Bovine Mammary Immune Response to an Experimental Intramammary Infection with a Staphylococcus aureus Strain Containing a Gene for Staphylococcal Enterotoxin C1

¹ Department of Animal Sciences, Washington State University, Pullman 99164
² Department of Veterinary Clinical Science, Washington State University, Pullman 99164
³ Department of Microbiology, Molecular Biology, and Biochemistry, University of Idaho, Moscow 83844
⁴ Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164

Staphylococcal enterotoxin C (SEC), a superantigen, is the most frequently expressed enterotoxin by bovine strains of Staphylococcus aureus causing mastitis. To examine the possible impact of SEC on the immune response of the bovine mammary gland, we monitored changes in lymphocyte subpopulations in mammary glands of four lactating cows after intramammary instillation of S. aureus strain Rn4220 transformed with a plasmid containing a gene coding for SEC1. Four other lactating cows received the same strain transformed with the plasmid without the SEC1 gene (positive control), and four cows were untreated (negative control). Mammary quarter milk samples for somatic cell count (SCC) analysis and determination of N-acetyl-ß-D-glucosimindase (NAGase) activity levels were collected daily for 21 d postinstillation. Flow cytometry utilizing three-color analysis was used to phenotype lymphocyte subpopulations isolated from milk samples collected on d 0, 4, 7, 11, 14, 18, and 21 postinstillation from all the cows. Milk from mammary gland halves (positive control and experimental) or all mammary quarters (negative control) was collected for flow cytometric analysis. Increased NAGase activity, SCC, and isolated S. aureus demonstrated that infection was established in mammary quarters intrammarily instilled with bacteria. There were no significant differences (P > 0.05) in the proportions of BoCD4 helper T lymphocytes or BoCD8 cytotoxic T lymphocytes between the two infected treatment groups. There was a significant day x treatment difference of the proportion of a T cell subpopulation that did not express BoCD2, but did express the ACT2 activation molecule and a significant treatment difference of a T cell subpopulation that expressed BoCD2, but not the ACT2 activation molecule (P < 0.05). Results do not support the hypothesis that the presence of the gene for SEC1 alters the mammary BoCD4 or BoCD8 T lymphocyte response to infection.

Key Words: Staphylococcus aureus • mastitis • staphylococcal enterotoxin C • superantigen • T lymphocytes • lymphocytes

Submitted on May 15, 2000
Accepted on April 19, 2001