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CRITICAL EVALUATION OF DIFFERENT METHODS FOR MEASURING THE FUNCTIONAL ACTIVITY OF ANTIBODIES AGAINST MALARIA BLOOD STAGE ANTIGENS

Antibodies are thought to be the primary immune effectors in the defense against erythrocytic stage Plasmodium falciparum. Thus, malaria vaccines directed to blood stages of infection are evaluated based on their ability to induce antibodies with anti-parasite activity. Such antibodies may have different effector functions (e.g., inhibition of invasion or inhibition of parasite growth/development) depending on the target antigen. We evaluated four methods with regards to their ability to differentiate between invasion and/or growth inhibitory activities of antibodies specific for two distinct blood stage antigens: AMA1 and MSP1₄₂. We conclude that antibodies induced by these vaccine candidates have different modes of action that vary not only by the antigen, but also by the strain of parasite being tested. Analysis based on parasitemia and viability was essential for defining the full range of anti-parasite activities in immune sera.

Received March 31, 2006. Accepted for publication May 9, 2006.

Acknowledgments: The authors thank Kathleen Moch and Jeffrey Snavely for culturing and providing blood stage parasites.

Financial support: This work was supported by the United States Agency for International Development, Project Number 936-6001, Award Number AAG-P-00-98-00006, Award Number AAG-P-00-98-00005, and by the United States Army Medical Research and Materiel Command.

Disclaimer: Research was conducted in compliance with the Animal Welfare Act and other federal statutes and regulations relating to animals and experiments involving animals and adheres to principles stated in the Guide for the Care and Use of Laboratory Animals, NRC Publication, 1996 edition. The authors’ views are private and are not to be construed as official policy of the Department of Defense or the US Army.

^* Address correspondence to Elke S. Bergmann-Leitner, Department of Immunology, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, MD 20910. E-mail: elke.bergmannleitner{at}us.army.mil

Authors’ addresses: Elke S. Bergmann-Leitner, Elizabeth H. Duncan, John Robert Burge, Farhat Khan, Evelina Angov, and Jeffrey A. Lyon, Department of Immunology, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, MD. Gregory E. Mullen and Carole A. Long, Malaria Vaccine Development Branch, NIAID/NIH, 12441 Parklawn Drive, Twinbrook 2, Rockville, MD 20852.

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