Development of novel Hepatitis C Virus DNA vaccines.

Format:

Book

Thesis/Dissertation

Author/Creator:

Kuhs, Krystle A. Lang.

Contributor:

University of Pennsylvania.

Language:

English

Subjects (All):

Immunology.

0982.

Local Subjects:

0982.

Physical Description:

132 pages

Contained In:

Dissertation Abstracts International 72-09B.

System Details:

Mode of access: World Wide Web.

text file

Summary:

Hepatitis C Virus (HCV) is a global pandemic affecting over 170 million individuals worldwide. Currently there is no vaccine for HCV however it is believed that an effective vaccine strategy must be able to induce strong intrahepatic HCV-specific T cell-based immunity. In this thesis, we have taken a multi-step approach to develop four distinct HCV DNA vaccines encoding consensus genotype 1 immunogens of HCV proteins, NS3/NS4A, NS4B, NS5A and NS5B. We show that these constructs are expressed in vitro and are able to induce potent peripheral HCV-specific IFN-gamma+ T cell responses in C57BL/6 mice. In order to test the ability of immunization to induce functional HCV-specific T cell-based immunity in the liver, a small animal model of HCV infection was developed. In this model, we show that we are able to induce liver selective expression of HCV viral proteins in C57BL/6 mice and that liver expression of HCV NS3 is able to mimic the immunological affects of HCV infection as evidenced by liver inflammation, upregulation of liver chemokine expression, splenomegaly and recruitment of T cells to the liver. Using this model, we have shown the all four constructs are able to induce potent HCV-specific T cell-based immunity within the liver both in the presence and absence of infection as shown by upregulation of IFN-gamma and clearance of HCV antigen expressing hepatocytes. While highly immunogenic in mice, given the both the immunological differences between mice and humans, as well as, the reported decrease in immunogenicity of DNA vaccines when moving into larger animal models, one HCV DNA vaccine construct was further tested in the non-human primate, rhesus macaque model. In this model, immunization was shown to induce HCV-specific T cell-based immunity strikingly similar to that seen in patients with resolving HCV infection and included strong proliferative responses, polyfunctional T cells and broadly reactive T cell responses able to recognize multiple epitopes spanning the length of the NS3 protein. Taken together our findings suggest that this type of immunization strategy merits further study in the context of future prophylactic HCV T cell-based vaccines.

Notes:

Source: Dissertation Abstracts International, Volume: 72-09, Section: B, page: 5205.

Adviser: David B. Weiner.

Thesis (Ph.D.)--University of Pennsylvania, 2011.

Local Notes:

School code: 0175.

ISBN:

9781124732824

Access Restriction:

Restricted for use by site license.