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In B cells, the processing of antigens in the context of MHC class II molecules is initiated by the binding of antigen to the B cell antigen receptor (BCR). The BCR serves two roles in antigen processing, signaling for enhanced processing and endocytosing bound antigen. CD19 is a B cell surface molecule which has been demonstrated to function in modifying signals generated through the BCR, regulating...
A single small segment (sequence recognition) or domain (conformation recognition) of a protein could act as an antigen (antigenic determinant) vs an antibody. Epitopes of the first kind being a continuous segment along the sequence (linear), generally bent with a typical non-ordered structure (turns and/or loops), can be predicted from the only knowledge of the primary structure. After reviewing...
Previous work from our group showed that the entrapment of HAV-related synthetic peptides into multilamellar liposomes yielded satisfactory immunoresponses when administered to mice. In the present work we report investigative results for several liposome formulations of a 20-mer peptide related to VP3 capsid protein of HAV. In this sense, the recently introduced surface plasmon resonance technique...
Recombinant, insect cell derived SIV Pr56 gag virus-like particles (VLPs) have been modified either by inserting HIV-1 Gp160 derived peptides into the Pr56 gag precursor or by integrating the complete HIV-1 gp120 in the particle membrane. To investigate the protective efficacy of these particulate antigens, rhesus macaques were immunized with VLPs both adjuvant-free...
The sequence H236-256 of the measles virus (MV) hemagglutinin (H) contains the sequential epitope of the neutralizing and protective monoclonal antibody (mAb) BH129 with the minimal epitope E 245 L-QL 249 . Using this mAb, we have recently developed 7mer mimotopes binding up to 135x better than the corresponding 7mer epitope H244-250. In this study, we combined...
MUC1 has been used as a target for immunotherapy and with oxidised mannan in mice there is selective delivery into the class I pathway and the induction of a T1 response. We have also been working in pig-to-human transplantation and of particular interest is the description in humans of natural Galα(1,3)Gal antibodies (Abs) which react with pig tissues. A peptide mimic (DAHWESWL) to the Galα(1,3)Gal...
We have designed a novel vaccine strategy which enables display of short peptides expressed from chimeras of the gene encoding the coat protein of the RNA bacteriophage MS2 and inserted foreign DNA. MS2 coat protein has a β-hairpin loop at the N-terminus which forms the most radially distinct feature of the mature capsid. The coat protein gene was modified to enable insertion of DNA at the central...
Although the induction of antibodies and T cells by synthetic peptides representing defined antigenic determinants is a routine laboratory procedure, their use as vaccines has not yet been generally realised. There are a number of reasons for this and paramount is the limitation of valency which affects not only immunogenicity, but also the coverage of the antigenic universe. This paper is a review...
An antigen delivery system based on hybrid recombinant parvovirus-like particles (VLPs) formed by the self-assembly of the capsid VP2 protein of porcine (PPV) or canine parvovirus (CPV) expressed in insect cells with the baculovirus system has been developed. PPV:VLPs containing a CD8 + epitope from the LCMV nucleoprotein evoked a potent CTL response and were able to protect mice against a...
Biodegradable polyester microspheres (MS) have shown potential for single-dose vaccines. This study examined the immunogenicity of diphtheria toxoid (Dtxd) microencapsulated in different types of poly(lactide) (PLA) and poly(lactide-co-glycolide) (PLGA) MS prepared by the methods of spray-drying and coacervation. We investigated the influence of polymer type (PLGA 50:50 of low M w ; PLA of...
Recent developments in peptide technology enable the use of random peptide libraries for identifying linear amino acid sequences (mimotopes) which can mimic conformational epitopes without necessarily exhibiting amino acid sequence homology with the native linear sequence. In this study a 15-mer random peptide library displayed on the surface of a filamentous phage has been used to characterise the...
The main antigenic site (site A) of foot-and-mouth disease virus (FMDV, strain C-S8c1) may be adequately reproduced by a 15-peptide with the amino acid sequence H-YTASARGDLAHLTTT-NH 2 (A15), corresponding to the residues 136-150 of the viral protein VP1. The effect of amino acid substitutions within A15 on its antigenicity towards monoclonal antibodies (MAb) raised against antigenic site A,...
Mice immunized with plasmid DNA encoding Nef regulatory protein of human immunodeficiency virus type 1 developed high levels of anti-Nef antibodies. After 4 intramuscular injections of 100 μg plasmid DNA, anti-Nef antibodies reached titers up to 2x10 4 . A significant specific antibody response was maintained for at least 16 months.Using a set of seven 31-66 mer synthetic peptides covering...
A new class of sequential oligopeptide carriers (SOC n ) for anchoring antigenic/immunogenic peptides has been constructed. The carrier, formed by the repetitive Lys-Aib-Gly moiety, is designed to display a predetermined 3D structure, so that the attached peptides would obtain a defined spatial orientation. Conformational analysis showed that SOC n adopt a distorted 3 10 ...
In recent years microspheres or microparticles produced from biodegradable polymers such as poly(d,l-lactide) (PLA) and poly(d,l-lactide-co-glycolide) (PLGA) containing encapsulated vaccine antigens have been investigated for administration via parenteral, oral, and intranasal routes. These microparticles allow the controlled release of vaccines with an aim to reduce the number of doses for primary...
We have previously found synthetic oligodeoxynucleotides (ODN) containing immunostimulatory CpG motifs to be a potent adjuvant to protein administered by intramuscular injection or intranasal inhalation to BALB/c mice. Herein we have further evaluated the potential of CpG ODN as a mucosal adjuvant to purified hepatitis B surface antigen (HBsAg) when administered alone or with cholera toxin (CT). CpG...
Three different approaches of using overlapping peptides have been compared to analyse the fine specificity of the antibody response to a protective epitope from measles virus (MV) fusion protein, spanning residues 397-420. Anti-peptide antibodies raised in BALB/c, CBA and C57BL/6 mice were shown to react with the homologous peptide and the MV by ELISA. Results from indirect ELISA using 15mer peptides...
A DNA vaccine encoding glycoprotein D (gD) of herpes simplex virus type 2 (pHSV-gD2) was injected via parenteral and mucosal routes to determine the optimal route of delivery for immune stimulation. Generation of distal mucosal immunity following parenteral vaccination was also evaluated. While all routes of DNA vaccine administration resulted in systemic cellular and humoral responses, the intra-muscular...
Mimicry epitopes that are recognized by T-cells can be identified through screening of synthetic peptide libraries. We have shown that these mimicry epitopes share sequence similarity with the corresponding natural epitopes and that mimicry sequences can be used for the definition of protein derived T-cell epitopes from databases. This can be done by either homology searching or pattern searching...
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