An HTLV-III peptide produced by recombinant DNA is immunoreactive with sera from patients with AIDS

• DOI: 10.1038/315151a0
• Publication Date: 2004-11-18T02:32:41Z
• Authors: N. T. Chang; J. Huang; J. Ghrayeb; S. McKinney; P. K. Chanda; T. W. Chang; S. Putney; M. G. Sarngadharan; F. Wong-Staal; R. C. Gallo
• Keywords: Acquired Immunodeficiency Syndrome; 0303 health sciences; Base Sequence; DNA, Recombinant; Antibodies, Viral; Deltaretrovirus; Peptide Fragments; 3. Good health; Molecular Weight; Viral Proteins; 03 medical and health sciences; Antibody Specificity; Humans; Amino Acid Sequence

Human T-cell lymphotropic retrovirus type III (HTLV-III), also called lymphadenopathy-associated virus (LAV), has been identified as the aetiological agent of acquired immune deficiency syndrome (AIDS). The sera of most patients with AIDS or AIDS-related complexes, and of asymptomatic individuals infected with HTLV-III, contain antibodies against antigens of HTLV-III. The characterization of these antibodies and their corresponding viral antigens is important not only for understanding immunity against HTLV-III and the pathology of AIDS, but also for the development of diagnostic methods and preventive vaccine for AIDS. Following the successful establishment of a long-term T-cell line permissive for HTLV-III replication, large quantities of virus have been produced, facilitating the purification of viral proteins and the development of mouse monoclonal antibodies against several viral antigens. More recently, the structure of HTLV-III proviral DNA has been elucidated. We now report the production, by genetic engineering methods, of a peptide encoded by a gene segment of HTLV-III. A 1.1-kilobase (kb) EcoRI DNA segment from an isolate of HTLV-III was inserted into a lpp and lac promoter-coupled expression vector, pIN-III-ompA. Escherichia coli transformants of this plasmid produced a peptide of relative molecular mass (Mr) 15,000 (15K) which was strongly immunoreactive with anti-HTLV-III antibodies present in sera from AIDS patients. Lysates of the clones expressing this 15K peptide inhibited the reactivity of the p31 virion protein with AIDS sera, suggesting that it is a fragment of the viral p31 protein. The peptide reacted with sera from all 20 AIDS patients but none of the 8 normal controls tested. These results suggest that the peptide may be useful for detecting anti-HTLV-III antibodies in blood samples.