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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 effectively inhibited B-cell proliferation.Purified CD19+ B cells were labeled with CFSE, stimulated with anti-IgM, anti-CD40 mAb and IL-21 (BCR method) in the absence (control; CTRL) or presence of TAC (6ng/ml) or SRL (2ng/ml or 6ng/ml) and flow cytometric analyses were performed after 6 days in culture. (A) A representative experiment: cells were gated on viable lymphocytes and analyzed for CFSE diluting proliferating cells. This scheme of analysis was used in all subsequent experiment, unless indicated otherwise. (B) The percentage of proliferating CD19+ B cells as obtained in A from 7 different experiments. (C) Absolute number of proliferating CD19+ B cells was calculated in each experiment by multiplying the recovered cell counts with the percentage of proliferating cells as in A (n = 7). Statistically significant (*p < 0.05) inhibition of B cell proliferation was observed with SRL at both subtherapeutic (2ng/ml) and therapeutic (6ng/ml) doses.
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pone.0129658.g001: SRL effectively inhibited B-cell proliferation.Purified CD19+ B cells were labeled with CFSE, stimulated with anti-IgM, anti-CD40 mAb and IL-21 (BCR method) in the absence (control; CTRL) or presence of TAC (6ng/ml) or SRL (2ng/ml or 6ng/ml) and flow cytometric analyses were performed after 6 days in culture. (A) A representative experiment: cells were gated on viable lymphocytes and analyzed for CFSE diluting proliferating cells. This scheme of analysis was used in all subsequent experiment, unless indicated otherwise. (B) The percentage of proliferating CD19+ B cells as obtained in A from 7 different experiments. (C) Absolute number of proliferating CD19+ B cells was calculated in each experiment by multiplying the recovered cell counts with the percentage of proliferating cells as in A (n = 7). Statistically significant (*p < 0.05) inhibition of B cell proliferation was observed with SRL at both subtherapeutic (2ng/ml) and therapeutic (6ng/ml) doses.

Mentions: After 6 days of stimulation, more than 70% B cells proliferated in the control cultures without the drugs (Fig 1A and 1B). TAC at clinically relevant dose of 6 ng/ml did not have any inhibitory effect on B cell proliferation. In contrast, in the SRL treated cultures, the percentage of proliferating cells was significantly reduced even at subtherapeutic 2 ng/ml. Concomitantly, the absolute numbers of proliferating CD19+ B cells were also significantly reduced in the SRL treated cultures when compared to those in the controls or the TAC treated cultures (Fig 1C).


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 effectively inhibited B-cell proliferation.Purified CD19+ B cells were labeled with CFSE, stimulated with anti-IgM, anti-CD40 mAb and IL-21 (BCR method) in the absence (control; CTRL) or presence of TAC (6ng/ml) or SRL (2ng/ml or 6ng/ml) and flow cytometric analyses were performed after 6 days in culture. (A) A representative experiment: cells were gated on viable lymphocytes and analyzed for CFSE diluting proliferating cells. This scheme of analysis was used in all subsequent experiment, unless indicated otherwise. (B) The percentage of proliferating CD19+ B cells as obtained in A from 7 different experiments. (C) Absolute number of proliferating CD19+ B cells was calculated in each experiment by multiplying the recovered cell counts with the percentage of proliferating cells as in A (n = 7). Statistically significant (*p < 0.05) inhibition of B cell proliferation was observed with SRL at both subtherapeutic (2ng/ml) and therapeutic (6ng/ml) doses.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0129658.g001: SRL effectively inhibited B-cell proliferation.Purified CD19+ B cells were labeled with CFSE, stimulated with anti-IgM, anti-CD40 mAb and IL-21 (BCR method) in the absence (control; CTRL) or presence of TAC (6ng/ml) or SRL (2ng/ml or 6ng/ml) and flow cytometric analyses were performed after 6 days in culture. (A) A representative experiment: cells were gated on viable lymphocytes and analyzed for CFSE diluting proliferating cells. This scheme of analysis was used in all subsequent experiment, unless indicated otherwise. (B) The percentage of proliferating CD19+ B cells as obtained in A from 7 different experiments. (C) Absolute number of proliferating CD19+ B cells was calculated in each experiment by multiplying the recovered cell counts with the percentage of proliferating cells as in A (n = 7). Statistically significant (*p < 0.05) inhibition of B cell proliferation was observed with SRL at both subtherapeutic (2ng/ml) and therapeutic (6ng/ml) doses.
Mentions: After 6 days of stimulation, more than 70% B cells proliferated in the control cultures without the drugs (Fig 1A and 1B). TAC at clinically relevant dose of 6 ng/ml did not have any inhibitory effect on B cell proliferation. In contrast, in the SRL treated cultures, the percentage of proliferating cells was significantly reduced even at subtherapeutic 2 ng/ml. Concomitantly, the absolute numbers of proliferating CD19+ B cells were also significantly reduced in the SRL treated cultures when compared to those in the controls or the TAC treated cultures (Fig 1C).

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