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dc.contributor.advisorMutwiri, George
dc.contributor.advisorWilson, Heather
dc.creatorChaffey, Alyssa 1992-
dc.date.accessioned2017-07-12T21:17:38Z
dc.date.available2017-07-12T21:17:38Z
dc.date.created2017-06
dc.date.issued2017-07-12
dc.date.submittedJune 2017
dc.identifier.urihttp://hdl.handle.net/10388/7959
dc.description.abstractImmunostimulatory adjuvants are substances added to vaccines to promote and direct a robust Th1, Th2, or Th17 immune response. Murine Th17 cells are produced and differentiated from naïve T cells in the presence of transforming growth factor (TGF)-β and interleukin (IL)-6 and once differentiated, Th17 cells produce cytokines IL-17A, IL-17F, IL-21 and IL-22. We investigated how immunostimulatory molecules such as poly[di(sodium carboxylatoethylphenoxy)-phosphazene (PCEP), Alum, CpG oligodeoxynucleotide (CpG ODN), Curdlan, Leptin and Lipopolysaccharide (LPS), alone or in combination influenced differentiation and/or activation of Th17 type immune cells in mice. In vitro studies showed that murine splenocytes stimulated with CpG showed significantly induced production of IL-12, a cytokine important for induction of Th1 type immune cells and IL-12 is known to be inhibitory for differentiation of Th17-type immune cells. Curdlan + Leptin +/- PCEP and PCEP + Curdlan induced significant expression of TGF-β. No immunostimulant combination induced both IL-6 and TGF-β, which we anticipated would be required for Th17 cell differentiation. When we investigated the cytokines induced by the immunostimulants 48 hours after injection in muscle tissue, we determined that Curdlan + Leptin significantly induced production of IL-17, likely from activation of already differentiated T cells. TGF-β was significantly induced in response to Curdlan and Leptin, alone and in combination but they were poor inducers of IL-6. PCEP+/- CpG or LPS significantly induced expression of IL-6 but not TGF-β. Finally, we immunized mice via intramuscular (i.m.) route with OVA in the presence of the immunostimulatory adjuvants and assessed cytokine production from OVA-restimulated splenocytes 5 weeks later. ELISA results indicated that OVA-specific IL-17 production was significantly induced in splenocytes from mice immunized with PCEP + OVA relative to the mice immunized with Curdlan + OVA, although it was insignificant with respect to the OVA immunization group presumably due to the highly variable responses. Using flow cytometric analysis, we observed that vaccination with PCEP + OVA and Curdlan + Leptin + OVA significantly induced the frequency of OVA-specific splenic CD4+IL-17+ cells. Curdlan + Leptin also significantly induced the frequency of OVA-specific CD4+Foxp3+ cells and CD4+IL-17+Foxp3+ double positive cells. Thus, we conclude that in vitro studies are poorly predictive of the type of adaptive response that may be induced when immunostimulatory adjuvants were used in a vaccine. Furthermore, vaccines formulated with PCEP and Curdlan + Leptin adjuvants promote Th17 cell differentiation and should be investigated as a combinational adjuvant for bacteria or fungal based immunizations.
dc.format.mimetypeapplication/pdf
dc.subjectAdjuvant
dc.subjectTh17
dc.titleIdentification of immunostimulatory adjuvant(s) that will promote a Th17-type of immune response
dc.typeThesis
dc.date.updated2017-07-12T21:17:38Z
thesis.degree.departmentSchool of Public Health
thesis.degree.disciplineVaccinology and Immunotherapeutics
thesis.degree.grantorUniversity of Saskatchewan
thesis.degree.levelMasters
thesis.degree.nameMaster of Science (M.Sc.)
dc.type.materialtext
dc.contributor.committeeMemberTikoo, Suresh
dc.contributor.committeeMemberSingh, Baljit
dc.contributor.committeeMemberBull, Harold
dc.creator.orcid0000-0002-2282-9210


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