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Use of Mutated Self-Cleaving 2A Peptides as a Molecular Rheostat to Direct Simultaneous Formation of Membrane and Secreted Anti-HIV Immunoglobulins

Yu, Kenneth K. and Aguilar, Kiefer and Tsai, Jonathan and Galimidi, Rachel and Gnanapragasam, Priyanthi and Yang, Lili and Baltimore, David (2012) Use of Mutated Self-Cleaving 2A Peptides as a Molecular Rheostat to Direct Simultaneous Formation of Membrane and Secreted Anti-HIV Immunoglobulins. PLoS ONE, 7 (11). Art. No. e50438 . ISSN 1932-6203. PMCID PMC3508920. https://resolver.caltech.edu/CaltechAUTHORS:20130117-154355776

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Abstract

In nature, B cells produce surface immunoglobulin and secreted antibody from the same immunoglobulin gene via alternative splicing of the pre-messenger RNA. Here we present a novel system for genetically programming B cells to direct the simultaneous formation of membrane-bound and secreted immunoglobulins that we term a “Molecular Rheostat”, based on the use of mutated “self-cleaving” 2A peptides. The Molecular Rheostat is designed so that the ratio of secreted to membrane-bound immunoglobulins can be controlled by selecting appropriate mutations in the 2A peptide. Lentiviral transgenesis of Molecular Rheostat constructs into B cell lines enables the simultaneous expression of functional b12-based IgM-like BCRs that signal to the cells and mediate the secretion of b12 IgG broadly neutralizing antibodies that can bind and neutralize HIV-1 pseudovirus. We show that these b12-based Molecular Rheostat constructs promote the maturation of EU12 B cells in an in vitro model of B lymphopoiesis. The Molecular Rheostat offers a novel tool for genetically manipulating B cell specificity for B-cell based gene therapy.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1371/journal.pone.0050438DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3508920/PubMed CentralArticle
ORCID:
AuthorORCID
Baltimore, David0000-0001-8723-8190
Additional Information:© 2012 Yu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: May 15, 2012; Accepted: October 22, 2012; Published: November 28, 2012. This work is supported by the following two sources of funding: 1. The Grand Challenges in Global Health Program, The Bill and Melinda Gates Foundation, award # 37866 (http://www.grandchallenges.org/CureInfec​tion/Challenges/ImmunologicalMethods/Pag​es/HIV.aspx), 2. The California HIV/AIDS Research Program, The Office of the President of the Universities of California, award # UCOP.D08-CT-305 (http://chrp.ucop.edu/funded_research/abs​tracts/2009_yu.html). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We wish to thank Dr. Zhixin Zhang and Dr. Max Cooper for sharing the EU12 cell line with us. We also thank Dr. Alex Balazs for providing the pHAGE series vectors. Author Contributions: Conceived and designed the experiments: KKY LY DB. Performed the experiments: KKY KA JT RG PG. Analyzed the data: KKY LY DB. Contributed reagents/materials/analysis tools: KKY KA JT RG PG. Wrote the paper: KKY KA DB.
Funders:
Funding AgencyGrant Number
Bill and Melinda Gates Foundation37866
California HIV/AIDS Research ProgramUCOP.D08-CT-305
Issue or Number:11
PubMed Central ID:PMC3508920
Record Number:CaltechAUTHORS:20130117-154355776
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20130117-154355776
Official Citation:Yu KK, Aguilar K, Tsai J, Galimidi R, Gnanapragasam P, et al. (2012) Use of Mutated Self-Cleaving 2A Peptides as a Molecular Rheostat to Direct Simultaneous Formation of Membrane and Secreted Anti-HIV Immunoglobulins. PLoS ONE 7(11): e50438. doi:10.1371/journal.pone.0050438
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:36458
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:18 Jan 2013 15:59
Last Modified:12 Feb 2020 00:21

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