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

Resultado de la investigación: Articlerevisión exhaustiva

9 Citas (Scopus)


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.

Idioma originalEnglish (US)
Páginas (desde-hasta)287-304
Número de páginas18
PublicaciónJournal of Virological Methods
EstadoPublished - oct. 1993
Publicado de forma externa

ASJC Scopus subject areas

  • Virology


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