Combating coxsackievirus B infections.

Coxsackieviruses B (CVB) are small, non-enveloped, single-stranded RNA viruses belonging to the Enterovirus genus of the Picornaviridae family. They are common worldwide and cause a wide variety of human diseases ranging from those having relatively mild symptoms to severe acute and chronic pathologies such as cardiomyopathy and type 1 diabetes. The development of safe and effective strategies to combat these viruses remains a challenge. The present review outlines current approaches to control CVB infections and associated diseases. Various drugs targeting viral or host proteins involved in viral replication as well as vaccines have been developed and shown potential to prevent or combat CVB infections in vitro and in vivo in animal models. Repurposed drugs and alternative strategies targeting miRNAs or based on plant extracts and probiotics and their derivatives have also shown antiviral effects against CVB. In addition, clinical trials with vaccines and drugs are underway and offer hope for the prevention or treatment of CVB-induced diseases.

AIM2 Co-immunization with VP1 Is Associated with Increased Memory CD8 T Cells and Mounts Long Lasting Protection against Coxsackievirus B3 Challenge.

The recurrent Coxsackievirus B3 (CVB3) infection is the most important cause of intractable myocarditis which often leads to chronic myocarditis and even dilated cardiomyopathy. Therefore, enhanced DNA vaccines capable of memory CD8 T cells are essential for long-lasting immunological protection against CVB3 infection. In this study, absent in melanoma 2 (AIM2) was used as an adjuvant to enhance the induction of memory CD8 T cells elicited by VP1 (viral capsid protein 1) vaccine. Mice were intramuscularly injected with 50 µg AIM2 plasmid and equal amount of VP1 plasmid (pAIM2/pVP1) vaccine 4 times at 2 week-intervals. We observed that the protection of pAIM2/pVP1 vaccine against CVB3 challenge was evidenced by significantly improved cardiac function, reduced myocardial injuries, and increased survival rate when compared with immunization with pVP1. Co-immunization with pAIM2/pVP1 robustly augmented T lymphocytes proliferation and CVB3-specific cytotoxic T lymphocyte responses. Importantly, 16 weeks after the last immunization, pAIM2/pVP1 co-immunization significantly enhanced the expression of Bcl-6, SOCS3, and Sca-1 which are critical for memory CD8 T cells as compared with pVP1 immunization. Notably, CD8 T cells that are likely vaccine-induced memory T cells were responsible for the protective efficacy of pAIM2/pVP1 vaccine by abolition of a CD8 T cell immune response following a lethal dose of CVB3 infection. Our results indicate that AIM2-adjuvanted vaccine could be a potential and promising approach to promote a long-lasting protection against CVB3-induced myocarditis.

Optimized production and purification of Coxsackievirus B1 vaccine and its preclinical evaluation in a mouse model.

Coxsackie B viruses are among the most common enteroviruses, causing a wide range of diseases. Recent studies have also suggested that they may contribute to the development of type 1 diabetes. Vaccination would provide an effective way to prevent CVB infections, and the objective of this study was to develop an efficient vaccine production protocol for the generation of novel CVB vaccines. Various steps in the production of a formalin-inactivated Coxsackievirus B1 (CVB1) vaccine were optimized including the Multiplicity Of Infection (MOI) used for virus amplification, virus cultivation time, type of cell growth medium, virus purification method and formulation of the purified virus. Safety and immunogenicity of the formalin inactivated CVB1 vaccine was characterized in a mouse model. Two of the developed methods were found to be optimal for virus purification: the first employed PEG-precipitation followed by gelatin-chromatography and sucrose cushion pelleting (three-step protocol), yielding 19-fold increase in virus concentration (0.06µg/cm2) as compared to gold standard method. The second method utilized tandem sucrose pelleting without a PEG precipitation step, yielding 83-fold increase in virus concentration (0.24µg/cm2), but it was more labor-intensive and cannot be efficiently scaled up. Both protocols provide radically higher virus yields compared with traditional virus purification protocols involving PEG-precipitation and sucrose gradient ultracentrifugation. Formalin inactivation of CVB1 produced a vaccine that induced a strong, virus-neutralizing antibody response in vaccinated mice, which protected against challenge with CVB1 virus. Altogether, these results provide valuable information for the development of new enterovirus vaccines.

The heart-protective mechanism of Qishaowuwei formula on murine viral myocarditis induced by CVB3.

AIM OF STUDY: The heart-protective effect and mechanism of Qishaowuwei formula (QSW), a Traditional Chinese Medicine formula composed of Radix Astragali, Radix Paeoniae Rubra and Fructus Schisandrae was investigated on murine model of viral myocarditis (VMC) induced by Coxsackievirus B3 (CVB3). MATERIALS AND METHODS: Mice were randomly divided into infected control group, QSW high dose group, QSW medium dose group, QSW low dose group and Vitamin C plus Ribavirin treatment group. 50 mice were included in each group. The day of virus inoculation was defined as day 0 and the drug treatment continued once a day for 14 days. Mice were sacrificed on days 3, 7, 14, 21 postinoculation (p.i.). The histopathological changes of myocardium, CVB3 RNA copies in the myocardium, cardiomycytic apoptosis, the serum level of superoxide dismutase (SOD) and maleic dialdehyde (MDA) and the phenotype of T lymphocytes subsets in peripheral blood was analyzed. RESULTS: QSW treatment significantly increase the survival rate (p<0.05) in VMC model. Histopathology and flow cytometry inspection revealed low ratio of cardiomyocytes necrosis and apoptosis in QSW treated mice with dose dependent manner. The cardiomyocytic ultra-structure observed by transmission electron microscope also supported the above results. The ameliorated tissue damage was consistent with reduced CVB3 copy numbers detected by real-time PCR in the myocardium of QSW treated mice. The antioxidant effect of QSW was proved by elevated activity of SOD and reduced level of MDA in the serum. Furthermore, the disturbed balance of CD4+ and CD8+ subsets in peripheral blood was restored. CONCLUSION: These results demonstrated QSW had potent protective effect against CVB3-induced heart injury and this effect might be mediated by its inhibition on viral replication, antioxidant activity and immunoregulation mechanism.

Immune up-regulation and tumor apoptosis by androstene steroids.

beta Androstenes steroid up-regulates immunity to increase resistance against lethal infection and lethal radiation, and mediates a rapid recovery of hematopoietic precursor cells after radiation injury. beta Androstenetriol increases the levels of the TH(1) cytokines, IL-2, IL-3, IFN gamma and counteracts hydrocortisone mediated immune suppression. In contrast, 17 alpha androstenediol inhibits proliferation and mediates apoptosis in tumor cells of murine and human origin. Its epimer 17beta androstenediol does not. The antiproliferative functions of 17 alpha androstenediol are not dependent on either the estrogen or androgen receptors. Our findings show that beta androstenes and analogs protect the host from lethal infection by DNA or RNA viruses such as, herpesvirus type 2, coxsackievirus B4, influenza, and arthropod borne viruses. These androstenes also protected the host from lethal bacterial infections by Enterococcus faecalis, Pseudomonas aeruginosa, and Klebsiella pneumonia and from parasites infections, i.e. Cryptosporidium parvum, and malaria. In vivo, the level of potency follows the order: dehydroepiandrosterone<<

[Experimental study of preventive effect of xin-kang oral liquid on acute viral myocarditis in mice].

OBJECTIVE: To study the preventive effect of Xin-Kang oral liquid on acute Coxsackie B3 viral myocarditis in mice. METHODS: The mice were infected Coxsackie virus B3 (CVB3m) through abdominal cavity after they were fed with the Xin-Kang oral liquid for 2d, and the uses of the drug continued to the 20th day. Histopathological changes of the myocardial cells were observed under microscope and measured by morphometric quantification. RESULTS: The detection rate of pathological change of grade III of heart surface in Xin-Kang oral liquid groups were lower than in virus control group, the difference between Xin-Kang medium dose group and virus control group being specially significant (P < 0.01), and the detection rate of severe myocardial damage was much lower. The average area of myocardial damage and area of myocardial damage/area of the whole heart in Xin-Kang oral liquid groups were markedly lower than those in virus control at the 5th day and the 20th day. CONCLUSION: These results demonstrate that the preventive effect of Xin-Kang oral liquid on acute Coxsackie B3 viral myocarditis is similar to that of interferon.