Biological determinants perpetuating the transmission dynamics of mosquito-borne flaviviruses.

Mosquito-borne flaviviruses present a major public health concern. Their transmission is sustained in a cycle between mosquitoes and vertebrate hosts. However, the dynamicity of the virus-mosquito-host triad has not been completely understood. Herein, we discussed determinants of viral, vertebrate host, and mosquito origins that ensure virus adaptability and transmission in the natural environment. In particular, we provided insights into how proteins and RNAs of flaviviruses, blood parameters and odours of humans, and gut microbiota, saliva, and hormones of mosquitoes coordinate with each other to perpetuate the virus transmission cycle. A better knowledge of mechanisms permitting flaviviruses dissemination in nature can provide opportunities for establishing new virus-controlling strategies and could guide future epidemic and pandemic preparedness.

Quantitative and qualitative study of enzyme-linked immunosorbent assay to detect IgG against Japanese encephalitis virus in swine sera.

This study was designed to investigate the application of indirect enzyme-linked immunoassay (ELISA) in detecting IgG against Japanese encephalitis virus in swine sera and the qualitative nature of this test. The attenuated strain SA14-14-2 of Japanese encephalitis virus (JEV) was inoculated into 9-day-old chicken embryos and virus was harvested, purified and suspended in 0.9% saline as JEV antigen. The control antigen was prepared by the same method as for the antigen. In the ELISA, the optimal concentrations of antigen coated and dilution factor were selected using chi2 test. Ninety-two swine sera negative to haemagglutination inhibition (HI) were tested by this assay and the positive threshold was determined. The results of this study indicate that indirect ELISA has high specificity, sensitivity and reproducability. Simultaneous testing of 74 serum samples from nine pig farms was carried out to compare the existing HI test and the indirect ELISA. The coincidence rate of the two assays was 85.1% (63/74) and no significant difference was observed between them (p > 0.05). This ELISA test can detect 46 swine serum samples qualitatively and the titre of eight swine serum samples through endpoint dilution quantitatively within one 96-well plate.

The development and application of the latex agglutination test to detect serum antibodies against Japanese encephalitis virus.

The attenuated SA14-14-2 strain of Japanese encephalitis virus (JEV) was cultured in BHK-21 cells. The viral supernatant was purified and concentrated with PEG (MW 20,000). A suitable concentration of JEV antigen was used to sensitize latex to prepare the latex antigen. The specificity, sensitivity and stability of the antigen were assessed. A latex agglutination test (LAT) was developed for rapidly detecting antibody against JEV infection. The LAT and haemagglutination inhibition (HI) assay were compared by simultaneously testing 35 porcine serum samples from five farms. Ninety per cent (20/23) of the samples were seropositive by both assays. No significant difference was found between the two methods (p > 0.05). Furthermore, when 1,613 porcine sera from 120 farms were tested by LAT, the number of positive sera was 652, while that of negative sera was 961, ranging from 20% to 50% positive throughout the year. These results indicate that LAT is an appropriate candidate method for epidemiological surveys for and diagnosis of Japanese encephalitis.

The co-administrating of recombinant porcine IL-2 could enhance protective immune responses to PRV inactivated vaccine in pigs.

Three candidate cytokines: recombinant porcine interleukin-2 (rpIL-2), rpIL-6 and the fusion protein rpIL6-IL2 were used as adjuvants in this study to investigate the enhanced immune responses to PRV inactivated vaccine (IAV) in pigs. In this natural host trial, we demonstrated that rpIL-2 showed potential adjuvant effects on PRV IAV, which was characterized not only in antigen-specific immune responses, but also in protection against PRV infection. The use of rpIL-2 resulted in significantly higher virus neutralizing (VN) antibody levels and CTL activities on PRV IAV vaccination. The increased PRV-specific secretion of pIL-4 and pIFN-gamma from PBMC of pigs also demonstrated the adjuvant effects of rpIL-2. In addition, the co-administration of the rpIL-2 also produced an improved protection to the viral challenge, demonstrated by significant reduction of the ratios of fever and viral excretion in nasal swabs. However, there was no additional effect of adjuvant induced enhancement of immune responses and protection against challenge with the use of rpIL-6 and rpIL6-IL2 in this study.