| Objective:This study was purposed to applicate the bone marrow indirect-Coombs test, and investigate its clinical significances in diagnosis of the Immuno-related Pancytopenia (IRP).Methods130patients with pancytopenia (22IRP,8Idiopathic cytopenia of undetermined significance, ICUS and15healthy persons were enrolled in this study. The bone marrow nucleated cell (CD15+GlyCoA+and CD34+cells) antibodies (IgM or IgG) were tested by floweytometry.2After incubation of the healthy persons’bone marrow nucleated cell (BMNC)with the bone marrow supernatant of30patients and their owns respectively for45min, the positive rate of bone marrow nucleated cell (CD15+、GlyCoA+and CD34+cells) antibodies(BMNC-Ab)were detected by flow cytometry, and the correlation analysis of positive rate with the clinical data of patients were also analyzed.3After incubation of the30patients’bone marrow nucleated cell (BMNC)with the bone marrow supernatant of their owns and healthy persons’respectively for45min, we analyzed the bone marrow Mononuclear Cells (CD15+、GlyCoA+and CD34+cells) antibody (BMNC-Ab) by flow cytometry, the clinical data of patients were also analyzed.Results1Among30patients with pancytopenia, there were16cases present direct-Coombs test positive before incubation, the others were negative,and15healthy persons present direct-Coombs test negative.2After incubation, there were16cases present indirect-Coombs test positive, the total positive rate were53.33%(16/30),4cases present positive while it’s negative before incubation, the total positive rate were28.57%(4/14),4cases were positive among8ICUS,the total positive rate were50%(4/8)①In the experiment group, the positive rate of CD15+cells IgG was0.34%, which was significantly higher than that in control group0.20%(P<0.05) ②In the experiment group, the positive rate of CD15+cells IgM was0.34%, which was significantly higher than that in control group0.23%(P<0.05);③The positive rate of GlyCoA+cells IgG was (0.83±0.75)%, which was significantly higher than that in control group (0.47±0.43)%(P<0.05);④The positive rate of GlyCoA+cells IgM (2.12±1.98)%, which was significantly higher than that in control group (0.68±0.64)%(P<0.01);⑤The positive rate of CD34+cells IgG was0.64%, which was significantly higher than that in control group0.00(P<0.05)⑥The positive rate of CD34+cells IgM was0.21%which was significantly higher than that in control group0.00%(P<0.05);⑦The positive rate of CD15+cells IgG was positive correlated with the ratio of CD5+B cell (r=0.576,P=0.002),, but there were no relativitives with other clinic indicators;⑧The positive rate of CD15+cells IgM was positive correlated with the ratio of CD5+B cell (r=0.412,P=0.037),but there were no relativitives with other clinic indicators;⑨The positive rate of GlyCoA+cells IgG was negative correlated with the ratio of the level of HB r=-0.549,P=0.002)、RET%(r=-0.475, P=0.009)、the ratio of bone marrow erythroid (r=-0.475, P=0.014). DC1/DC2(r=0.421,P=0.03), was negative correlated with the ratio of the level of IBIL (r=0.730, P=0.000)、CD5+B cell (r=0.684,P=0.000);⑩The positive rate of GlyCoA+cells IgM was negative correlated with the ratio of the level of HB (r=-0.488,P=0.010)、RET%(r=-0.399, P=0.029)、the ratio of bone marrow erythroid(r=-0.5045,P=0.012)、DC1/DC2(r=-0.661,P=0.001),was negative correlated with the ratio of the level of IBIL (r=0.430,P=0.018)、CD5+B cell(r=0.841,P=0.000)3After incubation, there were18cases present positive BMNC-Ab,the total positive rate were60%(18/30),12cases present positive while it’s negative before incubation, the total positive rate were85.71%(12/14),6cases were positive among8ICUS,the total positive rate were75%(6/8)①In the experiment group, the positive rate of CD15+cells IgG was (2.72±3.24) %, which was significantly higher than that in control group (1.03±1.15)%(P<0.05);②The positive rate of the positive rate of CD15+cells IgM was (0.97±0.77)%, which was higher than that in control group (1.03±1.15)%(P<0.05);③The positive rate of GlyCoA+cells IgG was (3.80±6.82)%, which was significantly higher than that in control group0.95±1.05)%(P<0.001);④The positive rate of GlyCoA+cells IgM was (3.14±1.78)%、which was significantly higher than that in control group (1.93±1.32)%(P<0.001);⑤The positive rate of CD34+cells IgG was1.09%, which was higher than that in control group0.00%(P<0.05);⑥The positive rate of CD34+cells IgM was1.44%,which was higher than that in control group0.00%(P<0.05);⑦The positive rate of CD15+cells IgG was positive correlated with the ratio of CD5+B cell (r=0.685,P=0.000), was negative correlated with the ratio of DC1/DC2(r=-0.524,P=0.002),but there were no relativitives with other clinic indicators;⑧The positive rate of CD15+cells IgM was positive correlated with the ratio of CD5+B cell (r=0.421, P=0.021), was negative correlated to ratio of DC1/DC2(r=-0.539,P=0.002),but there were no relativitiveswith other clinic indicators;⑨The positive rate of GlyCoA+cells IgG was negative correlated with the ratio of the level of HB (r=-0.657,P=0.000). the ratio of bone marrow erythroid (r=-0.734, P=0.000)、DC1/DC2(r=-0.368,P=0.041),was positive correlated with the ratio of the level of IBIL (r=0.541,P=0.002)、the ratio of CD5+B cell (r=0.498, P=0.004) and the RET%(r=0.514, P=0.003);⑩The positive rate of GlyCoA+cells IgM was negative correlated with the ratio of the level of HB (r=-0.368,P=0.042)、the ratio of bone marrow erythroid (r=-0.699, P=0.000)、DC1/DC2(r=-0.448,P=0.011),was negative correlated to ratio of the level of IBIL (r=0.383,P=0.033)、the ratio of CD5+B cell (r=0.374, P=0.038) and the RET%(r=0.560, P=0.001)Conclusion These studies show that the autoantibodies (IgG or IgM) in the bone marrow supernatant of some IRP and ICUS patients were detected by the bone marrow indirect-Coombs test,and was correlated with the prognosis of IRP; Simultaneously,through this experiment, we can screen out some ture IRP patients with negative BMNC-Ab from ICUS, and give right treatment, increase the cure rate... |
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