[Prediction and immunologic identification of antigenic epitopes in genus-specific outer membrane protein LipL41 of Leptospira interrogans].
To predict and screen the efficient antigenic epitopes in genus-specific envelope protein LipL41 of Leptospira interrogans and to determine the immunoreactive diversity of LipL41s from different genotypes.



Bioinformatic methods were applied to predict the T/B combined epitope candidates in LipL41/1 and LipL41/2 molecules. The nucleotide fragments encoding epitopes were amplified by PCR. Phage display system with SDS-PAGE was performed to obtain the recombinant PIIIs containing different T/B combined epitopes. Western Blot assays were performed to determine the immunoreactivity of recombinant PIIIs to various antisera including antiserum against rLipL41/1, rLipL41/2 and whole cell of L.interrogans strain Lai, and serum from patients with leptospirosis.



Based on the predicting data, eight common or differential combined epitopes in LipL41s were selected. The nucleotide fragments encoding the epitopes were obtained by PCR. All the T/B combined epitope fragments were correctly inserted into the N end of phage PIII protein and then successfully expressed. All the antisera were able to recognize each of the epitopes but the hybridization signal intensity was different. Among these epitopes, the common T/B combined epitopes LipL41/1-30 and LipL41/1-233 showed a stronger and stable hybridization signals.



All 8 selected T/B combined epitopes in the study are the efficient antigenic epitopes. The common T/B epitopes LipL41/1-30 and LipL41/1-233 can be first used in development of leptospiral MAP vaccine. The cross immunoreaction is between the differential T/B epitopes LipL41s-89,LipL41s-299 and the different antisera.
Jiang
Jiu-kun
JK
Department of Microbiology and Parasitology, College of Medicine, Zhejiang University, and State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Hangzhou 310058, China.
Lin
Xu-ai
XA
Yan
Jie
J
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology*; D001428:Bacterial Vaccines / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology*; D005838:Genotype; D008969:Molecular Sequence Data; D019151:Peptide Library
D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; C538745:LipL41 protein, Leptospira interrogans; D019151:Peptide Library