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The prototype virus HIV-1 LAV and highly cytopathic Zairian virus HIV-1 NDK belong to the genetic subtypes B and D and represent low and highly cytopathic phenotypes, respectively. Their neutralization pattern and serotype were studied with respect to differences in their genotypes and phenotypes. Sera from HIV-1-infected persons living in four geographically distant areas, Philadelphia (USA), Ribeirao...
The principal neutralizing domain, IGPGRAF sequence, from the V3-loop of HIV-1 was inserted in two positions on the surface of the protein that makes up the capside shell of the insect Flock House Virus. The hybrid proteins were expressed in insect cells via recombinant baculoviruses. Three different hybrids were used as immunogens: two with a single copy of the insert in different positions of the...
To develop a nucleic acid vaccine against dengue type-2 virus, the PreM and 92% of the envelope (E) genes were cloned into different eukaryotic plasmid expression vectors (pkCMVintPolyli and pVR1012). The resultant plasmid constructs (pD2ME and P1012D2ME) properly expressed the truncated E protein in vitro as evidenced by the expected protein size on SDS-PAGE and the ability of the protein to be...
The immunity against poliomyelitis in 1000 subjects 13-14 years old was evaluated. Neutralizing antibodies against poliovirus type 1, 2 and 3 were detected in 97.6%, 95.8% and 70% of samples, respectively. 31000 (0.3%) subjects were simultaneously seronegative to the three types. WHO does not suggest a protective level of International Units (IU), but our data indicate that such level is 0.45 IU...
Administration of vaccinia immune globulin (VIG), derived from vaccinated healthy adult volunteers, is the treatment-of-choice for patients suffering from severe complications following smallpox vaccination. The present study was aimed to determine the time interval after vaccination, at which the highest titer of neutralizing antibodies is obtained. Ninety-nine 18-year-old soldiers, immunized with...
It has been clearly demonstrated that cellular antigens (HLA, β2-microglobulin) are incorporated at the virion surface. The same epitope derived from β2-microglobulin is presented on all virus isolates. The peptide was identified by blocking the neutralizing capacity of a monoclonal antibody directed to R7V epitope: using this peptide for developing an ELISA, we have detected antibodies in nonprogressor...
To examine whether genetic immunization with Sin Nombre (SN) hantavirus genes could elicit immune responses, nine fragments spanning the envelope glycoprotein genes G1 and G2, and the complete N gene were cloned into a CMV expression vector. To ensure representation of all potential epitopes, adjacent fragments of the glycoprotein genes overlapped one another by 100 nucleotides. Vectors containing...
Measles remains a significant problem in both the developed and developing world, and new measles vaccination strategies need to be developed. This paper examines the strategy of utilizing transgenic plants expressing a measles antigen for the development of an oral sub-unit measles vaccine. A 1.8 kb fragment encompassing the coding region of the measles virus hemagglutinin (H) protein was cloned...
DNA encoding HIV-1 env is a poorly efficient B-cell immunogen and one probable explanation is that the numerous gp120 N-linked glycans gp120 may interfere with B-cell epitope presentation. The N306 glycan in gp120 shields HIV-1 from neutralizing antibodies. A DNA immunogen lacking the N306 glycosylation signal (T308A) was constructed to determine whether this glycan affected the immune response. Mice...
Successes for neutralizing antibodies (nAbs) against the human immunodeficiency virus (HIV) include potent cross-clade neutralization of primary virus isolates by human neutralizing monoclonal antibodies (nmAbs) targeting conserved envelope epitopes. Furthermore, passively administered combinations of human nmAbs prevented infection in primates, indicating that epitopes recognized by such nmAbs are...
Given the increasing incidence of HIV-1 infection world-wide, an affordable, effective vaccine is probably the only way that this virus will be contained. Accordingly, our group is developing an oral prime-boost strategy with the primary goal of eliciting broadly neutralizing antibodies against HIV-1 to provide sterilizing immunity for this virus. Our secondary goal is to elicit broadly cross-reactive...
Quantification of human immunodeficiency virus type 1 (HIV-1) neutralizing antibodies is considered to be an important parameter in evaluating candidate vaccines. Most previous studies have failed to detect vaccine-induced antibodies against primary isolates, which are more resistant to antibody mediated neutralization compared with laboratory isolates, in neutralization assays. In this study, sera...
Phase III efficacy trials of recombinant human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins were postponed. In Phase I and II trials these candidate vaccines had failed to induce neutralizing antibodies against virus which had been isolated by co-culture with human peripheral blood mononuclear cells (PBMC). The aim of the present study was to determine assay conditions for detecting...
The membrane-proximal external region (MPER) of HIV-1 gp41 is recognized by all three anti-HIV antibodies 2F5, 4E10 and Z13 that were directly derived from AIDS patients and have broader anti-HIV neutralizing activities. Thus, the MPER has been the focus of anti-HIV vaccine design and development. However, it has been unsuccessful to generate anti-HIV neutralizing antibodies targeting this region...
The human immunodeficiency virus type 1 (HIV-1) Tat is a key protein playing a major role in the infectivity of the virus. Thus, HIV-Tat based vaccines have been proposed as an attractive option to treat AIDS. Recently, we have shown that the recombinant detoxified adenylate cyclase (CyaA) from Bordetella pertussis carrying HIV-Tat (CyaA-E5-Tat), targets dendritic cells (DCs) and induces specific...
The study was designed to evaluate a novel cationic lipid DNA adjuvant (N3) and its function for HIV-1gp160/rev DNA plasmid delivered intranasally. The primary N3/HIV-DNA plasmid immunizations were boosted intranasally with a gp41 peptide in a anionic L3 adjuvant. This novel prime-boost strategy of mucosal immunization provided a broad HIV-1 envelope specific immunity, and recognition of viruses of...
To understand the cross-reactivity of antibodies directed against variable regions of human immunodeficiency virus-1 (HIV-1) envelope (Env), chimeric immunogens were prepared from different clades with modifications in variable regions, and the resulting neutralizing antibody response was evaluated. The V3-specific neutralization activity induced by a clade B immunogen was limited to clade B viruses...
The spike (S) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) is considered as a major antigen for vaccine design. We previously demonstrated that the receptor-binding domain (RBD: residues 318–510) of S protein contains multiple conformation-dependent neutralizing epitopes (Conf I to VI) and serves as a major target of SARS-CoV neutralization. Here, we further characterized the...
HIV-1 N-glycans are known to shield underlying epitopes towards the protective antibody repertoire. We previously described HIV-1 acute infection Env glycomutants designed from 3D-model in which the removal of clustered N-glycans did not disturb the envelope antigenicity, but increased the neutralization sensitivity. The potential of such immunogens to elicit neutralizing responses was estimated after...
DNA plasmids and recombinant adenovirus serotype-5 (rAd5) vectors are being studied in human clinical trials as HIV-1 vaccine candidates. Each elicits robust T-cell responses and modest antibody levels. Since protein immunization alone elicits antibody but not CD8 T-cell responses, we studied protein boosting of DNA and rAd5 HIV-1 vaccine vectors. A single Env protein immunization provided a marked...
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