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Vaccination strategies for the treat...

Conwell, Julie Victoria.

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Vaccination strategies for the treatment of acute infection by Pseudomonas aeruginosa.

紀錄類型:	書目-語言資料,印刷品 : Monograph/item
正題名/作者:	Vaccination strategies for the treatment of acute infection by Pseudomonas aeruginosa./
作者:	Conwell, Julie Victoria.
面頁冊數:	126 p.
附註:	Adviser: Neil R. Baker.
Contained By:	Dissertation Abstracts International63-07B.
標題:	Biology, Microbiology. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3059226
ISBN:	0493747591

Vaccination strategies for the treatment of acute infection by Pseudomonas aeruginosa.
Conwell, Julie Victoria.

Vaccination strategies for the treatment of acute infection by Pseudomonas aeruginosa. - 126 p.

Adviser: Neil R. Baker.

Thesis (Ph.D.)--The Ohio State University, 2002.

The focus of this research is the development of a vaccine for the prevention of acute infection by <italic>Pseudomonas aeruginosa</italic>. A DNA vaccine was developed using the flagellin protein as the target antigen. The gene encoding flagellin was cloned into the pVR1020 expression vector and injected into mice using the intramuscular and gene gun routes of injection. Significant antibody levels to the protein were seen after injections on days 0 and 14 regardless of route of injection. Mice immunized using the gene gun were protected by this vaccine in the burn mouse model but mice immunized intramuscularly were not protected. The antibodies generated from both routes of injection could inhibit motility and the gene gun antibodies were able to opsonize and enhance phagocytosis of <italic>P. aeruginosa</italic> by neutrophils.

ISBN: 0493747591Subjects--Topical Terms:

1017734
Biology, Microbiology.

Vaccination strategies for the treatment of acute infection by Pseudomonas aeruginosa.
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245 1 0 $a Vaccination strategies for the treatment of acute infection by Pseudomonas aeruginosa.
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502 $a Thesis (Ph.D.)--The Ohio State University, 2002.
520 $a The focus of this research is the development of a vaccine for the prevention of acute infection by <italic>Pseudomonas aeruginosa</italic>. A DNA vaccine was developed using the flagellin protein as the target antigen. The gene encoding flagellin was cloned into the pVR1020 expression vector and injected into mice using the intramuscular and gene gun routes of injection. Significant antibody levels to the protein were seen after injections on days 0 and 14 regardless of route of injection. Mice immunized using the gene gun were protected by this vaccine in the burn mouse model but mice immunized intramuscularly were not protected. The antibodies generated from both routes of injection could inhibit motility and the gene gun antibodies were able to opsonize and enhance phagocytosis of <italic>P. aeruginosa</italic> by neutrophils.
520 $a The protective epitopes of the flagellin protein were determined. Trypsin digests, cloning/bacterial expression, phage display and <italic>in vitro </italic> transcription/translation were used to show that a protective monoclonal antibody corresponded to a conformational epitope. Peptides predicted to be exposed on the surface of the flagellin protein were synthesized and used to immunize mice. The generated antibodies were able to recognize the synthesized peptides, but not the native protein, suggesting that the protein folds in a manner that creates conformational epitopes.
520 $a In order to generate a rapid antibody response, flagellin was targeted to dendritic cells. The N418 monoclonal antibody specific for the CD11c surface receptor on dendritic cells was chemically cross-linked to flagellin. A rapid antibody response was generated by day seven in animals injected with the conjugate. The antibodies were mainly of the IgGI isotype and could inhibit the motility of <italic>P. aeruginosa</italic>. The ability of the conjugate to bind to cultured dendritic cells was demonstrated by immunofluorescence and flow cytometry. It was also shown that the conjugate and N418 monoclonal antibody alone can protect animals against lethal infection by <italic>P. aeruginosa</italic> in the burn mouse model, suggesting that the dendritic cells are stimulated when the antibody binds to its surface receptor and this causes a generalized enhancement of the immune response that is protective against <italic>P. aeruginosa</italic>, and perhaps other pathogens.
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