Low toxicity and high immunogenicity of an inactivated vaccine candidate against COVID-19 in different animal models.

The ongoing COVID-19 pandemic is causing huge impact on health, life, and global economy, which is characterized by rapid spreading of SARS-CoV-2, high number of confirmed cases and a fatality/case rate worldwide reported by WHO. The most effective intervention measure will be to develop safe and effective vaccines to protect the population from the disease and limit the spread of the virus. An inactivated, whole virus vaccine candidate of SARS-CoV-2 has been developed by Wuhan Institute of Biological Products and Wuhan Institute of Virology. The low toxicity, immunogenicity, and immune persistence were investigated in preclinical studies using seven different species of animals. The results showed that the vaccine candidate was well tolerated and stimulated high levels of specific IgG and neutralizing antibodies. Low or no toxicity in three species of animals was also demonstrated in preclinical study of the vaccine candidate. Biochemical analysis of structural proteins and purity analysis were performed. The inactivated, whole virion vaccine was characterized with safe double-inactivation, no use of DNases and high purity. Dosages, boosting times, adjuvants, and immunization schedules were shown to be important for stimulating a strong humoral immune response in animals tested. Preliminary observation in ongoing phase I and II clinical trials of the vaccine candidate in Wuzhi County, Henan Province, showed that the vaccine is well tolerant. The results were characterized by very low proportion and low degree of side effects, high levels of neutralizing antibodies, and seroconversion. These results consistent with the results obtained from preclinical data on the safety.

Biochemical and antigenic characterization of the structural proteins and their post-translational modifications in purified SARS-CoV-2 virions of an inactivated vaccine candidate.

In the face of COVID-19 pandemic caused by the newly emerged SARS-CoV-2, an inactivated, Vero cell-based, whole virion vaccine candidate has been developed and entered into phase III clinical trials within six months. Biochemical and immunogenic characterization of structural proteins and their post-translational modifications in virions, the end-products of the vaccine candidate, would be essential for the quality control and process development of vaccine products and for studying the immunogenicity and pathogenesis of SARS-CoV-2. By using a panel of rabbit antisera against virions and five structural proteins together with a convalescent serum, the spike (S) glycoprotein was shown to be N-linked glycosylated, PNGase F-sensitive, endoglycosidase H-resistant and cleaved by Furin-like proteases into S1 and S2 subunits. The full-length S and S1/S2 subunits could form homodimers/trimers. The membrane (M) protein was partially N-linked glycosylated; the accessory protein 3a existed in three different forms, indicative of cleavage and dimerization. Furthermore, analysis of the antigenicity of these proteins and their post-translationally modified forms demonstrated that S protein induced the strongest antibody response in both convalescent and immunized animal sera. Interestingly, immunization with the inactivated vaccine did not elicit antibody response against the S2 subunit, whereas strong antibody response against both S1 and S2 subunits was detected in the convalescent serum. Moreover, vaccination stimulated stronger antibody response against S multimers than did the natural infection. This study revealed that the native S glycoprotein stimulated neutralizing antibodies, while bacterially-expressed S fragments did not. The study on S modifications would facilitate design of S-based anti-SARS-CoV-2 vaccines.

[Community structure and leaf trait diversity in a vulnerable species, Phoebe chekiangensis (Lauraceae).] / æ¸å±æ¤ç‰©æµ™æ±Ÿæ¥ ç¾¤è½ç»“æž„åŠå¶ç‰‡æ€§çŠ¶å¤šæ ·æ€§.

Phoebe chekiangensis, as one original species of 'Nanmu with golden tint', harbors limited natural distribution regions, which is recognized as a vulnerable species in China under nationwide protection. Under the background of global climate change, it is of scientific significance to study the community structure and diversity of the natural populations of P. chekiangensis. In this study, community structure, species diversity and the phenotypic variation leaf traits in the communities of thirteen residue natural populations were analyzed. A total of 235 vascular species from 162 genera and 87 families were found in 16 plots within 13 natural populations, including 221 seed plants from 151 genera and 79 families. The species diversity in tree layer of community in Kaihua, Zhejiang and Linan, Zhejiang was significantly lower than that of other communities due to severe disturbance, while intermediate disturbance promoted the species diversity of shrub layer in community in Jianning, Fujian, and slight disturbance was beneficial to natural regeneration of P. chekiangensis. Foliage phenotype was important for species identification within Phoebe genus. There existed substantial variation in foliage phenotypic diversity among and within populations. The mean coefficient of variation was 17.2%, ranging from 10.4% to 27.5%. The variation was greater among populations (53.6%) than within populations (17.0%). Mean phenotypic differentiation coefficient was 75.1% among populations, indicating that the variance among populations was the main source for the phenotypic variation of the species. Results from the cluster analysis indicated that the 13 natural populations were divided into two distinct groups based on the Euclidean distance (10 cm), with stochastic variation.

Halobaculum roseum sp. nov., isolated from underground salt deposits.

Two extremely halophilic archaea, strains D90T and D93, were isolated from underground salt deposits of Yunnan salt mine, China. Both strains were pleomorphic or short rods, non-motile, Gram-negative and required 1.7 M NaCl for growth, with optimum at 3.4 M. Mg2+ was not required for growth. Multiple copies of the 16S rRNA gene were obtained for both strains. Sequence similarity analysis of 16S rRNA genes revealed that strains D90T and D93 were closely related to Halobaculum magnesiiphilum MGY-184T and Halobaculum gomorrense DSM 9297T with the sequence similarity between 96.2-98.1 %. The sequence similarity of the rpoB' gene between strain D90T and Halobaculum gomorrenseJCM 9908T was 94.1 %. The DNA G+C contents of strains D90T and D93 were 65.9 and 67.6 mol%, respectively. The major polar lipids of both strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and glycolipid. The DNA-DNA relatedness value between strains D90T and D93 was 90.1±0.5 %, while that between strain D90T and Halobaculum gomorrenseJCM 9908T was 30.0±0.7 %. The comparison of physiological and biochemical characteristics, including the requirements of NaCl, Mg2+, pH, etc., differentiated strains D90T and D93 from Halobaculum magnesiiphilum MGY-184T and Halobaculum gomorrenseJCM 9908T. Therefore, strains D90T and D93 represent a novel species of the genus Halobaculum, for which the name Halobaculum roseum sp. nov. is proposed. The type strain is D90T (=CGMCC 1.15501T=JCM 31273T).