Thesis--PlanA (M.S.)--University of Wisconsin--Stout, 2002. / Includes bibliographical references.

No safe and effective vaccine exists against respiratory syncytial virus (RSV), the main viral cause of lower respiratory tract infections in young children. Proteosome-based adjuvants, derived from the outer membrane proteins (OMP) of Neisseria species are potent inducers of mucosal and systemic immunity in humans and animals. RSV subunit vaccines based on enriched RSV proteins (eRSV) were formulated with proteosomes (Pro) or its S. flexneri LPS-supplemented derivative, Protollin (Prl). Administered intranasally (IN) in BALB/c mice, the vaccines elicited systemic and mucosal RSV-specific antibodies and fully protected against RSV challenge without enhanced pulmonary pathology or evidence of a Th2-biased response (eg: eosinophil infiltation or antigen-specific 1L-5 production by restimulated splenocytes or lung cells). Restimulation of cells from Prl-eRSV immunized mice elicited F peptide-specific CD8+ T cells producing IFNgamma and supernatant IFNgamma, TNFalpha, 1L-2 and IL-10. The Prl-eRSV vaccine was also studied in C57Bl/6 mice, to exploit the TLR2, TLR4 and MyD88-deficient (-/-) animals available on this background. Protection was significantly impaired in both TLR4-/- and MyD88 -/- mice, but not in TLR2 -/- mice following Prl- eRSV immunization and challenge. These studies revealed a role for the LPS component of Protollin in both initial (innate) cytokine release as well as dendritic cell maturation and Th1 polarization. Although antibody levels were sustained in MyD88-/- mice, the IgG1/IgG2a ratio was markedly higher in the absence of this pathway. The MyD88-/- mice also displayed elevated levels of pulmonary eosinophils following challenge, with concomitant reduction of neutrophils compared to wt mice. Using CD1d-iNKT cell-deficient mice (CD1-/-) in our BALB/c model, we also identified a significant role for the lipid component of both the Pro- and Prl-based vaccines. Responses to both vaccines in CD1-/- animals elicited lower antibody titers and reduced restimulated splenocyte supernatant cytokines (IFNgamma, IL-17 and IL-10), with concomitant augmentation of neutrophil recruitment (Prl only). Pro- and Prl-eRSV vaccines may therefore exert their powerful adjuvant effects by exploiting both CD1d-iNKT and, in the case of the Prlbased formulations the TLR4-MyD88-dependant signalling pathway. These pathways not only promote stronger Th1 immune responses but also act to control pulmonary eosinophil (MyD88-dependent) and neutrophil (MyD88 and CD1d-NKT-dependent) recruitment in a murine RSV challenge model.

No description available.

Respiratory syncytial virus (RSV) is a pathogen associated with lower respiratory tract infection, and is a common cause of infant hospitalization worldwide. Despite efforts to create safe and cost-effective RSV therapeutics, there remains no vaccine, and antiviral drugs have been developed with limited success. Among the 11 proteins coded by the negative-sense single-stranded RNA genome of RSV, the phosphoprotein (P) and nucleoprotein (N) aid in the formation of an RNA-dependent RNA polymerase (RdRp) complex, which is essential for RSV virulence. The specificities of the N-P binding interaction have been researched extensively, which has provided researchers with a novel target for an RSV therapeutic. In this study, a recombinant peptide mimetic (P220-241) containing the final 21 C-terminal amino acids of RSV P fused to Maltose-Binding Protein (MBP), and a cell-penetrating peptide (CPP), was purified for the purpose of targeting this interaction. In addition to successfully entering cells, the peptide was shown to inhibit both RSV subtype A and subtype B infection in vitro, with a percent inhibition (PI) of infection as high as 95% at 20 μM. Additionally, P220-241 did not inhibit infection of parainfluenza virus type 2 (PIV-2), indicating this inhibition was not an artifact of the peptide acting as a pathogen-associated molecular pattern (PAMP). A series of three different assays demonstrated that P220-241 does not appear to have any cytotoxic effects in vitro. Finally, using both glutathione S-transferase (GST) pull-downs and in vitro immunoprecipitations, we demonstrated that P220-241 is able to bind the N protein, while also preventing binding of full-length P protein. Taken together, this study provides the framework for a novel method of targeting RSV protein-protein interactions using chimeric cell-penetrating peptide mimetics. / Thesis / Master of Science (MSc)

Thesis (Ph.D.)--Baylor University, 2009. / Includes bibliographical references (p. 107-129).

No description available.

The human respiratory syncytial virus (hRSV) infection is a global public health burden in children aged under 2 years and immunocompromised or elderly adults. The choice for prophylaxis and therapy of hRSV infection was constrained to Palivizumab and Ribavirin. Therefore, this study aimed to develop effective anti-hRSV infection agents, such as short interfering RNA (siRNA) and β-defensin-4 (β-D-4). Since there still is no compatible animal model to evaluate antiviral effect of anti-hRSV agents, we first attempted to establish suitable animal model. A clinical isolate of hRSV (KI-RSV-W) was adapted in BALB/c mice by serial passages. Old male mice (age > 8 months) were used for establishment of hRSV infection model, because the more efficient viral infection in lungs of the old male mice than that old female mice and young mice (age < 3 weeks). After the virus was propagated in old male mice for 20 passages, a virus variant KI-RSV-P70-4 exhibited more efficient infection/replication in the mice. Its viral load was about 100-fold higher than that of wild type strain KI-RSV-W. The infection of KI-RSV-P70-4 also caused more severe histopathological changes in lung tissues. Although KI-RSV-P70-4 could not result in death of the infected mice, both viral load and pathological change in lungs may be good indicators for evaluating antiviral effect. The mouse model and adapted hRSV strain solidly laid the foundation for evaluation of anti-hRSV agents. We then designed and evaluated anti-hRSV effect of siRNAs. A total of 25 siRNAs targeting 4 viral genes (M2-1, NS2, N and F) were designed and their anti-hRSV effect was assessed in vitro. The results showed that 6 siRNAs respectively targeting M2-1, N and F genes exhibited higher anti-hRSV effect than that of the positive control, whereas those targeting NS2 gene did not show significant antiviral effect. The 50% inhibitory concentrations (IC50) of three most potent siRNAs (M2-1-361, N889 and F-1143) were 0.51, 2.14 and 0.64 nM, respectively. Antiviral activity of β-D-4 against hRSV infection was evaluated in vitro and in vivo. In vitro experiments showed supreme antiviral effect with IC50 around 3.4 μg/ml when the virus was pretreated with β-D-4, but no significant inhibitory effect was observed when the cells were pretreated with β-D-4 or β-D-4 was maintained in the culture medium after viral infection. These results indicated that the inhibitory effect of β-D-4 was associated with direct interaction with the virus itself and blocked virus entry of the cells. Furthermore, a single dose (13.6 μg) of β-D-4 intratracheal (i.t.) administration in old male mice after the viral infection resulted in about 0.7 log reduce of viral load in lung tissues, while inoculation of premixed β-D-4 and the virus caused about 3 logs decrease of viral load in lungs. These results have demonstrated that β-D-4 may be an effective anti-hRSV agent. Taken together, old male BALB/c mice might be used to establish hRSV infection model. Three siRNAs and the β-D-4 were validated as the potent anti-hRSV agents, respectively. / published_or_final_version / Microbiology / Doctoral / Doctor of Philosophy

No description available.

Thesis (D.PT.)--Sage Colleges, 2009. / "May 2009." "A Capstone project for PTY 768 presented to the Faculty of the Physical Therapy Department Sage Graduate School in partial fulfillment of the requirements for the degree of Doctor of Physical Therapy." Includes bibliographical references.

Thesis (Ph. D.)--Ohio State University, 2006. / Available online via OhioLINK's ETD Center; full text release delayed at author's request until 2007 Mar 10