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Preliminary analysis of murine cytotoxic T cell responses to the proteins of the flavivirus Kunjin using vaccinia virus expression

Colin R. Parrish^1,

Ann Hill^2,>

¹ Department of Microbiology, Monash University, Clayton, Victoria 3168
and² Division of Cell Biology, John Curtin School of Medical Research, Australian National University, G.P.O. Box 334, Canberra, ACT 2601, Australia

A series of recombinant vaccinia viruses expressing various parts of the entire Kunjin virus (KUN) coding region was used to analyse the cytotoxic T (Tc) cell responses to KUN. CBA/H mice inoculated with KUN or West Nile virus were shown to develop responses to KUN or various vaccinia virus expression constructs in either primary cytotoxic assays, or after secondary stimulation of the Tc cells in vitro with KUN antigens. Tc cells from CBA mice showed the strongest response to target cells infected with recombinant vaccinia viruses expressing parts of the KUN NS3 and NS4A proteins, and only a weak response to the other structural or non-structural proteins. Further analysis of deleted versions of the NS3-NS4A region showed that the main epitope recognized was derived from a sequence of 99 amino acids spanning parts of NS3 and NS4A. No other major epitopes were detected by Tc cells from CBA mice in the remaining 3333 amino acids of the KUN polypeptide.

Present address: J. A. Baker Institute, New York State College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, U.S.A.

^> Present address: Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford OX3 9DU, U.K.

Received 12 November 1990; accepted 11 March 1991.

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