Screening and identification of HLA-A*02:01-restricted CTL epitopes in the glycoproteins Gn and Gc of Dabie bandavirus

ZHU Yuheng; LIU Jingxian; CUI Lunbiao; XIAO Zhongdang

doi:10.16016/j.2097-0927.202605008

您当前的位置：

首页 >

文章列表页 >

Screening and identification of HLA-A*02:01-restricted CTL epitopes in the glycoproteins Gn and Gc of Dabie bandavirus

Basic Medicine | 更新时间：2026-07-15

- Screening and identification of HLA-A*02:01-restricted CTL epitopes in the glycoproteins Gn and Gc of Dabie bandavirus
- Journal of Army Medical University Vol. 48, Issue 13, Pages: 1876-1885(2026)
- 作者机构：
  
  1.东南大学生物科学与医学工程学院，江苏南京
  2.江苏省疾病预防控制中心，江苏南京
- 作者简介：
  
  CUI Lunbiao， E-mail：lbcui@jscdc.cn
  XIAO Zhongdang， E-mail：zdxiao@seu.edu.cn
- 基金信息：
  
  the Scientific Research Fund of Jiangsu Provincial Health Department(MQ2025039)
- DOI：10.16016/j.2097-0927.202605008
  CLC： R341;R373.9;R392.3
- Received：07 May 2026，
  
  Revised：2026-06-16，
  
  Accepted：18 June 2026，
  
  Published：15 July 2026
- 稿件说明：
移动端阅览
朱誉恒，刘静娴，崔仑标，等.大别班达病毒糖蛋白Gn和Gc中HLA-A*02：01限制性CTL表位的筛选与鉴定［J］.陆军军医大学学报，2026，48（13）：1876-1885.DOI：10.16016/j.2097-0927.202605008.

ZHU Y H，LIU J X，CUI L B，et al.Screening and identification of HLA-A*02：01-restricted CTL epitopes in the glycoproteins Gn and Gc of Dabie bandavirus［J］.J Army Med Univ，2026，48（13）：1876-1885.DOI：10.16016/j.2097-0927.202605008.
朱誉恒，刘静娴，崔仑标，等.大别班达病毒糖蛋白Gn和Gc中HLA-A*02：01限制性CTL表位的筛选与鉴定［J］.陆军军医大学学报，2026，48（13）：1876-1885.DOI：10.16016/j.2097-0927.202605008. DOI：

ZHU Y H，LIU J X，CUI L B，et al.Screening and identification of HLA-A*02：01-restricted CTL epitopes in the glycoproteins Gn and Gc of Dabie bandavirus［J］.J Army Med Univ，2026，48（13）：1876-1885.DOI：10.16016/j.2097-0927.202605008. DOI：

摘要

目的

大别班达病毒（Dabie bandavirus，DBV）感染所致发热伴血小板减少综合征（severe fever with thrombocytopnia syndrome，SFTS）病死率高，目前尚无特效疫苗，而DBV糖蛋白前体（glycoprotein precursor，GP）是重要的免疫靶点，但其细胞毒性T淋巴细胞（cytotoxic T lymphocyte，CTL）表位尚不明确。本研究以DBV GP为研究对象，通过生物信息学工具预测结合动物体外实验验证的方法筛选并鉴定具有较强免疫原性的CTL表位，为DBV特异性细胞免疫机制研究及表位疫苗设计提供理论依据。

方法

以DBV HB29株糖蛋白n（glycoprotein n，Gn）和糖蛋白c（glycoprotein c，Gc）氨基酸序列为模板，利用IEDB、NetMHCpan 4.1、STMHCpan 3种生物信息学工具，预测长度为8~11个氨基酸的HLA-A*02：01限制性T细胞表位。经ToxinPred2、AlgPred 2.0评估毒性与致敏性，并通过20株国内DBV分离株序列比对分析保守性。选取在2种及以上工具中结合亲和力排名前2%的作为表位肽。将6~8周龄雌性HLA-A*02：01转基因小鼠随机分为2组（

=2）：Gn组和Gc组，分别以Gn（100 μg/只）或Gc（100 μg/只）联合弗氏佐剂于第0、14、28天进行3次免疫。末次免疫后7 d处死小鼠，取脾脏制备单细胞悬液。使用各表位肽（终浓度25 μg/mL）体外刺激脾细胞（5×10

个/孔），采用ELISPOT检测IFN-γ分泌水平，以每10

个脾细胞中斑点形成细胞（spot-forming cell，SFC）值100为阳性判断标准。为了避免CD4

T细胞分泌IFN-γ对结果的影响，通过磁珠阳性分选法获得CD4

T细胞耗竭的脾细胞（CD4

T cells-depleted splenocytes，CD4⁻SP），以相同条件再次进行ELISPOT检测，以每10

个CD4⁻SP中SFC值100为阳性判断标准。

结果

经3种工具交叉预测，共筛选出12条表位肽（Gn1~Gn6、Gc1~Gc6）。各表位肽经免疫原性预测均显示良好潜力，通过毒性及致敏性评估，并且在多地的分离株中表现出良好的保守性。分子对接模拟结果显示，12条表位肽与HLA-A*02：01分子均能形成稳定的对接复合物。ELISPOT结果显示：Gn5、Gn6、Gc1、Gc2、Gc3、Gc4、Gc5、Gc6能显著诱导免疫小鼠脾细胞分泌IFN-γ（SFC值均100，

均0.000 1）；在CD4⁻SP中，Gn5、Gn6、Gc3、Gc4仍能诱导强IFN-γ应答（SFC值均100，

均0.000 1）。

结论

本研究成功从DBV中鉴定出4条能特异性诱导CD8

T细胞产生IFN-γ应答的CTL表位（Gn5、Gn6、Gc3、Gc4），并且在国内DBV分离株中高度保守，为DBV多表位疫苗的研发提供了实验依据。

Abstract

Objective

Dabie bandavirus (DBV) infection causes severe fever with thrombocytopenia syndrome (SFTS)

which has a high fatality rate and for which no specific vaccine is currently available. The DBV glycoprotein precursor (GP) is an important immunological target

however

its cytotoxic T lymphocyte (CTL) epitopes remain unclear. This study aims to screen and identify CTL epitopes with strong immunogenicity from DBV GP using bioinformatics tools for prediction

combined with

in vitro

animal experiments for validation

providing a theoretical basis for research on DBV-specific cellular immune mechanisms and epitope-based vaccine design.

Methods

Using the amino acid sequences of glycoprotein n (Gn) and glycoprotein c (Gc) of the DBV HB29 strain as templates

3 bioinformatics tools

namely IEDB

NetMHCpan 4.1

and STMHCpan

were employed to predict HLA-A*02:01-restricted T-cell epitopes with a length of 8 to 11 amino acids. Toxicity and allergenicity were evaluated using ToxinPred2 and AlgPred 2.0

and conservation was analyzed through sequence alignment of 20 domestic DBV isolates. The peptides ranked in the top 2% for binding affinity by at least 2 of the 3 tools were selected as epitope peptides. Female HLA-A*02:01 transgenic mice

(6 to 8 weeks old) were randomly divided into a Gn group and a Gc group (

=2). The transgenic mice were immunized on days 0

and 28 for a total of 3 immunizations with Gn (100 μg/mouse) or Gc (100 μg/mouse) combined with Freund's adjuvant. In 7 d after the last immunization

the mice were sacrificed

and the spleens were harvested to prepare single-cell suspensions. Splenocytes (5×10⁵ cells/well) were stimulated

in vitro

with each epitope peptide at a final concentration of 25 μg/mL

and IFN-γ secretion was detected using ELISPOT assay. A spot-forming cell (SFC) value 100 per 10⁵ splenocytes was set as the positive criterion. To eliminate the potential interference of IFN-γ secreted by CD4⁺T cells

CD4⁺T cell-depleted splenocytes (CD4⁻SP) were obtained

via

magnetic bead-based positive selection

and ELISPOT assay was performed again under the same conditions

with an SFC value 100 per 10⁴ CD4⁻SP set as the positive criterion.

Results

A total of 12 epitope peptides (Gn1 to Gn6 and Gc1 to Gc6) were screened out through cross-prediction using the 3 tools. All epitope peptides showed promising potential in immunogenicity

passed toxicity and allergenicity evaluation

and exhibited good conservation among isolates from multiple regions. Molecular docking showed that all 12 epitope peptides could significantly induce IFN-γ secretion by splenocytes from immunized mice (all SFC values 100

all

0.000 1). In CD4⁻SP

Gn5

Gn6

Gc3 and Gc4 still induced strong IFN-γ responses (all SFC values 100

all

0.000 1).

Conclusion

Four CTL epitopes (Gn5

Gn6

Gc3 and Gc4) that can specifically induce IFN-γ responses in CD8⁺T cells were successfully identified from DBV. These epitopes are highly conserved among domestic DBV isolates

providing experimental evidence for the development of multi-epitope vaccines against DBV.

关键词

Keywords

references

ZHUANG L ， SUN Y ， CUI X M ， et al . Transmission of severe fever with thrombocytopenia syndrome virus by Haemaphysalis longicornis ticks， China ［J］. Emerg Infect Dis ， 2018 ， 24 （ 5 ）： 868 - 871 . DOI： 10.3201/eid2405.151435 http://dx.doi.org/10.3201/eid2405.151435 .

TRAN X C ， YUN Y ， VAN AN L ， et al . Endemic severe fever with thrombocytopenia syndrome， Vietnam ［J］. Emerg Infect Dis ， 2019 ， 25 （ 5 ）： 1029 - 1031 . DOI： 10.3201/eid2505.181463 http://dx.doi.org/10.3201/eid2505.181463 .

HE Z Q ， WANG B H ， LI Y ， et al . Severe fever with thrombocytopenia syndrome： a systematic review and meta-analysis of epidemiology， clinical signs， routine laboratory diagnosis， risk factors， and outcomes ［J］. BMC Infect Dis ， 2020 ， 20 （ 1 ）： 575 . DOI： 10.1186/s12879-020-05303-0 http://dx.doi.org/10.1186/s12879-020-05303-0 .

LI J Z ， WANG C P ， LI X ， et al . Direct transmission of severe fever with thrombocytopenia syndrome virus from farm-raised fur animals to workers in Weihai， China ［J］. Virol J ， 2024 ， 21 （ 1 ）： 113 . DOI： 10.1186/s12985-024-02387-x http://dx.doi.org/10.1186/s12985-024-02387-x .

KIM S ， JEON K ， CHOI H ， et al . Comparative analysis of the efficacy of vaccines using structural protein subunits of the severe fever with thrombocytopenia syndrome virus ［J］. Front Microbiol ， 2024 ， 15 ： 1348276 . DOI： 10.3389/fmicb. 2024.1348276 http://dx.doi.org/10.3389/fmicb.2024.1348276 .

KIM D ， LAI C J ， CHA I ， et al . Current progress of severe fever with thrombocytopenia syndrome virus （SFTSV） vaccine development ［J］. Viruses ， 2024 ， 16 （ 1 ）： 128 . DOI： 10.3390/v16010128 http://dx.doi.org/10.3390/v16010128 .

KWAK J E ， KIM Y I ， PARK S J ， et al . Development of a SFTSV DNA vaccine that confers complete protection against lethal infection in ferrets ［J］. Nat Commun ， 2019 ， 10 （ 1 ）： 3836 . DOI： 10.1038/s41467-019-11815-4 http://dx.doi.org/10.1038/s41467-019-11815-4 .

DU S W ， PENG R C ， XU W ， et al . Cryo-EM structure of severe fever with thrombocytopenia syndrome virus ［J］. Nat Commun ， 2023 ， 14 （ 1 ）： 6333 . DOI： 10.1038/s41467-023-41804-7 http://dx.doi.org/10.1038/s41467-023-41804-7 .

HOFMANN H ， LI X X ， ZHANG X A ， et al . Severe fever with thrombocytopenia virus glycoproteins are targeted by neutralizing antibodies and can use DC-SIGN as a receptor for pH-dependent entry into human and animal cell lines ［J］. J Virol ， 2013 ， 87 （ 8 ）： 4384 - 4394 . DOI： 10.1128/JVI.02628-12 http://dx.doi.org/10.1128/JVI.02628-12 .

MANZONI T B ， WESTOVER J B ， LUNDGREEN K A ， et al . Homologous and heterologous vaccination regimens with mRNA and rVSV platforms induce potent immune responses against SFTSV glycoprotein ［J］. Viruses ， 2025 ， 17 （ 8 ）： 1095 . DOI： 10.3390/v17081095 http://dx.doi.org/10.3390/v17081095 .

SUN H ， HU Q M ， LU S W ， et al . Current status of severe fever with thrombocytopenia syndrome in China （Review）［J］. Int J Mol Med ， 2025 ， 56 （ 5 ）： 169 . DOI： 10.3892/ijmm.2025.5610 http://dx.doi.org/10.3892/ijmm.2025.5610 .

JIANG D B ， MA Z L ， ZHANG J Q ， et al . Immunoreactivit y analysis of MHC-I epitopes derived from the nucleocapsid protein of SARS-CoV-2 via computation and vaccination ［J］. Vaccines ， 2024 ， 12 （ 11 ）： 1214 . DOI： 10.3390/vaccines12111214 http://dx.doi.org/10.3390/vaccines12111214 .

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 http://dx.doi.org/10.1093/nar/gkz1029 .

WANG G Z ， WAN H H ， JIAN X X ， et al . INeo-epp： a novel T-cell HLA class-I immunogenicity or neoantigenic epitope prediction method based on sequence-related amino acid features ［J］. Biomed Res Int ， 2020 ， 2020 ： 5798356 . DOI： 10.1155/2020/5798356 http://dx.doi.org/10.1155/2020/5798356 .

VITA R ， BLAZESKA N ， MARRAMA D ， et al . The immune epitope database （IEDB）： 2024 update ［J］. Nucleic Acids Res ， 2025 ， 53 （ D1 ）： D436 - D443 . DOI： 10.1093/nar/gkae1092 http://dx.doi.org/10.1093/nar/gkae1092 .

YE Z ， LI S H ， MI X ， et al . STMHCpan， an accurate Star-Transformer-based extensible framework for predicting MHC I allele binding peptides ［J］. Brief Bioinform ， 2023 ， 24 （ 3 ）： bbad164 . DOI： 10.1093/bib/bbad164 http://dx.doi.org/10.1093/bib/bbad164 .

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 http://dx.doi.org/10.1093/nar/gkaa379 .

CROOKS G E ， HON G ， CHANDONIA J M ， et al . WebLogo： a sequence logo generator ［J］. Genome Res ， 2004 ， 14 （ 6 ）： 1188 - 1190 . DOI： 10.1101/gr.849004 http://dx.doi.org/10.1101/gr.849004 .

SHEN Y M ， MAUPETIT J ， DERREUMAUX P ， et al . Improved PEP-FOLD approach for peptide and miniprotein structure prediction ［J］. J Chem Theory Comput ， 2014 ， 10 （ 10 ）： 4745 - 4758 . DOI： 10.1021/ct500592m http://dx.doi.org/10.1021/ct500592m .

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

SHARMA N ， NAOREM L D ， JAIN S ， et al . ToxinPred2： an improved method for predicting toxicity of proteins ［J］. Brief Bioinform ， 2022 ， 23 （ 5 ）： bbac174 . DOI： 10.1093/bib/bbac174 http://dx.doi.org/10.1093/bib/bbac174 .

SHARMA N ， PATIYAL S ， DHALL A ， et al . AlgPred 2.0： an improved method for predicting allergenic proteins and mapping of IgE epitopes ［J］. Brief Bioinform ， 2021 ， 22 （ 4 ）： bbaa294 . DOI： 10.1093/bib/bbaa294 http://dx.doi.org/10.1093/bib/bbaa294 .

CHEN C H ， LI J X ， ZHOU Y M ， et al . Latest advances and prospects in the pathogenesis， animal models， and vaccine research of severe fever with thrombocytopenia syndrome virus ［J］. Front Immunol ， 2025 ， 16 ： 1624290 . DOI： 10.3389/fimmu.2025.1624290 http://dx.doi.org/10.3389/fimmu.2025.1624290 .

GRIFONI A ， WEISKOPF D ， RAMIREZ S I ， et al . Targets of T cell responses to SARS-CoV-2 coronavirus in humans with COVID-19 disease and unexposed individuals ［J］. Cell ， 2020 ， 181 （ 7 ）： 1489 - 1501.e15 . DOI： 10.1016/j.cell.2020.05.015 http://dx.doi.org/10.1016/j.cell.2020.05.015 .

SETTE A ， CROTTY S . Adaptive immunity to SARS-CoV-2 and COVID-19 ［J］. Cell ， 2021 ， 184 （ 4 ）： 861 - 880 . DOI： 10.1016/j.cell.2021.01.007 http://dx.doi.org/10.1016/j.cell.2021.01.007 .

TARKE A ， SIDNEY J ， KIDD C K ， et al . Comprehensive analysis of T cell immunodominance and immunoprevalence of SARS-CoV-2 epitopes in COVID-19 cases ［J］. Cell Rep Med ， 2021 ， 2 （ 2 ）： 100204 . DOI： 10.1016/j.xcrm.2021. 100204 http://dx.doi.org/10.1016/j.xcrm.2021.100204 .

EICKHOFF C S ， TERRY F E ， PENG L D ， et al . Highly conserved influenza T cell epitopes induce broadly protective immunity ［J］. Vaccine ， 2019 ， 37 （ 36 ）： 5371 - 5381 . DOI： 10.1016/j.vaccine.2019.07.033 http://dx.doi.org/10.1016/j.vaccine.2019.07.033 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Alpha-2,3 sialylation promotes osteogenic and chondrogenic differentiation by regulating extracellular matrix homeostasis to participate in skeletal development

Impact of subjective cognitive decline on frailty in elderly patients with metabolic syndrome: the parallel mediating effects of physical activity and depression

FABP4 inhibitor BMS309403 attenuates hepatopulmonary injury in rats with hepatopulmonary syndrome through regulating the FABP4/PPAR-γ axis

MCC950 protects retinal photoreceptor cells against blast-induced injury by inhibiting the NLRP3/Caspase-1/GSDMD pathway in mice

Diminished small-world properties and compensatory reorganization of resting-state functional brain networks in patients with subacute spontaneous intracerebral hemorrhage: a graph-theoretic analysis of topological properties and their relationship with cognitive impairment

Related Author

No data

Related Institution

No data

⁰