Document Detail

Optimization of DNA-based vaccination in cows using green fluorescent protein and protein A as a prelude to immunization against staphylococcal mastitis.
MedLine Citation:
PMID:  12741542     Owner:  NLM     Status:  MEDLINE    
Staphylococcus aureus is a contagious pathogen that often results in chronic intramammary infections in dairy cows. Current vaccine formulations are ineffective in preventing this infection. The objective of this study was to stimulate an immune response in dairy cows through injection of plasmid DNA designed to express staphylococcal Protein A in transfected cells. Intramuscular and intradermal vaccination sites were evaluated using a plasmid containing the human cytomegalovirus (CMV) promoter/enhancer directing expression of green fluorescent protein (pcDNA3/GFP). DNA was delivered by needle and syringe, or by high-, intermediate-, or low-pressure jet injections (Ped-o-Jet and LectraJet). Five cows per treatment were injected with 0.5 mg of plasmid DNA at 6, 4, and 2 wk prepartum. Serum antibody levels determined by ELISA indicated that intradermal high-pressure jet injection elicited a greater immune response compared to needle and syringe injection. Differences in antibody production among low-pressure and needle and syringe treatment groups were not significant. An expression plasmid containing the CMV promoter/enhancer driving expression of the Fc-binding domain of S. aureus Protein A was coinjected into cows by vulvamucosal vaccination using the high-pressure Ped-o-Jet. Beginning 6 wk prepartum, groups of cows (n = 5) were injected three times at 2-wk intervals with DNA in saline, DNA in aluminum phosphate adjuvant, or served as noninjected controls. A cellular immune response to Protein A was detected in 4 of 10 animals, while cellular responses to GFP were not detected. Humoral responses to Protein A were observed in 6 of 10 animals and to GFP in 2 of 10 animals. Aluminum phosphate adjuvant appeared to enhance antibody production in response to Protein A. In experiment 3, a protein boost injection of Protein A was given to six animals approximately 5 mo postpartum. Three animals were nonvaccinated controls, and three were among those stimulated to produce antibody in response to the DNA-based vaccine. These results showed that Protein A specific antibodies remained elevated as compared to nonvaccinated controls and were stimulated in response to the protein boost. However, the magnitude of the response in animals previously vaccinated with DNA was not different than that observed in the nonvaccinated controls. We have shown that a humoral and cellular immune response to abbreviated Protein A can be raised in dairy cows using intravulvamucosal jet injection of a DNA-based vaccine.
E W Carter; D E Kerr
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of dairy science     Volume:  86     ISSN:  0022-0302     ISO Abbreviation:  J. Dairy Sci.     Publication Date:  2003 Apr 
Date Detail:
Created Date:  2003-05-13     Completed Date:  2003-06-27     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985126R     Medline TA:  J Dairy Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1177-86     Citation Subset:  IM    
Department of Animal Science, University of Vermont, Burlington 05405, USA.
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MeSH Terms
Antibodies / blood
COS Cells
Gene Expression
Green Fluorescent Proteins
Immunization, Secondary
Luminescent Proteins / genetics*,  immunology
Mastitis, Bovine / microbiology*,  prevention & control*
Staphylococcal Infections / prevention & control
Staphylococcal Protein A / genetics,  immunology*
Staphylococcal Vaccines* / administration & dosage
Vaccination / methods,  veterinary
Vaccines, DNA* / administration & dosage
Reg. No./Substance:
0/Antibodies; 0/Luminescent Proteins; 0/Staphylococcal Protein A; 0/Staphylococcal Vaccines; 0/Vaccines, DNA; 147336-22-9/Green Fluorescent Proteins

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