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Peptide microarray mapping of B cell epitopes on bovine leukemia virus and peptide ELISA analysis of conservation of epitopes in BLV infected Japanese cattle
© Bai et al. 2015
Published: 28 August 2015
The bovine leukemia virus (BLV), a retrovirus structurally and functionally related to the human T-lymphotropic viruses HTLV-1 and HTLV-2, is the etiological agent of bovine leucosis. B cell immune mechanisms may play a major role in protection against BLV infection. A battery of 157 synthetic peptides, 15-mer length, 4 amino acids overlapped, was used to mapping B cell epitopes on BLV envelope glycoprotein gp5l and capsid protein by peptide microarrays. Two susceptible cattle with the homozygote BoLA-DRB3 *1601 allele and two non-susceptible cattle with DRB3 *1501/*2703 alleles and DRB3 *1501/*0503 alleles were infected with BLV, and serums were collected from those cattle each of having challenged BLV before and after. Only one epitope A out of 157 synthetic peptides responds with all of four BLV positive serums not their negative serums. To demonstrate whether epitope A is common B cell epitope or not by our established peptide ELISA system that uses maleimide activated mariculture keyhole limpet hemocyanin (mcKLH) carrier protein. To found, there are 7 kinds of BLV positive serum no response with epitope A among 232 kinds of positive serum. Furthermore, we searched other peptides respond with BLV positive serum that contain 7 kinds of serum no responded with epitope A and found epitope B among peptides of non-specifically responded on peptide microarray. Epitope B strongly responded with all of 232 kinds of serum as estimated by peptide ELISA system. Our results shows that epitope B has a tendency to react some BLV negative serums, thus we will perform further experiments to determine the common B cell epitope just responses with positive serum by peptides that are 3 alanine substitutions as shifting one amino acid on epitope sequence.
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.