BLOCKING OF HUMAN-IMMUNODEFICIENCY-VIRUS INFECTION DEPENDS ON CELL-DENSITY AND VIRAL STOCK AGE

被引:58
作者
LAYNE, SP
MERGES, MJ
SPOUGE, JL
DEMBO, M
NARA, PL
机构
[1] NCI, FREDERICK CANC RES & DEV CTR, TUMOR CELL BIOL SECT, VIRUS BIOL SECT, FREDERICK, MD 21701 USA
[2] NATL LIB MED, NATL CTR BIOTECHNOL INFORMAT, BETHESDA, MD 20894 USA
关键词
D O I
10.1128/JVI.65.6.3293-3300.1991
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Quantitative infectivity assays were used to study how the blocking activity of soluble CD4 (sCD4) is affected by sCD4 concentration, target cell density, and viral stock age. During incubation with 20 nM sCD4, human immunodeficiency virus type 1 (HIV-1) stocks underwent irreversible inactivation. In contrast, inactivation with 2 nM sCD4 was almost entirely reversible. At lower sCD4 concentrations (less-than-or-equal-to 2 nM) and target cell densities of 6.25 x 10(4) ml-1, sCD4 blocking activity for HIV-1 gave a gp120-sCD4 association constant (K(assoc)) of 1.7 x 10(9) M-1, which agrees with chemical measurements. At the higher density of 1.6 x 10(7) cells ml-1, however, the blocking activity was 20-fold less. During incubation of HIV-1 stock optimized for infectivity by rapid harvest, sCD4 blocking activity increased 20-fold during a 3-h window. These results show that competitive blocking activity depends strongly on target cell density and virion age. Thus, unappreciated variations in HIV stocks and assay conditions may hinder comparisons of blockers from laboratory to laboratory, and the age of HIV challenge stocks may influence studies of drug and vaccine efficacy. The results also suggest that blocking of viral particles in lymphoid compartments will require very high competitive blocker concentrations, which may explain the refractory outcomes from sCD4-based drug trials in humans.
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页码:3293 / 3300
页数:8
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