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Longitudinal analysis of T and B cell phenotype and function in renal transplant recipients with or without rituximab induction therapy.

Kamburova EG, Koenen HJ, van den Hoogen MW, Baas MC, Joosten I, Hilbrands LB - PLoS ONE (2014)

Bottom Line: In addition, the ratio between the percentage of central memory CD4+ and CD4+ regulatory T cells was significantly higher up to 24 months post-transplant compared to pre-transplant levels.Addition of a single dose of rituximab resulted in a long-lasting B-cell depletion.Rituximab treatment had no effect on the T-cell phenotype and function post-transplant.

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine - Medical Immunology, Radboud University Medical Centre, Nijmegen, The Netherlands.

ABSTRACT

Background: Prevention of rejection after renal transplantation requires treatment with immunosuppressive drugs. Data on their in vivo effects on T- and B-cell phenotype and function are limited.

Methods: In a randomized double-blind placebo-controlled study to prevent renal allograft rejection, patients were treated with tacrolimus, mycophenolate mofetil (MMF), steroids, and a single dose of rituximab or placebo during transplant surgery. In a subset of patients, we analyzed the number and phenotype of peripheral T and B cells by multiparameter flow cytometry before transplantation, and at 3, 6, 12, and 24 months after transplantation.

Results: In patients treated with tacrolimus/MMF/steroids the proportion of central memory CD4+ and CD8+ T cells was higher at 3 months post-transplant compared to pre-transplant levels. In addition, the ratio between the percentage of central memory CD4+ and CD4+ regulatory T cells was significantly higher up to 24 months post-transplant compared to pre-transplant levels. Interestingly, treatment with tacrolimus/MMF/steroids resulted in a shift toward a more memory-like B-cell phenotype post-transplant. Addition of a single dose of rituximab resulted in a long-lasting B-cell depletion. At 12 months post-transplant, the small fraction of repopulated B cells consisted of a high percentage of transitional B cells. Rituximab treatment had no effect on the T-cell phenotype and function post-transplant.

Conclusions: Renal transplant recipients treated with tacrolimus/MMF/steroids show an altered memory T and B-cell compartment post-transplant. Additional B-cell depletion by rituximab leads to a relative increase of transitional and memory-like B cells, without affecting T-cell phenotype and function.

Trial registration: ClinicalTrials.gov NCT00565331.

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Subset distribution of circulating T cells in renal transplant recipients after treatment with tacrolimus, MMF and steroids over time.(A) Representative dot plots for a renal transplant recipient showing CD3+CD4+ and CD3+CD8+ T cells within the CD45+ lymphocyte population. Circulating CD4+ and CD8+ T cells can be characterized as naive (TN; CD27+CD45RO−), central memory (TCM; CD27+CD45RO+), effector memory (TEM; CD27−CD45RO+) and highly differentiated memory (TEMRA; CD27+CD45RO−) cells. Furthermore, CD4+ T cells can be characterized as regulatory T cells (TREGS; CD25hiFOXP3+). (B) Shown are the absolute numbers of CD4+ T cells and the percentages of TN, TCM and TEM within the CD4+ T-cell population for 14 triple immunosuppression-treated patients before transplantation (t = 0) and at 3, 6, 12 and 24 months after transplantation (n = 10 at 24 m). (C) As described under B, for CD8+ T cells. (D) The ratio between the percentage of CD4+ TCM or TEM and the percentage of TREGS. (E) Longitudinal analysis of the percentages of CXCR3+, CCR4+, and CCR6+ cells within the CD4+ T-cell population of 14 triple immunosuppression-treated patients (n = 10 at 24 m). Results are shown as box plots displaying the median, 25th and 75th percentiles as the box, and the 5th and 95th percentiles as whiskers. Significant differences are indicated compared to pre-transplant levels: *P<0.05, **P<0.01.
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pone-0112658-g001: Subset distribution of circulating T cells in renal transplant recipients after treatment with tacrolimus, MMF and steroids over time.(A) Representative dot plots for a renal transplant recipient showing CD3+CD4+ and CD3+CD8+ T cells within the CD45+ lymphocyte population. Circulating CD4+ and CD8+ T cells can be characterized as naive (TN; CD27+CD45RO−), central memory (TCM; CD27+CD45RO+), effector memory (TEM; CD27−CD45RO+) and highly differentiated memory (TEMRA; CD27+CD45RO−) cells. Furthermore, CD4+ T cells can be characterized as regulatory T cells (TREGS; CD25hiFOXP3+). (B) Shown are the absolute numbers of CD4+ T cells and the percentages of TN, TCM and TEM within the CD4+ T-cell population for 14 triple immunosuppression-treated patients before transplantation (t = 0) and at 3, 6, 12 and 24 months after transplantation (n = 10 at 24 m). (C) As described under B, for CD8+ T cells. (D) The ratio between the percentage of CD4+ TCM or TEM and the percentage of TREGS. (E) Longitudinal analysis of the percentages of CXCR3+, CCR4+, and CCR6+ cells within the CD4+ T-cell population of 14 triple immunosuppression-treated patients (n = 10 at 24 m). Results are shown as box plots displaying the median, 25th and 75th percentiles as the box, and the 5th and 95th percentiles as whiskers. Significant differences are indicated compared to pre-transplant levels: *P<0.05, **P<0.01.

Mentions: CD4+ and CD8+ T cells can be divided into naive, central and effector memory, and highly differentiated memory cells based on CD27 and CD45RO expression. [15]. The absolute numbers of CD4+ and CD8+ T cells in peripheral blood did not change during the use of triple drug immunosuppression after transplantation (Figure 1A–C). However, the percentages of central memory (TCM; CD27+CD45RO+) CD4+ and CD8+ T cells were significantly higher at 3 months after transplantation compared to before transplantation, while the percentages of effector memory (TEM; CD27−CD45RO+) CD4+ and CD8+ T cells, and regulatory (TREGS; CD25hiFOXP3+) CD4+ T cells were significantly decreased. The percentages of naive (TN; CD27+CD45RO−) and highly differentiated memory (TEMRA; CD27−CD45RO−) CD4+ and CD8+ T cells were comparable to pre-transplant levels. Next to the T-cell subset distribution, the ratio between memory and regulatory T cells might be an important determinant of the risk of rejection [16]. Interestingly, the CD4+ TCM/CD4+ TREGS ratio significantly increased from 3 months after transplantation and remained elevated compared to the pre-transplant ratio (Figure 1D). The CD4+ TEM/CD4+ TREGS ratio after transplantation was comparable to pre-transplant levels. Finally, we determined the expression of chemokine receptors associated with T helper (Th) 1 (CXCR3), Th2 (CCR4), and Th17 (CCR6) cells (Figure 1E). The percentage of CXCR3+ and CCR6+ CD4+ T cells increased during immunosuppressive treatment, whereas triple drug immunosuppression did not affect the percentage of CCR4+ CD4+ T cells.


Longitudinal analysis of T and B cell phenotype and function in renal transplant recipients with or without rituximab induction therapy.

Kamburova EG, Koenen HJ, van den Hoogen MW, Baas MC, Joosten I, Hilbrands LB - PLoS ONE (2014)

Subset distribution of circulating T cells in renal transplant recipients after treatment with tacrolimus, MMF and steroids over time.(A) Representative dot plots for a renal transplant recipient showing CD3+CD4+ and CD3+CD8+ T cells within the CD45+ lymphocyte population. Circulating CD4+ and CD8+ T cells can be characterized as naive (TN; CD27+CD45RO−), central memory (TCM; CD27+CD45RO+), effector memory (TEM; CD27−CD45RO+) and highly differentiated memory (TEMRA; CD27+CD45RO−) cells. Furthermore, CD4+ T cells can be characterized as regulatory T cells (TREGS; CD25hiFOXP3+). (B) Shown are the absolute numbers of CD4+ T cells and the percentages of TN, TCM and TEM within the CD4+ T-cell population for 14 triple immunosuppression-treated patients before transplantation (t = 0) and at 3, 6, 12 and 24 months after transplantation (n = 10 at 24 m). (C) As described under B, for CD8+ T cells. (D) The ratio between the percentage of CD4+ TCM or TEM and the percentage of TREGS. (E) Longitudinal analysis of the percentages of CXCR3+, CCR4+, and CCR6+ cells within the CD4+ T-cell population of 14 triple immunosuppression-treated patients (n = 10 at 24 m). Results are shown as box plots displaying the median, 25th and 75th percentiles as the box, and the 5th and 95th percentiles as whiskers. Significant differences are indicated compared to pre-transplant levels: *P<0.05, **P<0.01.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4231065&req=5

pone-0112658-g001: Subset distribution of circulating T cells in renal transplant recipients after treatment with tacrolimus, MMF and steroids over time.(A) Representative dot plots for a renal transplant recipient showing CD3+CD4+ and CD3+CD8+ T cells within the CD45+ lymphocyte population. Circulating CD4+ and CD8+ T cells can be characterized as naive (TN; CD27+CD45RO−), central memory (TCM; CD27+CD45RO+), effector memory (TEM; CD27−CD45RO+) and highly differentiated memory (TEMRA; CD27+CD45RO−) cells. Furthermore, CD4+ T cells can be characterized as regulatory T cells (TREGS; CD25hiFOXP3+). (B) Shown are the absolute numbers of CD4+ T cells and the percentages of TN, TCM and TEM within the CD4+ T-cell population for 14 triple immunosuppression-treated patients before transplantation (t = 0) and at 3, 6, 12 and 24 months after transplantation (n = 10 at 24 m). (C) As described under B, for CD8+ T cells. (D) The ratio between the percentage of CD4+ TCM or TEM and the percentage of TREGS. (E) Longitudinal analysis of the percentages of CXCR3+, CCR4+, and CCR6+ cells within the CD4+ T-cell population of 14 triple immunosuppression-treated patients (n = 10 at 24 m). Results are shown as box plots displaying the median, 25th and 75th percentiles as the box, and the 5th and 95th percentiles as whiskers. Significant differences are indicated compared to pre-transplant levels: *P<0.05, **P<0.01.
Mentions: CD4+ and CD8+ T cells can be divided into naive, central and effector memory, and highly differentiated memory cells based on CD27 and CD45RO expression. [15]. The absolute numbers of CD4+ and CD8+ T cells in peripheral blood did not change during the use of triple drug immunosuppression after transplantation (Figure 1A–C). However, the percentages of central memory (TCM; CD27+CD45RO+) CD4+ and CD8+ T cells were significantly higher at 3 months after transplantation compared to before transplantation, while the percentages of effector memory (TEM; CD27−CD45RO+) CD4+ and CD8+ T cells, and regulatory (TREGS; CD25hiFOXP3+) CD4+ T cells were significantly decreased. The percentages of naive (TN; CD27+CD45RO−) and highly differentiated memory (TEMRA; CD27−CD45RO−) CD4+ and CD8+ T cells were comparable to pre-transplant levels. Next to the T-cell subset distribution, the ratio between memory and regulatory T cells might be an important determinant of the risk of rejection [16]. Interestingly, the CD4+ TCM/CD4+ TREGS ratio significantly increased from 3 months after transplantation and remained elevated compared to the pre-transplant ratio (Figure 1D). The CD4+ TEM/CD4+ TREGS ratio after transplantation was comparable to pre-transplant levels. Finally, we determined the expression of chemokine receptors associated with T helper (Th) 1 (CXCR3), Th2 (CCR4), and Th17 (CCR6) cells (Figure 1E). The percentage of CXCR3+ and CCR6+ CD4+ T cells increased during immunosuppressive treatment, whereas triple drug immunosuppression did not affect the percentage of CCR4+ CD4+ T cells.

Bottom Line: In addition, the ratio between the percentage of central memory CD4+ and CD4+ regulatory T cells was significantly higher up to 24 months post-transplant compared to pre-transplant levels.Addition of a single dose of rituximab resulted in a long-lasting B-cell depletion.Rituximab treatment had no effect on the T-cell phenotype and function post-transplant.

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine - Medical Immunology, Radboud University Medical Centre, Nijmegen, The Netherlands.

ABSTRACT

Background: Prevention of rejection after renal transplantation requires treatment with immunosuppressive drugs. Data on their in vivo effects on T- and B-cell phenotype and function are limited.

Methods: In a randomized double-blind placebo-controlled study to prevent renal allograft rejection, patients were treated with tacrolimus, mycophenolate mofetil (MMF), steroids, and a single dose of rituximab or placebo during transplant surgery. In a subset of patients, we analyzed the number and phenotype of peripheral T and B cells by multiparameter flow cytometry before transplantation, and at 3, 6, 12, and 24 months after transplantation.

Results: In patients treated with tacrolimus/MMF/steroids the proportion of central memory CD4+ and CD8+ T cells was higher at 3 months post-transplant compared to pre-transplant levels. In addition, the ratio between the percentage of central memory CD4+ and CD4+ regulatory T cells was significantly higher up to 24 months post-transplant compared to pre-transplant levels. Interestingly, treatment with tacrolimus/MMF/steroids resulted in a shift toward a more memory-like B-cell phenotype post-transplant. Addition of a single dose of rituximab resulted in a long-lasting B-cell depletion. At 12 months post-transplant, the small fraction of repopulated B cells consisted of a high percentage of transitional B cells. Rituximab treatment had no effect on the T-cell phenotype and function post-transplant.

Conclusions: Renal transplant recipients treated with tacrolimus/MMF/steroids show an altered memory T and B-cell compartment post-transplant. Additional B-cell depletion by rituximab leads to a relative increase of transitional and memory-like B cells, without affecting T-cell phenotype and function.

Trial registration: ClinicalTrials.gov NCT00565331.

Show MeSH