Glycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.

<h4>Background</h4>The RV144 clinical trial showed for the first time that vaccination could provide modest but significant protection from HIV-1 infection. To understand the protective response, and to improve upon the vaccine's efficacy, it is important to define the structure of...

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Autores principales:	Bin Yu, Javier F Morales, Sara M O'Rourke, Gwen P Tatsuno, Phillip W Berman
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/34b4ba247fa246ce969cefb256a8b7db
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spelling	oai:doaj.org-article:34b4ba247fa246ce969cefb256a8b7db2021-11-18T07:07:56ZGlycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.1932-620310.1371/journal.pone.0043903https://doaj.org/article/34b4ba247fa246ce969cefb256a8b7db2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22928048/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>The RV144 clinical trial showed for the first time that vaccination could provide modest but significant protection from HIV-1 infection. To understand the protective response, and to improve upon the vaccine's efficacy, it is important to define the structure of the immunogens used in the prime/boost regimen. Here we examined the heterogeneity in net charge, attributable to glycoform variation, of the gp120 immunogens contained in the AIDSVAX B/E vaccine.<h4>Methodology/principal findings</h4>Isoelectric focusing and glycosidase digestion were used to assess variation in net charge of the gp120s contained in the AIDSVAX B/E vaccine used in the RV144 trial. We observed 16 variants of MN-rgp120 and 24 variants of A244-rgp120. Glycoform variation in gp120 produced in Chinese hamster ovary cells was compared to glycoform variation in gp120 produced in the 293F human embryonic kidney cell line, often used for neutralization assays. We found that gp120 variants produced in CHO cells were distinctly more acidic than gp120 variants produced in 293 cells. The effect of glycoform heterogeneity on antigenicity was assessed using monoclonal antibodies. The broadly neutralizing PG9 MAb bound to A244-rgp120, but not to MN-rgp120, whether produced in CHO or in 293. However, PG9 was able to bind with high affinity to MN-rgp120 and A244-rgp120 produced in 293 cells deficient in N-acetylglucosaminyltransferase I.<h4>Conclusions/significance</h4>MN- and A244-rgp120 used in the RV144 trial exhibited extensive heterogeneity in net charge due to variation in sialic acid-containing glycoforms. These differences were cell line-dependent, affected the antigenicity of recombinant envelope proteins, and may affect assays used to measure neutralization. These studies, together with recent reports documenting broadly neutralizing antibodies directed against carbohydrate epitopes of gp120, suggest that glycoform variation is a key variable to be considered in the production and evaluation of subunit vaccines designed to prevent HIV infection.Bin YuJavier F MoralesSara M O'RourkeGwen P TatsunoPhillip W BermanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 8, p e43903 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Bin Yu Javier F Morales Sara M O'Rourke Gwen P Tatsuno Phillip W Berman Glycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.
description	<h4>Background</h4>The RV144 clinical trial showed for the first time that vaccination could provide modest but significant protection from HIV-1 infection. To understand the protective response, and to improve upon the vaccine's efficacy, it is important to define the structure of the immunogens used in the prime/boost regimen. Here we examined the heterogeneity in net charge, attributable to glycoform variation, of the gp120 immunogens contained in the AIDSVAX B/E vaccine.<h4>Methodology/principal findings</h4>Isoelectric focusing and glycosidase digestion were used to assess variation in net charge of the gp120s contained in the AIDSVAX B/E vaccine used in the RV144 trial. We observed 16 variants of MN-rgp120 and 24 variants of A244-rgp120. Glycoform variation in gp120 produced in Chinese hamster ovary cells was compared to glycoform variation in gp120 produced in the 293F human embryonic kidney cell line, often used for neutralization assays. We found that gp120 variants produced in CHO cells were distinctly more acidic than gp120 variants produced in 293 cells. The effect of glycoform heterogeneity on antigenicity was assessed using monoclonal antibodies. The broadly neutralizing PG9 MAb bound to A244-rgp120, but not to MN-rgp120, whether produced in CHO or in 293. However, PG9 was able to bind with high affinity to MN-rgp120 and A244-rgp120 produced in 293 cells deficient in N-acetylglucosaminyltransferase I.<h4>Conclusions/significance</h4>MN- and A244-rgp120 used in the RV144 trial exhibited extensive heterogeneity in net charge due to variation in sialic acid-containing glycoforms. These differences were cell line-dependent, affected the antigenicity of recombinant envelope proteins, and may affect assays used to measure neutralization. These studies, together with recent reports documenting broadly neutralizing antibodies directed against carbohydrate epitopes of gp120, suggest that glycoform variation is a key variable to be considered in the production and evaluation of subunit vaccines designed to prevent HIV infection.
format	article
author	Bin Yu Javier F Morales Sara M O'Rourke Gwen P Tatsuno Phillip W Berman
author_facet	Bin Yu Javier F Morales Sara M O'Rourke Gwen P Tatsuno Phillip W Berman
author_sort	Bin Yu
title	Glycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.
title_short	Glycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.
title_full	Glycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.
title_fullStr	Glycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.
title_full_unstemmed	Glycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.
title_sort	glycoform and net charge heterogeneity in gp120 immunogens used in hiv vaccine trials.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/34b4ba247fa246ce969cefb256a8b7db
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Glycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.

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