IDENTIFICATION OF A CD36-RELATED THROMBOSPONDIN 1-BINDING DOMAIN IN HIV-1 ENVELOPE GLYCOPROTEIN GP120 - RELATIONSHIP TO HIV-1-SPECIFIC INHIBITORY FACTORS IN HUMAN SALIVA
R. Crombie et al., IDENTIFICATION OF A CD36-RELATED THROMBOSPONDIN 1-BINDING DOMAIN IN HIV-1 ENVELOPE GLYCOPROTEIN GP120 - RELATIONSHIP TO HIV-1-SPECIFIC INHIBITORY FACTORS IN HUMAN SALIVA, The Journal of experimental medicine, 187(1), 1998, pp. 25-35
Human and non-human primate salivas retard the infectivity of HIV-1 in
vitro and in vivo. Because thrombospondin 1 (TSP1), a high molecular
weight trimeric glycoprotein, is concentrated in saliva and can inhibi
t the infectivity of diverse pathogens in vitro, we sought to determin
e the role of TSP1 in suppression of HIV infectivity. Sequence analysi
s revealed a TSP1 recognition motif previously defined for the CD36 ge
ne family of cell adhesion receptors, in conserved regions nanking the
disulfide-linked cysteine residues of the V3 loop of HIV envelope gly
coprotein gp120, important for HIV binding to its high affinity cellul
ar receptor CD4. Using solid-phase in vitro binding assays, we demonst
rate direct binding of radiolabeled TSP1 to immobilized recombinant gp
120. Based on peptide blocking experiments, the TSP1-gp120 interaction
involves CSVTCG sequences in the type 1 properdin-like repeats of TSP
1, the known binding site for CD36. TSP1 and fusion proteins derived f
i-om CD36-related TSP1-binding domains were able to compete with radio
labeled soluble CD4 binding to immobilized gp120. In parallel, purifie
d TSP1 inhibited HIV-1 infection of peripheral blood mononuclear cells
and transformed T and promonocytic cell lines. Levels of TSP1 require
d for both viral aggregation and direct blockade of HIV-1 infection we
re physiologic, and affinity depletion of salivary TSP1 abrogated >70%
of the inhibitory effect of whole saliva on HIV infectivity. Characte
rization of TSP1-gp120 binding specificity suggests a mechanism for di
rect blockade of HIV infectivity that might be exploited to retard HIV
transmission that occurs via mucosal routes.