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Engineering human hematopoietic stem/progenitor cells to produce a broadly neutralizing anti-HIV antibody after in vitro maturation to human B lymphocytes

Luo, Xin M. and Maarschalk, Emily and O'Connell, Ryan M. and Wang, Pin and Yang, Lili and Baltimore, David (2009) Engineering human hematopoietic stem/progenitor cells to produce a broadly neutralizing anti-HIV antibody after in vitro maturation to human B lymphocytes. Blood, 113 (7). pp. 1422-1431. ISSN 0006-4971. https://resolver.caltech.edu/CaltechAUTHORS:20090828-115250714

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Image (JPEG) (Figure S1. Lentiviral transduction of HSPCs and testing of b12-encoding constructs in cell lines) - Supplemental Material
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Image (JPEG) (Figure S2. Differentiation of HSPCs and generation of myeloid cells early in Stage-2) - Supplemental Material
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Image (JPEG) (Figure S3. Effect of the transgene on B-cell development during Stage-2) - Supplemental Material
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Image (JPEG) (Figure S4. Effect of GFP on B-cell activation and differentiation into plasma cells during Stage-3) - Supplemental Material
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Image (JPEG) (Figure S5. Morphology of normal plasmablasts and plasma cells) - Supplemental Material
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Abstract

Broadly neutralizing anti-HIV antibodies are rare and have proved hard to elicit with any immunogen. We have tested in vitro the notion that such antibodies or other antiviral proteins could be made by lentivirus-mediated gene transfer into human hematopoietic stem/progenitor cells (HSPCs), followed by differentiation of the transduced cells into B cells, the most potent antibody-producing cells. To do this, we have developed a highly efficient system for in vitro maturation of secreting B lymphocytes and plasma cells from CD34^+ HSPCs. It is a 3-stage, in vitro culture system that supports normal human B-lineage development from HSPCs to antibody-secreting plasmablasts (~36%) and plasma cells (~20%). By transducing human cord blood CD34^+ cells with lentiviral vectors encoding a secretory monoclonal anti-HIV antibody, b12 (IgG1), we were able to program human B cells to produce in vitro up to 1.5 µg/mL of this broadly neutralizing antibody. Our results suggest that an HIV vaccine might be delivered by autologous transplantation of in vitro–programmed HSPCs, which would develop into antibody-secreting B cells in vivo and provide a continuous supply of anti-HIV neutralizing antibodies.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1182/blood-2008-09-177139DOIUNSPECIFIED
http://bloodjournal.hematologylibrary.org/cgi/content/abstract/113/7/1422PublisherUNSPECIFIED
ORCID:
AuthorORCID
Baltimore, David0000-0001-8723-8190
Additional Information:Copyright © 2009 by American Society of Hematology. Submitted September 3, 2008; accepted November 20, 2008. Prepublished online as Blood First Edition paper, December 4, 2008; DOI 10.1182/blood-2008-09-177139. The online version of this article contains a data supplement. The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked ‘‘advertisement’’ in accordance with 18 USC section 1734. The authors thank D. Burton (Scripps Research Institute, La Jolla, CA) for providing b12 variable regions, G. Crooks (University of Southern California) for MS5 cells, J. Klein (California Institute of Technology) for monomeric gp120MN, and M. Boldin (California Institute of Technology) for the THP-1 cell line with integrated FUW. The authors also thank A. West, P. Peiris, and D. Rao (California Institute of Technology) for Biacore analysis, pseudovirus neutralization assay and microscopy, respectively; and R. Diamond for the use of Flow Cytometry Core Facility at California Institute of Technology. This study was supported by the Bill and Melinda Gates Foundation (Seattle, WA) through the Grand Challenges in Global Health Initiative (D.B.). Contribution: X.M.L. designed the study; X.M.L. and E.M. performed the experiments; R.M.O. and P.W. prepared plasmids; D.B. and L.Y. supervised the study; and X.M.L. prepared the manuscript. Conflict-of-interest disclosure: The authors declare no competing financial interests.
Funders:
Funding AgencyGrant Number
Bill and Melinda Gates FoundationUNSPECIFIED
Issue or Number:7
Record Number:CaltechAUTHORS:20090828-115250714
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20090828-115250714
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:15410
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:14 Sep 2009 18:49
Last Modified:03 Oct 2019 00:57

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