The Effect Of HBV In The Formation Of Neutrophil Extracellar Traps(NETs)

As important immunological cells, neutrophils are necessary parts of host first defence line agaist pathogens. In 2004, neutrophil extracellar traps, thereafter use NETs for short, was reported for the first time by Brinkman. NETs, an special structure of activated neutrophils makes great contribution to the neutrophil defense. Morphology analysis revealed that NETs are mainly composed of reticular fibers and some attachment proteins. Then the special fluorometric analysis was used in order to have the deeper insight. It was confirmed that NETs was composed of double-strands DNA(ds DNA) and many adhesive compositions, such as myeloperoxidase(MPO)、histone、eutrophil elastase(NE). In fact, a lot of explorations have found that many biological and chemical factors can induce NETs formation, pathogens 、proinflammatory factor(IL- 8, TNF-α)、activated platelets、endothelial cells、phorbolmyristate acetate(PMA), and so on. Neutrophils were able to grape and clear pathogenic microorganisms through NETs, protecting human bodies from infection effectively. Chronic hepatitis B(CHB) is an common infection disease caused by HBV persistence, while a lot of CHB patients has high reproduction rate in cells. The immune system can’t remove viruses effectively is the main mechanism. Recently, another research reveals that HBV can infect neutrophils inducing infections beside liver, restrain the migration of neutrophils, and also neutrophils have changes in construction and function in CHB patients, all may influence the anti-infection function of neutrophils followed. All implied that HBV may have influence on neutrophils even on innate immunity, and also NETs maybe the central hub between them.Objective Neutrophil extracellar traps(NETs) is an important extracellar structure of activated neutrophils. Many explorations were played with circulating blood either HBV positive or HBV negative.1. Doing a comparison of the ability of two groups both HBV positive and HBV negative circulating blood neutrophils to release NETs, we are ready to elucidate the relationship between hepatitis B virus(HBV) infection and neutrophil NETs formation;2. Correlational analysis was done between serological markers of HBV and neutrophil NETs formation, then the judgment of HBV viral components to neutrophil NETs formation was needed;3. Correlational analysis was done DNA value of HBV and neutrophil NETs formation, then the judgment of HBV DNA value to neutrophil NETs formation was requirement;Method Fifty fresh human circulating blood samples with or without HBV infection were collected in Shandong Provincial hospital, those of anti-coagulant EDTA-K2,respectively.1. Five serological markers of HBV were measured by Chemiluminescence Microparticle Immuno Assay(CMIA);2. The DNA value of HBV were measured by PCR-fluorescent Probing method quantitatively;3. Neutrophils were isolated from fresh human circulating blood by density gradient centrifugation;4. As the primary part of NETs, released DNA was measured by commercial picogreen ds DNA kit quantitatively and the formation ability of NETs was presented by NETs ratio(NETs-DNA of stimulated neutrophils to NETs-DNA of natural neutrophils).All the indexes collected through tests were analyzed with proper statistical methods.Following the NETs ratio was calculated as said before, the NETs ratio is NETs of activated neutrophils to NETs of resting neutrophils.1. The neutrophil NETs ratio mean of HBV negative samples were higher than those of HBV positive, whose difference was statistically significant(P<0.05);2. By bivariate correlation analysis, the correlation of HBs Ag 、 HBe Ag 、 HBc Ab with neutrophil NETs release was negative(r<0,p<0.05), while the correlation of HBs Ab with neutrophil NETs ratio was positive, whose difference were all meaningful using statistical significance standard(r>0, p<0.05). However, the correlation of HBe Ab with neutrophil NETs ratio was of no significance(p>0.05);3. The correlation of HBV DNA value with neutrophil NETs ratio was of no significance(p>0.05);ResultsConclusion Stimulated with PMA, neutrophils improved NETs formation compared with resting one.1. HBV can inhibit NETs formation of neutrophils using HBV negative as control;2. HBs、HBe and HBc of HBV may take part in the negative regulation of NETs formation;...