Efficient Humoral and Cellular Immune Responses Induced by a Chimeric Virus-like Particle Displaying the Epitope of EV71 without Adjuvant / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To eliminate the side effects of aluminum adjuvant and His-tag, we constructed chimeric VLPs displaying the epitope of EV71 (SP70) without His-tagged. Then evaluating whether the VLPs could efficiently evoke not only humoral but also cellular immune responses against EV71 without adjuvant.</p><p><b>METHODS</b>The fusion protein was constructed by inserting SP70 into the MIR of truncated HBcAg sequence, expressed in E. Coli, and purified through ion exchange chromatography and density gradient centrifugation. Mice were immunized with the VLPs and sera were collected afterwards. The specific antibody titers, IgG subtypes and neutralizing efficacy were detected by ELISA, neutralization assay, and EV71 lethal challenge. IFN-γ and IL-4 secreted by splenocytes were tested by ELISPOT assay.</p><p><b>RESULTS</b>HBc-SP70 proteins can self-assemble into empty VLPs. After immunization with HBc-SP70 VLPs, the detectable anti-EV71 antibodies were effective in neutralizing EV71 and protected newborn mice from EV71 lethal challenge. There was no significant difference for the immune efficacy whether the aluminum adjuvant was added or not. The specific IgG subtypes were mainly IgG1 and IgG2b and splenocytes from the mice immunized produced high levels of IFN-γ and IL-4.</p><p><b>CONCLUSION</b>The fusion proteins without His-tagged was expressed and purified as soluble chimeric HBc-SP70 VLPs without renaturation. In the absence of adjuvant, they were efficient to elicit high levels of Th1/Th2 mixed immune response as well as assisted by aluminum adjuvant. Furthermore, the chimeric VLPs have potential to prevent HBV and EV71 infection simultaneously.</p>

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.

Expression and synergistic function of ENHANCIN-like gene of Agrotis Segetum granulovirus / 病毒学报

ENHANCIN is an enhancing protein chiefly found in insect baculoviruses. One ENHANCIN homologue was identified, by blast method, in Agrotis Segetum granulovirus (AgseGV) genome, named enhancin-like. Sequence analysis indicated that this gene includes the conserved domains, conserved in other ENHANCIN, and it has no signal peptide or a-transmembrane helix. A proline-rich domain, which is similar to those of mammals, is present at its C-terminal. To analyze the synergistic function of AgseGV enhancin-like gene, prokaryotic expression vectors of its whole gene and the 5'-truncated fragment (1, 017bp) were constructed. Expression product of truncated fragment was purified by chromatography, and then it was used to prepare antibody. The expression product of whole gene was identified by Western blot with specific antibody and anti-His-Tag antibody. Bioassay proved that the expression product of whole gene can increase the mortality with 16.25% to 3th instar larvae of Helicoverpa armigera (HaNPV: 1.17 x 10(2) PIBS/mL), while the truncated fragment has no obvious synergistic function.

Iodine nutrition survey of children aged 8 to 10 in coastal and non-coastal areas of Rizhao city Shandong province / 中国地方病学杂志

Objective To investigate the iodine nutritional status of children aged 8 to 10 in both costal and non-costal areas of Rizhao city Shandong province,and provide a basis for scientific supplementation of iodine.Methods In 2009 and 2010,townships of Lanshan and Donggang were chosen as survey points which situated close to the coast,the counties of Wulian and Juxian that were more than 50 kilometers away from the coast were selected as controls.The iodine concentration of drinking water in every village and water supply point was determined.At the county level,5 to 9 towns were selected according to their sub-area positions of east,west,south,north and center,4 villages were selected in each chosen township,8 - 15 households were selected in each chosen village,edible salt from the households was collected; 5 primary schools were selected in each chosen township,60 students aged 8 - 10 were selected to take thyroid examination in each chosen school.Twenty copies of urine samples were collected from the 60 students to detect the iodine concentration.Iodine in drinking water was tested by cerous sulfate catalytic spectrophotometry,in iodized salt by direct titration,in urine by arsenic-cerium contact method,and thyroids were examined by palpation.Results Totally 3483 copies of drinking water samples,1164 copies of edible salt samples,and 476 copies of urine samples were tested,and a total of 1200 children aged 8 to 10 were investigated; and the ratio of water iodine frequency distribution ≤ 10 μg/L (in costal and non-costal area) accounted for 90.03％ ( 1011/1123 ) and 91.10％ (2150/2360),respectively,the medians of water iodine were 5.6 and 4.2 μg/L,respectively; the iodine medians of edible salt were 29.03 and 29.99 mg/kg,respectively; the consumption rates of qualified iodized salt were 96.77％ (569/588) and 97.05％ (559/576),respectively; total goiter rates were 1.17％(7/600) and 1.33％(8/600),respectively; the medians of urinary iodine were 144.05 and 159.15 μg/L,respectively; the percentages of urinary iodine that bellow 100 μg/L were 26.58％ (63/237) and 22.59％ (54/239),respectively; between 100 - 300 μg/L were 66.75％ (158/237) and 64.02％ (153/239),respectively;above 300μg/L were 6.75％ (16/237)and13.39％ (32/239),respectively.ConclusionsThe costal of Rizhao is an iodine deficient area.The current popularity of household's consumption of qualified iodized salt is suitable.The levels of iodine nutrition of children aged 8 to 10 is appropriate,which met the national standard of elimination of iodine deficiency disorders (IDD).There is no iodine excess.There are no significant differences in levels of iodine nutrition between costal and non-costal areas on the premise that the same iodized salt is supplied in both the areas,and there is no need to make differentiated supply of iodized salt.