Evaluation of an attenuated vaccine candidate based on the genotype C of bovine parainfluenza virus type 3 in albino guinea pigs

Bovine parainfluenza virus type 3（BPIV3） is considered as one of the most important respiratory tract pathogens of both young and adult cattle, and widespread among cattle in the world. BPIV3 was first reported in China in 2008 and four strains of BPIV3 were isolated from Shandong Province, known as genotype C（BPIV3c）. Pathogen investigations had shown that BPIV3 c infection was very common among cattle in China. To date, BPIV3 can be classified into genotypes A, B and C based on genetic and phylogenetic analysis. Serological survey also demonstrates that BPIV3 infection is widespread in China, however, there is still no available vaccine for BPIV3 prevention in China nowadays. In the present study, the BPIV3 c strain SD0835 was continuously passaged on Madin-Darby bovine kidney（MDBK） cells for hundreds of times, and the pathogenicity of passage 209 was reduced in guinea pigs. The passage 209 of BPIV3 c strain SD0835 was used as a live vaccine candidate to immunize the guinea pigs. The vaccination results revealed that two vaccinations could induce excellent serum neutralizing antibody responses as well as proliferation of T lymphocytes. The vaccinated guinea pigs were well protected against challenge with a low passage of BPIV3 c strain SD0835. Additionally, the percentages of CD4~＋ and CD8~＋ T cell subsets of animals in vaccinated group increased after immunization; T cell subsets on day 2 after challenge in both groups decreased, and the decline of CD4~＋ and CD8~＋ T cell subsets levels of four guinea pigs in vaccinated group was relatively moderate, comparing with that of the control group. These data support further testing of the attenuated virus as an effective candidate vaccine.

The antipyretic effects of Chai Ge fever relief oral liquid

Backgroud:To verify the antipyretic effect and mechanism of Chai Ge fever relief oral liquid,and to provide evidence for the clinical application of Chai Ge fever relief oral liquid.Methods:By designing three groups of experimental models of yeast-induced rat fever model,endotoxin-induced rabbit fever model,and para-influenza virus-induced rabbit fever model,the antipyretic effect and mechanism of Chai Ge fever relief oral liquid were studied by measuring body temperature before and after treatment and the expressions of protein kinase A(PKA),protein kinase C,arginine vasopressin(AVP),prostaglandin E2 and cyclic adenosine monophosphate(cAMP)in hypothalamus after administration.Results:In the yeast-induced fever model of rats,the body temperature and PKA expression levels in different dose groups of Chai Ge fever relief oral liquid were significantly different from those of the model group 1–3 h after treatment.In the rabbit fever model induced by endotoxin,different dosage groups of Chai Ge fever relief oral liquid have obvious antipyretic effect on 1–3 h after the model is established,and can significantly reduce the expression of AVP and cAMP in hypothalamus.In the para-influenza virus-induced rabbit fever model,each dose group of Chai Ge fever relief oral liquid has obvious antipyretic effect on 1–2 h after the model is established.Conclusion:Chai Ge fever relief oral liquid has good antipyretic effect on fever models induced by yeast,endotoxin and parainfluenza by regulating the expression of PKA,AVP,and cAMP.

Parainfluenza Virus Types 1, 2, and 3 in Pediatric Patients with Acute Respiratory Infections in Beijing During 2004 to 2012

Background： Although human parainfluenza virus （HPIV） has been determined as an important viral cause of acute respiratory infections （ARIs） in infants and young children, data on long-term investigation are still lacking to disclose the infection pattern of HPIV in China. Methods： Nasopharyngeal aspirates were collected from 25,773 hospitalized pediatric patients with ARIs from January 2004 through December 2012 for respiratory virus screen by direct immuno-fluorescence assay. Results： Out of these specimens, 1675 （6.50%, 1675/25,773） showed HPIV positive, including 261 （1.01%, 261/25,773） for HPIVI, 28 （0.11%, 28/25,773） for HPIV2, and 1388 （5.39%, 1388/25373） for HPIV3, 2 of the samples were positive for both HPIV1 and HPIV3, and 36 were co-detected with other viruses. The positive rates of HPIVs were higher in those younger than 3 years old. HPIV3 was detected from all age groups, predominantly from patients under 3 years of age, and the highest frequency was found in those 6 months to 1-year old （352/4077, 8.63%）. HPlV3 was the dominant type in each of the years detected between May and July. HPIV1 showed a peak in every odd year, mainly in August or September. HPIV was detected most frequently from patients with upper respiratory infection （12.49%, 157/1257）, followed by bronchitis （ 11.13%, 176/2479）, asthma （9.31%, 43/462）, bronchiolitis （5.91%, 150/2536）, pneumonia （6.06%, 1034/17,068）, and those with underlying diseases （1.0%, 15/1506）. HPIV3 is the dominant type in these six disease groups referred above, especially in the asthma group. Conclusions： HPIV is one of the important viral causes of ARIs in infants and young children in Beijing based on the data from the hospitalized children covering a 9-year term. HPIV3 is the predominant type in all these years and in most of the disease groups. HPIVs with different types show different seasonality.

Sumoylation of Human Parainfluenza Virus Type 3 Phosphoprotein Correlates with A Reduction in Viral Replication

Human parainfluenza virus type 3(HPIV3), a member of the Paramyxoviridae family, can cause lower respiratory disease in infants and young children. The phosphoprotein(P) of HPIV3 is an essential cofactor of the viral RNA-dependent RNA polymerase large protein(L). P connects nucleocapsid protein(N) with L to initiate genome transcription and replication.Sumoylation influences many important pathways of the target proteins, and many viral proteins are also themselves sumoylated. In this study, we found that the P of HPIV3 could be sumoylated, and mutation of K492 and K532 to arginine(PK492 R/K532 R) failed to be sumoylated within P, which enhances HPIV3 minigenome activity. Biochemical studies showed that PK492 R/K532 Rhad no effect on its interactions with N, formation of homo-tetramers and formation of inclusion bodies.Finally, we found that incorporation of K492 R/K532 R into a recombinant HPIV3(rHPIV3-PK492 R/K532 R) increased viral production in culture cells, suggesting that sumoylation attenuates functions of P and down-regulates viral replication.