Adaptation of two primary human immunodeficiency virus type 1 isolates to growth in transformed T cell lines correlates with alterations in the responses of their envelope glycoproteins to soluble CD4.
Two sCD4-resistant, primary viruses (P-08 and P-17) were compared with two sCD4-sensitive, T cell line-adapted variants (C-08 and C-17) for their biochemical responses to sCD4. At 37 degrees C, neither primary virus shed gp120 within 8 hr at sCD4 concentrations of up to 500 nM, whereas C-08 and C-17 lost gp120 within minutes of addition of 5-10 nM sCD4. At 4 degrees C, however, P-17 and C-17 shed gp120 at similar rates in response to the same sCD4 concentration. Irrespective of the temperature, gp120 dissociation from both P-17 and C-17 was inhibited by CD4 MAbs 6H10 and 5A8, the latter of which blocks events subsequent to sCD4 binding. Binding of sCD4 to P-17 was greater at 4 degrees C than at 37 degrees C, whereas the converse was found for C-17. Consistent with this, P-17 was neutralized much more potently by sCD4 at 4 degrees C than at 37 degrees C, whereas C-17 was slightly more sensitive to sCD4 at 37 degrees C than at 4 degrees C. Resistance to neutralization by sCD4 is probably determined by kinetic parameters. We suggest that the acquisition of sCD4 neutralization sensitivity and the above biochemical responses to sCD4 are coincidental to the process by which some primary viruses adapt to growth in transformed T cells. Sequence data indicate that there are a limited number of amino acid differences between the Env glycoproteins of the primary viruses and their T cell line-adapted counterparts; the significance of the individual changes is under investigation, but both pairs of viruses have amino acid substitutions in a region of gp41 thought to contact gp120.
Moore
J P
JP
Aaron Diamond AIDS Research Center, New York University School of Medicine, New York 10016.
Burkly
L C
LC
Connor
R I
RI
Cao
Y
Y
Tizard
R
R
Ho
D D
DD
Fisher
R A
RA
D000220:Adaptation, Biological; D000595:Amino Acid Sequence; D001483:Base Sequence; D015704:CD4 Antigens / Q000378:metabolism*; D002461:Cell Line, Transformed; D003080:Cold Temperature; D015686:Gene Products, env / Q000378:metabolism*; D014644:Genetic Variation; D015699:HIV Envelope Protein gp120 / Q000378:metabolism; D015700:HIV Envelope Protein gp41 / Q000378:metabolism; D015497:HIV-1 / Q000254:growth & development*; D006358:Hot Temperature; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017421:Sequence Analysis; D012995:Solubility; D013601:T-Lymphocytes / Q000382:microbiology*; D014779:Virus Replication
D015704:CD4 Antigens; D015686:Gene Products, env; D015699:HIV Envelope Protein gp120; D015700:HIV Envelope Protein gp41