Primary regulatory T cell in-vitro amplification system and method

Abstract

The invention discloses a primary regulatory T cell in-vitro amplification system. The amplification system is prepared by adding anti-CD3/28 antibody, interleukin-2. Rapa and transforming growth factor-beta 1 into a basic culture medium, wherein the final concentration of the anti-CD3/28 antibody is 2 microgram/ml, the final concentration of the interleukin-2 is 1000U/ml, the final concentrationof the Rapa is 100-200 nanogram/ml, and the final concentration of the transforming growth factor-beta 1 is 4-8 nanogram/ml. Compared with an amplification system which uses the Rapa alone, the Rapa and transforming growth factor-beta 1 combined amplification system is good in stability and high in Foxp3+regulatory T cell purity.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an in vitro expansion system and expansion method of primary regulatory T cells. Background technique [0002] CD4 + CD25 + Regulatory T cells (Tregs) are the most important immunosuppressive cells in the body, which can effectively suppress immune responses and regulate immune balance. Their research and application in the fields of transplant tolerance induction and autoimmune disease treatment are widely focus on. [0003] In 1995, Sakaguchi et al. found that mice lacked CD4 + CD25 + Regulatory T cells can trigger a variety of autoimmune diseases, and the reinfusion of these regulatory T cells can inhibit the occurrence of diseases. Regulatory T cell subsets are developed and differentiated in the thymus or peripheral CD4+ naive T cells, and can inhibit the proliferation and differentiation of effector T cells through direct cell-to-cell contact, secretion...

AI Technical Summary

Problems solved by technology

[0005] However, the current in vitro culture of primary regulatory T cells still has the following problems: ① Regulatory T cells show immune incompetence in vitro, and it is difficult to expand with conventional culture medium, so it is necessary to add activating factors anti-CD3, CD28 antibodies (anti- CD3 / 28Ab) and interleukin-2 (IL-2) to stimulate cell proliferation; ②Immunomagnetic bead sorting method can obtain high-purity primary regulatory T cells, but there are still a small amount of effector T cells remaining. The secreted inflammatory factors will lead to down-regulation of the transcription factor Foxp3 expression of regulatory T cells, which will significantly reduce the purity of regulatory T cells in the expansion system; ③ Studies have shown that rapamycin (Rapamycin, Rapa) is a commonly used clinical immunosuppressant , can selectively inhibit the proliferation of effector T cells, which is beneficial to the expansion of regulatory T cells in vitro and the stable expression of Foxp3, but the actual effect is limited

Images