Title:
METHOD FOR SCREENING CTL EPITOPES FROM SELF-CONSTRUCTED SLA-2-HB01-PCDH/ST2 CELL LINE
Document Type and Number:
WIPO Patent Application WO/2023/103266
Kind Code:
A1
Abstract:
A method for screening CTL epitopes from a self-constructed SLA-2-HB01-pCDH/sT2 cell line. An exogenous SLA-2-HB01 gene is transfected to sT2 cells to establish a stable SLA-2 gene expression model. Positive epitope peptides such as EB155 are loaded on the surfaces of cells, an SLA-2-peptides-β2m complex expressed on the surfaces of the cells is detected by means of FACs, and that SLA-2-HB01-pCDH/sT2 has the function of presenting exogenous polypeptide epitopes is determined. Results show that: As63, EB155, and Hu62 peptides can all be presented by an SLA-2-HB01-pCDH/sT2 cell line, a series of constructed sT2 for expressing the SLA-2-HB01 gene has the function of presenting antigens and can be used for screening polypeptide epitopes of swine viruses on a cellular level, and the antigen presentation function of a cell line is not affected when a Flag tag is added to a C-terminal of an SLA-2 heavy chain molecule.
Inventors:
GAO FENGSHAN (CN)
Application Number:
PCT/CN2022/089951
Publication Date:
June 15, 2023
Filing Date:
April 28, 2022
Export Citation:
Assignee:
UNIV DALIAN (CN)
International Classes:
G01N33/68; C12N5/077
Foreign References:
| CN114181895A | 2022-03-15 | |||
| CN112480238A | 2021-03-12 |
Other References:
PARK CHAN-SU, IM SUN-A, SONG SUKGIL, KIM KYUNGJAE, LEE CHONG-KIL: "Identification of HLA-A2-restricted immunogenic peptides derived from a xenogenic porcine major histocompatibility complex", XENOTRANSPLANTATION, WILEY-BLACKWELL PUBLISHING, INC., US, vol. 21, no. 5, 1 September 2014 (2014-09-01), US , pages 465 - 472, XP093070771, ISSN: 0908-665X, DOI: 10.1111/xen.12119
FENG LEI, SUN MING-WEI, JIANG PING, LI ZI-BIN, GAO HUA, ZHAI XIAO-XIN, HAN YONG, ZHANG ZONG-HUI, GAO FENG-SHAN: "Purification, crystallization and preliminary X-ray crystallographic studies of swine MHC class I complexed with an FMDV CTL epitope Hu64", RESEARCH IN VETERINARY SCIENCE, BRITISH VETERINARY ASSOCIATION, LONDON., GB, vol. 119, 1 August 2018 (2018-08-01), GB , pages 205 - 208, XP093070773, ISSN: 0034-5288, DOI: 10.1016/j.rvsc.2018.06.018
GAO FENGSHAN, JIANG PING, FANG QINGMEI, LI YUNGANG, LI XINSHENG, WANG QINFU, CHI YAN, XIA CHUN: "Screening of Reconstructed Porcine SLA-I Single Chain Complex and Its Restricted Epitope of Foot-and-Mouth Disease Virus VP1 Protein", PROGRESS IN NATURAL SCIENCE, vol. 17, no. 6, 1 June 2007 (2007-06-01), pages 724 - 730, XP093070778
GAO FENG-SHAN, FENG LEI, JIANG PING, LI ZI-BIN, GAO HUA, ZHAI XIAO-XIN, ZHANG ZONG-HUI, HU XIAO: "Expression, purification, crystallization and preliminary X-ray diffraction analysis of swine leukocyte antigen 2 complexed with a CTL epitope AS64 derived from Asia1 serotype of foot-and-mouth disease virus", BMC VETERINARY RESEARCH, vol. 14, no. 1, 1 December 2018 (2018-12-01), pages 407, XP093070781, DOI: 10.1186/s12917-018-1742-3
ZHANG NIANZHI, QI JIANXUN, FENG SIJIA, GAO FENG, LIU JUN, PAN XIAOCHENG, CHEN RONG, LI QIRUN, CHEN ZHAOSAN, LI XIAOYING, XIA CHUN,: "Crystal Structure of Swine Major Histocompatibility Complex Class I SLA-1*0401 and Identification of 2009 Pandemic Swine-Origin Influenza A H1N1 Virus Cytotoxic T Lymphocyte Epitope Peptides", JOURNAL OF VIROLOGY, THE AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 85, no. 22, 15 November 2011 (2011-11-15), US , pages 11709 - 11724, XP093070785, ISSN: 0022-538X, DOI: 10.1128/JVI.05040-11
FENG LEI, SUN MING-WEI, JIANG PING, LI ZI-BIN, GAO HUA, ZHAI XIAO-XIN, HAN YONG, ZHANG ZONG-HUI, GAO FENG-SHAN: "Purification, crystallization and preliminary X-ray crystallographic studies of swine MHC class I complexed with an FMDV CTL epitope Hu64", RESEARCH IN VETERINARY SCIENCE, BRITISH VETERINARY ASSOCIATION, LONDON., GB, vol. 119, 1 August 2018 (2018-08-01), GB , pages 205 - 208, XP093070773, ISSN: 0034-5288, DOI: 10.1016/j.rvsc.2018.06.018
GAO FENGSHAN, JIANG PING, FANG QINGMEI, LI YUNGANG, LI XINSHENG, WANG QINFU, CHI YAN, XIA CHUN: "Screening of Reconstructed Porcine SLA-I Single Chain Complex and Its Restricted Epitope of Foot-and-Mouth Disease Virus VP1 Protein", PROGRESS IN NATURAL SCIENCE, vol. 17, no. 6, 1 June 2007 (2007-06-01), pages 724 - 730, XP093070778
GAO FENG-SHAN, FENG LEI, JIANG PING, LI ZI-BIN, GAO HUA, ZHAI XIAO-XIN, ZHANG ZONG-HUI, HU XIAO: "Expression, purification, crystallization and preliminary X-ray diffraction analysis of swine leukocyte antigen 2 complexed with a CTL epitope AS64 derived from Asia1 serotype of foot-and-mouth disease virus", BMC VETERINARY RESEARCH, vol. 14, no. 1, 1 December 2018 (2018-12-01), pages 407, XP093070781, DOI: 10.1186/s12917-018-1742-3
ZHANG NIANZHI, QI JIANXUN, FENG SIJIA, GAO FENG, LIU JUN, PAN XIAOCHENG, CHEN RONG, LI QIRUN, CHEN ZHAOSAN, LI XIAOYING, XIA CHUN,: "Crystal Structure of Swine Major Histocompatibility Complex Class I SLA-1*0401 and Identification of 2009 Pandemic Swine-Origin Influenza A H1N1 Virus Cytotoxic T Lymphocyte Epitope Peptides", JOURNAL OF VIROLOGY, THE AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 85, no. 22, 15 November 2011 (2011-11-15), US , pages 11709 - 11724, XP093070785, ISSN: 0022-538X, DOI: 10.1128/JVI.05040-11
Attorney, Agent or Firm:
DALIAN EAST PATENT AGENT LTD. (CN)
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