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Infection and Immunity, October 2000, p. 5595-5602, Vol. 68, No. 10
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Identification of Vaccine Candidates for Experimental Visceral Leishmaniasis by Immunization with Sequential Fractions of a cDNA Expression Library

Peter C. Melby,1,2,* Gary B. Ogden,3 Hector A. Flores,1,2 Weiguo Zhao,1 Christopher Geldmacher,3,dagger Natalie M. Biediger,1,3 Sunil K. Ahuja,1,2 Jose Uranga,1,3 and Maria Melendez1,3

Medical Service, Audie L. Murphy Division, South Texas Veterans Health Care System,1 Departments of Medicine and Microbiology, The University of Texas Health Sciences Center,2 and Department of Biological Sciences, St. Mary's University,3 San Antonio, Texas

Received 24 March 2000/Returned for modification 14 June 2000/Accepted 12 July 2000

Visceral leishmaniasis caused by the intracellular parasite Leishmania donovani is a significant public health problem in many regions of the world. Because of its large genome and complex biology, developing a vaccine for this pathogen has proved to be a challenging task and, to date, protective recombinant vaccine candidates have not been identified. To tackle this difficult problem, we adopted a reductionist approach with the intention of identifying cDNA sequences in an L. donovani amastigote cDNA library that collectively or singly conferred protection against parasite challenge in a murine model of visceral leishmaniasis. We immunized BALB/c mice with plasmid DNA isolated and pooled from 15 cDNA sublibraries (~2,000 cDNAs/sublibrary). Following systemic challenge with L. donovani, mice immunized with 6 of these 15 sublibraries showed a significantly reduced (35- to 1,000-fold) hepatic parasite burden. Because of the complexity and magnitude of the sequential fractionation-immunization-challenge approach, we restricted our attention to the two sublibraries that conferred the greatest in vivo protection. From one of these two sublibraries, we identified several groups of cDNAs that afforded protection, including a set of nine novel cDNAs and, surprisingly, a group of five cDNAs that encoded L. donovani histone proteins. At each fractionation step, the cDNA sublibraries or the smaller DNA fractions that afforded in vivo protection against the parasite also induced in vitro parasite-specific T helper 1 immune responses. Our studies demonstrate that immunization with sequential fractions of a cDNA library is a powerful strategy for identifying anti-infective vaccine candidates.

* Corresponding author. Mailing address: Department of Medicine, Division of Infectious Diseases, The University of Texas Health Science Center, 7703 Floyd Curl Dr., Mailstop 7881, San Antonio, TX 78229-3900. Phone: (210) 567-4614. Fax: (210) 567-4670. E-mail: melby{at}uthscsa.edu.

dagger Present address: AG Brocker, Max Planck Institute for Immunology, 79104 Freiburg, Germany.

Infection and Immunity, October 2000, p. 5595-5602, Vol. 68, No. 10
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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