Expression of HIV-1 envelope glycoproteins by Semliki Forest virus vectors.
We have used Semliki Forest virus (SFV) vectors to express both the human immunodeficiency virus type 1 (HIV-1) envelope precursor gp160 and the cleaved external portion gp120. Expression of the foreign gene in this system is by transfection of recombinant SFV RNA, or by infection with a recombinant SFV virus that has a wide host range. pSFV1-gp120 or pSFV1-gp160 were expressed in baby hamster kidney (BHK) cells and two human cell lines: HeLa cervical carcinoma and MOLT-4 CD4+ T cells. After SFV1-gp120 infection of HeLa cells, 3.3 micrograms of gp120 was secreted into the media by 1 million cells in a 24-hr period. The secreted envelope glycoprotein was recognized by anti-gp120 monoclonal antibodies directed against both linear and conformation-dependent epitopes in different regions of the molecule. The recombinant gp120 also bound to a soluble form of the CD4 receptor. Syncytium formation was observed when MOLT-4 cells were infected with SFV1-gp160. The gp160 expressed by BHK cells induced syncytia during cocultivation with C8166 CD4+ T cells. These data indicate that SFV vectors can be used to produce the HIV-1 envelope glycoproteins to high levels, and that these proteins are correctly processed, folded, and transported to the cell surface. Furthermore, they exhibit functional activity as indicated by their ability to bind to soluble receptor and induce cell-to-cell fusion.
Paul
N L
NL
Chester Beatty Laboratories, Institute of Cancer Research, London, England.
Marsh
M
M
McKeating
J A
JA
Schulz
T F
TF
Liljeström
P
P
Garoff
H
H
Weiss
R A
RA
D000818:Animals; D002478:Cells, Cultured; D006224:Cricetinae; D015967:Gene Expression Regulation, Viral; D015686:Gene Products, env / Q000096:biosynthesis* / Q000235:genetics; D005822:Genetic Vectors; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis* / Q000235:genetics; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D012672:Semliki forest virus / Q000235:genetics / Q000254:growth & development
D015686:Gene Products, env; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D011498:Protein Precursors; D011994:Recombinant Proteins