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Infection and Immunity, August 2004, p. 4707-4715, Vol. 72, No. 8
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.8.4707-4715.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Novel Phage Display-Based Subtractive Screening To Identify Vaccine Candidates of Brugia malayi

Munirathinam Gnanasekar,1,{dagger} Kakaturu V. N. Rao,1,{dagger},{ddagger} Yi-Xun He,1 Pankaj K. Mishra,2 Thomas B. Nutman,3 Perumal Kaliraj,2 and Kalyanasundaram Ramaswamy1*

Department of Biomedical Sciences, University of Illinois College of Medicine, Rockford, Illinois 61107;,1 Center for Biotechnology, Anna University, Chennai-600 025, India,2 Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 208923

Received 10 September 2003/ Returned for modification 24 January 2004/ Accepted 13 April 2004

This study describes a novel phage display method based on an iterative subtraction strategy to identify candidate vaccine antigens of Brugia malayi. A cDNA library of the infective larval stage of B. malayi expressed on the surface of T7 phage was sequentially screened with sera samples from human subjects showing different manifestations of the disease. Antigens that selectively and specifically bind to immune sera were then enriched using a multi-step panning procedure. This strategy identified five antigens, four of which were previously reported (ALT-2, TPX-2, VAH and COX-2) and the other one was a novel cuticular collagen (Col-4). Sera from immune individuals specifically recognized all the five antigens. However, ALT-2 appeared to be the most predominantly recognized antigen by the immune sera. Therefore, it was decided to evaluate the vaccine potential of recombinant ALT-2 (rALT-2) in a mouse and jird model. The results presented show that immunization with rALT-2 conferred over 73% protection against a challenge infection in the jird model and over 64% protection in the mouse model. The present study suggests that phage display-based cDNA screening may be a powerful tool to identify candidate vaccine antigens of infectious agents.

* Corresponding author. Mailing address: Department of Biomedical Sciences, College of Medicine, University of Illinois, Rockford, IL 61107. Phone: (815) 395-5696. Fax: (815) 395-5666. E-mail: ramswamy{at}uic.edu.

Editor: J. F. Urban, Jr.

{dagger} These authors contributed equally to the study.

{ddagger} Present address: Abbott Diagnostics Division, Abbott Laboratories, Abbott Park, IL 60064.

Infection and Immunity, August 2004, p. 4707-4715, Vol. 72, No. 8
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.8.4707-4715.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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