1.Indirect immunofluorescence on human B lymphoma cell line Raji and promyelocytic line HL60 for detection of DNA-associated autoantibodies to cell membrane in systemic lupus erythematosus
Yue ZHAO ; Jinli RU ; Liyun ZHANG ; Jing LUO ; Zhiqin Lü ; Huaming ZHAO
Chinese Journal of Microbiology and Immunology 2011;31(4):361-365
Objective To compare the significance of DNA-associated autoantibodies to cell membrane(cmDNA)in systemic lupus erythematosus(SLE)detected with indirect immunofluorescence on human B lymphoma cell line Raji and pmmyelocytic line HL60.Methods Indirect immunofluorescence assay both on cell line Raji and HL60 was used to measure anti-cmDNA antibodies in sera of 306 SLE patients.192 patients with other rheumatic diseases and 50 healthy controls.Results Indirect immunofluorescence assay on cell line Raji was used to measure anti-cmDNA antibodies.72.5% SLE and 10.4% other rheumatic diseases were positive for anti-cmDNA,but negative in 50 blood donors(P<0.01).Indirect immunofluorescence assay on cell line HL60 was used to measure anti-cmDNA antibodies,76.1% SLE and 16.7% other rheumatic diseases were positive for anti-cmDNA,but negative in 50 blood donors(P<0.01).The sensitivity of anti-cmDNA were 72.5%and 76.1%,respectively.The specificity of anti-cmDNA was 91.7% and 86.8%,respectively.There was no significant difference in sensitivity and spocificity(P>0.05).The methods of culture,freeze and resuscitation on the two cells were similar.but cell line Raji was easier to resuscitate than cell line HL60.Observing with fluorescence microscope.we find that cmDNA was expressed on the both cells and the staining was stronger on cellline Raji than HL60.Conclusion Anti-cmDNA antibody has high positivity which is one of the most valuable marker in the diagnosis of SLE.We recommend to measure anti-cmDNA antibodies with indirect immunofluorescence assay on cell line Raji rather than HL60.
2.The detection of DNA-associated autoantibodies to cell membrane and its combination with other autoantibodies in the diagnosis of systemic lupus erythematosus
Hua WEI ; Zhiqin Lü ; Jinli RU ; Xiaofeng LI ; Ke XU ; Hongyan WEN
Chinese Journal of Rheumatology 2010;14(3):185-187,后插2
Objective To evaluate the diagnostic value of autoantibodies to cell membrane associated with DNA (mDNA) in systemic lupus erythematosus (SLE) and the combination with other autoantibodies in the diagnosis of SLE. Method The anti-mDNA antibody had the characteristic pattern of perip-heral membrane fluorescence on cultured HL60. The same serum samples were detected for other antibo-dies of SLE. Pearson's Chi-square test was used for statistical analysis. Results This pattern was observed in 145 of 205 serum samples of SLE patients , but in 5 of 55 the serum samples of rheumatoid arthritis , in 10 of 45 primary Sjogren syndrome's patients and in 4 of 35 PM/DM and absent in 50 blood donors. The sensitivity and specificity of anti-mDNA antibody to SLE was 70.7% and 86.7%. The sensitivity and specificity of combined anti-mDNA antibody and ANA was 94.6% and 76.7%. The sensitivity and specificity of combined anti-mDNA antibody and anti-dsDNA antibody was 76.8% and 95.5%. The sensitivity and specificity of combined anti-mDNA antibody and anti-Sm antibody was 79.6% and 100%. The sensitivity and specificity of combined anti-mDNA antibody and AnuA was 93.0% and 100%. Conclusion This novel rapid immunofluorescence method can be a useful diagnostic test for SLE patients. Due to its high sensitivity and specificity, it is better than other diagnostic tests such as anti-dsDNA antibody and anti-Sm antibody for the diagnosis of SLE.
3.Role of chloride channels on apoptosis of CNE-2Z cells induced by arsenic trioxide
Ruiling Lü ; Hong GAO ; Zhiqin DENG ; Haibo WANG ; Xiechou LIANG ; Qiuchan TAN ; Linyan ZHU ; Liwei WANG ; Lixin CHEN
Chinese Journal of Pathophysiology 2017;33(4):647-654
AIM: To investigate the role of chloride channels in the apoptosis of human poorly differentiated nasopharyngeal carcinoma CNE-2Z cells induced by arsenic trioxide (As2O3).METHODS: The apoptotic rates of CNE-2Z cells induced by As2O3 for 24 h or 48 h were monitored by flow cytometry.The technique of whole-cell patch clamp was used to record the currents activated by As2O3 in the CNE-2Z cells.The inhibition of As2O3-induced apoptosis by chloride channel blocker DIDS in the CNE-2Z cells was analyzed by flow cytometry.RESULTS: As2O3 at 5 μmol/L induced apoptosis of CNE-2Z cells in time-dependent manner.The currents with outward rectification were activated when the cells were exposed to 5 μmol/L As2O3.No obvious time-and voltage-dependent inactivation of the currents was observed.The reverse potential of the currents was close to the equilibrium potential for chloride.The activated currents were inhibited by the chloride channel blockers NPPB and DIDS.The 47% hypertonic solution inhibited the activated currents completely.Chloride channel blocker DIDS inhibited the apoptosis of CNE-2Z cells induced by As2O3.CONCLUSION: As2O3 activates volume-sensitive chloride channels, and chloride channels may play an important role in the apoptosis of CNE-2Z cells induced by As2O3.