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Characteristics of ovine cytotoxic lymphocytes

Thesis/Dissertation ·
OSTI ID:6859756
Experiments were conducted to examine characteristics of the effector cells responsible for cell-mediated cytotoxicity in the sheep. Conditions for the production and assay of ovine T cell growth factor (TCGF) activity were evaluated. Peripheral blood leukocytes (PBL) were stimulated with concanavalin A (Con A) in the presence of 2% autologous serum or serum-free media. A 28 h proliferation assay with 2.5 x 10/sup 4/ h Con A blasts per well was optimal for detection of TCGF. Peak TCGF activity occurred with a 30-37kD molecular weight fraction. Ovine PBL were used for in vitro generation of genetically-restricted cytotoxic T lymphocytes (CTL). Peripheral blood leukocytes from sheep that had been previously inoculated with live vaccinia virus were stimulated by being cultured in vitro on glutaraldehyde-fixed vaccinia-infected autologous skin fibroblasts. Cytotoxic T lymphocyte activity was assessed in a 6 h /sup 51/Cr-release assay on autologous and allogeneic fibroblasts targets. Killing was restricted to virus-infected autologous targets. In vitro generation of both anti-vaccinia and anti-TNP CTL activity could be enhanced by the addition of TCGF containing media from ConA-stimulated PBL.
Research Organization:
Colorado State Univ., Fort Collins (USA)
OSTI ID:
6859756
Country of Publication:
United States
Language:
English