Safety,immunogenicity, and protection provided by unadjuvanted and adjuvanted formulations of a recombinant plant-derived virus-like particle vaccine candidate for COVID-19 in nonhuman primates

Although antivirals are important tools to control severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, effective vaccines are essential to control the current coronavirus disease 2019 (COVID-19) pandemic. Plant-derived virus-like particle (VLP) vaccine candidates have previously demonstrated immunogenicity and efficacy against influenza. Here, we report the immunogenicity and protection induced in rhesus macaques by intramuscular injections of a VLP bearing a SARS-CoV-2 spike protein (CoVLP) vaccine candidate formulated with or without Adjuvant System 03 (AS03) or cytidine-phospho-guanosine (CpG) 1018. Although a single dose of the unadjuvanted CoVLP vaccine candidate stimulated humoral and cell-mediated immune responses, booster immunization (at 28 days after priming) and adjuvant administration significantly improved both responses, with higher immunogenicity and protection provided by the AS03-adjuvanted CoVLP. Fifteen micrograms of CoVLP adjuvanted with AS03 induced a polyfunctional interleukin-2 (IL-2)-driven response and IL-4 expression in CD4 T cells. Animals were challenged by multiple routes (i.e., intratracheal, intranasal, and ocular) with a total viral dose of 10⁶ plaque-forming units of SARS-CoV-2. Lower viral replication in nasal swabs and bronchoalveolar lavage fluid (BALF) as well as fewer SARS-CoV-2-infected cells and immune cell infiltrates in the lungs concomitant with reduced levels of proinflammatory cytokines and chemotactic factors in the BALF were observed in animals immunized with the CoVLP adjuvanted with AS03. No clinical, pathologic, or virologic evidence of vaccine-associated enhanced disease was observed in vaccinated animals. The CoVLP adjuvanted with AS03 was therefore selected for vaccine development and clinical trials.     

LncRNA FGF14‐AS2 represses growth of prostate carcinoma cells via modulating miR‐96‐5p/AJAP1 axis

ObjectiveThis investigation devoted to lncRNA FGF14 antisense RNA 2 (FGF14‐AS2) in prostate carcinoma progression.MethodsThe levels of lncRNA FGF14‐AS2, miR‐96‐5p, and Adherens junction‐associated protein‐1 (AJAP1) in prostate carcinoma were tested by Western blot and qRT‐PCR. How these two genes interacted was confirmed by RNA immunoprecipitation and dualluciferase gene methods. The effect of FGF14‐AS2/miR‐96‐5p/AJAP1 axis in prostate carcinoma progression was determined by MTT, Transwell, and nude mice tumor model.ResultsFGF14‐AS2 was a downregulated lncRNA in prostate carcinoma tissue and cells. FGF14‐AS2 could restrain miR‐96‐5p expression while miR‐96‐5p hampered AJAP1. FGF14‐AS2 could effectively decrease the biological behaviors of prostate carcinoma cells, while knock‐down of FGF14‐AS2 triggered opposite results. Moreover, miR‐96‐5p mimic presented a cancer promoter role in prostate carcinoma cells. AJAP1 expression level could affect levels of proteins related to epithelial‐mesenchymal transition. In vivo experiment suggested that overexpressing FGF14‐AS2 could reverse the promotion of silenced AJAP1 on prostate carcinoma cell metastasis, thus to inhibit tumor growth.ConclusionlncRNA FGF14‐AS2 was a downregulated lncRNA in prostate carcinoma and influenced cell proliferation and metastasis. The influence relied on modulating miR‐96‐5p and its target gene AJAP1.     

The protective effects of Acanthopanax senticosus Harms aqueous extracts against oxidative stress: Role of Nrf2 and antioxidant enzymes

Ethnopharmacological relevance

Acanthopanax senticosus (Rupr.et Maxim.) Harms, classified into the family of Araliaceae, is used in a variety of diseases in traditional Chinese system of medicine including hypertension, ischemic heart disease and hepatitis.

Materials and methods

Different doses (75 mg/kg, 150 mg/kg and 300 mg/kg) of aqueous extracts of Acanthopanax senticosus Harms were evaluated for the antioxidant activity against oxidative stress in mice induced by tert-butyl hydroperoxide (t-BHP) through observating histopathology of the liver and detecting antioxidant enzyme activity, concentration of antioxidant, and related gene and protein expression.

Results

Acanthopanax senticosus Harms aqueous extracts (ASE) attenuated the morphological injury of liver induced by t-BHP and increased the activity of antioxidant enzymes and the ratio of GSH/GSSG in serum and liver homogenates. Medium and high doses of ASE also elevated the gene expression of NF-E2-related factor-2 (Nrf2), but not CuZnSOD, MnSOD, catalase (CAT), glutathione peroxidase (GPx) and GCLC. Protein expression results showed that Nrf2 and the antioxidant enzymes were all increased significantly by medium and high doses of ASE.

Conclusion

The present results indicated that ASE protect against oxidative stress which may be generated via the induction of Nrf2 and related antioxidant enzymes.     

Lead and mercury mutagenesis: Role of H2O2, superoxide dismutase,and xanthine oxidase

It has been suggested that reactive oxygen intermediates (ROIs) may have a role in the genotoxic effects of lead (Pb²⁺) and mercury (Hg²⁺), but there have not been any definitive studies demonstrating a causal relationship between the induction of ROIs by these metals and mutagenesis. We previously demonstrated, using the transgenic Chinese hamster ovary cell line AS52, that low concentrations (0.1–1 μM) of Pb²⁺ and Hg²⁺ are mutagenic. In the present study, using a novel histochemical computer-enhanced image analysis technique, we demonstrate that Pb²⁺ and Hg²⁺ induce the formation of H₂O₂ in AS52 cells by at least two distinct mechanisms. One is characterized by the rapid induction of H₂O₂ following treatment of cells with concentrations of Pb²⁺ or Hg²⁺ below 0.8 and 1 μM, respectively, while the second occurs in AS52 cells treated with concentrations of Pb²⁺ or Hg²⁺ greater than 0.8 and 1 μM, respectively. Pb²⁺ and Hg²⁺ (0.1–1 μM) had no effect on the activities of partially purified catalase, glutathione peroxidase, or glutathione reductase, important enzymes involved with antioxidant defense, but these metals stimulated the activities of copper-zinc superoxide dismutase (CuZn-SOD) and xanthine oxidase (XO). Allopurinol (50 μM), a specific inhibitor of xanthine oxidase, inhibited the induction of H₂O₂ by Pb²⁺ (0.8–1 μM) and Hg²⁺ (1 μM) and also inhibited Pb²⁺- and Hg²⁺-induced mutagenesis. These results demonstrate that Pb²⁺ and Hg²⁺ disrupt the redox status of AS52 cells by enhancing the activities of CuZn-SOD and XO. Furthermore, the results of these studies also demonstrate that there is a causal relationship between the induction of H₂O₂ by these metals and mutagenesis. Environ. Mol. Mutagen. 31:352–361, 1998. © 1998 Wiley-Liss, Inc.     

Anti-nociceptive effects induced by intrathecal injection of BmK AS, a polypeptide from the venom of Chinese-scorpion Buthus martensi Karsch, in rat formalin test

AIM OF THE STUDY: Asian scorpion Buthus martensi Karsch (BmK) is widely used to treat neurological symptoms, especially chronic pain, in traditional Chinese medicine for thousands of years. BmK AS, a polypeptide from BmK venom, could produce peripheral potent anti-nociceptive effects in rats. In the present study, spinal anti-nociceptive effects of BmK AS were investigated in rat formalin test. MATERIALS AND METHODS: Spinal anti-nociceptive activity of BmK AS was studied using formalin test in rats. BmK AS in doses of 0.02, 0.1 and 0.5 microg was administered intrathecally before formalin injection 10 min. The suppression by intrathecal injection of BmK AS on formalin-induced spontaneous nociceptive behaviors and spinal c-Fos expression were investigated. RESULTS: Intrathecal injection of BmK AS markedly reduced formalin-evoked biphasic spontaneous nociceptive behaviors in a dose-dependent manner. Formalin-induced c-Fos expression could be dose-dependently inhibited by BmK AS in superficial (I-II), the nucleus proprius (III and IV) and deep (V-VI) dorsal horn laminae, but not in the ventral gray laminae (VII-X) of lumbar spinal cord. The suppression by BmK AS on c-Fos expression in superficial laminaes was much stronger than that in deep laminaes. CONCLUSION: The present study demonstrates that BmK AS is capable of producing remarkable anti-nociceptive effects not only in periphery but also in spinal cord.     

强直性脊柱炎患者HLA-B27和免疫指标检测的临床意义

目的:探讨HLA-B27和免疫指标在强直性脊柱炎(AS)患者的临床意义.方法:采用流式细胞术检测147例AS患者、95例正常对照者的HLA-B27、CD3 T、CD4 T、CD8 T、CD19 B,用免疫比浊法检测IgA、IgG和IgM.结果:AS患者HLA-B27、CD3 T、CD4 T淋巴细胞的百分率及CD4 /CD8 的比值、血清IgA、IgG均明显高于正常对照组(P＜0.01);CD8 T淋巴细胞的百分率低于正常对照组(P＜0.05);CD19 B淋巴细胞的百分率高于正常对照组(P＜0.05);IgM与正常对照组相比无显著性差异.结论:HLA-B27、CD3 T、CD4 T与AS密切相关,提示,HLA-B27与CD3 T、CD4 T、CD4 /CD8 的比值可作为AS辅助诊断指标.     

Modelling the relative cost-effectiveness of the RTS,S/AS01 malaria vaccine compared to investment in vector control or chemoprophylaxis

BackgroundThe World Health Organization has recommended a 4-dose schedule of the RTS,S/AS01 (RTS,S) vaccine for children in regions of moderate to high P. falciparum transmission. Faced with limited supply and finite resources, global funders and domestic malaria control programs will need to examine the relative cost-effectiveness of RTS,S and identify target areas for vaccine implementation relative to scale-up of existing interventions.MethodsUsing an individual-based mathematical model of P. falciparum, we modelled the cost-effectiveness of RTS,S across a range of settings in sub-Saharan Africa, incorporating various rainfall patterns, insecticide-treated net (ITN) use, treatment coverage, and parasite prevalence bands. We compare age-based and seasonal RTS,S administration to increasing ITN usage, switching to next generation ITNs in settings experiencing insecticide-resistance, and introduction of seasonal malaria chemoprevention (SMC) in areas of seasonal transmission.ResultsFor RTS,S to be the most cost-effective intervention option considered, the maximum cost per dose was less than $9.30 USD in 90.9% of scenarios. Nearly all (89.8%) values at or above $9.30 USD per dose were in settings with 60% established bed net use and / or with established SMC, and 76.3% were in the highest PfPR_2-10 band modelled (40%). Addition of RTS,S to strategies involving 60% ITN use, increased ITN usage or a switch to PBO nets, and SMC, if eligible, still led to significant marginal case reductions, with a median of 2,653 (IQR: 1,741 to 3,966) cases averted per 100,000 people annually, and 82,270 (IQR: 54,034 to 123,105) cases averted per 100,000 fully vaccinated children (receiving at least three doses).ConclusionsUse of RTS,S results in reductions in malaria cases and deaths even when layered upon existing interventions. When comparing relative cost-effectiveness, scale up of ITNs, introduction of SMC, and switching to new technology nets should be prioritized in eligible settings.     

Extending the range of Plasmodium falciparum transmission blocking antibodies

Recent work demonstrating that asymptomatic carriers of P. falciparum parasites make up a large part of the infectious reservoir highlights the need for an effective malaria vaccine. Given the historical challenges of vaccine development, multiple parasite stages have been targeted, including the sexual stages required for transmission. Using flow cytometry to efficiently screen for P. falciparum gamete/zygote surface reactivity, we identified 82 antibodies that bound live P. falciparum gametes/zygotes. Ten antibodies had significant transmission-reducing activity (TRA) in a standard membrane feeding assay and were subcloned along with 9 nonTRA antibodies as comparators. After subcloning, only eight of the monoclonals obtained have significant TRA. These eight TRA mAbs do not recognize epitopes present in any of the current recombinant transmission-blocking vaccine candidates, Pfs230D1M, Pfs48/45.6C, Pf47 D2 and rPfs25. One TRA mAb immunoprecipitates two surface antigens, Pfs47 and Pfs230, that are expressed by both gametocytes and gametes/zygotes. These two proteins have not previously been reported to associate and the recognition of both by a single TRA mAb suggests the Pfs47/Pfs230 complex is a new vaccine target. In total, Pfs230 was the dominant target antigen, with five of the eight TRA mAbs and 8 of 11 nonTRA gamete/zygote surface reactive mAbs interacting with Pfs230. Of the three remaining TRA mAbs, two recognized non-reduced, parasite-produced Pfs25 and one bound non-reduced, parasite-produced Pfs48/45. None of the TRA mAbs bound protein on an immunoblot of reduced gamete/zygote extract and two TRA mAbs were immunoblot negative, indicating none of the new TRA epitopes are linear. The identification of eight new TRA mAbs that bind epitopes not included in any of the constructs currently under advancement as transmission-blocking vaccine candidates may provide new targets worthy of further study.