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CD4+Foxp3+ regulatory T cells prolong drug-induced disease remission in (NZBxNZW) F1 lupus mice.

Weigert O, von Spee C, Undeutsch R, Kloke L, Humrich JY, Riemekasten G - Arthritis Res. Ther. (2013)

Bottom Line: The influence of CTX on the numbers, frequencies and proliferation of endogenous Treg and Tcon was analyzed in lymphoid organs by flow cytometry.Apart from abrogating the proliferation of Tcon, we found that treatment with CTX induced also a significant inhibition of Treg proliferation and a decline in Treg numbers in lymphoid organs.The additional clinical amelioration was associated with an increase in the Treg frequency in the peripheral blood indicating a compensation of CTX-induced Treg deficiency by the Treg transfer.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: The ability to ameliorate murine lupus renders regulatory T cells (Treg) a promising tool for the treatment of systemic lupus erythematosus (SLE). In consideration to the clinical translation of a Treg-based immunotherapy of SLE, we explored the potential of CD4+Foxp3+ Treg to maintain disease remission after induction of remission with an established cyclophosphamide (CTX) regimen in lupus-prone (NZBxNZW) F1 mice. As a prerequisite for this combined therapy, we also investigated the impact of CTX on the biology of endogenous Treg and conventional CD4+ T cells (Tcon).

Methods: Remission of disease was induced in diseased (NZBxNZW) F1 mice with an established CTX regimen consisting of a single dose of glucocorticosteroids followed by five day course with daily injections of CTX. Five days after the last CTX injection, differing amounts of purified CD4+Foxp3+CD25+ Treg were adoptively transferred and clinical parameters, autoantibody titers, the survival and changes in peripheral blood lymphocyte subsets were determined at different time points during the study. The influence of CTX on the numbers, frequencies and proliferation of endogenous Treg and Tcon was analyzed in lymphoid organs by flow cytometry.

Results: Apart from abrogating the proliferation of Tcon, we found that treatment with CTX induced also a significant inhibition of Treg proliferation and a decline in Treg numbers in lymphoid organs. Additional adoptive transfer of 1.5×10⁶ purified Treg after the CTX regimen significantly increased the survival and prolonged the interval of remission by approximately five weeks compared to mice that received only the CTX regimen. The additional clinical amelioration was associated with an increase in the Treg frequency in the peripheral blood indicating a compensation of CTX-induced Treg deficiency by the Treg transfer.

Conclusions: Treg were capable to prolong the interval of remission induced by conventional cytostatic drugs. This study provides valuable information and a first proof-of-concept for the feasibility of a Treg-based immunotherapy in the maintenance of disease remission in SLE.

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Regulatory T cells (Treg) prolong drug-induced disease remission. (A-D) Changes in clinical parameters and survival of (NZBxNZW) F1 mice with active disease after induction of remission with glucocorticosteroid (GC)/cyclophosphamide (CTX) and after an additional adoptive transfer of 1.5 × 106 Treg/mouse (Treg) compared to age-matched mice that received only GC/CTX and PBS (Control). (A-C) Average proteinuria score (A), leukocyturia score (B) and levels of antibodies against ds-DNA (C) determined at the indicated time points during the study. Error bars indicate standard error of the mean (SEM) (*P <0.05; Treg vs Control at the indicated time point). (D) Survival time is presented as a Kaplan-Meier curve (P = 0.01; Treg vs Control). (A-D) Data are the summary of two to three independent experiments, with eight to ten mice per group in total.
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Figure 3: Regulatory T cells (Treg) prolong drug-induced disease remission. (A-D) Changes in clinical parameters and survival of (NZBxNZW) F1 mice with active disease after induction of remission with glucocorticosteroid (GC)/cyclophosphamide (CTX) and after an additional adoptive transfer of 1.5 × 106 Treg/mouse (Treg) compared to age-matched mice that received only GC/CTX and PBS (Control). (A-C) Average proteinuria score (A), leukocyturia score (B) and levels of antibodies against ds-DNA (C) determined at the indicated time points during the study. Error bars indicate standard error of the mean (SEM) (*P <0.05; Treg vs Control at the indicated time point). (D) Survival time is presented as a Kaplan-Meier curve (P = 0.01; Treg vs Control). (A-D) Data are the summary of two to three independent experiments, with eight to ten mice per group in total.

Mentions: Treatment with GC/CTX alone induced remission of disease shown by the transient decrease in proteinuria (day -10 vs day -4, P <0.001) (Figure 3A). However, leukocyturia was not affected by the GC/CTX treatment (Figure 3B). In addition, a transient decrease in the levels of antibodies against dsDNA was observed (day -10 vs day - 4, P <0.001) (Figure 3C). Proteinuria and anti-dsDNA antibodies returned to pre-treatment levels approximately three weeks after the last CTX injection (Figure 3A,C, control).


CD4+Foxp3+ regulatory T cells prolong drug-induced disease remission in (NZBxNZW) F1 lupus mice.

Weigert O, von Spee C, Undeutsch R, Kloke L, Humrich JY, Riemekasten G - Arthritis Res. Ther. (2013)

Regulatory T cells (Treg) prolong drug-induced disease remission. (A-D) Changes in clinical parameters and survival of (NZBxNZW) F1 mice with active disease after induction of remission with glucocorticosteroid (GC)/cyclophosphamide (CTX) and after an additional adoptive transfer of 1.5 × 106 Treg/mouse (Treg) compared to age-matched mice that received only GC/CTX and PBS (Control). (A-C) Average proteinuria score (A), leukocyturia score (B) and levels of antibodies against ds-DNA (C) determined at the indicated time points during the study. Error bars indicate standard error of the mean (SEM) (*P <0.05; Treg vs Control at the indicated time point). (D) Survival time is presented as a Kaplan-Meier curve (P = 0.01; Treg vs Control). (A-D) Data are the summary of two to three independent experiments, with eight to ten mice per group in total.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3672693&req=5

Figure 3: Regulatory T cells (Treg) prolong drug-induced disease remission. (A-D) Changes in clinical parameters and survival of (NZBxNZW) F1 mice with active disease after induction of remission with glucocorticosteroid (GC)/cyclophosphamide (CTX) and after an additional adoptive transfer of 1.5 × 106 Treg/mouse (Treg) compared to age-matched mice that received only GC/CTX and PBS (Control). (A-C) Average proteinuria score (A), leukocyturia score (B) and levels of antibodies against ds-DNA (C) determined at the indicated time points during the study. Error bars indicate standard error of the mean (SEM) (*P <0.05; Treg vs Control at the indicated time point). (D) Survival time is presented as a Kaplan-Meier curve (P = 0.01; Treg vs Control). (A-D) Data are the summary of two to three independent experiments, with eight to ten mice per group in total.
Mentions: Treatment with GC/CTX alone induced remission of disease shown by the transient decrease in proteinuria (day -10 vs day -4, P <0.001) (Figure 3A). However, leukocyturia was not affected by the GC/CTX treatment (Figure 3B). In addition, a transient decrease in the levels of antibodies against dsDNA was observed (day -10 vs day - 4, P <0.001) (Figure 3C). Proteinuria and anti-dsDNA antibodies returned to pre-treatment levels approximately three weeks after the last CTX injection (Figure 3A,C, control).

Bottom Line: The influence of CTX on the numbers, frequencies and proliferation of endogenous Treg and Tcon was analyzed in lymphoid organs by flow cytometry.Apart from abrogating the proliferation of Tcon, we found that treatment with CTX induced also a significant inhibition of Treg proliferation and a decline in Treg numbers in lymphoid organs.The additional clinical amelioration was associated with an increase in the Treg frequency in the peripheral blood indicating a compensation of CTX-induced Treg deficiency by the Treg transfer.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: The ability to ameliorate murine lupus renders regulatory T cells (Treg) a promising tool for the treatment of systemic lupus erythematosus (SLE). In consideration to the clinical translation of a Treg-based immunotherapy of SLE, we explored the potential of CD4+Foxp3+ Treg to maintain disease remission after induction of remission with an established cyclophosphamide (CTX) regimen in lupus-prone (NZBxNZW) F1 mice. As a prerequisite for this combined therapy, we also investigated the impact of CTX on the biology of endogenous Treg and conventional CD4+ T cells (Tcon).

Methods: Remission of disease was induced in diseased (NZBxNZW) F1 mice with an established CTX regimen consisting of a single dose of glucocorticosteroids followed by five day course with daily injections of CTX. Five days after the last CTX injection, differing amounts of purified CD4+Foxp3+CD25+ Treg were adoptively transferred and clinical parameters, autoantibody titers, the survival and changes in peripheral blood lymphocyte subsets were determined at different time points during the study. The influence of CTX on the numbers, frequencies and proliferation of endogenous Treg and Tcon was analyzed in lymphoid organs by flow cytometry.

Results: Apart from abrogating the proliferation of Tcon, we found that treatment with CTX induced also a significant inhibition of Treg proliferation and a decline in Treg numbers in lymphoid organs. Additional adoptive transfer of 1.5×10⁶ purified Treg after the CTX regimen significantly increased the survival and prolonged the interval of remission by approximately five weeks compared to mice that received only the CTX regimen. The additional clinical amelioration was associated with an increase in the Treg frequency in the peripheral blood indicating a compensation of CTX-induced Treg deficiency by the Treg transfer.

Conclusions: Treg were capable to prolong the interval of remission induced by conventional cytostatic drugs. This study provides valuable information and a first proof-of-concept for the feasibility of a Treg-based immunotherapy in the maintenance of disease remission in SLE.

Show MeSH
Related in: MedlinePlus