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Objective To investigate the change of CD35 expression on neutrophils in the peripheral blood and the relationship between the change and disease activity in patient with myeloperoxidase antineutrophil cytoplasmic antibody-associated vasculitis (MPO-AAV).Methods Forty untreated patients with active MPO-AAV(patient group)and forty healthy volunteers (control group) were enrolled into this study,and Bermingham vasculitis activity score (BVAS) for every patient was recorded.Flow cytometry (FCM) was employed to detect the CD35 and MPO expression on the neurtrophil,and enzyme linked immunosorbent assay (ELISA) was taken to test the levels of autoantibody against MPO-Antineutrophil cytoplasmic antibody (MPO-ANCA),fragment a from the activated complement factor B (Ba) and MPO in peripheral blood from both group.All test results were compared between the 2 groups by t test,Non-parametric test,Spearman correlation analysis.In addition,the relations among the laboratory results and the relationship between BVAS and the laboratory results were analyzed respectively.Results Compared with the control group,the expression level,which was represented as mean flourscence indensity (MFI),of CD35 and neutrophil membrane MPO on peripheral blood neutrophils was significantly increased [(2 014±968) vs (1 454±511),t=3.024,P=0.002 and (709±244) vs (580±158),t=2.806,P＜0.01,respectively],and the MPO expression level in neutrophils was significantly lower [(1 525±1 033) vs (3 196±2 126),t=-4.468,P＜0.01].Ba and MPO levels in serum of the patient group was significantly higher than that in the control group [37.89(26.17,63.14) μg/L vs 27.99(18.64,46.52) μg/L,Z=-2.521,P=0.012 and 546.16(450.55,729.96) U/L vs 327.93(279.02,365.10) U/L,Z=7.121,P＜0.01,respectively].In patient group,the expression level of CD35 had a significant positive relationship with peripheral blood neutrophil count (r=0.573,P＜0.01),serum Ba (r=0.433,P=0.005) and BVAS (r=0.368,P=0.020),respectively,whereas,there was a negative correlation between the MPO expressed on the neutrophils and that in the neutrophils (r=-0.458,P=0.003),and a positive relationship between MPO-ANCA and BVAS (r=0.351,P=0.026).Conclusion There is significant increased expression of CD35 on the neutrophil of patient with MPO-AAV,which might protect the neutrophil from destruction by the activated complement alternative pathway,and more neutrophils consequently contribute to the MPO-AAV pathogenesis.Inhibition of CD35 expression might become one of the potential new pathways for the treatment of MPO-AAV.
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Objective To study complement receptor typeⅠ(CR1)activity of erythrocytes in pa-tients with SLE.Methods Using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP),CR1rosette(RBC-CR1R)and immuno-complex rosette(RBC-ICR)on red cell,the ery-throcyte complement receptor I type(ECR1)genomic density polymorphism(HH type,HL type,LL type)and erythrocyte CR1immune activity were determined in32patients with SLE and in48normal individuals.Results It was found that HH type rate of ECR1density polymorphism in patients with active SLE was significantly lower(10/16,62.5%)than that(13/16,81.3%)in patients with stable SLE.The level of CR1immune activity in HH type was significantly higher than that in HL,LL type of SLE,and significantly dif-ferent from that in48normal individuals(P
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Objective To evaluate of red blood cell as giving instruction in whole white blood cell immunological activity by new nature experimental system of hemaimmune reaction rood map. Methods Plasma 0.3ml were added to whole blood cells (including: red blood cell and white blood cells) or white blood cells 0.2ml, and incubated for 1h at 37℃. The content of IL-8 and IL-12 was determined by enzyme linked immunadsorbent assay (ELISA) method. The expression level of CD4, CD8, CD35 and CXCR4 on white blood cells was determined by method of Flow Cytometry. Results The content of IL-8 (5.96?4.26) and IL-12 (9.84?2.23) in whole blood and plasma nature group was significantly lower than that (13.59?3.69?B?pg~ -1 ?ml~ -1 ) and (15.09?9.86?B?pg~ -1 ?ml~ -1 ) in white blood cell and plasma isolation group (P
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Objective To study the changes in the expressions of CXCR4 of leucocytes as modulated by erythrocytes from cord blood. Methods 0.2ml suspension of whole blood cells (or 0.2ml suspension of leucocytes) was added to 0.3ml of auto plasma, and 0.2ml suspension of S180 (5?10~6/ml) was added as stimuli. The suspension was incubated at 37 ℃ for 1 hour. Normal saline instead of S180 was used as control The expression changes in CD35 on erythrocytes and CXCR4 on leucocytes were determined with flow cytometry assay. The data could be grouped under four heads: (1) cancer cells 0.2ml were added to whole blood cells 0.2ml and plasma 0.3ml; (2) NS 0.2ml were added to whole blood cells 0.2ml and plasma 0.3ml; (3) cancer cells 0.2ml were added to white blood cells 0.2ml and plasma 0.3ml; (4) NS 0.2ml were added to white blood cells and plasma 0.3ml. Results When activated by S180, the expression of CD35 on cord erythrocytes was decline not so low as that of adult′; while the expression of CXCR4 on cord leucocytes was increased not so high as that of adults′. When activated by S180, the expression of CD35 on adult erythrocytes was significantly decreased than that of group of not activating (40.21%?11.52% and 28.65%?8.08% respectively), while the expression of CXCR4 on adult erythrocytes was significantly increased (40.62%?17.89% and 64.18%?11.69% respectively, P
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Objective To investigate the role of CD35 on erythrocytes played in antigen activating immunological reaction of lymphocytes. Methods Suspensions of lymphocytes(1?10~6/ml)and erythrocytes (1?10~8/ml) were respectively separated from anticoaguted whole blood of healthy adults with the lymphocyte separation medium. Using inactivated ascites carcinoma cell S180(1?10~6/ml) as activation antigen and autologous plasma as reactive medium, the role of erythrocytes in regulating the anti-neoplasm immunological reaction of lymphocytes was appraised. The expression of CD25 on lymphocytes was detected and compared using flow cytometry assay with CD35 on erythrocytes blocked by monoclonal antibody or complements in plasma were destroyed by EDTA. Results The expression of CD25 on lymphocytes (18.22?4.27%) was significantly decreased when CD35 on erythrocytes was blocked (P