A bioinformatic approach towards designing a human papillomavirus vaccine based on L1 capsid protein sequence of HPV45

The cost of HPV vaccination is relatively expensive in low- to middle-income countries, hindering the introduction of HPV vaccination in these areas, although infection cases are high. In this study we designed a vaccine candidate based on L1 protein from Human Papillomavirus Subtype 45 (HPV45). Explorations of L1 HPV45 sequences from NCBI and Uniprot databases generated a consensus sequence, which was then optimised to improve its antigenicity character, whilst retaining the same epitope sites as observed in the consensus. Characteristics of the designed molecule was assessed, to ascertain its potential immunogenicity and good physicochemical characters. The study showed no major difference between our designed protein and either the Indonesian L1 HPV45 sequence (GenBank: QRG45832.1) (apart from two amino acids, N379 and G383), or the consensus sequence (apart from three amino acids, N81, T329, and H392). These differences do not seem to affect the 3D-structural similarities of the proteins. The designed protein is a non-allergenic, 60 kDa protein, with pI 8.61. It is relatively thermostable, with aliphatic index 75.26. The GRAVY score suggested that the protein is soluble in water. Pichia was selected as the expression host, because, unlike in E. coli, the protein has longer half-life and do not form inclusion bodies in the yeast.

Comparison of Consensus Sequences and Ancestor Sequences of HIV-1 nef and vif Genes for Development of Korean-Specfic HIV-1 Vaccine

There have been attempts to use consensus sequences or ancestor sequences for development vaccines against viruses with high diversity and variation. In this study, we generated and compared consensus sequences and ancestor sequences of nef and vif genes of HIV-1 isolated from Koreans. Phylogenetic analyses revealed that majorities of the Korean isolates were clustered to form the Korean clade within subtype B (KcB) where foreign isolates were not included. Consensus sequences inferred from the KcB as well as from all Korean isolates were almost identical but significantly different from subtype B consensus sequence or HIV-1 consensus sequence. The genetic distances from one of the Korean isolates to the other Korean isolates were much longer than to the consensus or ancestor sequences deduced from Korean isolates but similar to those of subtype B or HIV-1. Moreover, the genetic distances from the Korean isolates to the consensus sequences were shorter than to the ancestor sequences both in nef and vif genes. Thus, the consensus sequences may be useful in developing Korean-specific HIV-1 vaccine.

Interferon consensus sequence binding protein: Not essential for interferon alpha-mediated antiviral response to vesicular-stomatitis virus infection in HL-60 cells

BACKGROUND: The role of the interferon consensus sequence binding protein (ICSBP), a member of interferon regulatory factor family, in protecting against a vesicular stomatitis virus (VSV) infection has not been firmly elucidated. Thus, it was investigated utilizing the human promyelocytic leukemia HL-60 cells which do not express ICSBP. METHODS: HL-60 cells were stably transfected with plasmid containing cDNA for either ICSBP or DNA binding domain (DBD) and tested for their VSV-susceptibilities. The susceptibility of each transfectant group to a VSV infection was determined by a plaque assay at 1 h, 24 h, and 48 h post-infection in the presence (500 IU/ml) or absence of interferon alpha(IFN alpha). RESULTS: In the absence of IFN alpha, the three groups showed similar sensitivities to a VSV infection. However, when pre-treated with IFN, the viral titers in both the ICSBP and control clones steadily decreased over 48 h of incubation, indicating the existence of IFN alpha-mediated protection against VSV infection. The IFN alpha-treated ICSBP clones appeared to be more resistant to infection compared with the control clones, although the difference was not great . On the contrary, the viral titers in the IFN alpha-treated DBD clones increased at 24 h then decreased by 48 h. CONCLUSION: The expression of truncated ICSBP (DBD) does not appear to underlie the impaired protection against a VSV infection in the DBD clones, since even the control clones lacking ICSBP were protected from a VSV infection. This suggests that ICSBP does not play a critical role in the IFN alpha-mediated anti-VSV response of HL-60 cells, although it appears to confer some resistance to a VSV infection.

Establishment of consensus sequence of PreS/S and Enhll/CP/PreC of hepatitis B virus prevailing in Chongqing,China / 重庆医科大学学报

Objective :To establish the consensus sequence of PreS/S and Enhll/CP/PreC in Chongqing,China. Methods :The genes of PreS/S and Enhll/CP/PreC of HBV in 15 AsC were sequenced. The genotype and serotype of HBV were determined. The HBV strain prevailing in Chongqing was found and its consensus sequence was established. Results: Nine strains were genotype B/serolype adw2, 3 were genotype B/serotype aywl and 3 were genotype B/serotype adrq+ . There were 6 and 1 site differences between the consensus sequences of PreS/S and Enhll/CP/PreC of HBV with genotype B/serotype adw2 and that in Southeast China. Conclu-sions:The consensus sequence of PreS/S and Enhll/CP/PreC of hepatitis B virus prevailing in Chongqing was established.