HIV-1 pseudotype virus containing a Cocal virus genome and an HIV envelope: construction, assay and use

Susan Gregory, Ronald Collman, William James, Siamon Gordon, Francisco Gonzalez-Scarano, Neal Nathansona

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

A method is described for the production and assay of pseudotype viruses between human immunodeficiency virus type 1 (HIV-1) and Cocal virus (COV), containing an HIV-1 envelope and a COV genome (COV(HIV)). COV(HIV) pseudotype virus is a useful tool for the investigation of a variety of questions regarding HIV entry into susceptible cells, including steps in virus binding, fusion, and internalization, and the role of molecules which inhibit entry. COV, a rhabdovirus closely related to vesicular stomatitis virus (VSV), replicated and caused cytopathic effect in primary cultures of human peripheral blood lymphocytes (PBLs) and monocyte-derived macrophages (MDM), and in human cell lines of lymphocytoid or monocytoid origin, making it an ideal candidate for pseudotype production. 174XCEM cells, which were permissive for selected macrophage-tropic strains as well as most lymphocyte-tropic strains of HIV-1, were used to produce stocks of putative pseudotype virus. To neutralize parental COV in these stocks, a rabbit antiserum was produced which had a neutralization index of > 107 at a dilution of 1:100. Using these methods, pseudotype viruses were produced with a titer of about 104 PFU per ml; these same stocks contained HIV-1 at a titer of about 105 TCD50 per ml and COV at a titer of about 108 PFU per ml. CD4-expressing HeLa cells were used to assay pseudotype stocks made with lymphocyte-tropic strains of HIV-1. The authenticity of the pseudotype stocks was validated by several controls, including their failure to register on congenic CD4-negative HeLa cells and their inhibition by monoclonal anti-CD4 antibodies such as Leu 3a.

Original languageEnglish (US)
Pages (from-to)287-304
Number of pages18
JournalJournal of Virological Methods
Volume44
Issue number2-3
DOIs
Publication statusPublished - Oct 1993

    Fingerprint

Keywords

  • Cocal virus
  • Human immunodeficiency virus
  • Pseudotype virus
  • Rhabdovirus
  • Vesiculovirus
  • Virus entry

ASJC Scopus subject areas

  • Virology

Cite this

Gregory, S., Collman, R., James, W., Gordon, S., Gonzalez-Scarano, F., & Nathansona, N. (1993). HIV-1 pseudotype virus containing a Cocal virus genome and an HIV envelope: construction, assay and use. Journal of Virological Methods, 44(2-3), 287-304. https://doi.org/10.1016/0166-0934(93)90064-X