Production of a recombinant human T-cell leukemia virus type-I trans-activator (tax1) antigen and its utilization for generation of monoclonal antibodies against various epitopes on the tax1 antigen
✍ Scribed by Yuetsu Tanaka; Atsushi Yoshida; Hideki Tozawa; Hisatoshi Shida; Hiroshi Nyunoya; Kunitada Shimotohno
- Publisher
- John Wiley and Sons
- Year
- 1991
- Tongue
- French
- Weight
- 967 KB
- Volume
- 48
- Category
- Article
- ISSN
- 0020-7136
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✦ Synopsis
A 42-kDa recombinant protein, PX141, consisting of the trans-activator protein encoded by human T-cell leukemia virus (HTLV-I) (tax, antigen) and the amino-terminal fusion peptide of I 2 amino acid residues of the a-peptide encoded by the plasmid pUC I 9 was produced. In order to investigate the immunogenicity of the tax, antigen, mice were immunized with the purified PX141 and 4 anti-tax, monoclonal antibodies (MAbs) designated T A X Y -I , TAXY-6, TAXY-7 and TAXY-8 were generated, and their reactivity was characterized along with another anti-tax, MAb, Lt-4. lmmunoblot assays showed that all the MAbs reacted with the PX141, the native tax, antigen expressed in various HTLV-I-infected cell lines and the gp68 of MT-2 cells expressing the tax, amino acids 94-353. lmmunoblot assays using recombinant, truncated tax, antigens, XD59 (expressing amino acids 180-338) and XD128 (expressing amino acids 1-47 and 286353) showed that: ( I ) TAXY-I and Lt-4 did not react with either antigen; (2) TAXY-6 and TAXY-8 reacted with only XD128; and (3) TAXY-7 reacted with both. In addition, TAXY-I, but not the other MAbs, reacted with a putative tax antigen of an STLV-I-infected cell line, designated RfM26-I. Competitive binding assays showed that TAXY-6 and TAXY-8 did not compete against each other. Sera from HTLV-I-infected humans interfered with the binding of all of these anti-tax, MAbs. These results indicate that the tax, antigen and the PX141 express at least 5 distinct epitopes recognized by human and mouse antibodies.