1.The relationship between the changes of peripheral blood natural killer cells and HLA-Cw alleles in liver cirrhotic patients with chronic HBV infection
Haiying LI ; Xuemei JIANG ; Shijun CHEN ; Jingwen LIU ; Yong AN ; Qingang ZHANG ; Lixin ZHANG ; Shuchun GAO
Chinese Journal of Microbiology and Immunology 2008;28(7):643-646
Objective To investigate the relationship between the frequency of peripheral blood natural killer cells (NK) and HLA-Cw alleles in liver cirrhotic patients with chronic HBV infection and a-cute hepatitis B patients. Methods Thirty liver cirrhotic patients and 30 patients with acute hepatitis B were included in our study, and 41 healthy individuals were enrolled as controls. The numbers of circulating NK cells and activated NK ceils were analyzed by flow cytometry. HLA-Cw genotyping was conducted with polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO). Results The numbers of circu-lating NK cells and activated NK cells in liver cirrhotic patients were 13.22% ± 4.61% and 45.68% ± 14.64%, which was lower than that in healthy subjects (P < 0.05). The numbers of circulating NK cells and activated NK cells in acute hepatitis B patients were 22.62% ± 3.70% and 65.28%± 14.45%, which was higher than that in healthy subjects(P < 0. 05). There were statistically significant differences between the two groups(P < 0.01). The allele frequency of HLA-Cw * 15 in the patients with cirrhosis was signifi-cantly higher than that in the healthy (P < 0.05), and there was a significant negative correlation between the frequency of HLA-Cw * 15 and the numbers of activated NK cells in liver cirrhosis(r =4). 862, P < 0.05). No statistically significance was found between the group of acute hepatitis B and healthy subjects a- bout HLA-Cw(P > 0. 05). Conclusion The function of NK cells in liver cirrhotic patients is low, HLA-Cw * 15 gene may be one of the causes of effecting the antiviral function of NK ceils to induce the persistence of hepatitis B virus(HBV) infection.
2.Preparation of monoclonal antibodies against VP2 protein of Asia I type foot-and-mouth disease virus and establishment of a competitive ELISA for the detection of antibodies.
Min XIANG ; Keshan ZHANG ; Shun LU ; Lijun CAI ; Yong LUO ; Jianmin ZHANG ; Hua HE ; Qingang WANG ; Bin WU
Chinese Journal of Biotechnology 2008;24(9):1664-1669
Monoclonal antibodies against FMDV vp2 protein were prepared and a competitive ELISA based on the monoclonal antibodies and vp2 protein was established. Balb/c mice were immunized with Escherichia coli expressed fusion protein. The splenocytes from immunized mice were fused with myeloma cells SP2/0. The hybridism cells were screened by indirect ELISA and limited dilution method. Two hybndoma cell Iines secreting mAbs against Asia I type foot-and-mouth disease were obtained. The titer and relative affinity of mAbs were determined by ELISA. Specificity of mAbs was analyzed by Western blotting. The ELISA titers of the ascites induced by the two hybridism cells were above 100 x 2(9).A competitive ELISA for the use of FMDV antibody detection was established using E. coli expressed fusion protein as coating antigen and HRP-labled mAb as detecting antibody. Clinical tests showed the method had 89.0 percent agreement with UBI Kit to detection of FMDV antibodies and 86.5 percent agreement with LPB- ELISA kit (Ceditest kit) for detection of antibodies against Foot-and-Mouth Disease Virus respectively.
Animals
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Antibodies, Monoclonal
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biosynthesis
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immunology
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Antibodies, Viral
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blood
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Capsid Proteins
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immunology
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Enzyme-Linked Immunosorbent Assay
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methods
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Female
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Foot-and-Mouth Disease
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immunology
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virology
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Foot-and-Mouth Disease Virus
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immunology
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Mice
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Mice, Inbred BALB C
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Swine