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Am. J. Trop. Med. Hyg., 78(3), 2008, pp. 468-471
Copyright © 2008 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT


Miniaturization of a High-throughput pLDH-based Plasmodium falciparum Growth Inhibition Assay for Small Volume Samples from Preclinical and Clinical Vaccine Trials

Elke S. Bergmann-Leitner*, Elizabeth H. Duncan, John Robert Burge, Michele Spring, AND Evelina Angov
US Military Malaria Vaccine Program, Division of Malaria Vaccine Development, Walter Reed Army Institute of Research, Silver Spring, Maryland

 

ABSTRACT

To date, no immune correlates for blood stage–specific immunity against Plasmodium falciparum malaria parasites have been identified. Growth and/or invasion inhibition assays using sera from Phase 2a/b trials will aid in determining whether correlations with protective immunity can be established for these assays. A major constraint in the ability to evaluate functional antibody activities from populations in endemic areas is the relatively limited availability of sufficient sample quantity. For this reason, we developed a miniaturized and high-throughput method to measure growth inhibitory activity by quantification of parasite lactate dehydrogenase (pLDH) in a 384-microtiter plate format. This culture method can be extended beyond the pLDH-based readout to other techniques commonly used to determine growth/invasion inhibition.

Received November 1, 2007. Accepted for publication December 15, 2007.

Acknowledgments: The authors thank Ann V. Stewart for critical reading of the manuscript and helpful discussions.

Financial support: This work was supported by the US Agency for International Development, Project Number 936-6001, Award Number AAG-P-00-98-00006, Award Number AAG-P-00-98-00005, and the US 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. Serum samples from adults participating in the Phase 1/2a malaria vaccine trial at WRAIR (ClinicalTrials.gov identifier NCT00385047) were obtained under a human use protocol approved by the Walter Reed Army Institute of Research Human Use Review Committee and the US Army’s Human Subjects Research and Review Board of the Surgeon General. All volunteers gave informed consent before screening and enrollment into the study, and serum samples obtained from the volunteers were labeled with an alphanumeric code to maintain anonymity. 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, 503 Robert Grant Ave. 3W76, Silver Spring, MD 20910. E-mail: elke.bergmannleitner{at}us.army.mil

Authors’ addresses: Elke S. Bergmann-Leitner, Elizabeth H. Duncan, John R. Burge, Michele Spring, and Evelina Angov, WRAIR, 503 Robert Grant Ave. 3W76, Silver Spring, MD 20910. Telephone: 301-319-9278, Fax: 301-319-7358, E-mail: elke.bergmannleitner{at}us.army.mil.






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