Cargando…

Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus

Eradication of human immunodeficiency virus type 1 (HIV-1) by vaccination with epitopes that produce broadly neutralizing antibodies is the ultimate goal for HIV prevention. However, generating appropriate immune responses has proven difficult. Expression of broadly neutralizing antibodies by vagina...

Descripción completa

Detalles Bibliográficos
Autores principales: Marcobal, Angela, Liu, Xiaowen, Zhang, Wenlei, Dimitrov, Antony S., Jia, Letong, Lee, Peter P., Fouts, Timothy R., Parks, Thomas P., Lagenaur, Laurel A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Mary Ann Liebert, Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5067876/
https://www.ncbi.nlm.nih.gov/pubmed/26950606
http://dx.doi.org/10.1089/aid.2015.0378
_version_ 1782460722794463232
author Marcobal, Angela
Liu, Xiaowen
Zhang, Wenlei
Dimitrov, Antony S.
Jia, Letong
Lee, Peter P.
Fouts, Timothy R.
Parks, Thomas P.
Lagenaur, Laurel A.
author_facet Marcobal, Angela
Liu, Xiaowen
Zhang, Wenlei
Dimitrov, Antony S.
Jia, Letong
Lee, Peter P.
Fouts, Timothy R.
Parks, Thomas P.
Lagenaur, Laurel A.
author_sort Marcobal, Angela
collection PubMed
description Eradication of human immunodeficiency virus type 1 (HIV-1) by vaccination with epitopes that produce broadly neutralizing antibodies is the ultimate goal for HIV prevention. However, generating appropriate immune responses has proven difficult. Expression of broadly neutralizing antibodies by vaginal colonizing lactobacilli provides an approach to passively target these antibodies to the mucosa. We tested the feasibility of expressing single-chain and single-domain antibodies (dAbs) in Lactobacillus to be used as a topical microbicide/live biotherapeutic. Lactobacilli provide an excellent platform to express anti-HIV proteins. Broadly neutralizing antibodies have been identified against epitopes on the HIV-1 envelope and have been made into active antibody fragments. We tested single-chain variable fragment m9 and dAb-m36 and its derivative m36.4 as prototype antibodies. We cloned and expressed the antibody fragments m9, m36, and m36.4 in Lactobacillus jensenii-1153 and tested the expression levels and functionality. We made a recombinant L. jensenii 1153-1128 that expresses dAb-m36.4. All antibody fragments m9, m36, and m36.4 were expressed by lactobacilli. However, we noted the smaller m36/m36.4 were expressed to higher levels, ≥3 μg/ml. All L. jensenii-expressed antibody fragments bound to gp120/CD4 complex; Lactobacillus-produced m36.4 inhibited HIV-1(BaL) in a neutralization assay. Using a TZM-bl assay, we characterized the breadth of neutralization of the m36.4. Delivery of dAbs by Lactobacillus could provide passive transfer of these antibodies to the mucosa and longevity at the site of HIV-1 transmission.
format Online
Article
Text
id pubmed-5067876
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Mary Ann Liebert, Inc.
record_format MEDLINE/PubMed
spelling pubmed-50678762016-11-01 Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus Marcobal, Angela Liu, Xiaowen Zhang, Wenlei Dimitrov, Antony S. Jia, Letong Lee, Peter P. Fouts, Timothy R. Parks, Thomas P. Lagenaur, Laurel A. AIDS Res Hum Retroviruses Alternative Approaches to HIV Prevention Eradication of human immunodeficiency virus type 1 (HIV-1) by vaccination with epitopes that produce broadly neutralizing antibodies is the ultimate goal for HIV prevention. However, generating appropriate immune responses has proven difficult. Expression of broadly neutralizing antibodies by vaginal colonizing lactobacilli provides an approach to passively target these antibodies to the mucosa. We tested the feasibility of expressing single-chain and single-domain antibodies (dAbs) in Lactobacillus to be used as a topical microbicide/live biotherapeutic. Lactobacilli provide an excellent platform to express anti-HIV proteins. Broadly neutralizing antibodies have been identified against epitopes on the HIV-1 envelope and have been made into active antibody fragments. We tested single-chain variable fragment m9 and dAb-m36 and its derivative m36.4 as prototype antibodies. We cloned and expressed the antibody fragments m9, m36, and m36.4 in Lactobacillus jensenii-1153 and tested the expression levels and functionality. We made a recombinant L. jensenii 1153-1128 that expresses dAb-m36.4. All antibody fragments m9, m36, and m36.4 were expressed by lactobacilli. However, we noted the smaller m36/m36.4 were expressed to higher levels, ≥3 μg/ml. All L. jensenii-expressed antibody fragments bound to gp120/CD4 complex; Lactobacillus-produced m36.4 inhibited HIV-1(BaL) in a neutralization assay. Using a TZM-bl assay, we characterized the breadth of neutralization of the m36.4. Delivery of dAbs by Lactobacillus could provide passive transfer of these antibodies to the mucosa and longevity at the site of HIV-1 transmission. Mary Ann Liebert, Inc. 2016-11-01 2016-11-01 /pmc/articles/PMC5067876/ /pubmed/26950606 http://dx.doi.org/10.1089/aid.2015.0378 Text en © Angela Marcobal et al., 2016; Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons Attribution Noncommercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.
spellingShingle Alternative Approaches to HIV Prevention
Marcobal, Angela
Liu, Xiaowen
Zhang, Wenlei
Dimitrov, Antony S.
Jia, Letong
Lee, Peter P.
Fouts, Timothy R.
Parks, Thomas P.
Lagenaur, Laurel A.
Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus
title Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus
title_full Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus
title_fullStr Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus
title_full_unstemmed Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus
title_short Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus
title_sort expression of human immunodeficiency virus type 1 neutralizing antibody fragments using human vaginal lactobacillus
topic Alternative Approaches to HIV Prevention
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5067876/
https://www.ncbi.nlm.nih.gov/pubmed/26950606
http://dx.doi.org/10.1089/aid.2015.0378
work_keys_str_mv AT marcobalangela expressionofhumanimmunodeficiencyvirustype1neutralizingantibodyfragmentsusinghumanvaginallactobacillus
AT liuxiaowen expressionofhumanimmunodeficiencyvirustype1neutralizingantibodyfragmentsusinghumanvaginallactobacillus
AT zhangwenlei expressionofhumanimmunodeficiencyvirustype1neutralizingantibodyfragmentsusinghumanvaginallactobacillus
AT dimitrovantonys expressionofhumanimmunodeficiencyvirustype1neutralizingantibodyfragmentsusinghumanvaginallactobacillus
AT jialetong expressionofhumanimmunodeficiencyvirustype1neutralizingantibodyfragmentsusinghumanvaginallactobacillus
AT leepeterp expressionofhumanimmunodeficiencyvirustype1neutralizingantibodyfragmentsusinghumanvaginallactobacillus
AT foutstimothyr expressionofhumanimmunodeficiencyvirustype1neutralizingantibodyfragmentsusinghumanvaginallactobacillus
AT parksthomasp expressionofhumanimmunodeficiencyvirustype1neutralizingantibodyfragmentsusinghumanvaginallactobacillus
AT lagenaurlaurela expressionofhumanimmunodeficiencyvirustype1neutralizingantibodyfragmentsusinghumanvaginallactobacillus