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Infection and Immunity, September 2001, p. 5565-5572, Vol. 69, No. 9
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.9.5565-5572.2001

Multistage Multiantigen Heterologous Prime Boost Vaccine for Plasmodium knowlesi Malaria Provides Partial Protection in Rhesus Macaques

William O. Rogers,1,* J. Kevin Baird,1,dagger Anita Kumar,1 John A. Tine,2,Dagger Walter Weiss,1 João C. Aguiar,1 Kalpana Gowda,1 Robert Gwadz,3 Sanjai Kumar,1 Mark Gold,4 and Stephen L. Hoffman1,§

Malaria Program, Naval Medical Research Center, Silver Spring, Maryland 209101; Virogenetics Corporation, Troy, New York 121802; and Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health,3 and Armed Forces Radiobiology Research Institute,4 Bethesda, Maryland 20889

Received 28 March 2001/Returned for modification 17 May 2001/Accepted 4 June 2001

A nonhuman primate model for malaria vaccine development allowing reliable, stringent sporozoite challenge and evaluation of both cellular and antibody responses is needed. We therefore constructed a multicomponent, multistage DNA vaccine for the simian malaria species Plasmodium knowlesi including two preerythrocytic-stage antigens, the circumsporozoite protein (PkCSP) and sporozoite surface protein 2 (PkSSP2), and two blood stage antigens, apical merozoite antigen 1 (PkAMA1) and merozoite surface protein 1 (PkMSP1p42), as well as recombinant canarypox viruses encoding the four antigens (ALVAC-4). The DNA vaccine plasmids expressed the corresponding antigens in vitro and induced antiparasite antibodies in mice. Groups of four rhesus monkeys received three doses of a mixture of the four DNA vaccine plasmids and a plasmid encoding rhesus granulocyte-monocyte colony-stimulating factor, followed by boosting with a single dose of ALVAC-4. Three groups received the priming DNA doses by different routes, either by intramuscular needle injection, by intramuscular injection with a needleless injection device, the Biojector, or by a combination of intramuscular and intradermal routes by Biojector. Animals immunized by any route developed antibody responses against sporozoites and infected erythrocytes and against a recombinant PkCSP protein, as well as gamma interferon-secreting T-cell responses against peptides from PkCSP. Following challenge with 100 P. knowlesi sporozoites, 1 of 12 experimental monkeys was completely protected and the mean parasitemia in the remaining monkeys was significantly lower than that in 4 control monkeys. This model will be important in preclinical vaccine development.

* Corresponding author. Mailing address: Malaria Program, Naval Medical Research Center, 503 Robert Grant Ave., Silver Spring, MD 20910. Phone: (301) 319-7574. Fax: (301) 319-7460. E-mail: Rogersb{at}nmrc.navy.mil.

dagger Present address: Naval Medical Research Unit 2, American Embassy Jakarta, FPO AP 96520-8132.

Dagger Present address: Center for Comparative Functional Genomics, University at Albany, State University of New York, Rensselaer, NY 12144.

§ Present address: Celera Genomics, Rockville, MD 20850.

Infection and Immunity, September 2001, p. 5565-5572, Vol. 69, No. 9
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.9.5565-5572.2001


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