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Infection and Immunity, August 2004, p. 4464-4470, Vol. 72, No. 8
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.8.4464-4470.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Production of the Subdomains of the Plasmodium falciparum Apical Membrane Antigen 1 Ectodomain and Analysis of the Immune Response

P. V. Lalitha,^1, Lisa A. Ware,¹ Arnoldo Barbosa,¹ Sheetij Dutta,¹ J. Kathleen Moch,² J. David Haynes,¹ Bader B. Fileta,³ Charles E. White,⁴ and David E. Lanar^1*

Department of Immunology,¹ Department of Biometrics,⁴ Walter Reed Army Institute of Research, and Naval Medical Research Center, Silver Spring, Maryland 20910-7500,² Department of Clinical Investigations, Walter Reed Army Medical Center, Washington, DC 20307³

Received 1 October 2003/ Returned for modification 29 November 2003/ Accepted 30 March 2004

The apical membrane antigen 1 of Plasmodium falciparum is one of the leading candidate antigens being developed as a vaccine to prevent malaria. This merozoite transmembrane protein has an ectodomain that can be divided into three subdomains (D I, D II, and D III). We have previously expressed a major portion of this ectodomain and have shown that it can induce antibodies that prevent merozoite invasion into red blood cells in an in vitro growth and invasion assay. To analyze the antibody responses directed against the individual subdomains, we constructed six different genes that express each of the domains separately (D I, D II, or D III) or in combination with another domain (D I+II, D II+III, or D I+III). These proteins were purified and used to immunize rabbits to raise construct-specific antibodies. We demonstrated that D I+II induced a significant amount of the growth-inhibitory antibodies active in the growth and invasion assay.

Editor: W. A. Petri, Jr.

Present address: School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India.

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