Subramonia Pillai

The pulmonary immune response of Balb/c mice vaccinated with the fusion protein of respiratory syncytial virus

We have investigated the efficacy of vaccination with the purified fusion (F) protein of respiratory syncytial virus (RSV) on aluminium hydroxide adjuvant in Balb/c mice. The purpose of the study was to define the role of the local pulmonary mononuclear cell (PMC) infiltrate in the clearance of virus from the lower respiratory tract. Balb/c mice immunized with F protein were able to inhibit the replication of virus in the lungs as early as 4 days after intranasal challenge. In contrast, unimmunized mice required 8 days. Examination of humoral immune mechanisms demonstrated that vaccination with the purified protein induced moderate titres of serum neutralizing antibody. In addition, immunization induced low to moderate levels of antigen-dependent killer cell activity. To examine the immunological events responsible for virus clearance in vivo, PMC infiltrates were isolated after virus challenge and tested directly for protective capacity. After virus challenge, the F protein-immune mice were able to recall the cytolytic cells to the pulmonary tissues. The results further suggested that the local antigen-dependent killer activity was mediated by cytolytic T cells of the CD8 phenotype. Adoptive transfer studies were also conducted to identify further the role the PMC infiltrate had in protective immunity. Adoptive transfer of F protein-educated PMC into naive syngeneic recipients suggested that the pulmonary infiltrates contained the cellular constituents necessary for protective immunity. Both humoral and cellular immune elements were present.(ABSTRACT TRUNCATED AT 250 WORDS)

Induction of pneumococcal polysaccharide-specific mucosal immune responses by oral immunization

Liposome and cholera toxin (CT) are considered to be effective antigen delivery vehicles and adjuvants for mucosal vaccines. The effect of these antigen delivery systems on adjuvant responses to mucosally administered pneumococcal polysaccharide (Pnup) was investigated in this study. Both mucosal (e.g. oral) and systemic (i.p.) immunization of mice with purified preparations of Pnup type 23F induced antigen-specific IgM responses in sera. Interestingly, oral immunization of as little as 10 micrograms of Pnup type 23F was sufficient to induce systemic IgM responses. Pnup-specific IgM antibodies peaked by day 7 and no booster responses were evident after a second dose on day 14. In order to examine whether IgG and IgA Pnup-specific immune responses are induced by mucosal immunization, the mucosal adjuvant CT was mixed with Pnup type 23 as an oral vaccine. Co-oral administration of CT and Pnup type 23F resulted in the induction of Pnup-specific faecal IgA antibodies. These results were confirmed by detecting antigen-specific IgA-spot-forming cells in mononuclear cell suspensions prepared from the intestine of immunized mice. These findings suggest that oral immunization with Pnup in the presence of mucosal adjuvants, such as CT, could induce Pnup-specific IgA responses whereas Pnup alone did not. In an attempt to further enhance antigen-specific antibody responses, Pnup type 23F was encapsulated in liposomes and used as mucosal vaccine. However, immunogenicity of Pnup was not improved.

Synthetic Peptide Representing a T-Cell Epitope of CRM197 Substitutes as Carrier Molecule in a Haemophilus Influenzae Type B (HIB) Conjugate Vaccine

The cross-reactive material (CRM197) of diphtheria toxin is considered to be advantageous as a carrier molecule in the formulation of a Haemophilus influenzae type b conjugate vaccine. In order to more precisely understand the function of the CRM197 in the vaccine, we have begun mapping the T-cell epitopes of the protein. A peptide which represents a segment of the primary sequence of CRM197 has been identified and found to stimulate diphtheria toxoid or CRM197-primed murine T-lymphocytes. In addition, the peptide is capable of priming T-cells in vivo for a subsequent in vitro T-cell response to itself or to the intact CRM197 molecule. The ability of the peptide to replace CRM197 as a carrier molecule was examined by immunizing mice with PRP, PRP-CRM197 conjugate, or PRP covalently coupled to the peptide. Antibodies to PRP were only detected in the PRP-CRM197 or PRP-peptide immunized groups. Both conjugates elicited primary and secondary antibody responses. Thus, a synthetic peptide representing a defined T-cell epitope of CRM197 has been functionally demonstrated based on its ability to act as a carrier molecule in a PRP conjugate vaccine.

Augmentation by Interleukins of the Antibody Response to a Conjugate Vaccine against Haemophilus influenzae B

Interleukins have been recognized as potential adjuvants for use during vaccination. The immunogenicity of some poorly immunogenic bacterial capsular polysaccharides have been improved by conjugation to a protein carrier. Augmentation of the immune response to these glycoconjugates, however, may be realized in the presence of interleukins. The antibody response to one such vaccine which comprises a oligosaccharide derived from the capsule of Haemophilus influenzae type b coupled to CRM197 (HbOC) can be augmented in this manner. A suboptimal dose (0.1 microgram) of HbOC and varying concentrations of IL-1 alpha or IL-1 beta (10(2) - 5 x 10(5) U) were injected intramuscularly at 0 and 2 weeks into Swiss Webster mice. Vaccines were also formulated with and without aluminum phosphate. Antibody to the oligosaccharide was determined by Farr assay. In 3/3 experiments, IL-1 alpha enhanced primary and secondary antibody responses whereas with IL-1 beta, only a slight increase in the primary antibody response was seen but enhanced secondary responses were observed. Thus, IL-1 alpha and to some extent IL-1 beta enhanced the primary and secondary antibody responses to a glycoconjugate vaccine.