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Differential Effects of Tacrolimus versus Sirolimus on the Proliferation, Activation and Differentiation of Human B Cells.

Traitanon O, Mathew JM, La Monica G, Xu L, Mas V, Gallon L - PLoS ONE (2015)

Bottom Line: SRL effectively blocked B cell differentiation into plasma cells (CD19(+)CD138(+) and Blimp1(+)/Pax5(low) cells) even at low dose (2 ng/ml), and totally eliminated them at 6 ng/ml.SRL decreased absolute B cell counts, but the residual responding cells acquired an activated phenotype (CD25(+)/CD69(+)) and increased the expression of HLA-DR.Thus, SRL and TAC have different effects on B lymphocytes.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine-Nephrology, Northwestern University, Chicago, IL, United States of America; Comprehensive Transplant Center, Northwestern University, Chicago, IL, United States of America; Department of Medicine-Nephrology, Thammasart University Hospital, Pathumthani, Thailand.

ABSTRACT
The direct effect of immunosuppressive drugs calcineurin inhibitor (Tacrolimus, TAC) and mTOR inhibitor (Sirolimus, SRL) on B cell activation, differentiation and proliferation is not well documented. Purified human B cells from healthy volunteers were stimulated through the B Cell Receptor with Anti-IgM + anti-CD40 + IL21 in the absence / presence of TAC or SRL. A variety of parameters of B cell activity including activation, differentiation, cytokine productions and proliferation were monitored by flow cytometry. SRL at clinically relevant concentrations (6 ng/ml) profoundly inhibited CD19(+ )B cell proliferation compared to controls whereas TAC at similar concentrations had a minimal effect. CD27(+) memory B cells were affected more by SRL than naïve CD27- B cells. SRL effectively blocked B cell differentiation into plasma cells (CD19(+)CD138(+) and Blimp1(+)/Pax5(low) cells) even at low dose (2 ng/ml), and totally eliminated them at 6 ng/ml. SRL decreased absolute B cell counts, but the residual responding cells acquired an activated phenotype (CD25(+)/CD69(+)) and increased the expression of HLA-DR. SRL-treated stimulated B cells on a per cell basis were able to enhance the proliferation of allogeneic CD4(+)CD25(-) T cells and induce a shift toward the Th1 phenotype. Thus, SRL and TAC have different effects on B lymphocytes. These data may provide insights into the clinical use of these two agents in recipients of solid organ transplants.

No MeSH data available.


Related in: MedlinePlus

SRL-treated B cells enhances the proliferation and differentiation of CD4+ T cells.Purified CD19+ B cells were pre-stimulated for 6 days with anti-IgM, anti-CD40 mAb and IL-21 in the absence (CTRL) or presence of 6ng/ml TAC or SRL. These cultured B cells were used as stimulators in 6-day MLRs of allogeneic CFSE-labelled CD4+CD25− T cell responders. (A) Level of proliferation differentially induced by pre-cultured B cells as detected by CFSE dilution in the allogeneic CD4 responder cells (representative experiment on the left, and compiled data from 8 independent experiments on the right). (B) Percentage of responding T cells positive for memory marker (CD45RO) and activation markers (CD62L, CD25, CD69, CD95) after co-culture with pre-stimulated B cells (n = 8); (C) Mean ± SD (n = 4) percentage of responding proliferating T cells expressing intracellular cytokines (top row) and transcription factors (bottom row). Taken together the data indicated that B cells that proliferated in presence of SRL on a per cell basis were capable of inducing alloreactive T cell proliferation towards a Th1 phenotype. *p < 0.05. ** p < 0.01.
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pone.0129658.g006: SRL-treated B cells enhances the proliferation and differentiation of CD4+ T cells.Purified CD19+ B cells were pre-stimulated for 6 days with anti-IgM, anti-CD40 mAb and IL-21 in the absence (CTRL) or presence of 6ng/ml TAC or SRL. These cultured B cells were used as stimulators in 6-day MLRs of allogeneic CFSE-labelled CD4+CD25− T cell responders. (A) Level of proliferation differentially induced by pre-cultured B cells as detected by CFSE dilution in the allogeneic CD4 responder cells (representative experiment on the left, and compiled data from 8 independent experiments on the right). (B) Percentage of responding T cells positive for memory marker (CD45RO) and activation markers (CD62L, CD25, CD69, CD95) after co-culture with pre-stimulated B cells (n = 8); (C) Mean ± SD (n = 4) percentage of responding proliferating T cells expressing intracellular cytokines (top row) and transcription factors (bottom row). Taken together the data indicated that B cells that proliferated in presence of SRL on a per cell basis were capable of inducing alloreactive T cell proliferation towards a Th1 phenotype. *p < 0.05. ** p < 0.01.

Mentions: Since the SRL-treated B cells had higher proportion of HLA-DR expressing cells, the antigen presenting function of these cells was tested. B cells were stimulated in the absence or presence of TAC or SRL for 6 days, and were used as stimulators in primary MLRs with CFSE-labeled allogeneic CD4+CD25− responder T cells. B cells grown in presence of SRL were able to induced significantly more T cell proliferation than B cells from control or TAC-treated cultures (p<0.01) (Fig 6A). Phenotypic analysis of the proliferating T cells further revealed that SRL-treated B cells also induced higher proportion of cells expressing activation markers CD25 and CD69 but not CD95 (Fig 6B). No difference in the percentage of memory (CD45RO+) T cells was observed among the three groups.


Differential Effects of Tacrolimus versus Sirolimus on the Proliferation, Activation and Differentiation of Human B Cells.

Traitanon O, Mathew JM, La Monica G, Xu L, Mas V, Gallon L - PLoS ONE (2015)

SRL-treated B cells enhances the proliferation and differentiation of CD4+ T cells.Purified CD19+ B cells were pre-stimulated for 6 days with anti-IgM, anti-CD40 mAb and IL-21 in the absence (CTRL) or presence of 6ng/ml TAC or SRL. These cultured B cells were used as stimulators in 6-day MLRs of allogeneic CFSE-labelled CD4+CD25− T cell responders. (A) Level of proliferation differentially induced by pre-cultured B cells as detected by CFSE dilution in the allogeneic CD4 responder cells (representative experiment on the left, and compiled data from 8 independent experiments on the right). (B) Percentage of responding T cells positive for memory marker (CD45RO) and activation markers (CD62L, CD25, CD69, CD95) after co-culture with pre-stimulated B cells (n = 8); (C) Mean ± SD (n = 4) percentage of responding proliferating T cells expressing intracellular cytokines (top row) and transcription factors (bottom row). Taken together the data indicated that B cells that proliferated in presence of SRL on a per cell basis were capable of inducing alloreactive T cell proliferation towards a Th1 phenotype. *p < 0.05. ** p < 0.01.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4472515&req=5

pone.0129658.g006: SRL-treated B cells enhances the proliferation and differentiation of CD4+ T cells.Purified CD19+ B cells were pre-stimulated for 6 days with anti-IgM, anti-CD40 mAb and IL-21 in the absence (CTRL) or presence of 6ng/ml TAC or SRL. These cultured B cells were used as stimulators in 6-day MLRs of allogeneic CFSE-labelled CD4+CD25− T cell responders. (A) Level of proliferation differentially induced by pre-cultured B cells as detected by CFSE dilution in the allogeneic CD4 responder cells (representative experiment on the left, and compiled data from 8 independent experiments on the right). (B) Percentage of responding T cells positive for memory marker (CD45RO) and activation markers (CD62L, CD25, CD69, CD95) after co-culture with pre-stimulated B cells (n = 8); (C) Mean ± SD (n = 4) percentage of responding proliferating T cells expressing intracellular cytokines (top row) and transcription factors (bottom row). Taken together the data indicated that B cells that proliferated in presence of SRL on a per cell basis were capable of inducing alloreactive T cell proliferation towards a Th1 phenotype. *p < 0.05. ** p < 0.01.
Mentions: Since the SRL-treated B cells had higher proportion of HLA-DR expressing cells, the antigen presenting function of these cells was tested. B cells were stimulated in the absence or presence of TAC or SRL for 6 days, and were used as stimulators in primary MLRs with CFSE-labeled allogeneic CD4+CD25− responder T cells. B cells grown in presence of SRL were able to induced significantly more T cell proliferation than B cells from control or TAC-treated cultures (p<0.01) (Fig 6A). Phenotypic analysis of the proliferating T cells further revealed that SRL-treated B cells also induced higher proportion of cells expressing activation markers CD25 and CD69 but not CD95 (Fig 6B). No difference in the percentage of memory (CD45RO+) T cells was observed among the three groups.

Bottom Line: SRL effectively blocked B cell differentiation into plasma cells (CD19(+)CD138(+) and Blimp1(+)/Pax5(low) cells) even at low dose (2 ng/ml), and totally eliminated them at 6 ng/ml.SRL decreased absolute B cell counts, but the residual responding cells acquired an activated phenotype (CD25(+)/CD69(+)) and increased the expression of HLA-DR.Thus, SRL and TAC have different effects on B lymphocytes.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine-Nephrology, Northwestern University, Chicago, IL, United States of America; Comprehensive Transplant Center, Northwestern University, Chicago, IL, United States of America; Department of Medicine-Nephrology, Thammasart University Hospital, Pathumthani, Thailand.

ABSTRACT
The direct effect of immunosuppressive drugs calcineurin inhibitor (Tacrolimus, TAC) and mTOR inhibitor (Sirolimus, SRL) on B cell activation, differentiation and proliferation is not well documented. Purified human B cells from healthy volunteers were stimulated through the B Cell Receptor with Anti-IgM + anti-CD40 + IL21 in the absence / presence of TAC or SRL. A variety of parameters of B cell activity including activation, differentiation, cytokine productions and proliferation were monitored by flow cytometry. SRL at clinically relevant concentrations (6 ng/ml) profoundly inhibited CD19(+ )B cell proliferation compared to controls whereas TAC at similar concentrations had a minimal effect. CD27(+) memory B cells were affected more by SRL than naïve CD27- B cells. SRL effectively blocked B cell differentiation into plasma cells (CD19(+)CD138(+) and Blimp1(+)/Pax5(low) cells) even at low dose (2 ng/ml), and totally eliminated them at 6 ng/ml. SRL decreased absolute B cell counts, but the residual responding cells acquired an activated phenotype (CD25(+)/CD69(+)) and increased the expression of HLA-DR. SRL-treated stimulated B cells on a per cell basis were able to enhance the proliferation of allogeneic CD4(+)CD25(-) T cells and induce a shift toward the Th1 phenotype. Thus, SRL and TAC have different effects on B lymphocytes. These data may provide insights into the clinical use of these two agents in recipients of solid organ transplants.

No MeSH data available.


Related in: MedlinePlus