ovalbumin and Papillomavirus-Infections

Topics: Adenylate Cyclase Toxin; Adjuvants, Immunologic; Animals; Antigens, Neoplasm; Cancer Vaccines; Disease Models, Animal; DNA-Binding Proteins; Drug Carriers; Female; Immunologic Memory; Injections, Intradermal; Melanoma; Mice, Inbred C57BL; Neoplasm Proteins; Oncogene Proteins, Viral; Ovalbumin; Ovarian Neoplasms; Papillomavirus E7 Proteins; Papillomavirus Infections; Papillomavirus Vaccines; T-Lymphocytes; Treatment Outcome; Uterine Cervical Neoplasms; Vaccines, Synthetic

Protein-based vaccines have emerged as a potentially promising approach for the generation of antigen-specific immune responses. However, due to their low immunogenicity, there is a need for innovative approaches to enhance protein-based vaccine potency. One approach to enhance protein-based vaccine potency is the employment of toll-like receptor ligands, such as CpG oligonucleotides, to activate the antigen-specific T cell immune responses. Another approach involves employing a method capable of improving the delivery of protein-based vaccine intramuscularly to lead to the slow release of the protein, resulting in improved vaccine potency. In the current study, we aimed to determine whether intramuscular injection of protein-based vaccines in conjunction with CpG followed by electroporation can lead to increased delivery of the protein-based vaccine into muscle cells, resulting in enhanced protein-based vaccine potency. We found that intramuscular injection followed by electroporation can effectively transduce the protein-based vaccine into the muscle cells. Furthermore, we found that intramuscular vaccination with OVA protein in combination with CpG followed by electroporation generates the best OVA-specific CD8+ T cell immune responses as well as the best protective and therapeutic antitumor effects in vaccinated mice. CD8+ T cells were found to play an important role in the observed protective antitumor effects generated by the vaccination. Similar results were observed using the HPV-16 E7 protein-based vaccination system. Thus, our data indicate that intramuscular administration of protein-based vaccines in conjunction with CpG followed by electroporation can significantly enhance the antigen-specific CD8+ T cell immune responses. The clinical implications of the study are discussed.

Topics: Adjuvants, Immunologic; Animals; CD8-Positive T-Lymphocytes; Electroporation; Female; Injections, Intramuscular; Mice; Mice, Inbred C57BL; Oligodeoxyribonucleotides; Ovalbumin; Papillomavirus E7 Proteins; Papillomavirus Infections; Vaccination; Vaccines, Subunit

Adjuvant System 04 (AS04) combines the TLR4 agonist MPL (3-O-desacyl-4'-monophosphoryl lipid A) and aluminum salt. It is a new generation TLR-based adjuvant licensed for use in human vaccines. One of these vaccines, the human papillomavirus (HPV) vaccine Cervarix, is used in this study to elucidate the mechanism of action of AS04 in human cells and in mice. The adjuvant activity of AS04 was found to be strictly dependent on AS04 and the HPV Ags being injected at the same i.m. site within 24 h of each other. During this period, AS04 transiently induced local NF-kappaB activity and cytokine production. This led to an increased number of activated Ag-loaded dendritic cells and monocytes in the lymph node draining the injection site, which further increased the activation of Ag-specific T cells. AS04 was also found to directly stimulate those APCs in vitro but not directly stimulate CD4(+) T or B lymphocytes. These AS04-induced innate responses were primarily due to MPL. Aluminum salt appeared not to synergize with or inhibit MPL, but rather it prolonged the cytokine responses to MPL at the injection site. Altogether these results support a model in which the addition of MPL to aluminum salt enhances the vaccine response by rapidly triggering a local cytokine response leading to an optimal activation of APCs. The transient and confined nature of these responses provides further supporting evidence for the favorable safety profile of AS04 adjuvanted vaccines.

Topics: Adjuvants, Immunologic; Aluminum Hydroxide; Animals; B7-2 Antigen; CD4-Positive T-Lymphocytes; CD40 Antigens; Cell Line; Cytokines; Dendritic Cells; Female; Human papillomavirus 16; Human papillomavirus 18; Humans; Immunity, Innate; Lipid A; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; NF-kappa B; Ovalbumin; Papillomavirus Infections; Papillomavirus Vaccines; Toll-Like Receptor 4; Transfection

We have recently reported that the sublingual (s.l.) mucosa is an efficient site for inducing systemic and mucosal immune responses. In this study, the potential of s.l. immunization to induce remote Ab responses and CD8(+) cytotoxic responses in the female genital tract was examined in mice by using a nonreplicating Ag, OVA, and cholera toxin (CT) as an adjuvant. Sublingual administration of OVA and CT induced Ag-specific IgA and IgG Abs in blood and in cervicovaginal secretions. These responses were associated with large numbers of IgA Ab-secreting cells (ASCs) in the genital mucosa. Genital ASC responses were similar in magnitude and isotype distribution after s.l., intranasal, or vaginal immunization and were superior to those seen after intragastric immunization. Genital, but not blood or spleen, IgA ASC responses were inhibited by treatment with anti-CCL28 Abs, suggesting that the chemokine CCL28 plays a major role in the migration of IgA ASC progenitors to the reproductive tract mucosa. Furthermore, s.l. immunization with OVA induced OVA-specific effector CD8(+) cytolytic T cells in the genital mucosa, and these responses required coadministration of the CT adjuvant. Furthermore, s.l. administration of human papillomavirus virus-like particles with or without the CT adjuvant conferred protection against genital challenge with human papillomavirus pseudovirions. Taken together, these findings underscore the potential of s.l. immunization as an efficient vaccination strategy for inducing genital immune responses and should impact on the development of vaccines against sexually transmitted diseases.

Topics: Adjuvants, Immunologic; Administration, Sublingual; Animals; Antibodies, Bacterial; Antibodies, Viral; Antibody-Producing Cells; Cell Differentiation; Cells, Cultured; Cholera Toxin; Disease Models, Animal; Female; Human papillomavirus 16; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mucous Membrane; Ovalbumin; Papillomavirus Infections; T-Lymphocytes, Cytotoxic; Uterine Cervical Neoplasms; Virion

Previous reports have shown that transcutaneous immunization (TCI) with proteins or peptides in combination with adjuvants efficiently induces specific cellular and humoral immune responses. However, depending on the kind of skin pretreatment, induction of cellular immune responses was restricted to generation of either specific cytotoxic T lymphocytes (CTLs) or T-helper (Th) cells. In this study, we induced antigen-specific CTL responses together with the appropriate Th responses by TCI of C57BL/6 mice. We applied ovalbumin protein or an ovalbumin-derived fusion peptide containing a CTL and Th epitope together with a combination of cholera toxin (CT) and CpG oligodeoxynucleotide (CpG) onto cold wax-depilated and hydrated bare skin. TCI with the ovalbumin fusion peptide induced more robust CTL and Th responses than that with ovalbumin protein. The fusion peptide in combination with the nontoxic CT derivative CTA1-D2D1 and CpG induced an antigen-specific CTL response, albeit less efficiently than in combination with complete CT. Further, we compared the potency of HPV-16 E7 oncoprotein-derived peptides containing single (CTL) or multiple (CTL + Th + B cell) epitopes to induce effective CTL responses. Strong E7-specific CTL responses were detected only after TCI with the E7 multiepitope peptide. This peptide was also shown to protect mice against tumor growth after challenge with HPV-16 E7-positive tumor cells. TCI with E7 protein and CT/CpG led to formation of an E7-specific humoral immune response.

Topics: Administration, Cutaneous; Animals; Antibody Formation; Epitopes; Female; Human papillomavirus 16; Humans; Immunization; Lymphoma, T-Cell; Mice; Mice, Inbred C57BL; Ovalbumin; Papillomavirus Infections; Peptides; T-Lymphocytes, Cytotoxic; T-Lymphocytes, Helper-Inducer; Thymoma; Transfection