Characterization of granulocyte colony stimulating factor for in vitro induction of regulatory T cells for cellular immune intervention in transplant medicine.

Objectives: The application of regulatory T cells in the field of solid-organ and hematopoietic stem cell transplantation is under investigation to develop novel cellular strategies for tolerance induction. Establishing in vitro procedures to induce and expand regulatory T cells seeks to overcome the limiting small number of this rare T cell population. The present study is based on growing evidence that granulocyte colony stimulating factor exerts immune regulatory function in the adaptive immune system and may induce regulatory T cells in vivo.

Materials And Methods: We analyzed the effect of recombinant granulocyte colony stimulating factor to directly convert CD4+CD25- T cells into regulatory T cells in vitro. Marker molecules were analyzed by quantitative reverse transcriptase-polymerase chain reaction and fluorescent-activated cell sorter analyses. Functional assays were performed to investigate the suppressive capacity of granulocyte colony stimulating factor stimulated T cells.

Results: Kinetic analyses of Foxp3 gene expression uncovered increased levels early after in vitro stimulation with granulocyte colony stimulating factor. However, protein analyses for the master transcription factor Foxp3 and other regulatory T cells revealed that granulocyte colony stimulating factor did not directly induce a regulatory T cell phenotype. Moreover, functional analyses demonstrated that granulocyte colony stimulating factor stimulation in vitro does not result in a suppressive, immune regulatory T cell population.

Conclusions: Granulocyte colony stimulating factor does not induce regulatory T cells with a specific phenotype and suppressive potency in vitro. Therefore, granulocyte colony stimulating factor does not qualify for developing protocols aimed at higher regulatory T cell numbers for adoptive transfer strategies in solid organ and hematopoietic stem cell transplantation.