Incorporation of membrane-bound, mammalian-derived immunomodulatory proteins into influenza whole virus vaccines boosts immunogenicity and protection against lethal challenge

<p>Abstract</p> <p>Background</p> <p>Influenza epidemics continue to cause morbidity and mortality within the human population despite widespread vaccination efforts. This, along with the ominous threat of an avian influenza pandemic (H5N1), demonstrates the need for a much improved, more sophisticated influenza vaccine. We have developed an in vitro model system for producing a membrane-bound Cytokine-bearing Influenza Vaccine (CYT-IVAC). Numerous cytokines are involved in directing both innate and adaptive immunity and it is our goal to utilize the properties of individual cytokines and other immunomodulatory proteins to create a more immunogenic vaccine.</p> <p>Results</p> <p>We have evaluated the immunogenicity of inactivated cytokine-bearing influenza vaccines using a mouse model of lethal influenza virus challenge. CYT-IVACs were produced by stably transfecting MDCK cell lines with mouse-derived cytokines (GM-CSF, IL-2 and IL-4) fused to the membrane-anchoring domain of the viral hemagglutinin. Influenza virus replication in these cell lines resulted in the uptake of the bioactive membrane-bound cytokines during virus budding and release. <it>In vivo </it>efficacy studies revealed that a single low dose of IL-2 or IL-4-bearing CYT-IVAC is superior at providing protection against lethal influenza challenge in a mouse model and provides a more balanced Th₁/Th₂humoral immune response, similar to live virus infections.</p> <p>Conclusion</p> <p>We have validated the protective efficacy of CYT-IVACs in a mammalian model of influenza virus infection. This technology has broad applications in current influenza virus vaccine development and may prove particularly useful in boosting immune responses in the elderly, where current vaccines are minimally effective.</p

Efficient priming of CD4 T cells by Langerin-expressing dendritic cells targeted with porcine epidemic diarrhea virus spike protein domains in pigs

<p>Each cryotolerance index was calculated as follows:.</p><p>(m.p.: membrane permeability; o.a.m.: outer acrosome membrane; GMFI: Geometric mean of fluorescence intensity; arbitrary units).</p

Abstract 799: Obesity-associated modulation of the immune microenvironment of the omental fat band during peritoneal seeding of ovarian cancer

Abstract Ovarian cancer is the leading cause of death by gynecological malignancy in the Western world, the fourth most deadly cancer in women. Typically originating from the layer of epithelial cells surrounding the ovary, cancer cells exfoliate to disseminate throughout the peritoneal cavity. The omental fat band (OFB) has been described as a secondary lymphoid organ composed of heavily vascularized fatty tissue interspersed with immune cell aggregates, or “milky spots”. It is also the site of preferential tumor cell adhesion and invasion within the peritoneal cavity, and is removed at the time of surgical debulking in ovarian cancer patients as a preventative measure. Several studies have suggested that obesity is a risk factor for ovarian cancer, but it is unclear whether the obese state directly influences peritoneal dissemination of ovarian cancer. Here, we set out to elucidate the obesity-related changes in the immune microenvironment of the OFB in the homeostatic state as well as following cancer cell adhesion and outgrowth. Our lab has developed a spontaneously transformed murine ovarian surface epithelial (MOSE) cell model that mimics the progressive stages of ovarian cancer following long-term passaging. Importantly, these cells can be used to study the immune microenvironment of tumor cells in an immunocompetent host. EGFP-expressing MOSE-L (late) cells that had been selected in vivo for an aggressive phenotype were injected i.p. into 6 month-old C57Bl/6 mice and changes to the OFB immune microenvironment were evaluated after 6, 24 hrs and 4 weeks. Furthermore, mice fed a high- or low-fat diet (40 or 5% calories as fat, respectively) were used to determine the effects of diet-induced obesity on cancer cell seeding and outgrowth. Immunofluorescence microscopy, real-time PCR and flow cytometry were used to determine tumor cell attachment and growth and identify changes in OFB immune cell populations. Early seeding (24 hrs post-injection) of EGFP-MOSE-L cells at the OFB was increased in high-fat fed mice. Preliminary data from mRNA expression profiling identified genes in the OFB that were i) obesity-associated, ii) a direct response to tumor cell seeding or iii) were exacerbated in the obese microenvironment following tumor cell seeding. This suggests that obesity-associated signaling events could be polarizing the OFB towards a pro-tumorigenic microenvironment. There were also distinct differences in the immune cell populations within the OFB as function of both obesity and time post-tumor cell seeding. These results indicate that ovarian cancer cells preferentially populate the OFB early during peritoneal dissemination and are capable of escaping immune surveillance mechanisms. This study sheds light on the obesity-induced changes in immunomodulatory events associated with tumor seeding and outgrowth in the peritoneal cavity. Supported by NIH CA118846 to EMS and PCR Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 799. doi:10.1158/1538-7445.AM2011-799</jats:p

Virology Journal

Background Influenza epidemics continue to cause morbidity and mortality within the human population despite widespread vaccination efforts. This, along with the ominous threat of an avian influenza pandemic (H5N1), demonstrates the need for a much improved, more sophisticated influenza vaccine. We have developed an in vitro model system for producing a membrane-bound Cytokine-bearing Influenza Vaccine (CYT-IVAC). Numerous cytokines are involved in directing both innate and adaptive immunity and it is our goal to utilize the properties of individual cytokines and other immunomodulatory proteins to create a more immunogenic vaccine. Results We have evaluated the immunogenicity of inactivated cytokine-bearing influenza vaccines using a mouse model of lethal influenza virus challenge. CYT-IVACs were produced by stably transfecting MDCK cell lines with mouse-derived cytokines (GM-CSF, IL-2 and IL-4) fused to the membrane-anchoring domain of the viral hemagglutinin. Influenza virus replication in these cell lines resulted in the uptake of the bioactive membrane-bound cytokines during virus budding and release. In vivo efficacy studies revealed that a single low dose of IL-2 or IL-4-bearing CYT-IVAC is superior at providing protection against lethal influenza challenge in a mouse model and provides a more balanced Th1/Th2 humoral immune response, similar to live virus infections. Conclusion We have validated the protective efficacy of CYT-IVACs in a mammalian model of influenza virus infection. This technology has broad applications in current influenza virus vaccine development and may prove particularly useful in boosting immune responses in the elderly, where current vaccines are minimally effective.Published versio

Virology Journal

Background Potent and safe adjuvants are needed to improve the efficacy of parenteral and mucosal vaccines. Cytokines, chemokines and growth factors have all proven to be effective immunomodulatory adjuvants when administered with a variety of antigens. We have previously evaluated the efficacy of membrane-anchored interleukins (IL) such as IL-2 and IL-4 co-presented as Cytokine-bearing Influenza Vaccines (CYT-IVACs) using a mouse model of influenza challenge. Findings Here, we describe studies evaluating the parenteral and mucosal adjuvanticity of membrane-bound IL-12 and IL-23 CYT-IVACs in young adult mice. Mucosal immunization using IL-12 and IL-23 bearing whole influenza virus vaccine (WIV) was more effective at eliciting virus-specific nasal IgA and reducing viral lung burden following challenge compared to control WIV vaccinated animals. Both IL-12 and IL-23 bearing WIV elicited the highest anti-viral IgA levels in serum and nasal washes. Conclusions This study highlights for the first time the mucosal adjuvant potential of IL-12 and IL-23 CYT-IVAC formulations in eliciting mucosal immune responses and reducing viral lung burden. The co-presentation of immunomodulators in direct context with viral antigen in whole inactivated viral vaccines may provide a means to significantly lower the dose of vaccine required for protection.Published versio

Regulation of Cytoskeleton Organization by Sphingosine in a Mouse Cell Model of Progressive Ovarian Cancer

biomolecule