ABSTRACT
A broad coverage influenza vaccine against multiple viral strains based on the viral nucleoprotein (NP) is a goal pursued by many laboratories. If the goal is to formulate the vaccine with recombinant NP it is essential to count on adjuvants capable of inducing cellular immunity. This work have studied the effect of the monophosphoryl lipid A and trehalose dimycolate, known as the Ribi Adjuvant System (RAS), in the immune response induced in mice immunized with recombinant NP. The NP was formulated with RAS and used to immunize BALB/c mice. Immunizations with NP-RAS increased the humoral and cellular immune responses compared to unadjuvanted NP. The predominant antibody isotype was IgG2a, suggesting the development of a Th1 response. Analysis of the cytokines from mice immunized with NP-RAS showed a significant increase in the production of IFN-g and a decreased production of IL-10 and IL-4 compared to controls without RAS. These results are similar to those usually obtained using Freund's adjuvant, known to induce Th1 and CTL responses when co-administered with purified proteins, and suggest that a similar approach may be possible to enhance the performance of a T-cell vaccine containing NP.
Subject(s)
Cell Wall Skeleton/administration & dosage , Cord Factors/administration & dosage , Influenza, Human/immunology , Lipid A/analogs & derivatives , RNA-Binding Proteins/immunology , Th1 Cells/immunology , Viral Core Proteins/immunology , Animals , Antibodies, Viral/immunology , Cell Wall Skeleton/immunology , Cord Factors/immunology , Female , Humans , Immunity, Cellular , Immunization , Influenza Vaccines/administration & dosage , Influenza Vaccines/genetics , Influenza Vaccines/immunology , Influenza, Human/prevention & control , Influenza, Human/virology , Interferon-gamma/immunology , Interleukin-10/immunology , Interleukin-4/immunology , Lipid A/administration & dosage , Lipid A/immunology , Mice , Mice, Inbred BALB C , Nucleocapsid Proteins , RNA-Binding Proteins/administration & dosage , RNA-Binding Proteins/genetics , T-Lymphocytes, Cytotoxic/immunology , Viral Core Proteins/administration & dosage , Viral Core Proteins/geneticsABSTRACT
In the present work, immunogenicity of recombinant in vitro assembled hepatitis C virus core particles, HCcAg.120-VLPs, either alone or in combination with different adjuvants was evaluated in BALB/c mice. HCcAg.120-VLPs induced high titers of anti-HCcAg.120 antibodies and virus-specific cellular immune responses. Particularly, HCcAg.120-VLPs induced specific delayed type hypersensitivity, and generated a predominant T helper 1 cytokine pro file in immunized mice. In addition, HCcAg.120-VLPs prime splenocytes proliferate in vitro against different HCcAg.120-specific peptides, depending on either the immunization route or the adjuvant used. Remarkably, immunization with HCcAg.120-VLPs/Montanide ISA888 formulation resulted in a significant control of vaccinia virus titer in mice after challenge with a recombinant vaccinia virus expressing HCV core protein, vvCore. Animals immunized with this formulation had a marked increase in the number of IFN-gamma producing spleen cells, after stimulation with P815 cells infected with vvCore. These results suggest the use of recombinant HCV core particles as components of therapeutic or preventive vaccine candidates against HCV.
Subject(s)
Hepacivirus/immunology , Hepatitis C/immunology , Interferon-gamma/biosynthesis , Interleukin-4/biosynthesis , Peptide Fragments/immunology , Spleen/immunology , Viral Core Proteins/immunology , Adjuvants, Immunologic/administration & dosage , Animals , Female , Hepatitis C/prevention & control , Humans , Mice , Mice, Inbred BALB C , Peptide Fragments/administration & dosage , Spleen/cytology , Th2 Cells/immunology , Viral Core Proteins/administration & dosageABSTRACT
In the present work, immunogenicity of recombinant in vitro assembled hepatitis C virus core particles, HCcAg.120-VLPs, either alone or in combination with different adjuvants was evaluated in BALB/c mice. HCcAg.120-VLPs induced high titers of anti-HCcAg.120 antibodies and virus-specific cellular immune responses. Particularly, HCcAg.120-VLPs induced specific delayed type hypersensitivity, and generated a predominant T helper 1 cytokine pro file in immunized mice. In addition, HCcAg.120-VLPs prime splenocytes proliferate in vitro against different HCcAg.120-specific peptides, depending on either the immunization route or the adjuvant used. Remarkably, immunization with HCcAg.120-VLPs/Montanide ISA888 formulation resulted in a significant control of vaccinia virus titer in mice after challenge with a recombinant vaccinia virus expressing HCV core protein, vvCore. Animals immunized with this formulation had a marked increase in the number of IFN-γ producing spleen cells, after stimulation with P815 cells infected with vvCore. These results suggest the use of recombinant HCV core particles as components of therapeutic or preventive vaccine candidates against HCV.
Subject(s)
Animals , Female , Humans , Mice , Hepacivirus/immunology , Hepatitis C/immunology , Interferon-gamma/biosynthesis , /biosynthesis , Peptide Fragments/immunology , Spleen/immunology , Viral Core Proteins/immunology , Adjuvants, Immunologic/administration & dosage , Hepatitis C/prevention & control , Mice, Inbred BALB C , Peptide Fragments/administration & dosage , Spleen/cytology , /immunology , Viral Core Proteins/administration & dosageABSTRACT
Previously, the P64k meningococcal protein, an antigen of 64 kDa expressed in Escherichia coli, has been extensively characterized. We have successfully conjugated several synthetic peptides and meningococcal group C polysaccharide to P64k. In three out of four model peptides, the murine humoral immune response against the homologous peptide, evaluated after three doses of conjugate, was higher in the animals immunized with the coupled peptide than in those that received free peptide. The fourth and largest was immunogenic by itself. Similarly, the antigroup C polysaccharide levels reached by conjugated polysaccharide were significantly higher than those produced against unconjugated polysaccharide. As a carrier for one of the peptides, P64k was compared with bovine serum albumin (BSA) and tetanus toxoid (TT), being able to induce slightly higher or similar antipeptide antibody levels than these well-establish protein carriers. Our results suggest that recombinant P64k protein could be a readily available immunological carrier, as efficient as other commonly used large carrier molecules.