A variable-length PCR target protein of Ehrlichia chaffeensis contains major species-specific antibody epitopes in acidic serine-rich tandem repeats.
Ehrlichia chaffeensis and E. canis have a small subset of tandem repeat (TR)-containing proteins that elicit strong host immune responses and are associated with host-pathogen interactions. In a previous study, we molecularly characterized a highly conserved 19-kDa major immunoreactive protein (gp19) of E. canis and identified the corresponding TR-containing ortholog variable-length PCR target (VLPT) protein in E. chaffeensis. In this study, the native 32-kDa VLPT protein was identified and the immunodeterminants defined in order to further understand the molecular basis of the host immune response to E. chaffeensis. Synthetic and/or recombinant polypeptides corresponding to various regions of VLPT were used to localize major antibody epitopes to the TR-containing region. Major antibody epitopes were identified in three nonidentical repeats (R2, R3, and R4), which reacted strongly with antibodies in sera from an E. chaffeensis-infected dog and human monocytotropic ehrlichiosis patients. VLPT-R3 and VLPT-R2 reacted most strongly with antibody, and the epitope was further localized to a nearly identical proximal 17-amino-acid region common between these repeats that was species specific. The epitope in R4 was distinct from that of R2 and R3 and was found to have conformational dependence. VLPT was detected in supernatants from infected cells, indicating that the protein was secreted. VLPT was localized on both reticulate and dense-core cells, and it was found extracellularly in the morula fibrillar matrix and associated with the morula membrane.
Luo
Tian
T
Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555-0609, USA.
Zhang
Xiaofeng
X
Wakeel
Abdul
A
Popov
Vsevolod L
VL
McBride
Jere W
JW
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial; D018469:Ehrlichia chaffeensis / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000939:Epitopes / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D015964:Gene Expression Regulation, Bacterial; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012694:Serine / Q000737:chemistry*; D013045:Species Specificity; D020080:Tandem Repeat Sequences
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D000939:Epitopes; D012694:Serine