Antagonistic activity of HIV-1 T helper peptides flanked by an unrelated carrier protein.
Antagonism is the ability of a modified antigenic peptide (altered peptide ligand, APL) to prevent CD4 T cell activation by the original peptide. Here we show that antagonistic activity can be conferred to peptides of HIV envelope glycoprotein gp120 and reverse transcriptase p66 by adding flanking polypeptide sequences at the C or at the N terminus by genetic engineering, rather than by introducing substitutions by synthesis. The glutathione S-transferase (GST)-peptide system has been used to produce molecules that display the peptide at the appropriate end of the GST carrier. When the gp120 peptide 191-205 (pep24) was expressed at the C terminus of GST (GST-24), antigenicity of specific human CD4 T cells was maintained. In contrast, when the peptide was expressed at the N terminus of GST (24-GST), antigenicity was abolished and antagonistic activity was introduced. Similar results were obtained with a p66-derived peptide at the C terminus of the GST carrier. Antagonism was (1) specific; proliferation of a CD4 T cell line from the same donor responding to the envelope glycoprotein of another retrovirus, HTLV-1, was not affected; (2) reversible; proliferative response was rescued in T cells exposed to antigen-presenting cells (APC) pulsed with the antagonist; (3) dominant; T cells cultured with APC pulsed with the agonist and with APC pulsed with the antagonist did not proliferate. The carrier could be cleaved by proteolysis while the antagonistic activity was preserved. Thus a minimal sequence that confers antagonistic activity can be engineered or synthesized with peptides to antagonize undesired CD4 responses as an alternative to the use of APL.
Fenoglio
D
D
Department of Immunology, San Martino Hospital, University of Genoa, Italy.
Li Pira
G
G
De Berardinis
P
P
Saverino
D
D
Terranova
M P
MP
Ombra
M N
MN
Bracci
L
L
Lozzi
L
L
Viotti
C
C
Guardiola
J
J
Manca
F
F
D000595:Amino Acid Sequence; D005982:Glutathione Transferase / Q000276:immunology; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D054303:HIV Reverse Transcriptase / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*
D015699:HIV Envelope Protein gp120; D010455:Peptides; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D054303:HIV Reverse Transcriptase