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* Division of Immunology and Genetics, John Curtin School of Medical Research, Australian National University, Canberra, Australia; and
Department of Microbiology and Immunology, University of Melbourne, Victoria, Australia
The quality of virus-specific CD8+ CTL immune responses generated by mucosal and systemic poxvirus prime-boost vaccines were evaluated in terms of T cell avidity and single-cell analysis of effector gene expression. Intranasal (I.N.) immunization regimes generated higher avidity CTL responses specific for HIV KdGag197–205 (amino acid sequence AMQMLKETI; H-2Kd binding) compared with i.m. immunization regime. Single-cell RT-PCR of KdGag197–205-specific mucosal and systemic CTL revealed that the cytokine and granzyme B expression profiles were dependent on both the route and time after immunization. The I.N./i.m.-immunized group elicited elevated number of CTL-expressing granzyme B mRNA from the genitomucosal sites compared with the i.m./i.m. regime. Interestingly, CTL generated after both I.N. or i.m. immunization demonstrated expression of Th2 cytokine IL-4 mRNA that was constitutively expressed over time, although lower numbers were observed after I.N./I.N. immunization. Results suggest that after immunization, Ag-specific CTL expression of IL-4 may be an inherent property of the highly evolved poxvirus vectors. Current observations indicate that the quality of CTL immunity generated after immunization can be influenced by the inherent property of vaccine vectors and route of vaccine delivery. A greater understanding of these factors will be crucial for the development of effective vaccines in the future.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 This work was supported by National Health and Medical Research Council Program Grant 299907 and a National Health and Medical Research Council R.D. Wright Fellowship (to S.J.T.).
2 Address correspondence and reprint requests to Dr. Charani Ranasinghe, Division of Immunology and Genetics, John Curtin School of Medical Research, Australian National University, P.O. Box 334, 2601 Canberra, Australia. E-mail address: Charani.Ranasinghe{at}anu.edu.au
3 Abbreviations used in this paper: FPV, fowlpox virus; VV, vaccinia virus; I.N., intranasal; DEPC, diethyl pyrocarbonate; CT, cycle threshold.
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