The Application of Novel Identified CD8 Regulatory Precursors in Inducing Immune Tolerance After Allo-HSCT

Healthy donor samples will be collected at the onset of the project, and peripheral blood specimens from patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT) will be serially assessed at 90 days, 180 days, 365 days, and 2 years post-transplantation. In this study, flow cytometry will be utilized to analyse the proportions and absolute numbers of CD8Trp in the peripheral blood of healthy donors and transplant recipients by combining markers such as CD3、CD4、CD8、HLA-DR、CD27、CD45RA、CCR7、CD28、ICOS、FOXP3、TCF1.

Healthy donor samples will be collected at the onset of the project, and peripheral blood specimens from patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT) will be serially assessed at 90 days, 180 days, 365 days, and 2 years post-transplantation. Peripheral blood T cells will be isolated and subjected to in vitro stimulation using either CD3/CD28 beads or the pharmacological agents phorbol 12-myristate 13-acetate (PMA) and ionomycin. Following stimulation, the production of cytokines will be assessed, including but not limited to IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ, CD107A , Perforin and Granzyme B.

Healthy donor samples will be collected at the onset of the project, and peripheral blood specimens from patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT) will be serially assessed at 90 days, 180 days, 365 days, and 2 years post-transplantation. The activation markers CD25, CD38, CD69, CD137, CD71, and HLA-DR, among others, will be detected via flow cytometry to evaluate the activation status of T cells in the peripheral blood of both healthy donors and transplant recipients.

Healthy donor samples will be collected at the onset of the project, and peripheral blood specimens from patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT) will be serially assessed at 90 days, 180 days, 365 days, and 2 years post-transplantation. This study will employ flow cytometry to quantify the proportions of various T-cell subpopulations in the peripheral blood of healthy donors and transplant recipients, including recent thymic emigrants (RTE), naive T cells, central memory T cells (TCM), effector memory T cells (TEM), terminally differentiated effector memory T cells (TEMRA), and regulatory T cells (Tregs).

Healthy donor samples will be collected at the onset of the project, and peripheral blood specimens from patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT) will be serially assessed at 90 days, 180 days, 365 days, and 2 years post-transplantation. T-cell exhaustion will be evaluated through the analysis of a range of specific markers, including PD-1, TIGIT, LAG-3, TIM-3, and associated molecules.

Healthy donor samples will be collected at the onset of the project, and peripheral blood specimens from patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT) will be serially assessed at 90 days, 180 days, 365 days, and 2 years post-transplantation. The functional status of thymic output will be comprehensively evaluated by detecting CD31, CD38, and CD103 markers in peripheral blood using flow cytometry. Additionally, the level of sjTREC (Signal Joint T-Cell Receptor Excision Circle) will be measured to further illustrate the functional capacity of thymic output.