汉滩病毒Gc的细胞免疫反应性评估和验证

谢明阳; 孙报增; 马思婕; 杨端; 罗程; 王越越; 曹宇宸; 康文龙; 张赟; 杨琨; 姜东伯

doi:10.16016/j.2097-0927.202208161

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汉滩病毒Gc的细胞免疫反应性评估和验证

基础医学 | 更新时间：2026-01-09

- 汉滩病毒Gc的细胞免疫反应性评估和验证
- Evaluation and validation on cellular immunoreactivity of Hantaan virus Gc
- 陆军军医大学学报 2023年45卷第19期页码：2007-2017
- 作者机构：
  
  1.710032 西安，空军军医大学基础医学院免疫学教研室
  2.335000 江西鹰潭，中国人民武装警察部队江西总队鹰潭支队
- 作者简介：
  
  [姜东伯，E-mail: superjames1991@foxmail.comJIANG Dongbo, E-mail: superjames1991@foxmail.com
- 基金信息：
  
  国家自然科学金面上项目(82073154);空军军医大学提升计划课题(2020XXXX03)
- DOI：10.16016/j.2097-0927.202208161
  中图分类号： R373.32;R392.2;R394-33
- 收稿：2022-08-06，
  
  修回：2023-06-26，
  
  纸质出版：2023-10-15
- 稿件说明：
移动端阅览
谢明阳, 孙报增, 马思婕, 等. 汉滩病毒Gc的细胞免疫反应性评估和验证[J]. 陆军军医大学学报, 2023,45(19):2007-2017.

Mingyang XIE, Baozeng SUN, Sijie MA, et al. Evaluation and validation on cellular immunoreactivity of Hantaan virus Gc[J]. Journal of Army Medical University, 2023, 45(19): 2007-2017.
谢明阳, 孙报增, 马思婕, 等. 汉滩病毒Gc的细胞免疫反应性评估和验证[J]. 陆军军医大学学报, 2023,45(19):2007-2017. DOI： 10.16016/j.2097-0927.202208161.

Mingyang XIE, Baozeng SUN, Sijie MA, et al. Evaluation and validation on cellular immunoreactivity of Hantaan virus Gc[J]. Journal of Army Medical University, 2023, 45(19): 2007-2017. DOI： 10.16016/j.2097-0927.202208161.

摘要

目的

筛选、鉴定并验证汉滩病毒包膜糖蛋白C末端(Hantaan virus glycoprotein C-terminal，HTNV Gc)上免疫反应性表位。

方法

在预测覆盖率超过全球98%人群主要组织相容性复合物(major histocompatibility complex，MHC)基因基础上，通过免疫表位数据库(Immune Epitope Database

IEDB)、丹麦技术大学(Technical University of Denmark

DTU)等免疫信息学前沿算法系统分析HTNV Gc上9肽和15肽表位的亲和力，使用Vaxijen工具分析免疫原性，使用Blastp工具分析种间种内保守性；双向层次聚类比较MHC-Ⅰ类和Ⅱ类限制性表位的交叉免疫反应性；使用分子对接模拟出优势表位与MHC对接的可能结果；通过酶联免疫斑点(ELISpot)实验验证计算机分析结果。

结果

将多种生物信息学算法结果整合后，获得高亲和力、种间种内保守和强免疫原性的9肽和15肽表位，分别为8和80个，其中人鼠共同优势表位8个；聚类分析证明H-2基因型与HLA基因型存在部分相似；分子对接结果模拟出下优势表位与多个MHC基因型对接良好，且与亲和力分析结果一致；在非优势表位对比下，优势表位组小鼠脾细胞分泌IFN-γ的量更高。

结论

使用多种生物信息学算法成功筛选出HTNV Gc上优势表位，并通过ELISpot实验鉴定了上述表位的细胞免疫反应性。

Abstract

Objective

To screen

identify and verify the immunoreactive epitopes on Hantaan virus glycoprotein C-terminal (HTNV Gc).

Methods

On the basis of predicting the major histocompatibility complex (MHC) gene in more than 98% of the global population

the affinity of 9-peptide and 15-peptide epitopes on HTNV Gc was systematically analyzed by bioinformatics cutting-edge algorithms such as Immune Epitope Database (IEDB) and Technical University of Denmark (DTU). Their immunogenicity was analyzed by using the Vaxijen tool

and the interspecies intraspecific conservation was analyzed with the Blastp tool. Bidirectional hierarchical clustering was employed to compare the cross-immunoreactivity of MHC-class Ⅰ and class Ⅱ restricted epitopes. Possible results of dominant epitopes docking with MHC were simulated by using molecular docking. Enzyme-linked immunospot (ELISpot) assay was performed to validate the results of computer analyses.

Results

After integrating the results of various bioinformatics algorithms

we obtained 8 9-peptide epitopes and 80 15-peptide epitopes with high affinity

intraspecific conservation and strong immunogenicity

including 8 human-mouse common dominant epitopes. Cluster analysis showed that H-2 genotype had certain similarity as HLA genotype. The results of molecular docking revealed that dominant epitopes had good docking with various MHC genotypes

which were consistent with the results of affinity analysis. The amount of IFN-γ secreted from the spleen cells was higher in the mice with dominant epitopes than those with non-dominant epitopes.

Conclusion

With the aids of various bioinformatics algorithms

we successfully screen out the dominant epitopes on HTNV Gc

and validate the cellular immunoreactivity of these epitopes through ELISpot assay.

关键词

Keywords

references

WANG Q W, YUE M, YAO P P, et al . Epidemic trend and molecular evolution of HV family in the main hantavirus epidemic areas from 2004 to 2016, in P.R. China[J ] . Front Cell Infect Microbiol, 2021, 10: 584814. DOI: 10.3389/fcimb.2020.584814.

The problem with neoantigen prediction[J ] . Nat Biotechnol, 2017, 35(2): 97. DOI: 10.1038/nbt.3800 http://dx.doi.org/10.1038/nbt.3800 .

MUYANGWA M, MARTYNOVA E V, KHAIBOULLINA S F, et al . Hantaviral proteins: structure, functions, and role in hantavirus infection[J ] . Front Microbiol, 2015, 6: 1326. DOI: 10.3389/fmicb.2015.01326.

丁国永, 王志玉, 高璐, 等. 埃博拉病毒包膜糖蛋白研究进展[J]. 病毒学报, 2013, 29(2): 233-237.

DING G Y, WANG Z Y, GAO L, et al . Research progress on Ebola virus glycoprotein[J ] . Chin J Virol, 2013, 29(2): 233-237.

刘硕, 张黎, 王玉琳, 等. 埃博拉病毒包膜糖蛋白变异及抗原表位研究进展[J]. 病毒学报, 2019, 35(5): 801-812. DOI: 10.13242/j.cnki.bingduxuebao.003581.

LIU S, ZHANG L, WANG Y L, et al . Research progress in the envelope glycoprotein variation and epitope of Ebola virus[J ] . China Ind Econ, 2019, 35(5): 801-812. DOI: 10.13242/j.cnki.bingduxuebao.003581.

JIANG D B, ZHANG J P, CHENG L F, et al . Hantavirus Gc induces long-term immune protection via LAMP-targeting DNA vaccine strategy[J ] . Antivir Res, 2018, 150: 174-182. DOI: 10.1016/j.antiviral.2017.12 .011..

MA R X, CHENG L F, YING Q K, et al . Screening and identification of an H-2Kb-restricted CTL epitope within the glycoprotein of Hantaan virus[J ] . Front Cell Infect Microbiol, 2016, 6: 151. DOI: 10.3389/fcimb.2016.00151.

TANG K, CHENG L F, ZHANG C M, et al. Novel identified HLA-A*0201-restricted Hantaan virus glycoprotein cytotoxic T-cell epitopes could effectively induce protective responses in HLA-A2.1/K(b) transgenic mice may associate with the severity of hemorrhagic fever with renal syndrome[J]. Front Immunol, 2017, 8: 1797.

GREENBAUM J, SIDNEY J, CHUNG J, et al . Functional classification of class Ⅱ human leukocyte antigen (HLA) molecules reveals seven different supertypes and a surprising degree of repertoire sharing across supertypes[J ] . Immunogenetics, 2011, 63(6): 325-335. DOI: 10.1007/s00251-011-0513-0.

SERRIS A, STASS R, BIGNON E A, et al . The Hantavirus surface glycoprotein lattice and its fusion control mechanism[J ] . Cell, 2020, 183(2): 442-456. e16. DOI: 10.1016/j.cell.2020.08.023.

VITA R, MAHAJAN S, OVERTON J A, et al . The immune epit ope database (IEDB): 2018 update[J ] . Nucleic Acids Res, 2019, 47(D1): D339-D343. DOI: 10.1093/nar/gky1006.

KIM Y, SIDNEY J, PINILLA C, et al . Derivation of an amino acid similarity matrix for peptide: MHC binding and its application as a Bayesian prior[J ] . BMC Bioinformatics, 2009, 10: 394. DOI: 10.1186/1471-2105-10-394.

REYNISSON B, ALVAREZ B, PAUL S, et al . NetMHCpan-4.1 and NetMHCⅡpan-4.0: improved predictions of MHC antigen presentation by concurrent motif deconvolution and integration of MS MHC eluted ligand data[J ] . Nucleic Acids Res, 2020, 48(W1): W449-W454. DOI: 10.1093/nar/gkaa379.

RAMMENSEE H G, BACHMANN J, EMMERICH N P N, et al . SYFPEITHI: database for MHC ligands and peptide motifs[J ] . Immunogenetics, 1999, 50(3): 213-219. DOI: 10.1007/s002510050595.

RECHE P A, GLUTTING J P, ZHANG H, et al . Enhancement to the RANKPEP resource for the prediction of peptide binding to MHC molecules using profiles[J ] . Immunogenetics, 2004, 56(6): 405-419. DOI: 10.1007/s00251-004-0709-7.

JENSEN K K, ANDREATTA M, MARCATILI P, et al . Improved methods for predicting peptide binding affinity to MHC class Ⅱ molecules[J ] . Immunology, 2018, 154(3): 394-406. DOI: 10.1111/imm.12889.

GONZALEZ-GALARZA F F, MCCABE A, SANTOS E J M D, et al . Allele frequency net database (AFND) 2020 update: gold-standard data classification, open access genotype data and new query tools[J ] . Nucleic Acids Res, 2020, 48(D1): D783-D788. DOI: 10.1093/nar/gkz1029.

DOYTCHINOVA I A, FLOWER D R. VaxiJen: a server for prediction of protective antigens, tumour antigens and subunit vaccines[J]. BMC Bioinform, 2007, 8(1): 1-7. DOI: 10.1186/1471-2105-8-4.

ZHANG J, MADDEN T. PowerBLAST: a new network BLAST application for interactive or automated sequence analysis and annotation[J]. Genome Res, 1997, 7(6): 649-656. DOI: 10.1101/gr.7.6.649.

CHEN C J, CHEN H, ZHANG Y, et al . TBtools: an integrative toolkit developed for interactive analyses of big biological data[J ] . Mol Plant, 2020, 13(8): 1194-1202. DOI: 10.1016/j.molp.2020.06.009.

ZHOU P, JIN B W, LI H, et al . HPEPDOCK: a web server for blind peptide-protein docking based on a hierarchical alg orithm[J ] . Nucleic Acids Res, 2018, 46(W1): W443-W450. DOI: 10.1093/nar/gky357.

ATANASOVA M, PATRONOV A, DIMITROV I, et al . EpiDOCK: a molecular docking-based tool for MHC class Ⅱ binding prediction[J ] . Protein Eng Des Sel, 2013, 26(10): 631-634. DOI: 10.1093/protein/gzt018.

NGUYEN Q P, DENG T Z, WITHERDEN D A, et al . Origins of CD4 + circulating and tissue-resident memory T-cells[J ] . Immunology, 2019, 157(1): 3-12. DOI: 10.1111/imm.13059..

KAECH S M, WHERRY E J, AHMED R. Effector and memory T-cell differentiation: implications for vaccine development[J]. Nat Rev Immunol, 2002, 2(4): 251-262. DOI: 10.1038/nri778.

LIU J Y, MAO Q Y, WU X, et al . Considerations for the feasibility of neutralizing antibodies as a surrogate endpoint for COVID-19 vaccines[J ] . Front Immunol, 2022, 13: 814365.

SLOTA M, LIM J B, DANG Y S, et al . ELISpot for measuring human immune responses to vaccines[J ] . Expert Rev Vaccines, 2011, 10(3): 299-306. DOI: 10.1586/erv.10.169.

HILL P C, BROOKES R H, FOX A, et al . Longitudinal assessment of an ELISPOT test for Mycobacterium tuberculosis infection[J ] . PLoS Med, 2007, 4(6): e192. DOI: 10.1371/journal.pmed.0040192..

KESTER K E, CUMMINGS J F, OCKENHOUSE C F, et al . Phase 2a trial of 0, 1, and 3 month and 0, 7, and 28 day immunization schedules of malaria vaccine RTS, S/AS02 in malaria-naive adults at the Walter Reed Army Institute of Research[J ] . Vaccine, 2008, 26(18): 2191-2202. DOI: 10.1016/j.vaccine.2008.02.048.

ZHU F C, LI Y H, GUAN X H, et al . Safety, tolerability, and immunogenicity of a recombinant adenovirus type-5 vectored COVID-19 vaccine: a dose-escalation, open-label, non-randomised, first-in-human trial[J ] . Lancet, 2020, 395(10240): 1845-1854. DOI: 10.1016/S0140-6736(20)31208-3.

FOLEGATTI P M, EWER K J, ALEY P K, et al . Safety and immunogenicity of the ChAdOx1 nCoV-19 vaccine against SARS-CoV-2: a preliminary report of a phase 1/2, single-blind, randomised controlled trial[J ] . Lancet, 2020, 396(10249): 467-478. DOI: 10.1016/S0140-6736(20)31604-4.

MICHEL-TODÓ L, BIGEY P, RECHE P A, et al . Design of an epitope-based vaccine en semble for animal trypanosomiasis by computational methods[J ] . Vaccines, 2020, 8(1): 130. DOI: 10.3390/vaccines8010130.

RUSSO G, RECHE P, PENNISI M, et al . The combination of artificial intelligence and systems biology for intelligent vaccine design[J ] . Expert Opin Drug Discov, 2020, 15(11): 1267-1281. DOI: 10.1080/17460441.2020.1791076.

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