Date of Degree

2011

Document Type

PhD diss.

Degree Name

PhD (Doctor of Philosophy)

Department

Pharmacy

First Advisor

Aliasger K. Salem

Abstract

Cancer is a collection of diseases that is characterized by an uncontrolled proliferation of aberrant cells. The conventional treatment strategies including chemotherapy and radiotherapy prove detrimental to healthy cells and are unable to combat primary and secondary metastases. Hence immunotherapy is gaining momentum in cancer clinics and translational research labs as an alternative safer and more efficacious approach to tumor therapy. Vaccines based on antigen alone lack the ability to optimally activate the antigen presenting cells (APCs) including dendritic cells (DCs) to generate significantly greater antigen-specific T cell responses. A typical strategy to overcome this limitation has been the use of adjuvants in order to improve the immunogenicity of the vaccines. A newer class of adjuvants called toll like receptor (TLR) ligands recognize pathogen associated molecular patterns (PAMP) and thus elicit a Th-1 polarized response. The hypothesis of the current research is that co-delivery of antigen and adjuvant in a microparticulate carrier will elicit a strong cell-mediated immune response conferring anti-tumor immunity. Furthermore, we proposed that a combination of TLR ligands when co-delivered with an antigen will be able to mount a synergistic anti-tumor response in comparison to the delivery of antigen alone. There were three primary objectives of the study; 1) Fabrication, process design optimization and characterization of antigen and adjuvant co-loaded microparticles (MP), 2) Study the ability of the MP fabricated in step 1 in eliciting a potent immune response in a murine model by various routes of administration and 3) Study the effect of the various treatment as prophylactic and therapeutic cancer vaccines in a murine tumor model.

Objective 1 was achieved by optimizing various process parameters including PLGA type and concentration, surfactant concentration and modification of a previously reported double emulsification solvent evaporation technique. Process optimization lead to the development of the following 5 treatment groups, a) Empty PLGA MP, b) OVA PLGA MP, c) OVA + CpG PLGA MP, d) OVA + poly I: C PLGA MP and e) OVA + CpG + poly I:C tri-component PLGA MP. The optimized microparticles exhibited a mean particle size of 1.5 πm with a smooth spherical surface and a desirable biphasic release pattern of the entrapped components. Results from step 2 indicated the ability of the co-delivery and the tri-component systems to generate several fold higher immunostimulatory response in comparison to the group treated with antigen alone. Of the two routes of administration tested, the intraperitonial (i.p) route gave the strongest response followed by intramuscular (i.m) route. A synergistic antibody response response was observed upon vaccination with the tri-component model. Prophylactic vaccination studies showed that the co-delivery and tri-component system affording the strongest protection against aggressive tumor growth. The therapeutic studies also indicated enhanced tumor protection and 2-fold improvement in survival times when the mice vaccinated with the co-delivery and tri-component MP carriers in comparison to vaccination with microparticles loaded with antigen alone. The carriers also showed strong evidence for the generation of anti-tumor immunogenic memory. In summary, the current study has laid an interesting premise for the use of immunotherapy using a combination of antigen and TLR adjuvants alone or in conjunction with chemotherapy as a treatment strategy for tumor therapy. This research is expected to lead to the utilization of PLGA delivery systems as novel carriers for cancer immunotherapy.

Pages

xx, 193

Bibliography

182-193

Copyright

Copyright 2011 Yogita Krishnamachari