Distribution of epstein-barr virus antigenic sites on the carboxyl terminal end of ribonucleotide reductase against nasopharyngeal carcinoma serum antibodies using an immunoabsorption method.
In an attempt to clone and express proteins from the Epstein-Barr virus (EBV) cDNA library to be used as antigens in an enzyme-linked immunosorbent assay (ELISA) format to test against the antibodies found in the sera of nasopharyngeal carcinoma (NPC) patients, we have isolated and characterized three clones. All three clones expressed the same polypeptides of different lengths, which belong to the carboxyl terminal end of the large subunit of ribonucleotide reductase (RR) of the EBV genome. All three clones were found to be immunogenic and could be used in an IgA and IgG ELISA against the NPC sera with various degrees of sensitivity and specificity. Because the clones varied in length, this difference provides a simple system to determine where most of the antibody epitopes lies on the protein. We designed an immunoabsorption assay and a mathematical model to help map the segment of the polypeptide most immunogenic to 43 NPC patients. Results were unexpected: 77% of the patients were most immunogenic to region z, which was the smallest fragment among the three fragments studied. Fragment z was only 33 amino acids in length. Only 14% and 19% of patients showed the most immunogenic region in segment x and y, respectively. This variation could be due to major histocompatibility complex antigens. The patients could be divided into three groups based on the immunoabsorption assays, in which each group responded to a different immunodominant segment in the RR antigen. The largest group responded to an immunodominant segment, which was only 33 amino acids long. This domain was coded for by the gene fragment from nucleotide 78,129 to nucleotide 78,227 of the EBV genome. This segment of the protein would be suitable for further epitope mapping studies.
Gan
Y Y
YY
School of Science, Nanyang Technological University, Singapore. ganyy@nievax.nie.ac.sg
Hu
R
R
Chai
D
D
Tan
T T
TT
Gan
Y H
YH
Chan
S H
SH
Tsao
S Y
SY
Gan
L H
LH
D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000097:blood / Q000378:metabolism; D000956:Antigens, Viral / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D002277:Carcinoma / Q000276:immunology* / Q000821:virology; D018604:Epitope Mapping; D004854:Herpesvirus 4, Human / Q000201:enzymology / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007163:Immunosorbent Techniques; D008969:Molecular Sequence Data; D009303:Nasopharyngeal Neoplasms / Q000276:immunology* / Q000821:virology; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012264:Ribonucleotide Reductases / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins; D012264:Ribonucleotide Reductases