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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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Changes in lymphocyte subsets during induction of remission and sequential regulatory T cell (Treg) transfer. Average absolute numbers of total lymphocytes (A) and average percentage of CD4+ T cells (B) and CD19+ B cells (C) among lymphocytes in the peripheral blood of (NZBxNZW) F1 mice with active disease before and 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). Data are the summary of two independent experiments with six to eight mice per group in total. Error bars indicate standard error of the mean (SEM).
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Figure 4: Changes in lymphocyte subsets during induction of remission and sequential regulatory T cell (Treg) transfer. Average absolute numbers of total lymphocytes (A) and average percentage of CD4+ T cells (B) and CD19+ B cells (C) among lymphocytes in the peripheral blood of (NZBxNZW) F1 mice with active disease before and 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). Data are the summary of two independent experiments with six to eight mice per group in total. Error bars indicate standard error of the mean (SEM).

Mentions: Treatment with GC/CTX alone induced a late and transient increase in the numbers of total lymphocytes in the peripheral blood (Figure 4A). Consistent with prior data, the percentage of CD4+ T cells among total lymphocytes transiently increased after the CTX regimen (day -10 vs day -4, P <0.001) (Figure 4B). In contrast, there was a dramatic decrease in the percentage of CD19+ B cells among lymphocytes (P <0.001) that persisted throughout the observation time (Figure 4C). However, no differences in total lymphocytes, CD4+ T cells or CD19+ B cells after additional adoptive transfer of Treg could be observed compared to mice that received only GC/CTX (Figure 4 A-C).


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)

Changes in lymphocyte subsets during induction of remission and sequential regulatory T cell (Treg) transfer. Average absolute numbers of total lymphocytes (A) and average percentage of CD4+ T cells (B) and CD19+ B cells (C) among lymphocytes in the peripheral blood of (NZBxNZW) F1 mice with active disease before and 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). Data are the summary of two independent experiments with six to eight mice per group in total. Error bars indicate standard error of the mean (SEM).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Changes in lymphocyte subsets during induction of remission and sequential regulatory T cell (Treg) transfer. Average absolute numbers of total lymphocytes (A) and average percentage of CD4+ T cells (B) and CD19+ B cells (C) among lymphocytes in the peripheral blood of (NZBxNZW) F1 mice with active disease before and 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). Data are the summary of two independent experiments with six to eight mice per group in total. Error bars indicate standard error of the mean (SEM).
Mentions: Treatment with GC/CTX alone induced a late and transient increase in the numbers of total lymphocytes in the peripheral blood (Figure 4A). Consistent with prior data, the percentage of CD4+ T cells among total lymphocytes transiently increased after the CTX regimen (day -10 vs day -4, P <0.001) (Figure 4B). In contrast, there was a dramatic decrease in the percentage of CD19+ B cells among lymphocytes (P <0.001) that persisted throughout the observation time (Figure 4C). However, no differences in total lymphocytes, CD4+ T cells or CD19+ B cells after additional adoptive transfer of Treg could be observed compared to mice that received only GC/CTX (Figure 4 A-C).

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