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Infection and Immunity, April 2006, p. 2428-2435, Vol. 74, No. 4
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.4.2428-2435.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Monoclonal Antibody Detection of CD46 Clustering beneath Neisseria gonorrhoeae Microcolonies

Nathan J. Weyand,1* Shaun W. Lee,1,{dagger} Dustin L. Higashi,1 Daniel Cawley,2 Paul Yoshihara,2 and Magdalene So1

Department of Molecular Microbiology & Immunology, Oregon Health and Science University, Portland, Oregon 97201-3098,1 Monoclonal Antibody Core Facility, Vaccine and Gene Therapy Institute, Oregon Health and Science University, Beaverton, Oregon 970062

Received 18 November 2005/ Accepted 18 January 2006

CD46 (membrane cofactor protein), a complement-regulatory protein that participates in innate and acquired immunity, also serves as a receptor for viral and bacterial pathogens. CD46 isoforms terminate in one of two cytoplasmic tails, Cyt1 or Cyt2, which differ in signaling and trafficking properties. Dissecting the functions of the two cytoplasmic tails in these cellular processes has been hampered by the absence of specific reagents. Here we report the construction of Cyt1- and Cyt2-specific monoclonal antibodies (MAbs). These MAbs recognize unique epitopes within the tails and can be used for immunofluorescence microscopy, immunoblotting, and immunoprecipitation. Studies of Neisseria gonorrhoeae-infected cells with the CD46 tail MAbs demonstrate the differential recruitment of Cyt1 and Cyt2 to the cortical plaque.

* Corresponding author. Mailing address: Department of Molecular Microbiology & Immunology, L220, Oregon Health and Science University, Portland, OR 97201-3098. Phone: (503) 494-2436. Fax: (503) 494-6862. E-mail: weyandn{at}ohsu.edu.

Editor: D. L. Burns

{dagger} Present address: Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093-0721.

Infection and Immunity, April 2006, p. 2428-2435, Vol. 74, No. 4
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.4.2428-2435.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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