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 Xin, H.
Right arrow Articles by Cutler, J. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Xin, H.
Right arrow Articles by Cutler, J. E.

 Previous Article  |  Next Article 

Infection and Immunity, July 2006, p. 4310-4321, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00234-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Hybridoma Passage In Vitro May Result in Reduced Ability of Antimannan Antibody To Protect against Disseminated Candidiasis

Hong Xin and Jim E. Cutler*

Department of Pediatrics, Louisiana State University Health Sciences Center and Research Institute for Children, Children's Hospital, New Orleans, Louisiana 70118

Received 10 February 2006/ Returned for modification 20 March 2006/ Accepted 19 April 2006

We previously reported the enhanced resistance of monoclonal antibodies B6.1 (an immunoglobulin M [IgM]) and C3.1 (an IgG3) against experimental candidiasis. Both MAbs recognize the same fungal epitope. We have since found that a highly passaged B6.1 hybridoma (hp-B6.1) resulted in antibody that has little protective potential. The potential clinical applicability of the antibody and our interest in understanding antibody protection against candidiasis led us to investigate an explanation for this phenomenon. Antibody genetic structure of hp-B6.1, the original hybridoma clone (ori-B6.1) stored frozen since 1995, a subclone of hp-B6.1 that produces protective antibody, the IgG3-producing hybridoma, and a nonprotective IgG1-producing hybridoma were compared. Variable region gene sequences of heavy (VH) and light chains showed genetic instability of VH chains with only the hp-B6.1; the VH sequences from ori-B6.1 and the subclone were, however, identical. Activation-induced cytidine deaminase levels were greatest in the B6.1 hybridomas, which may explain the instability. The constant region CH3 domain remained unchanged, implying normal N-glycation and complement-fixing potential, and antibody binding affinities appeared unchanged. Complement fixation assays surprisingly showed that ori-B6.1 antibody fixes C3 more rapidly than does hp-B6.1 antibody. The VH region primary structure may affect complement activation, which could explain our result. Indeed, antibody from the hp-B6.1 subclone fixed complement like antibody from ori-B6.1. These results show that the greatest protection occurs when antimannan antibodies possess the dual abilities of recognizing the appropriate carbohydrate epitope and rapidly fixing complement; loss of the latter property results in the loss of protective potential by the antibody.

* Corresponding author. Mailing address: Research Institute for Children, Children's Hospital, 200 Henry Clay Ave., New Orleans, LA 70118. Phone: (504) 896-2711. Fax: (504) 896-2720. E-mail: jcutler{at}chnola-research.org.

Editor: A. Casadevall

Infection and Immunity, July 2006, p. 4310-4321, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00234-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Xin, H., Dziadek, S., Bundle, D. R., Cutler, J. E. (2008). Synthetic glycopeptide vaccines combining {beta}-mannan and peptide epitopes induce protection against candidiasis. Proc. Natl. Acad. Sci. USA 105: 13526-13531 [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»