Study Of IgG Subclass Of The Patients With Immuno-related Hemocytopenia

Objective:To investigate the immunoglobulin G(Ig G)sublass on the membrane of bone marrow hematopoietic cells and Ig G levels in bone marrow supernatant of patients with immuno-related Immuno-related hemtocytopenia(IRH),for exploring advantage subclass of Ig G in IRH patients.Method:1.Newly diagnosed IRH patients and ICUS patients were enrolled in this study, as well as healthy donors as controls.We carried on study as following :(1) The categories of auto-antibodies(Ig M and Ig G) on the membrane of bone marrow hemmatopoietic cells(CD15+,CD34+ and Gly Co A+ cells)were assayed by Flow cytometry(FCM).(2) The categories of auto-antibodies(Ig G1 and Ig G3) on the membrane of bone marrow hemmatopoietic cells(CD15+and Gly Co A+ cells)were assayed by Flow cytometry(FCM).(3) Bone marrow nucleated cells antibodies(Ig G1、Ig G2、Ig G3 and Ig G4)were also detected by immunohistochemical staining.2.BMMNC-Ab Ig G+ IRH patients, remitted IRH patients,ICUS patients and healthy donors were enrolled in this study. We carried on study as following :(1)The Ig G1、Ig G2、Ig G3 and Ig G4 levels in bone marrow supernatant were detected by Cytometric Bead Array(CBA);(2)The ratio of CD5+CD19+/CD19+ in blood were tested by Flow cytometry(FCM);(3)the m RNA expression of Ig G1 in peripheral blood were detected by RT-PCR.Result:1. The positive rate of antibodies(Ig G) on bone marrow hemmatopoietic cells of IRH patients was 58.69(27/46), which was significantly higher than that in ICUS group(0%) and healthy control group(0%).2. The positive rate of Ig G1 on CD15+ cells was(3.13±1.80)% in IRH group,which was significantly higher than that in healthy control group. The positive rate of Ig G3 on CD15+ cells was(2.73±2.09)% in IRH group,which was significantly higher than that in healthy control group. The positive rate of Ig G1 on Gly Co A+ cells was( 3.97±2.21)% in IRH group,which was significantly higher than that in healthy control group. The positive rate of Ig G3 on Gly Co A+ cells was(2.24±1.82)% in IRH group,which was significantly higher than that in healthy control group.3.Ig G subclass(Ig G1、Ig G2、Ig G3 and Ig G4)on bone marrow nucleated cells in IRH patients, ICUS patients and healthy donors were detected by Immunohistochemistry. 11/15 IRH patients’ Ig G1 positive cells≥3(count 100 bone marrow nucleated cells). There were rarely Ig G1+ cells in ICUS patients and healthy donors’ bone marrow smears. Ig G2,Ig G3 and Ig G4 positive bone marrow nucleated cells were rarely detected in three groups by immunohistochemistry.4.We detected Ig G subclass(Ig G1,Ig G2,Ig G3 and Ig G4) in bone marrow supernatant by Cytometric Bead Array. The Ig G1 level in IRH group was(6998.51±2498.38)mg/L which was higher than remission group(4702.27±1235.25)mg/L, healthy control group(5613.20±1640.93) mg/L and ICUS group(5166.20±1761.64) mg/L, P 〈 0.05. The Ig G3 level in IRH group was(637.63±252.93)mg/Lwhich was higher than remission group(415.47±196.60)mg/L,healthy control group(422.85±212.69) mg/L and ICUS group(485.70±197.29) mg/L, P〈0.05. Ig G2 levels in four groups were no statistical difference. Ig G4 levels in these four groups were no statistical difference. The total Ig G level in IRH group was(10903.29±2576.10)mg/L which was higher than remission group(8468.27±1957.58)mg/L,P〈0.05.5.The ratio of CD5+CD19+/CD19+ in IRH group was(23.34±9.36)% which were significance higher than healthy control group(13.67±5.10)% and ICUS group(12.40±3.63)%(P<0.01). Ig G1 level had a positive correlation with the proportion of CD5+ B lymphocytes(r=0.392, P =0.043).6.There was an inverse correlation between the Ig G1 level and the count of neutrophil(r =-0.459, P = 0.016) in IRH patients. There was an inverse correlation between the level of Ig G3 and hemoglobin(Hb)(r =-0.389, P = 0.045) in IRH patients.7. In IRH group,the m RNA expression of Ig G1 in peripheral blood is 5.27±4.83 which is higher than healthy control 1.58±1.50,P<0.05. Conclusion:1.There are auto-antibodies Ig G on the membrane of bone marrow hematopoietic cells of IRH patients,which can effect on bone marrow hematopoietic cells leading to loss of hematopoietic cells, and then reduce the blood cells. By FCM,the ratio of Ig G1 and Ig G3 on CD15+ cells and Gly Co A+ cells increased significantly in BMMNC-Ab Ig G positive IRH patients. By immunohistochemistry, the mainly Ig G subclass on the bone marrow nucleated cells is Ig G1,which suggesting that Ig G1 and Ig G3 participate in the destruction of hematopoietic.2.Ig G1 and Ig G3 level in bone marrow supernatant increased significantly in BMMNC-Ab Ig G positive IRH patients. Ig G1 level had a positive correlation with the proportion of CD5+ B lymphocytes.There was an inverse correlation between the Ig G1 level and the count of neutrophil in IRH patients. There was an inverse correlation between the level of Ig G3 and hemoglobin in IRH patients. The m RNA expression of Ig G1 is higher than healthy control. Above suggested that the production of Ig G1 increased which has closely associated with hematopoietic function.