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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

B-cell stimulation in the presence of SRL resulted in a population shift toward an activated phenotype.Purified CD19+ B cells were stimulated with anti-IgM, anti-CD40 mAb and IL-21 and multi-color flow cytometric analyses were performed on day 6 as described in Fig 1. The figures A, B and C show the expression of various surface markers on stimulated B cells (Percentage of positive cells/total proliferating CD19+ cells) p < 0.05, **p < 0.01. CTRL, control; TAC6, 6 ng/ml TAC; SRL2, 2 ng/ml SRL; SRL6, 6 ng/ml SRL.
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pone.0129658.g002: B-cell stimulation in the presence of SRL resulted in a population shift toward an activated phenotype.Purified CD19+ B cells were stimulated with anti-IgM, anti-CD40 mAb and IL-21 and multi-color flow cytometric analyses were performed on day 6 as described in Fig 1. The figures A, B and C show the expression of various surface markers on stimulated B cells (Percentage of positive cells/total proliferating CD19+ cells) p < 0.05, **p < 0.01. CTRL, control; TAC6, 6 ng/ml TAC; SRL2, 2 ng/ml SRL; SRL6, 6 ng/ml SRL.

Mentions: Subsequently, we studied the phenotypic make-up of the proliferating B cells in the control, TAC and SRL treated cultures by flow cytometry (Fig 2). Both the control and TAC-treated cultures had similar subset profile; the only significant difference was a decreased percentage of immature CD24+ B cells with TAC, suggesting that TAC did not favor or inhibited their expansion. In contrast, residual B cells that proliferated in the presence of SRL had an entirely different subset profile. There were significantly lower percentages of immature CD24+ and CD38+ B cells [25, 26], lesser naïve CD27-IgD+ cells (especially at 6ng/ml SRL; p = 0.022), and reduced CD27+ total memory B cells or even memory B cells that differentiated into plasma cells (CD27hi B cells) (p<0.05, p<0.01 compared to TAC or control group, respectively). However, no significant difference was observed among the three different cultures in the percentages of memory B cells expressing the trafficking receptor CD62L (CD27+CD62L+) or the complement receptor 2 (CD21). The B cells that proliferated in presence of SRL, however, had significantly higher percentage of cells that expressed CD23, a low affinity IgE receptor present on mature/activated B cells [27–29] and a subset of cells with the follicular B cell phenotype (CD21+CD23+ B cells) [30].


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)

B-cell stimulation in the presence of SRL resulted in a population shift toward an activated phenotype.Purified CD19+ B cells were stimulated with anti-IgM, anti-CD40 mAb and IL-21 and multi-color flow cytometric analyses were performed on day 6 as described in Fig 1. The figures A, B and C show the expression of various surface markers on stimulated B cells (Percentage of positive cells/total proliferating CD19+ cells) p < 0.05, **p < 0.01. CTRL, control; TAC6, 6 ng/ml TAC; SRL2, 2 ng/ml SRL; SRL6, 6 ng/ml SRL.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0129658.g002: B-cell stimulation in the presence of SRL resulted in a population shift toward an activated phenotype.Purified CD19+ B cells were stimulated with anti-IgM, anti-CD40 mAb and IL-21 and multi-color flow cytometric analyses were performed on day 6 as described in Fig 1. The figures A, B and C show the expression of various surface markers on stimulated B cells (Percentage of positive cells/total proliferating CD19+ cells) p < 0.05, **p < 0.01. CTRL, control; TAC6, 6 ng/ml TAC; SRL2, 2 ng/ml SRL; SRL6, 6 ng/ml SRL.
Mentions: Subsequently, we studied the phenotypic make-up of the proliferating B cells in the control, TAC and SRL treated cultures by flow cytometry (Fig 2). Both the control and TAC-treated cultures had similar subset profile; the only significant difference was a decreased percentage of immature CD24+ B cells with TAC, suggesting that TAC did not favor or inhibited their expansion. In contrast, residual B cells that proliferated in the presence of SRL had an entirely different subset profile. There were significantly lower percentages of immature CD24+ and CD38+ B cells [25, 26], lesser naïve CD27-IgD+ cells (especially at 6ng/ml SRL; p = 0.022), and reduced CD27+ total memory B cells or even memory B cells that differentiated into plasma cells (CD27hi B cells) (p<0.05, p<0.01 compared to TAC or control group, respectively). However, no significant difference was observed among the three different cultures in the percentages of memory B cells expressing the trafficking receptor CD62L (CD27+CD62L+) or the complement receptor 2 (CD21). The B cells that proliferated in presence of SRL, however, had significantly higher percentage of cells that expressed CD23, a low affinity IgE receptor present on mature/activated B cells [27–29] and a subset of cells with the follicular B cell phenotype (CD21+CD23+ B cells) [30].

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