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Ex vivo generation of human alloantigen-specific regulatory T cells from CD4(pos)CD25(high) T cells for immunotherapy.

Peters JH, Hilbrands LB, Koenen HJ, Joosten I - PLoS ONE (2008)

Bottom Line: Efficient increases in cell numbers were obtained.Primary allogeneic stimulation appeared a prerequisite in the generation of alloantigen-specific Treg, while secondary allogeneic or polyclonal stimulation with anti-CD3 plus anti-CD28 monoclonal antibodies enriched alloantigen-specificity and cell yield to a similar extent.This approach is of particular interest for recipients of HLA mismatched transplants.

View Article: PubMed Central - PubMed

Affiliation: Department of Bloodtransfusion and Transplantation Immunology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.

ABSTRACT

Background: Regulatory T cell (Treg) based immunotherapy is a potential treatment for several immune disorders. By now, this approach proved successful in preclinical animal transplantation and auto-immunity models. In these models the success of Treg based immunotherapy crucially depends on the antigen-specificity of the infused Treg population. For the human setting, information is lacking on how to generate Treg with direct antigen-specificity ex vivo to be used for immunotherapy.

Methodology/principal findings: Here, we demonstrate that in as little as two stimulation cycles with HLA mismatched allogeneic stimulator cells and T cell growth factors a very high degree of alloantigen-specificity was reached in magnetic bead isolated human CD4(pos)CD25(high) Treg. Efficient increases in cell numbers were obtained. Primary allogeneic stimulation appeared a prerequisite in the generation of alloantigen-specific Treg, while secondary allogeneic or polyclonal stimulation with anti-CD3 plus anti-CD28 monoclonal antibodies enriched alloantigen-specificity and cell yield to a similar extent.

Conclusions/significance: The ex vivo expansion protocol that we describe will very likely increase the success of clinical Treg-based immunotherapy, and will help to induce tolerance to selected antigens, while minimizing general immune suppression. This approach is of particular interest for recipients of HLA mismatched transplants.

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Related in: MedlinePlus

Schematic overview of expansion strategies.Treg were expanded in two cycles, in which alloantigen and polyclonal stimulation was alternated, resulting in four distinct strategies: two subsequent cycles with alloantigen stimulation; primary cycle with alloantigen stimulus and secondary cycle with polyclonal stimulus; primary cycle with polyclonal stimulus and secondary cycle with alloantigen stimulus; and two subsequent cycles with polyclonal stimulation.
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pone-0002233-g001: Schematic overview of expansion strategies.Treg were expanded in two cycles, in which alloantigen and polyclonal stimulation was alternated, resulting in four distinct strategies: two subsequent cycles with alloantigen stimulation; primary cycle with alloantigen stimulus and secondary cycle with polyclonal stimulus; primary cycle with polyclonal stimulus and secondary cycle with alloantigen stimulus; and two subsequent cycles with polyclonal stimulation.

Mentions: With the objective to obtain the highest numbers of functionally active Treg with optimal direct alloantigen-specificity, we devised four expansion strategies, employing polyclonal and alloantigen-specific stimulation in two subsequent cycles of Treg expansion (Figure 1). The cells that were obtained with these expansion strategies were compared in terms of absolute cell numbers, phenotype, suppressive capacity, antigen-specificity and anergy. Prior to analysis of the four selected Treg expansion strategies, individual expansion cycles were optimized, with regard to strength and mode of stimulation.


Ex vivo generation of human alloantigen-specific regulatory T cells from CD4(pos)CD25(high) T cells for immunotherapy.

Peters JH, Hilbrands LB, Koenen HJ, Joosten I - PLoS ONE (2008)

Schematic overview of expansion strategies.Treg were expanded in two cycles, in which alloantigen and polyclonal stimulation was alternated, resulting in four distinct strategies: two subsequent cycles with alloantigen stimulation; primary cycle with alloantigen stimulus and secondary cycle with polyclonal stimulus; primary cycle with polyclonal stimulus and secondary cycle with alloantigen stimulus; and two subsequent cycles with polyclonal stimulation.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376059&req=5

pone-0002233-g001: Schematic overview of expansion strategies.Treg were expanded in two cycles, in which alloantigen and polyclonal stimulation was alternated, resulting in four distinct strategies: two subsequent cycles with alloantigen stimulation; primary cycle with alloantigen stimulus and secondary cycle with polyclonal stimulus; primary cycle with polyclonal stimulus and secondary cycle with alloantigen stimulus; and two subsequent cycles with polyclonal stimulation.
Mentions: With the objective to obtain the highest numbers of functionally active Treg with optimal direct alloantigen-specificity, we devised four expansion strategies, employing polyclonal and alloantigen-specific stimulation in two subsequent cycles of Treg expansion (Figure 1). The cells that were obtained with these expansion strategies were compared in terms of absolute cell numbers, phenotype, suppressive capacity, antigen-specificity and anergy. Prior to analysis of the four selected Treg expansion strategies, individual expansion cycles were optimized, with regard to strength and mode of stimulation.

Bottom Line: Efficient increases in cell numbers were obtained.Primary allogeneic stimulation appeared a prerequisite in the generation of alloantigen-specific Treg, while secondary allogeneic or polyclonal stimulation with anti-CD3 plus anti-CD28 monoclonal antibodies enriched alloantigen-specificity and cell yield to a similar extent.This approach is of particular interest for recipients of HLA mismatched transplants.

View Article: PubMed Central - PubMed

Affiliation: Department of Bloodtransfusion and Transplantation Immunology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.

ABSTRACT

Background: Regulatory T cell (Treg) based immunotherapy is a potential treatment for several immune disorders. By now, this approach proved successful in preclinical animal transplantation and auto-immunity models. In these models the success of Treg based immunotherapy crucially depends on the antigen-specificity of the infused Treg population. For the human setting, information is lacking on how to generate Treg with direct antigen-specificity ex vivo to be used for immunotherapy.

Methodology/principal findings: Here, we demonstrate that in as little as two stimulation cycles with HLA mismatched allogeneic stimulator cells and T cell growth factors a very high degree of alloantigen-specificity was reached in magnetic bead isolated human CD4(pos)CD25(high) Treg. Efficient increases in cell numbers were obtained. Primary allogeneic stimulation appeared a prerequisite in the generation of alloantigen-specific Treg, while secondary allogeneic or polyclonal stimulation with anti-CD3 plus anti-CD28 monoclonal antibodies enriched alloantigen-specificity and cell yield to a similar extent.

Conclusions/significance: The ex vivo expansion protocol that we describe will very likely increase the success of clinical Treg-based immunotherapy, and will help to induce tolerance to selected antigens, while minimizing general immune suppression. This approach is of particular interest for recipients of HLA mismatched transplants.

Show MeSH
Related in: MedlinePlus