This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Persson, K. E. M.
Right arrow Articles by Beeson, J. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Persson, K. E. M.
Right arrow Articles by Beeson, J. G.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, May 2006, p. 1665-1673, Vol. 44, No. 5
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.5.1665-1673.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Development and Optimization of High-Throughput Methods To Measure Plasmodium falciparum-Specific Growth Inhibitory Antibodies

Kristina E. M. Persson,1 Chee T. Lee,2 Kevin Marsh,3 and James G. Beeson1,3*

The Walter and Eliza Hall Institute of Medical Research, Victoria 3050, Australia,1 Department of Medicine, University of Melbourne, Royal Melbourne Hospital, Victoria 3050, Australia,2 Centre for Geographic Medicine Research, Coast, Kenya Medical Research Institute, Kilifi, Kenya3

Received 28 November 2005/ Returned for modification 13 February 2006/ Accepted 27 February 2006

Antibodies that inhibit replication of Plasmodium falciparum in erythrocytes are thought to be important both in acquired immunity to malaria and as mediators of immunity generated by candidate blood-stage vaccines. However, several constraints have limited the study of these functional antibodies in population studies and vaccine trials. We report the development and optimization of high-throughput growth inhibition assays with improved sensitivity that use minimal volumes of test serum. The major inhibitory activity of serum from exposed donors was antibody mediated, but nonspecific inhibitory factors were found in untreated serum. Culture volumes could be effectively reduced to 25 µl to limit amounts of test serum or inhibitors used in assays. Performing inhibition assays over two cycles of parasite replication gave greater sensitivity than single-cycle assays, and a simple two-cycle inhibition assay was developed that yielded highly reproducible results. Determination of parasite growth by flow cytometry was most suitable for high-throughput assays using small culture volumes and was more sensitive than parasite lactate dehydrogenase assays and less prone to error and variation than microscopy. We evaluated and optimized methods to remove antimalarials and nonspecific inhibitory factors from serum that are suitable for use with small volumes of samples that are typically obtained from clinical studies. Both microdialysis and immunoglobulin purification by ammonium sulfate precipitation were effective and practical. These methods should facilitate evaluation of vaccine trials and clinical studies of immunity and are also suitable for testing drugs and other compounds for antimalarial activity.

* Corresponding author. Mailing address: The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3050, Australia. Phone: 61 3 9345 2555. Fax: 61 3 9347 0842. E-mail: beeson{at}wehi.edu.au.

Journal of Clinical Microbiology, May 2006, p. 1665-1673, Vol. 44, No. 5
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.5.1665-1673.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Murhandarwati, E. E. H., Wang, L., Black, C. G., Nhan, D. H., Richie, T. L., Coppel, R. L. (2009). Inhibitory Antibodies Specific for the 19-Kilodalton Fragment of Merozoite Surface Protein 1 Do Not Correlate with Delayed Appearance of Infection with Plasmodium falciparum in Semi-Immune Individuals in Vietnam. Infect. Immun. 77: 4510-4517 [Abstract] [Full Text]  
  • Kadekoppala, M., O'Donnell, R. A., Grainger, M., Crabb, B. S., Holder, A. A. (2008). Deletion of the Plasmodium falciparum Merozoite Surface Protein 7 Gene Impairs Parasite Invasion of Erythrocytes. Eukaryot Cell 7: 2123-2132 [Abstract] [Full Text]  
  • Arnot, D. E., Cavanagh, D. R., Remarque, E. J., Creasey, A. M., Sowa, M. P. K., Morgan, W. D., Holder, A. A., Longacre, S., Thomas, A. W. (2008). Comparative Testing of Six Antigen-Based Malaria Vaccine Candidates Directed Toward Merozoite-Stage Plasmodium falciparum. CVI 15: 1345-1355 [Abstract] [Full Text]  
  • Bergmann-Leitner, E. S., Duncan, E. H., Burge, J. R., Spring, M., Angov, E. (2008). Miniaturization of a High-throughput pLDH-based Plasmodium falciparum Growth Inhibition Assay for Small Volume Samples from Preclinical and Clinical Vaccine Trials. Am J Trop Med Hyg 78: 468-471 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»