PLGA-Polymer Encapsulating Tumor Antigen and CpG DNA Administered into the Tumor Microenvironment Elicits a Systemic Antigen-Specific IFN-γ Response and Enhances Survival

Kevin P. Nikitczuk, Rene S. Schloss, Martin L. Yarmush, Edmund C. Lattime

Department of Biomedical Engineering, Rutgers University, Piscataway, USA.
The Cancer Institute of New Jersey and Departments of Surgery, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, USA.

DOI: 10.4236/jct.2013.41035                

Abstract

Critical to the generation of an effective therapeutic antitumor immune response is the elicitation of effective antigen presentation coupled with overcoming tumor-immune escape mechanisms. Towards this end, we aimed to understand the therapeutic effectiveness of a polymer based vaccine approach at enhancing the anti-tumor responses in a tumor-bearing mouse model. While we and others have previously demonstrated the effectiveness of PLGA based systems in delivering antigen etc., studies scarcely focus on understanding the immunological mechanisms of polymer based therapies in tumor bearing treatment models. Considering tumors modulate the immune system and consequently the efficacy of therapies, understanding treatment mechanisms in the presence of tumor will help lead to more efficacious treatment options. We demonstrate here that a poly(lactic-co-glycolic acid) (PLGA) based delivery system encapsulating tumor antigen (OVA) and the TLR9 agonist CpG motif DNA administered into the tumor microenvironment initiates an effective type 1 mediated (IFN-γ producing) anti-tumor response in a syngeneic murine model of T cell lymphoma (E.G7-OVA). Although E.G7-OVA tumors spontaneously generate antigen specific CTLs in draining lymph nodes (LN), tumors progress rapidly. Modulation of the tumor microenvironment via local PLGA based therapy led to the generation of a systemic antigen specific Th1 response, absent in the non-polymer delivery method, subsequently associated with reduced tumor growth and prolongation of survival. These studies provide further insight into the use of a PLGA-based therapeutic approach at modulating the tumor microenvironment and highlight the need for analyzing the treatment effects in a tumor bearing model.

Keywords

Cytokine; Immune Response; Immunomodulation; Macrophage; Microencapsulation; Vaccine

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	G. Raimondi, M. S. Turner, A. W. Thomson and P. A. Morel, “Naturally Occurring Regulatory T Cells: Recent Insights in Health and Disease,” Critical Reviews in Immunology, Vol. 27, No. 1, 2007, pp. 61-95.
[2]	P. Neeson and Y. Paterson, “Effects of the Tumor Microenvironment on the Efficacy of Tumor Immunotherapy,” Immunological Investigations, Vol. 35, No. , 2006, pp. 359-394.
[3]	J. R. Wilczynski, M. Radwan and J. Kalinka, “The Characterization and Role of Regulatory T Cells in Immune Reactions,” Frontiers in Bioscience, Vol. 13, No. 6, 2008, pp. 2266-2274.
[4]	A. F. Ochsenbein, “Principles of Tumor Immunosurveillance and Implications for Immunotherapy,” Cancer Gene Therapy, Vol. 9, No. 12, 2002, pp. 1043-1055.
[5]	A. S. Yang and E. C. Lattime, “Tumor-Induced Interleukin 10 Suppresses the Ability of Splenic Dendritic Cells to Stimulate CD4 and CD8 T-Cell Responses,” Cancer Research, Vol. 63, No. 9, 2003, pp. 2150-2157.
[6]	M. O. Lasaro and H. C. Ertl, “Targeting Inhibitory Pathways in Cancer Immunotherapy,” Current Opinion in Immunology, Vol. 22, No. 3, 2010, pp. 385-390.
[7]	T. L. Whiteside, “Inhibiting the Inhibitors: Evaluating Agents Targeting Cancer Immunosuppression,” Expert Opinion on Biological Therapy, Vol. 10, No. 7, 2010, pp. 1019-1035.
[8]	A. S. Yang, C. E. Monken and E. C. Lattime, “Intratumoral Vaccination with Vaccinia-Expressed Tumor Antigen and Granulocyte Macrophage Colony-Stimulating Factor Overcomes Immunological Ignorance to Tumor Antigen,” Cancer Research, Vol. 63, No. 20, 2003, pp. 6956-6961.
[9]	R. Audran, K. Peter, J. Dannull, et al., “Encapsulation of Peptides in Biodegradable Microspheres Prolongs Their MHC Class-I Presentation by Dendritic Cells and Macrophages in Vitro,” Vaccine, Vol. 21, No. 11-12, 2003, 1250-1255.
[10]	H. Shen, A. L. Ackerman, V. Cody, et al., “Enhanced and Prolonged Cross-Presentation Following Endosomal Escape of Exogenous Antigens Encapsulated in Biodegradable Nanoparticles,” Immunology, Vol. 117, No. 1, 2006, pp. 78-88.
[11]	S. Hamdy, A. Haddadi, R. W. Hung and A. Lavasanifar, “Targeting Dendritic Cells with Nano-Particulate PLGA Cancer Vaccine Formulations,” Advanced Drug Delivery Reviews, Vol. 63, no. 10-11, 2011, pp. 943-955.
[12]	Y. Waeckerle-Men, E. U. Allmen, B. Gander, et al., “Encapsulation of Proteins and Peptides into Biodegradable Poly(D,L-lactide-co-glycolide) Microspheres Prolongs and Enhances Antigen Presentation by Human Dendritic Cells,” Vaccine, Vol. 24, No. 11, 2006, pp. 1847-1857.
[13]	K. Nikitczuk, E. Lattime, R. Schloss and M. Yarmush, “Analysis of Dendritic Cell Stimulation Utilizing a Multi-Faceted Nanopolymer Deliver System and the Immune Modulator 1-Methyl Tryptophan,” Nano LIFE, Vol. 1, No. 3, 2010, pp. 1-12.
[14]	A. Heit, F. Schmitz, T. Haas, et al., “Antigen Co-Encapsulated with Adjuvants Efficiently Drive Protective T Cell Immunity,” European Journal of Immunology, Vol. 37, No. 8, 2007, pp. 2063-2074.
[15]	M. Diwan, P. Elamanchili, H. Lane, et al. “Biodegradable Nanoparticle Mediated Antigen Delivery to Human Cord Blood Derived Dendritic Cells for Induction of Primary T Cell Responses,” Journal of Drug Targeting, Vol. 11, No. 8-10, 2003, pp. 495-507.
[16]	P. Elamanchili, C. M. Lutsiak, S. Hamdy, et al., “Pathogen-Mimicking Nanoparticles for Vaccine Delivery to Dendritic Cells,” Journal of Immunotherapy, Vol. 30, No. 4, 2007, pp. 378-395.
[17]	M. W. Moore, F. R. Carbone and M. J. Bevan, “Introduction of Soluble Protein into the Class I Pathway of Antigen Processing and Presentation,” Cell, Vol. 54, No. 6, 1988, pp. 777-785.
[18]	K. D. Newman, P. Elamanchili, G. S. Kwon and J. Samuel, “Uptake of Poly(D,L-lactic-co-glycolic acid) Microspheres by Antigen-Presenting Cells in Vivo,” Journal of Biomedical Materials Research, Vol. 60, No. 3, 2002, pp. 480-486.
[19]	A. M. Krieg, “Therapeutic Potential of Toll-Like Receptor 9 Activation,” Nature Reviews. Drug Discovery, Vol. 5, No. 6, 2006, pp. 471-484.
[20]	A. M. Krieg, “CpG Motifs in Bacterial DNA and Their Immune Effects,” Annual Review of Immunology, Vol. 20, 2002, pp. 709-760.
[21]	A. M. Krieg, “Development of TLR9 Agonists for Cancer Therapy,” The Journal of Clinical Investigation, Vol. 117, No. 5, 2007, pp. 1184-1194.
[22]	M. Mueller, E. Schlosser, B. Gander and M. Groettrup, “Tumor Eradication by Immunotherapy with Biodegradable PLGA Microspheres—An Alternative to Incomplete Freund ’s adjuvant,” International Journal of Cancer, Vol. 129, No. 2, 2011, pp. 407-416.
[23]	T. Sato, P. McCue, K. Masuoka, et al. “Interleukin 10 Production by Human Melanoma,” Clinical Cancer Research: An Official Journal of the American Association for Cancer Research, Vol. 2, No. 8, 1996; 1383-1390.
[24]	B. K. Halak, H. C. Maguire Jr. and E. C. Lattime, “Tumor-Induced Interleukin-10 Inhibits Type 1 Immune Responses Directed at a Tumor Antigen as Well as a Non-Tumor Antigen Present at the Tumor Site,” Cancer Research, Vol. 59, No. 4, 1999; 911-917.
[25]	T. F. Gajewski, Y. Meng and H. Harlin, “Immune Suppression in the Tumor Microenvironment,” Journal of Immunotherapy, Vol. 29, No. , 2006, pp. 233-240.
[26]	D. I. Gabrilovich, H. L. Chen, K. R. Girgis et al., “Production of Vascular Endothelial Growth Factor by Human Tumors Inhibits the Functional Maturation of Dendritic Cells,” Nature Medicine, Vol. 2, No. 10, 1996, pp. 1096-1103.
[27]	V. C. Liu, L. Y. Wong, T. Jang, et al., “Tumor Evasion of the Immune System by Converting CD4+CD25-T Cells into CD4+CD25+T Regulatory Cells: Role of Tumor-Derived TGF-Beta,” Journal of Immunology, Vol. 178, No. 5, 2007, pp. 2883-2892.
[28]	K. A. Athanasiou, G. G. Niederauer and C. M. Agrawal, “Sterilization, Toxicity, Biocompatibility and Clinical Applications of Polylactic Acid/Polyglycolic Acid Copolymers,” Biomaterials, Vol. 17, No. 2, 1996, pp. 93-102.
[29]	J. Tel, A. J. Lambeck, L. J. Cruz, et al., “Human Plasmacytoid Dendritic Cells Phagocytose, Process, and Present Exogenous Particulate Antigen,” Journal of Immunology, Vol. 184, No. 8, 2010, pp. 4276-4283.
[30]	W. Ma, T. Smith, V. Bogin, et al., “Enhanced Presentation of MHC Class Ia, Ib and Class II-Restricted Peptides Encapsulated in Biodegradable Nanoparticles: A Promising Strategy for Tumor Immunotherapy,” Journal of Translational Medicine, Vol. 9, 2011, pp. 34.
[31]	P. Elamanchili, M. Diwan, M. Cao and J. Samuel, “Characterization of Poly(D,L-lactic-co-glycolic acid) Based Nanoparticulate System for Enhanced Delivery of Antigens to Dendritic Cells,” Vaccine, Vol. 22, No. 19, 2004, pp. 2406-2412.
[32]	Z. Zhang, S. Tongchusak, Y. Mizukami, et al., “Induction of Anti-Tumor Cytotoxic T Cell Responses through PLGA-Nanoparticle Mediated Antigen Delivery,” Biomaterials, Vol. 32, No. 14, 2011, pp. 3666-3678.
[33]	D. T. O’Hagan, H. Jeffery and S. S. Davis, “Long-Term Antibody Responses in Mice Following Subcutaneous Immunization with Ovalbumin Entrapped in Biodegradable Microparticles,” Vaccine, Vol. 11, No. 9, 1993, pp. 965-969.
[34]	Y. Men, C. Thomasin, H. P. Merkle, et al., “A Single Administration of Tetanus Toxoid in Biodegradable Microspheres Elicits T cell and Antibody Responses Similar or Superior to Those Obtained with Aluminum Hydroxide,” Vaccine, Vol. 13, No. 7, 1995, pp. 683-689.
[35]	H. C. Ertl, I. Varga, Z. Q. Xiang, et al., “Poly(DL-lac-tide-co-glycolide) Microspheres as Carriers for Peptide Vaccines,” Vaccine, Vol. 14, No. 9, 1996, pp. 879-885.
[36]	K. J. Maloy, A. M. Donachie, D. T. O’Hagan and A. M. Mowat, “Induction of Mucosal and Systemic Immune Responses by Immunization with Ovalbumin Entrapped in Poly(lactide-co-glycolide) Microparticles,” Immunology, Vol. 81, No. 4, 1994, pp. 661-667.
[37]	A. Moore, P. McGuirk, S. Adams et al., “Immunization with a Soluble Recombinant HIV Protein Entrapped in Biodegradable Microparticles Induces HIV-Specific CD8+ Cytotoxic T Lymphocytes and CD4+ Th1 Cells,” Vaccine, Vol. 13, No. 18, 1995, pp. 1741-1749.
[38]	R. Kennedy and E. Celis, “Multiple Roles for CD4+ T Cells in Anti-Tumor Immune Responses,” Immunological Reviews, Vol. 222, 2008, pp. 129-144.
[39]	Z. Qin, J. Schwartzkopff, F. Pradera, et al., “A Critical Requirement of Interferon Gamma-Mediated Angiostasis for Tumor Rejection by CD8+T Cells,” Cancer Research, Vol. 63, No. 14, 2003, pp. 4095-4100.
[40]	G. L. Beatty and Y. Paterson, “Regulation of Tumor Growth by IFN-Gamma in Cancer Immunotherapy,” Immunologic Research, Vol. 24, No. 2, 2001, pp. 201-210.
[41]	W. H. Schmiegel, J. Caesar, H. Kalthoff, et al., “Antiproliferative Effects Exerted by Recombinant Human Tumor Necrosis Factor-Alpha (TNF-Alpha) and Interferon-Gamma (IFN-Gamma) on Human Pancreatic Tumor Cell Lines,” Pancreas, Vol. 3, No. 2, 1988, pp. 180-188.
[42]	Y. Waeckerle-Men and M. Groettrup, “PLGA Microspheres for Improved Antigen Delivery to Dendritic Cells as Cellular Vaccines,” Advanced Drug Delivery Reviews, Vol. 57, No. 3, 2005, pp. 475-482.
[43]	J. E. Eyles, Z. C. Carpenter, H. O. Alpar, E. D. Williamson, “Immunological Aspects of Polymer Microsphere Vaccine Delivery Systems,” Journal of Drug Targeting, Vol. 11, No. 8-10, 2003, pp. 509-514.