Systemic lupus erythematosus (SLE) is a systemic autoimmune disease characterized by polyclonal B-cell activation, autoantibody production and immune complex-mediated glomerulonephritis (GN). NZB/W F1 mice spontaneously develop SLE-like symptoms and have been widely used as an animal model for SLE. Even though the etiologic cause of autoimmunity in both human and murine lupus is not clearly understood, mounting evidence indicates the involvement of autoreactive B cells and T cells. Blockade of costimulatory pathways using CTLA4Ig fusion protein and anti-CD40 ligand monoclonal antibody (mAb) has been able to suppress autoantibody production and inhibit lupus nephritis in NZB/W F1 mice. In spite of these successes, protein- or mAb-based therapies are expensive, deliveries are problematic, and bolus injections result in transient and toxic levels of these reagents. In comparison, gene therapy offers an advantageous solution by achieving long-term, stable transgene expression and convenient gene delivery. Among the vectors used in gene therapy, recombinant adeno-associated virus (rAAV) has been proven to be a promising one because of its non-pathogenicity and non-immunogenicity.
In this project, the hypothesis that rAAV serotype 8 (rAAV8)-mediated immunomodulator gene transfer was able to prevent and suppress lupus development in NZB/W F1 mice was tested. First, the transgene expression pattern following systemic delivery was studied. rAAV8-mediated gene delivery was able to achieve long-lasting and high-level transgene expression following a single intraperitoneal or intrasplenic injection. Second, rAAV8-mediated costimulatory blockade gene transfer prevented autoantibody production, delayed proteinuria onset, prolonged survival and protected kidneys from immune-complex induced tissue damage. Moreover, it successfully suppressed CD4+ T cell activation and also the transition from naïve to memory T cells. Among all early prevention strategies tested in newborn mice, the combination of murine CTLA4Ig (mCTLA4Ig) and murine CD40Ig (mCD40Ig) was more efficient than both single vectors, suggesting a synergistic effect between these two costimulatory blockades. Surprisingly, the late prevention in young lupus-prone mice achieved results similar to those observed in the early prevention. Furthermore, rAAV-mediated costimulatory blockade gene transfer did not suppress host humoral response to foreign antigens. Finally, in the treatment of lupus mice with high autoantibody titers, mCTLA4Ig alone, or in combination with mCD40Ig, was able to decrease autoantibody titers, delay proteinuria onset, and prolong survival. The therapeutic effects were evidenced to be dose-dependent. In general, this study indicates that rAAV8-mediated costimulatory blockade gene transfer is capable of preventing and suppressing lupus development, thereby suggesting the successful application of this autoimmune disease gene therapy both in research and in clinical settings to promote public health.
| Identifer | oai:union.ndltd.org:PITT/oai:PITTETD:etd-02242005-151614 |
| Date | 23 March 2005 |
| Creators | Ye, Xiaojing |
| Contributors | Paula Clemens, Xiao Xiao, Paul D. Robbins, M.Ilyas Kamboh |
| Publisher | University of Pittsburgh |
| Source Sets | University of Pittsburgh |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.pitt.edu/ETD/available/etd-02242005-151614/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Pittsburgh or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
Page generated in 0.0023 seconds