Human immunodeficiency virus type 1 envelope gene structure and diversity in vivo and after cocultivation in vitro

Kenro Kusumi, Brian Conway, Scott Cunningham, Amy Berson, Christine Evans, Astrid K N Iversen, David Colvin, Michael V. Gallo, Steven Coutre, Eugene G. Shpaer, Donald V. Faulkner, Anthony DeRonde, Sarah Volkman, Carolyn Williams, Martin S. Hirsch, James I. Mullins

Research output: Contribution to journalArticle

117 Citations (Scopus)

Abstract

Nested-primer polymerase chain reaction (PCR) has been applied to the molecular cloning of 4.6-kb half-genome fragments of human immunodeficiency virus type 1 (HIV-1) taken directly from the peripheral blood mononuclear cells (PBMC) of an individual with neurological symptoms of HIV-1 infection. In a similar manner, gp120-coding portions of the envelope gene were cloned after PBMC from the same blood sample were cocultivated with uninfected PBMC for 28 days. The complete 1.6-kb nucleotide sequence of the gp120 gene was determined from each of 35 clones examined. Two of 13 (15%) PBMC-derived gp120 genes and 3 of 22 (14%) coculture-derived gp120 genes were defective as a result of frameshifts and an in-frame stop codon(s). Mean diversity between individual gp120-coding sequences in PBMC was fivefold greater (3.24%) than after coculture (0.65%). A predominant sequence or "strain" was found after coculture that was distinct from the diverse viral genotypes detected in vivo and therefore was selectively amplified during in vitro propagation. Multiple distinct third variable (V3) regions encoding the principal neutralizing domain of the envelope protein were detected in PBMC-derived genes, suggesting the presence of immunologic diversity of HIV env genes in vivo not reflected in the cocultured virus sample. The large size of the HIV fragments generated in this study will permit analysis of the diversity of immunologic reactivity, gene function, and pathogenicity of HIV genomes present within infected individuals, including the functional significance of the loss of diversity that occurs upon coculture.

Original languageEnglish (US)
Pages (from-to)875-885
Number of pages11
JournalJournal of Virology
Volume66
Issue number2
StatePublished - Feb 1992
Externally publishedYes

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Coculture Techniques
Human immunodeficiency virus 1
mononuclear leukocytes
HIV-1
Blood Cells
coculture
Genes
genes
HIV
Genome
env Genes
Terminator Codon
Molecular Cloning
Virus Diseases
genome
stop codon
Virulence
In Vitro Techniques
neutralization
signs and symptoms (animals and humans)

ASJC Scopus subject areas

  • Immunology

Cite this

Kusumi, K., Conway, B., Cunningham, S., Berson, A., Evans, C., Iversen, A. K. N., ... Mullins, J. I. (1992). Human immunodeficiency virus type 1 envelope gene structure and diversity in vivo and after cocultivation in vitro. Journal of Virology, 66(2), 875-885.

Human immunodeficiency virus type 1 envelope gene structure and diversity in vivo and after cocultivation in vitro. / Kusumi, Kenro; Conway, Brian; Cunningham, Scott; Berson, Amy; Evans, Christine; Iversen, Astrid K N; Colvin, David; Gallo, Michael V.; Coutre, Steven; Shpaer, Eugene G.; Faulkner, Donald V.; DeRonde, Anthony; Volkman, Sarah; Williams, Carolyn; Hirsch, Martin S.; Mullins, James I.

In: Journal of Virology, Vol. 66, No. 2, 02.1992, p. 875-885.

Research output: Contribution to journalArticle

Kusumi, K, Conway, B, Cunningham, S, Berson, A, Evans, C, Iversen, AKN, Colvin, D, Gallo, MV, Coutre, S, Shpaer, EG, Faulkner, DV, DeRonde, A, Volkman, S, Williams, C, Hirsch, MS & Mullins, JI 1992, 'Human immunodeficiency virus type 1 envelope gene structure and diversity in vivo and after cocultivation in vitro', Journal of Virology, vol. 66, no. 2, pp. 875-885.
Kusumi, Kenro ; Conway, Brian ; Cunningham, Scott ; Berson, Amy ; Evans, Christine ; Iversen, Astrid K N ; Colvin, David ; Gallo, Michael V. ; Coutre, Steven ; Shpaer, Eugene G. ; Faulkner, Donald V. ; DeRonde, Anthony ; Volkman, Sarah ; Williams, Carolyn ; Hirsch, Martin S. ; Mullins, James I. / Human immunodeficiency virus type 1 envelope gene structure and diversity in vivo and after cocultivation in vitro. In: Journal of Virology. 1992 ; Vol. 66, No. 2. pp. 875-885.
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