ABSTRACT
Objective To investigate the significant of peripheral CD4+CD69+T lymphocytes in patients with autoimmune hemolytic anemia (AIHA)/Evans syndrome (ES).Methods In this study peripheral blood samples from 32 patients with AIHA/ES (15 hemolytic episode patients,17 remission patients) and 13 healthy controls were collected.Patients with AIHA/ES were recruited in Tianjin Medical University General Hospital from October 2015 to May 2016.The percentages of CD69+ T lymphocytes were analyzed by flow cytometry.The expression of CD69 mRNA in CD4+T lymphocytes which was sorted from peripheral blood by magnetic activated cell sorting (MACS) was detected using real-time PCR.Soluable CD69 was measured by ELISA.Results In hemolytic episode patients,the ratio of CD3+CD69+/CD3+T lymphocytes [(3.08 ± 1.48)％] was significantly higher than that in healthy controls [(1.28 ± 0.83)％,P＜0.01] and in remission group[(1.96± 1.33)％,P＜0.05].The absolute count of CD3+CD69+T lymphocytes in hemolytic episode group [(2.94± 1.81)× 107/L] was higher than that in healthy controls [(1.48± 1.42)× 107/L,P＜0.05].The ratio of CD3+CD4+CD69+/CD3+CD4+T cells in hemolytic episode group [(2.16± 1.56)％] was significantly higher than that in remission group [(1.16±0.62)％,P＜0.05] and healthy controls[(0.94±0.78)％,P＜0.05].The quantity of CD3+CD4+CD69+T lymphocytes in hemolytic episode group[(1.04±0.98)× 107/L] was higher than in healthy controls [(0.44± 0.38) × 107 / L,P＜0.05].The ratio of CD3+CD8+CD69+/CD3+ CD8+T lymphocyte in hemolytic episode group [(4.87±2.56)％] was significantly higher than that in healthy controls[(1.83± 1.27)％,P＜0.01].The quantity of CD3+CDs+CD69+T lymphocytes in three groups did not show significant difference.The ratio of CD3+CD4+CD69+/CD3+CD4+T lymphocytes in hemolytic episode group was negatively correlated with hemoglobin (Hb) (P＜0.01),positively correlated with the percentage of reticulocytes (Ret％)(P=0.01)total bilirubin(TBil),indirect bilirubin(IBil) (P＜0.01) and not correlated with absolute reticulocytes count,lactic dehydrogenase (LDH),complement 3(C3),complement 4 (C4).The ratio of CD3+CD4+CD69+/CD3+CD4+T lymphocytes in remission group was negatively correlated with Hb (P＜0.05).In hemolytic episode patients CD69 mRNA (32.26±35.11) was significantly higher than that in remission group(6.05±5.87)(P＜0.05)and healthy controls (1.76± 1.85)(P＜0.01).CD69 mRNA in remission group was significantly higher than healthy controls (P＜0.05).Serum CD69 in hemolytic episode patients [(494.21 ± 16.06) ng/L] was significantly higher than that in healthy controls [(441.39± 104.6) ng/L,P＜0.05].Conclusion Our findings suggest that the proportion of CD4+CD69+ T lymphocytes increase in AIHA/ES patients,which is correlated with the severity of disease.