Blood donors with HBV core anti-body: a follow-up study / 中国输血杂志

【Objective】 To evaluate the role of anti-HBc detection in current blood screening strategy by the follow-up of repeated donors with antibody to hepatitis B virus core antigen. 【Methods】 Plasma samples were collected randomly from Dalian Blood Center. to test anti-HBc(dual reagents) and anti-HBs via ELISA. The re-donation of eligible donors who were anti-HBc+ and donors reactive to HBV detection were followed up. 【Results】 A total of 1 291 plasma samples were collected randomly from May 2017 to March 2018, among which 405 samples(31.4%)were anti-HBc+. The median age of anti-HBc+ group was observed much higher than that of anti-HBc-group (39 vs 31 years old) (P0.05). Among the 405 anti-HBc+ donors, 3 donors were OBI (0.7%), of which one was screened out in second donation. No HBV DNA was detected out in 3 OBI cases. 【Conclusion】 Although anti-HBc detection is not suitable in blood screening currently, it is of great value in the assessment of blood donor re-entry for HBV reactive donors in blood screening due to the high anti-HBc prevalence among blood donors.

Prevalence of Hepatitis B Virus (HBV) Among Blood Donors in Eastern Saudi Arabia: Results From a Five-Year Retrospective Study of HBV Seromarkers

BACKGROUND: Transfusion-transmissible hepatitis B virus (HBV) infection is a major problem worldwide. Recently, confirmatory nucleic acid tests (NATs) for HBV DNA have been employed in several countries. We assessed the prevalence and yearly trends of HBV infection in blood donors in the Eastern Province of Saudi Arabia, screening for HBV surface antigen (HBsAg), antibody against HBV core antigen (anti-HBc), and HBV DNA. METHODS: Between 2011 and 2015, a total of 22,842 donors were screenedfor HBsAg, anti-HBc, and HBV DNA using the HBsAg Qualitative II kit (Abbott, Ireland Diagnostics Division, Sligo, Ireland), ARCHITECT Anti-hepatitis B core antigen antibody (HBc) II Assay kit (Abbott GmbH & Co. KG, Wiesbaden, Germany), and NAT Procleix Ultrio Elite Assay kit (Grifols Diagnostic Solutions Inc., Los Angeles, CA, USA), respectively. RESULTS: A total of 739 (3.24%) donors were HbsAg(+), anti-HBc(+), or HBV DNA(+); 63 (0.28%) were HbsAg(+), anti-HBc(+), and HBV DNA(+). Twelve (0.05%) were anti-HBc(+) and HBV DNA(+) but HBsAg(−); they were considered to have occult infection. Further, 664 (2.91%) were HBsAg(−) but anti-HBc(+), indicating chronic or resolving infection. HBV prevalence increased significantly from 2011 to 2012, increased marginally till 2013, and showed a decreasing trend from 2013 (P>0.05). CONCLUSIONS: The five-year prevalence of HBV infection among blood donors in the Eastern Province of Saudi Arabia (3.24%) is lower than that reported for other regions in the country. The occult HBV infection rate of 0.05% emphasizes the importance of NATs in isolating potential infectious blood units.

Expression and purification of the chimeric virus-like particles displaying enterovirus 71 epitopes / 中华实验和临床病毒学杂志

Objective@#To study the expression and purification of the chimeric virus-like particles displaying epitopes of EV71 as a candidate of enterovirus 71 gene recombined vaccinet.@*Methods@#The fusion protein hepatitis B core (HBc)-SP70 was constructed by inserting SP70 into the main immunogenic region of truncated hepatitis B core antigen (HBcAg) sequence, expressed in E. Coli, and purified through sonication, ion exchange chromatography, CsCl cushion centrifugation and density gradient centrifugation. Then its antigenicity was detected by ELISA and Western blot assay.@*Results@#Recombinant plasmid pHBc-SP70 was successfully constructed. And the soluble fusion protein was efficiently expressed induced by IPTG. The purity of the chimeric virus-like particles (VLPs) was up to 90% after the purification process described in method . The purified fusion protein HBc-SP70 could be spontaneously folded and assembled into empty virus-like particles and react with the monoclonal antibodies against HBc and SP70.@*Conclusions@#The chimeric VLPs displaying epitopes of EV71 were efficiently expressed and purified in E. Coli. with excellent antigenicity, which laid a foundation for evaluation of the immune effect evoked by this EV71 gene recombined vaccine.

Study of a virus-like particle vaccine containing N-terminal epitopes of human papilloma virus L2 protein / 中国免疫学杂志

Objective:To prepare a virus-like particle (VLP),containing Hepatitis B virus core antigen (HBcAg) and N-terminal peptides of the L2 protein of human papilloma virus (HPV),and investigate the immunogenicity of the VLP in mice and the protection against different strains of HPV .Methods:A fusion gene was synthesized to insert a DNA fragment ,coding for the N-terminal epitopes of the L2 protein of HPV16,into the HBcAg coding sequence;HBc-L2 fusion protein was highly expressed in E.coli using the pET9a and BL21(DE3) expression system;the purified fusion protein was used to immunize BALB/c mice and antibody titers against the L2 epitopes in mouse sera were determined by indirect ELISA;the levels of neutralizing antibodies against both HPV 16 and 18 were also analyzed.Results:HBc-L2 fusion protein was expressed in E.coli and purified,with the purity >80%,by ammonium sulfate pre-cipitation and CL-4B gel filtration;analysis of the purified fusion protein ,using size exclusion chromatography with multi-angle laser light scattering detection ( SEC-MALS) and electron microscope ,revealed that HBc-L2 was assembled into a stable VLP structure auto-matically following its expression;immunization of BALB/c mice with the purified VLPs resulted in high antibody titers in mouse sera against the L2 epitopes;furthermore,it was demonstrated that the sera from the immunized mice had neutralization activities against both HPV16 and HPV18.Conclusion:The immunogenicity of the L2 epitopes was highly enhanced by the construction of HBc-L2 fusion protein and the formation of the VLP structure;the fusion protein was also capable of inducing protections against different serotypes of HPV,therefore,it could be a potential HPV vaccine with a broad coverage and low production cost .

Establishment of the Quantitative Immunohistochemical Examination of DHBcAg in Duck Liver / 现代检验医学杂志

Objective To develop a quantitative immunohistochemistry assay for duck hepatitis B virus core antigen (DHB-cAg)in duck liver tissue.Methods By comparison with no repair antigen and repair antigen with high pressure,microwave and trypsin,the best solution of antigen retrieval was determined.By optimizing the parameter of image acquisition and de-ducting blank area,mean density of yellow areas was calculated using Image-Pro Plus 6.0 software.Using the assay devel-oped to determine the level of DHBcAg in liver tissue from duck infected by DHBV,anti-DHBV activity of DHBcMAb-TAT PTD conj ugate was examined.Results SABC method with no repair antigen was selected,which was better than other methods.DHBcAg expression in duck liver tissue could be objectively and accurately quantified by setting Image-Pro Plus 6.0 software parameters and calculating mean density of yellow areas.By comparison with the differences between mean densityat baseline of treatment and end of treatment,it was showed that DHBcMAb-TATPTD conjugate treatment dose-de-pendently reduced the levels of DHBcAg in liver tissue,which show that the assay developed could effectively evaluate the anti-DHBV activity of agent.Conclusion The immunohistochemistry assay developed in this study can objectively and accu-rately evaluate the level of DHBcAg in duck liver tissue.

Levels of Interferon ? and Interleukin-10 in Sera and in Culture Supernatants of Peripheral Blood Mononuclear Cells from Children with Chronic Hepatitis B Virus Carriers and Their Clinical Significance / 实用儿科临床杂志

Objective To explore the levels of interferon ?(IFN-?) and interleukin-10(IL-10) in sera and in hepatitis B virus(HBV) core antigen(HBcAg) stimulated culture supernatants of peripheral blood mononuclear cells(PBMC) from chronic HBV carriers children and their clinical significance.Methods Five milliliter peripheral blood was collected from 21 HBeAg-positive children(group A),20 HBeAg-negative children(group B) and 19 normal controls(group C),sera and PBMC was isolated,after 72 h culture of PBMC with HBcAg or not.The levels of IFN-? and IL-10 in sera and in HBcAg stimulated or not stimulated culture supernatants of PBMC were detected.Results The level of sera IFN-? had no difference between each group;the sera level of IL-10 in group A and B were significantly higher than that of group C.The level of IFN-? in HBcAg stimulated culture supernatants of PBMC from group B and C were higher than that from group A;while IL-10 levels in HBcAg stimulated culture supernatants from group A and B were higher than that from group C.Conclusions The level of IL-10 and the lower reactivity to HBcAg of PBMC play roles in the mechanisms of chronic HBV carriers children.