Liang Wang1, Jian Gao2, Xi Lan1,3, Hui Zhao1, Xiaoqian Shang1, Fengming Tian1, Hao Wen1, Jianbing Ding1,2, Li Luo1, Xiumin Ma1,2. 1. Clinical Medical Research Institute, State Key Laboratory of Pathogenesis, Prevention, Treatment of Central Asian High Incidence Diseases, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, China. 2. College of Basic Medicine of Xinjiang Medical University, Urumqi 830011, China. 3. Shenzhen Hospital, Southern Medical University, Shenzhen 518000, China.
Abstract
BACKGROUND: Identification of combined T-cell and B-cell reactive Eg95 antigens for the potential development of a multi-epitope vaccine against Echinococcus granulosus (EG), the causative agent of cystic echinococcosis (CE). METHODS: This study involved the recombinant expression of Eg95 along with associated immune rabbit antiserum preparation. Bioinformatics technology was used to facilitate the analysis of Eg95 molecules. PCR was subsequently used to amplify genetic sequences of the epitopes encoding the T-cell and B-cell reactive peptide fragments. SDS-PAGE was used to assess the expression levels of three proteins. Eg95 serum and patient antiserum, which were assessed using Western blot in order to identify suitable antigenic epitope peptides. ELISA detection assay facilitated comparison of the immune reactivity of the short peptide epitopes. The assay results could be used to determine an EG epitope-based vaccine candidate list from suitably reactive Eg95 epitopes. RESULTS: Eg95 molecules have 3 T-B table. The phage display systems were successfully built using the M13KE carrier. Expression of the three fusion protein peptides were detected. Western blot showed Eg95 antiserum against EG facilitated identification of the three T-cell and B-cell reactive epitopes. After the reaction intensities analyzed by the ELISA, both of the short peptide epitopes Eg95-2 and Eg95-3 showed strong signal strength and associated antigenicity when combined with patient serum and rabbit anti-rEg95 serum. CONCLUSIONS: This study used bioinformatics methods to construct successfully a T-cell and B-cell epitope phage display system for the Eg95 antigen from EG. The two epitopes of Eg95-2 and Eg95-3 demonstrated strong antigenicity with potential applications for peptide vaccine development. 2019 Annals of Translational Medicine. All rights reserved.
BACKGROUND: Identification of combined T-cell and B-cell reactive Eg95 antigens for the potential development of a multi-epitope vaccine against Echinococcus granulosus (EG), the causative agent of cystic echinococcosis (CE). METHODS: This study involved the recombinant expression of Eg95 along with associated immune rabbit antiserum preparation. Bioinformatics technology was used to facilitate the analysis of Eg95 molecules. PCR was subsequently used to amplify genetic sequences of the epitopes encoding the T-cell and B-cell reactive peptide fragments. SDS-PAGE was used to assess the expression levels of three proteins. Eg95 serum and patient antiserum, which were assessed using Western blot in order to identify suitable antigenic epitope peptides. ELISA detection assay facilitated comparison of the immune reactivity of the short peptide epitopes. The assay results could be used to determine an EG epitope-based vaccine candidate list from suitably reactive Eg95 epitopes. RESULTS: Eg95 molecules have 3 T-B table. The phage display systems were successfully built using the M13KE carrier. Expression of the three fusion protein peptides were detected. Western blot showed Eg95 antiserum against EG facilitated identification of the three T-cell and B-cell reactive epitopes. After the reaction intensities analyzed by the ELISA, both of the short peptide epitopes Eg95-2 and Eg95-3 showed strong signal strength and associated antigenicity when combined with patient serum and rabbit anti-rEg95 serum. CONCLUSIONS: This study used bioinformatics methods to construct successfully a T-cell and B-cell epitope phage display system for the Eg95 antigen from EG. The two epitopes of Eg95-2 and Eg95-3 demonstrated strong antigenicity with potential applications for peptide vaccine development. 2019 Annals of Translational Medicine. All rights reserved.
Authors: Liina Kinkar; Teivi Laurimäe; Gerardo Acosta-Jamett; Vanessa Andresiuk; Ibrahim Balkaya; Adriano Casulli; Robin B Gasser; Joke van der Giessen; Luis Miguel González; Karen L Haag; Houria Zait; Malik Irshadullah; Abdul Jabbar; David J Jenkins; Eshrat Beigom Kia; Maria Teresa Manfredi; Hossein Mirhendi; Selim M'rad; Mohammad Rostami-Nejad; Myriam Oudni-M'rad; Nora Beatriz Pierangeli; Francisco Ponce-Gordo; Steffen Rehbein; Mitra Sharbatkhori; Sami Simsek; Silvia Viviana Soriano; Hein Sprong; Viliam Šnábel; Gérald Umhang; Antonio Varcasia; Urmas Saarma Journal: Int J Parasitol Date: 2018-05-19 Impact factor: 3.981
Authors: Christiaan R de Vries; Qingquan Chen; Sally Demirdjian; Gernot Kaber; Arya Khosravi; Dan Liu; Jonas D Van Belleghem; Paul L Bollyky Journal: Curr Opin Biotechnol Date: 2020-12-11 Impact factor: 9.740