Retrospective Comparison of Lymphocyte Subsets in the Different Stage of Patients with Chronic Graft-Versus-Host Disease after HSCT for Possibility of Early Diagnosis.
Authors: Xing Haizhou , Du Xin , Weng Jianyu , Lu Zesheng , Zhang Jianjun , Zhou Maohua 
Published: October 2019

Abstract
Chronic graft-versus-host disease (cGVHD) continue to cause significant morbidity and mortality in patients undergoing hematopoietic stem cell transplantation (HSCT). It remains the most frequent complication following allogeneic hematopoietic stem cell transplantation, occurring in 30% to 70% of long-term survivors.[Lee SJ, Blood 2002 ]. An experienced clinician may be able to make a diagnosis of chronic GVHD based on classic manifestations such as lichenoid or sclerodermatous skin changes. If the findings are not typical, a tissue biopsy is warranted to aid the diagnosis and to determine if infection is present (A.L. Gilman Gilman and J. Serody seminhematol.2005). Recently, there are many research in lymphocyte subsets on cGVHD by using flowcytometry.It helps us to understand the mechsim about it, and could be help for diagnosis, staging, and response criteria in this disorder. However, the change of lymphocytes subsets is still unknown in the different stage of cGVHD. We are here to study the lymphocyte subsets in the peripheral blood with different stage of cGVHD after allo-HSCT. Peripheral blood mononuclear cells were isolated from anticoagulated venous blood using lymphocyte separation medium according to manufacturer’s nstructions.Freshly isolated peripheral blood mononuclear cells were analyzed. Fluorescein sothiocyanate conjugated monoclonal antibodies (Becton Dickinson, mAb) By using FACScalibur, (Becton Dickinson).The data of the two-group comparison was done on the analysis of variance and Students t test.According to the National Institutes of Health Consensus Development Project on Criteria for Clinical Trials in chronic Graft-versus-Host Disease in 2005, we analysis the diffierent groups with non-cGVHD, mild, moderate and severe cGVHD. The proportions of the lymphocyte subsets are different with the different stage of cGVHD. The more serious of cGVHD, the more proportions of the total lymphoctyes. B lymphocyte has significant difference with different stage of cGVHD, although the percentage of the total lymphocytes is 2.14%∼16.14% (P=0.0023). There is significant difference on the percent of CD8+ CD45RO+ cell among the non-cGVHD, mild cGVHD, moderate cGVHD or severe cGVHD(44.56%,46.59%,56.77% and 61.68%, respectively. P=0.0475). But the percent of CD4+CD25+ cell is higher than that of CD8+CD25+ cell (31.86%∼43.75%/4.50%∼7.08%)The percent of CD69+CD4+,CD69+CD8+ (CD95+CD4+, CD95+CD8+, CD28+ CD4+, CD28+CD8+ cell are not significant different within the different statue cGVHD. We can draw a conclusion as below: It may help for clinician to early diagnosis,stage using routine monoclone antibodies by flowcytometry. the percents of the lymphocyte subsets are different with the different status of cGVHD. With the more serious of cGVHD, the more proportions of the total lymphocytes; CD4+CD25+ cell ; CD3+CD8+ cell and B cell increase significantly, the percent of CD45RO+CD8+ cell and CD3+CD4+ cell drease at the same time. It may help for early diagnoses and treatment for the cGVHD.