Production and characterization of a fragment containing the HIV-gp120 binding region of CD4 using a bovine papilloma virus (BPV) vector.
We have used a bovine papilloma virus (BPV) based mammalian cell expression vector consisting of the complete BPV genome and a human cytomegalovirus transcription unit for the production of soluble CD4. Mouse C-127 cells were transfected with vector DNA together with a selectable G418 resistance plasmid. Surviving clones were selected for high production using a solid phase ELISA based on the immobilization of supernatant-derived CD4 onto nitrocellulose paper and subsequent detection with anti-CD4 antibodies. The expressed protein was shown to bind HIV-gp120 and efficiently block HIV-1 infection in vitro. The possibility to use the above system for rapid production of defined glycoprotein fragments harboring defined functional regions, for the further elucidation of the functional role of CD4 in antigen presentation and cell to cell contact, and for possible intervention during HIV infection is discussed.
Gidlund
M
M
Department of Immunology, Karolinska Institute, Stockholm, Sweden.
Sydow
M
M
Stenbeck
A
A
Flodby
P
P
Fossum
H
H
Matsuda
S
S
Lundin
K
K
Wigzell
H
H
Lind
P
P
D052688:Bovine papillomavirus 1 / Q000235:genetics*; D015704:CD4 Antigens / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D002460:Cell Line; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics*; D015699:HIV Envelope Protein gp120 / Q000378:metabolism; D015497:HIV-1 / Q000187:drug effects / Q000502:physiology; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D012995:Solubility; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects
D015704:CD4 Antigens; D015699:HIV Envelope Protein gp120; D011993:Recombinant Fusion Proteins