Expression of HTLV-I Env and Tax recombinant peptides in yeast: identification of immunogenic domains.
A peptide library of HTLV-I Env and Tax proteins was constructed in yeast in order to characterize which domains of these proteins are immunogenic in HTLV-I-infected individuals. Five yeast colonies (A to E) were selected using HTLV-I positive plasma, and one yeast colony (F) was selected using rabbit anti-Tax sera. Plasmid DNA present in each positive clone was recovered and sequenced. Overlapping clones A to E covered the C-terminal part of the gp46 exterior glycoprotein (aa 197 to 305) and were all glycosylated. Clone F encoded the C-terminal 25 aa of Tax (aa 329-353). Recombinant peptides were recognized by more than 40% of the HTLV-I positive human sera, confirming that they are major immunodominant domains. We studied the antibody response to each recombinant peptide in patients with TSP/HAM and asymptomatic carriers. Higher absorbance values were obtained with sera from TSP/HAM patients than from asymptomatic carriers, but the difference between the two groups was not statistically significant. Our study confirms that the COOH-terminal region of gp46 is highly immunogenic in HTLV-I-infected individuals and demonstrates a new immunogenic epitope of the Tax protein.
Noraz
N
N
Inserm U271, Lyon, France.
Benichou
S
S
Madaule
P
P
Tiollais
P
P
Vernant
J C
JC
Desgranges
C
C
D000595:Amino Acid Sequence; D001483:Base Sequence; D003429:Cross Reactions / Q000276:immunology; D000939:Epitopes / Q000276:immunology; D015686:Gene Products, env / Q000276:immunology*; D016356:Gene Products, tax / Q000276:immunology*; D006031:Glycosylation; D015481:HTLV-I Antibodies / Q000097:blood; D015490:HTLV-I Infections / Q000276:immunology*; D015482:HTLV-II Antibodies / Q000276:immunology; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D015682:Retroviridae Proteins, Oncogenic / Q000276:immunology*; D012441:Saccharomyces cerevisiae
D000939:Epitopes; D015686:Gene Products, env; D016356:Gene Products, tax; D015481:HTLV-I Antibodies; D015482:HTLV-II Antibodies; D010455:Peptides; D011994:Recombinant Proteins; D015682:Retroviridae Proteins, Oncogenic; C055266:gp46 protein, Human T-cell leukemia virus type I