Abstract
We show that the pertussis toxin B oligomer (PTX-B), and the PTX mutant PT9K/129G, which is safely administered in vivo, inhibit both transcription and secretion of TGF-beta elicited by HIV-1 Tat in NK cells. Tat-induced TGF-beta mRNA synthesis is also blocked by the ERK1 inhibitor PD98059, suggesting that ERK1 is needed for TGF-beta production. Moreover, Tat strongly activates the c-Jun component of the multimolecular complex AP-1, whereas TGF-beta triggers c-Fos and c-Jun. Of note, treatment of NK cells with PTX-B or PT9K/129G inhibits Tat- and TGF-beta-induced activation of AP-1. TGF-beta enhances starvation-induced NK cell apoptosis, significantly reduces transcription of the antiapoptotic protein Bcl-2, and inhibits Akt phosphorylation induced by oligomerization of the triggering NK cell receptor NKG2D. All these TGF-beta-mediated effects are prevented by PTX-B or PT9K/129G through a PI3K-dependent mechanism, as demonstrated by use of the specific PI3K inhibitor, LY294002. Finally, PTX-B and PT9K/129G up-regulate Bcl-x(L), the isoform of Bcl-x that protects cells from starvation-induced apoptosis. It is of note that in NK cells from patients with early HIV-1 infection, mRNA expression of Bcl-2 and Bcl-x(L) was consistently lower than that in healthy donors; interestingly, TGF-beta and Tat were detected in the sera of these patients. Together, these data suggest that Tat-induced TGF-beta production and the consequent NK cell failure, possibly occurring during early HIV-1 infection, may be regulated by PTX-B and PT9K/129G.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Adjuvants, Immunologic / antagonists & inhibitors
-
Adjuvants, Immunologic / physiology
-
Amino Acid Substitution / genetics
-
Apoptosis / genetics
-
Apoptosis / immunology*
-
Cells, Cultured
-
Down-Regulation / genetics
-
Down-Regulation / immunology*
-
Enzyme Activation / immunology
-
Gene Products, tat / antagonists & inhibitors
-
Gene Products, tat / physiology*
-
Glycine / genetics
-
HIV Infections / immunology
-
HIV Infections / pathology
-
HIV Infections / virology
-
HIV-1 / immunology
-
Humans
-
Killer Cells, Natural / cytology
-
Killer Cells, Natural / immunology*
-
Killer Cells, Natural / metabolism
-
Lysine / genetics
-
Pertussis Toxin / genetics*
-
Pertussis Toxin / immunology*
-
Pertussis Toxin / toxicity
-
Phosphorylation
-
Protein Serine-Threonine Kinases / antagonists & inhibitors
-
Protein Serine-Threonine Kinases / metabolism
-
Protein Subunits / genetics
-
Protein Subunits / immunology
-
Protein Subunits / toxicity
-
Proto-Oncogene Proteins / antagonists & inhibitors
-
Proto-Oncogene Proteins / metabolism
-
Proto-Oncogene Proteins c-akt
-
Proto-Oncogene Proteins c-bcl-2 / antagonists & inhibitors
-
Proto-Oncogene Proteins c-bcl-2 / genetics
-
Proto-Oncogene Proteins c-bcl-2 / metabolism
-
RNA, Messenger / metabolism
-
Transcription Factor AP-1 / antagonists & inhibitors
-
Transcription Factor AP-1 / metabolism
-
Transcription, Genetic / immunology
-
Transforming Growth Factor beta / antagonists & inhibitors*
-
Transforming Growth Factor beta / biosynthesis*
-
Transforming Growth Factor beta / metabolism
-
Transforming Growth Factor beta / physiology
-
Vaccines, Subunit / genetics
-
Vaccines, Subunit / immunology
-
Vaccines, Subunit / toxicity
-
bcl-X Protein
-
tat Gene Products, Human Immunodeficiency Virus
Substances
-
Adjuvants, Immunologic
-
BCL2L1 protein, human
-
Gene Products, tat
-
Protein Subunits
-
Proto-Oncogene Proteins
-
Proto-Oncogene Proteins c-bcl-2
-
RNA, Messenger
-
Transcription Factor AP-1
-
Transforming Growth Factor beta
-
Vaccines, Subunit
-
bcl-X Protein
-
tat Gene Products, Human Immunodeficiency Virus
-
Pertussis Toxin
-
AKT1 protein, human
-
Protein Serine-Threonine Kinases
-
Proto-Oncogene Proteins c-akt
-
Lysine
-
Glycine