Thesis
In vitro evaluation of the biological responses of canine macrophages challenged with poly (lactic-co-glycolic acid) nanoparticles containing monophosphoryl lipid A
Washington State University
Master of Science (MS), Washington State University
2016
Handle:
https://hdl.handle.net/2376/102201
Abstract
Poly (lactic-co-glycolic acid) nanoparticles have been considerably studied as a promising biodegradable delivery system to induce effective immune responses and to improve stability, safety, and cost effectiveness of vaccines and therapeutic drugs. The current study aims at evaluating the immunological effects of poly (lactic-co-glycolic acid) nanoparticles, co-encapsulating ovalbumin, and the adjuvant monophosphoryl lipid A on canine macrophages in vitro. The different nanoparticle formulations were prepared following the emulsion-solvent evaporation technique. Peripheral blood mononuclear cells were isolated from canine whole blood by magnetic separation and further cultured to generate macrophages. The uptake of nanoparticle constructs by macrophages in vitro was demonstrated by transmission electron microscopy and confocal microscopy. Macrophage viability and morphology were evaluated by trypan blue exclusion and light microscopy. The mRNA expression of Interleukin-10, Interleukin-12, and tumor necrosis factor alpha by macrophages was determined by quantitative real time polymerase chain reaction. The mean size of the nanoparticles was 207.14 nm (SD ±158.74 nm). An appropriate yield of monocyte-derived macrophages was obtained. Incubation of nanoparticles with macrophages demonstrated effective uptake and macrophage viability was not altered. Poly (lactic-co-glycolic acid) nanoparticles containing the model-antigen ovalbumin and the adjuvant monophosphoryl lipid A strongly induced expression of pro-inflammatory and pro-immune cytokines by macrophages. Therefore, the efficient nanoparticle internalization, lack of toxic effects, and stimulation of pro-inflammatory and pro-immune cytokine expression, indicate that poly (lactic-co-glycolic acid) nanoparticles containing the model-antigen ovalbumin with addition of monophosphoryl lipid A represent a good model for further development of antigen carriers capable of stimulating effective immune responses in dogs such as in cancer immunotherapy.
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Details
- Title
- In vitro evaluation of the biological responses of canine macrophages challenged with poly (lactic-co-glycolic acid) nanoparticles containing monophosphoryl lipid A
- Creators
- Delphine Guldner
- Contributors
- Cleverson D. Souza (Degree Supervisor)
- Awarding Institution
- Washington State University
- Academic Unit
- Veterinary Medicine, College of
- Theses and Dissertations
- Master of Science (MS), Washington State University
- Publisher
- Washington State University; [Pullman, Washington] :
- Identifiers
- 99900525196801842
- Language
- English
- Resource Type
- Thesis