25- T effector cells were cultured alone and in the presence of different ratios of CD4+CD25++ Treg. The graph depicts the percentage of suppression by CD4+CD25++ Treg in co-culture either in the absence (black line) or the presence of abatacept (grey line). A summary of six experiments is shown and all values are expressed as mean+SD. (B) The graph displays the suppression of proliferation at the 1:1 ratio of T effector cells and Tregs. (C) Proliferation of CD4+CD25- T effector cells alone was measured by thymidine incorporation in the absence or presence of abatacept (n=6). (D-F) SFMC from RA patients were stimulated with α-CD3 (white bars) or influenza vaccine (grey bars) in the presence of 10 μg/ml abatacept or a control compound. Proliferation of SFMC was measured by thymidine incorporation following 72 hours (α-CD3) or 6 days (influenza) in culture. All values are expressed as mean+SD. (D) All patients are displayed, (α-CD3: n=15 patients; Influenza: n=7). (E) Only ACPA-positive patients are displayed, (α-CD3: n=9 patients; Influenza: n=5). (F) Only ACPA-negative patients are displayed, (α-CD3: n=6 patients; Influenza: n=2).
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Figure 5: Abatacept reduces T cell proliferation in vitro. (A-C) SFMC from RA patients (n=6) were sorted into CD4+CD25- T effector cells as well as CD4+CD25++ Treg and were co-cultured with CD3-APC for 6 days in the presence of plate-bound α-CD3, either in the presence or absence of 10 μg/ml abatacept. (A) Synovial CD4+CD25- T effector cells were cultured alone and in the presence of different ratios of CD4+CD25++ Treg. The graph depicts the percentage of suppression by CD4+CD25++ Treg in co-culture either in the absence (black line) or the presence of abatacept (grey line). A summary of six experiments is shown and all values are expressed as mean+SD. (B) The graph displays the suppression of proliferation at the 1:1 ratio of T effector cells and Tregs. (C) Proliferation of CD4+CD25- T effector cells alone was measured by thymidine incorporation in the absence or presence of abatacept (n=6). (D-F) SFMC from RA patients were stimulated with α-CD3 (white bars) or influenza vaccine (grey bars) in the presence of 10 μg/ml abatacept or a control compound. Proliferation of SFMC was measured by thymidine incorporation following 72 hours (α-CD3) or 6 days (influenza) in culture. All values are expressed as mean+SD. (D) All patients are displayed, (α-CD3: n=15 patients; Influenza: n=7). (E) Only ACPA-positive patients are displayed, (α-CD3: n=9 patients; Influenza: n=5). (F) Only ACPA-negative patients are displayed, (α-CD3: n=6 patients; Influenza: n=2).
Mentions: First we studied the effect of abatacept on synovial T cells in vitro by investigating suppression in the presence of abatacept. Treg co-culture experiments were performed but no difference in suppressive capacity was seen in cultures with abatacept compared to control cultures (Figure 5A and B). However, we observed a significant reduction of CD25-negative T effector cell proliferation in the presence of abatacept (Figure 5C).

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