Regulation of Murine Class I Genes by Interferons is Controlled by Regions Located Both 5' and 3' to the Transcription Initiation Site

Interferons regulate the expression of a large number of mammalian genes, including the major histocompatibility antigen genes. To investigate the mechanisms involved in interferon action, we have analyzed the ability of murine H-2Ld and H-2Dd DNA sequences to control the responses to interferon. The results indicate that interferon regulation of class I gene expression is complex and involves at least two mechanisms that are dependent on class I sequences located upstream and downstream to the transcription initiation site. In transfected mouse L cells, both of these regions are required for full enhancement of class I gene expression, with the major portion of the response controlled by the sequences located 3' to the transcription initiation site. The fine-mapping analysis of the 5' region-encoded response also suggests that recombinant alpha and gamma interferons may exert their effects on class I gene expression by using different cis-acting regulatory sequences

Improving the reliability of the jpHMM recombination prediction in HIV

Accurate classification of HIV and the identification of recombinants, including precise breakpoint definitions, is of crucial importance for epidemiological monitoring and the design of potential drugs. Recently we developed jpHMM, a new method to detect recombinations in HIV-l genomes. jpHMM predicts phylogenetic recombination breakpoints in a query sequence and assigns to each segment of the sequence one of the major HIV-l subtypes. For the user the reliability of the predicted breakpoint positions and parental subtypes is most important. For this reason we extended the output of jpHMM to include the information on regions where the model is 'uncertain' about the parental subtype and an interval estimate of the breakpoint. This information is determined using the posterior probabilities of the subtypes at each query sequence position

A jumping profile Hidden Markov Model and applications to recombination sites in HIV and HCV genomes

BACKGROUND: Jumping alignments have recently been proposed as a strategy to search a given multiple sequence alignment A against a database. Instead of comparing a database sequence S to the multiple alignment or profile as a whole, S is compared and aligned to individual sequences from A. Within this alignment, S can jump between different sequences from A, so different parts of S can be aligned to different sequences from the input multiple alignment. This approach is particularly useful for dealing with recombination events. RESULTS: We developed a jumping profile Hidden Markov Model (jpHMM), a probabilistic generalization of the jumping-alignment approach. Given a partition of the aligned input sequence family into known sequence subtypes, our model can jump between states corresponding to these different subtypes, depending on which subtype is locally most similar to a database sequence. Jumps between different subtypes are indicative of intersubtype recombinations. We applied our method to a large set of genome sequences from human immunodeficiency virus (HIV) and hepatitis C virus (HCV) as well as to simulated recombined genome sequences. CONCLUSION: Our results demonstrate that jumps in our jumping profile HMM often correspond to recombination breakpoints; our approach can therefore be used to detect recombinations in genomic sequences. The recombination breakpoints identified by jpHMM were found to be significantly more accurate than breakpoints defined by traditional methods based on comparing single representative sequences

CATNAP: a tool to compile, analyze and tally neutralizing antibody panels

CATNAP (Compile, Analyze and Tally NAb Panels) is a new web server at Los Alamos HIV Database, created to respond to the newest advances in HIV neutralizing antibody research. It is a comprehensive platform focusing on neutralizing antibody potencies in conjunction with viral sequences. CATNAP integrates neutralization and sequence data from published studies, and allows users to analyze that data for each HIV Envelope protein sequence position and each antibody. The tool has multiple data retrieval and analysis options. As input, the user can pick specific antibodies and viruses, choose a panel from a published study, or supply their own data. The output superimposes neutralization panel data, virus epidemiological data, and viral protein sequence alignments on one page, and provides further information and analyses. The user can highlight alignment positions, or select antibody contact residues and view position-specific information from the HIV databases. The tool calculates tallies of amino acids and N-linked glycosylation motifs, counts of antibody-sensitive and -resistant viruses in conjunction with each amino acid or N-glycosylation motif, and performs Fisher's exact test to detect potential positive or negative amino acid associations for the selected antibody. Website name: CATNAP (Compile, Analyze and Tally NAb Panels). Website address: http://hiv.lanl.gov/catnap

Development of Pan-filovirus vaccine against Ebola and Marburg virus challenges

Filoviruses such as Ebola (EBOV) and Marburg (MARV) viruses cause deadly viral hemorrhagic fever in humans with high case fatality rates. To date, no licenced therapeutic or vaccine has been clinically approved to prevent infection. Several vaccine candidates are under development against the few most common filoviruses targeting the virus glycoprotein (GP). However, protective antibodies induced by such GP vaccines are usually limited to the same species. In contrast, T-cell vaccines offer an opportunity to design a single pan-filovirus vaccine protecting against all members of the Filoviridae family. In this study FILOcepX vaccines were constructed targeting the four most conserved regions among the viral proteomes with the aim to induce protective T-cell responses against different filoviruses. BALB/c mice were immunized with FILOcep 1 and 2 vaccines vectored by non-replicating engineered simian adenovirus and poxvirus MVA. Groups of 20 BALB/c mice were primed and boosted with either the FILOcep1 and FILOcep2 vaccines or control ChAdOx1- and MVA-vectored vaccines. Four animals in each group were sacrificed after 1 week of boosting to detect T-cell response for the FILOcepX antigen. High frequency T cells specific responses were detected in mice receiving the test vaccines by IFN-γ ELISPOT kits. Of the remaining 16 animals in each group, 8 were challenged with mouse-adapted EBOV and 8 were challenged with mouse adapted MARV in Containment Level 4 laboratory. All the mice in the control group either died or had to be euthanized between 4 and 6 days post challenge. On the other hand all the FILOcepX vaccinated mice maintained their normal body mass and survived till the end of the scheduled protocol on day 29 post challenge. These FILOcepX vaccines provided 100% protection against the lethal challenges with filoviruses of two different genera. Further evaluation the efficacy of this vaccine in non-human primates (NHPs) is warranted

jpHMM at GOBICS: a web server to detect genomic recombinations in HIV-1

Detecting recombinations in the genome sequence of human immunodeficiency virus (HIV-1) is crucial for epidemiological studies and for vaccine development. Herein, we present a web server for subtyping and localization of phylogenetic breakpoints in HIV-1. Our software is based on a jumping profile Hidden Markov Model (jpHMM), a probabilistic generalization of the jumping-alignment approach proposed by Spang et al. The input data for our server is a partial or complete genome sequence from HIV-1; our tool assigns regions of the input sequence to known subtypes of HIV-1 and predicts phylogenetic breakpoints. jpHMM is available online at

Epigraph hemagglutinin vaccine induces broad cross-reactive immunity against swine H3 influenza virus

Influenza A virus infection in swine impacts the agricultural industry in addition to its zoonotic potential. Here, we utilize epigraph, a computational algorithm, to design a universal swine H3 influenza vaccine. The epigraph hemagglutinin proteins are delivered using an Adenovirus type 5 vector and are compared to a wild type hemagglutinin and the commercial inactivated vaccine, FluSure. In mice, epigraph vaccination leads to significant cross-reactive antibody and T-cell responses against a diverse panel of swH3 isolates. Epigraph vaccination also reduces weight loss and lung viral titers in mice after challenge with three divergent swH3 viruses. Vaccination studies in swine, the target species for this vaccine, show stronger levels of cross-reactive antibodies and T-cell responses after immunization with the epigraph vaccine compared to the wild type and FluSure vaccines. In both murine and swine models, epigraph vaccination shows superior cross-reactive immunity that should be further investigated as a universal swH3 vaccine

Novel Conserved-region T-cell Mosaic Vaccine With High Global HIV-1 Coverage Is Recognized by Protective Responses in Untreated Infection

An effective human immunodeficiency virus type 1 (HIV-1) vaccine is the best solution for halting the acquired immune deficiency syndrome epidemic. Here, we describe the design and preclinical immunogenicity of T-cell vaccine expressing novel immunogens tHIVconsvX, vectored by DNA, simian (chimpanzee) adenovirus, and poxvirus modified vaccinia virus Ankara (MVA), a combination highly immunogenic in humans. The tHIVconsvX immunogens combine the three leading strategies for elicitation of effective CD8+ T cells: use of regions of HIV-1 proteins functionally conserved across all M group viruses (to make HIV-1 escape costly on viral fitness), inclusion of bivalent complementary mosaic immunogens (to maximize global epitope matching and breadth of responses, and block common escape paths), and inclusion of epitopes known to be associated with low viral load in infected untreated people (to induce field-proven protective responses). tHIVconsvX was highly immunogenic in two strains of mice. Furthermore, the magnitude and breadth of CD8+ T-cell responses to tHIVconsvX-derived peptides in treatment-naive HIV-1+ patients significantly correlated with high CD4+ T-cell count and low viral load. Overall, the tHIVconsvX design, combining the mosaic and conserved-region approaches, provides an indisputably better coverage of global HIV-1 variants than previous T-cell vaccines. These immunogens delivered in a highly immunogenic framework of adenovirus prime and MVA boost are ready for clinical development