CN107022006A - 乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列 - Google Patents
乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列 Download PDFInfo
- Publication number
- CN107022006A CN107022006A CN201710191364.6A CN201710191364A CN107022006A CN 107022006 A CN107022006 A CN 107022006A CN 201710191364 A CN201710191364 A CN 201710191364A CN 107022006 A CN107022006 A CN 107022006A
- Authority
- CN
- China
- Prior art keywords
- hepatitis
- leu
- ser
- pro
- antigen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56966—Animal cells
- G01N33/56977—HLA or MHC typing
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
- G01N33/5761—Hepatitis B
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
- G01N33/5761—Hepatitis B
- G01N33/5762—Hepatitis B core antigen
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
- G01N33/5761—Hepatitis B
- G01N33/5764—Hepatitis B surface antigen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/53—DNA (RNA) vaccination
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2730/00—Reverse transcribing DNA viruses
- C12N2730/00011—Details
- C12N2730/10011—Hepadnaviridae
- C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
- C12N2730/10122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2730/00—Reverse transcribing DNA viruses
- C12N2730/00011—Details
- C12N2730/10011—Hepadnaviridae
- C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
- C12N2730/10134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Communicable Diseases (AREA)
- Virology (AREA)
- Organic Chemistry (AREA)
- Epidemiology (AREA)
- Genetics & Genomics (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Mycology (AREA)
- Zoology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明公开了一组乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列,分别是由HLA‑A*0101、A*0301、A*3001、A*3101、A*3201、A*3303、A*6801等分子限制性的抗原肽,可以特异性地与细胞毒性胸腺依赖性淋巴细胞结合,刺激后者活化、增殖和分化,从而发挥抗乙型肝炎病毒的免疫效应作用。这些抗原肽可以用来制备乙型肝炎病毒感染的治疗性和预防性疫苗,也可以用来制备检测乙型肝炎病毒特异性细胞毒型胸腺依赖性淋巴细胞的检测试剂等,在乙型肝炎的预防、治疗和诊断中有着潜在的应用价值。
Description
技术领域
本发明属于医学免疫学和感染病学领域,涉及多种乙型肝炎病毒的抗原表位肽序列。
背景技术
乙型肝炎病毒(Hepatitis B virus,HBV)感染是最常见感染性疾病之一,严重危及人类的健康。世界卫生组织2015年7月统计数据显示,预计全球1/3的人口感染过HBV,约2.48亿HBV携带者,后期可发展为肝硬化和原发性肝细胞癌。每年约78万HBV感染的患者因疾病进展为肝硬化、肝衰竭或是肝癌而导致死亡。流行病学调查显示在中国大陆地区约8亿人感染过HBV,乙肝表面抗原(hepatitis B surface antigen,HBsAg)携带率高达10.34%,即我国目前约有1.2亿人为HBV携带者,占全球慢性感染人数的一半。另外,我国也是一个肝癌大国,约占全球肝癌人数的55%以上。
HBV基因组由不完全双链环状DNA组成,长的为负链,含3020-3320个碱基,短的为正链,含1700-2800个碱基。包括四个开放阅读框(ORF),分别称为C、X、P和S编码区。C区由pre-C和C基因组成,分别由不同的起始密码子调控,在相同的终止密码子处结束,pre-C和C基因共同编码pre-C蛋白,Pre-C蛋白再经过切割加工后形成核心抗原(HBeAg)。S区由S基因、PreS1和PreS2基因组成,由3个不同的起始密码子启动翻译,在同一个终止密码子处终止翻译,分别编码表面抗原(HBsAg)蛋白、PreS1蛋白和PreS2蛋白。P区基因编码的HBV的聚合酶蛋白(HBpol)。X区的基因编码X蛋白(HBx),包含154个氨基酸,是最小的一个开放阅读框。
细胞毒性T细胞(Cytotoxic T lymphocyte,CTL)是介导适应性免疫应答的核心细胞,在抗感染、抗肿瘤以及超敏反应和自身免疫病的发生中起着至关重要的作用,其细胞膜上的T细胞受体(T cell receptor,TCR)能够特异性识别并结合抗原递呈细胞表面MHC I类分子与抗原肽的复合物,即MHC/抗原肽复合体分子。CTL表位是指与MHC I类分子结合的抗原肽,是抗原分子中能被TCR特异性识别的线性片段或空间构象性结构,是引起免疫应答反应的基本抗原单位,在CTL活化过程中扮演着关键角色。
MHC系统指主要组织相容性复合体(major histocompatibility complex,MHC),是脊椎动物基因组中一组紧密连锁的基因群,编码表达MHC I类和II类蛋白分子。HLA(human leukocyte antigen)是人类的MHC系统,是人体最复杂的基因群,在人群中具有高度多态性。HLA在抗原识别、抗原呈递等机体免疫过程中发挥重要作用,是影响人体免疫反应的主要因素。HLAI类分子主要负责将内源性HBV抗原呈递给CD8+CTL,活化的CTLs通过分泌穿孔素和颗粒酶等使病毒感染的肝细胞凋亡,同时分泌特定的细胞因子抑制HBV的复制。因此,动态监测HBV抗原特异性CD8+T细胞的数量和功能可以准确反映乙肝感染者针对HBV的特异性免疫功能状态。由于不同人的HLA分子型别不同,其对HBV不同抗原的加工、处理和提呈能力也不相同,从而引起不同程度的HBV抗原特异性免疫应答反应。根据乙肝患者不同的HLA分子型别,选择其所提呈的HBV特异性抗原肽,动态监测该HBV抗原肽特异性的CD8+T细胞的数量和功能,对HBV感染者的疾病进程监测、诊断与治疗方案的制定、疗效观察和预后转归的判断等都有重要意义,是实现乙肝精准医学治疗的重要技术手段。同时,利用这些HLA-A分子高亲和力结合的HBV特异性抗原肽,还可以制备多肽疫苗或基因疫苗,预防和治疗HBV感染。
目前,国内外报道的与HLA-A分子呈高亲和力结合的HBV特异性抗原肽中,HLA-A*0201、A*0203、A*0206、A*1101和A*2402等分子限制性的抗原肽序列被报道和鉴定的较多。而中国人群中频率相对较高的HLA-A*0101、A*0301、A*3001、A*3101、A*3201、A*3303和A*6801等分子的限制性HBV抗原肽则较少报道。本专利通过生物信息学方法,利用六种在线表位预测数据库对HLA-A等位基因型限制的HBV特异性抗原表位肽进行预测,获得一组分别与HLA-A*0101、A*0301、A*3001、A*3101、A*3201、A*3303和A*6801等HLA-A分子呈高亲和力结合的HBV特异性抗原肽序列,为制备HBV感染的治疗和预防性疫苗以及检测HBV抗原特异性T细胞的试剂等提供候选抗原表位肽。
发明内容
技术问题:本发明提供一种能制备HBV感染的治疗和预防性疫苗以及研制HBV抗原特异性T细胞检测试剂的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列。
技术方案:本发明的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列,为如下所示序列中的任一种:
进一步的,本发明中,该序列为乙肝病毒核心抗原HBcAg、表面抗原HBsAg、聚合酶蛋白HBpol或X蛋白HBx的抗原肽序列。
进一步的,本发明中,该序列能分别HLA-A*0101、A*0301、A*3001、A*3101、A*3201、A*3303、A*6801分子呈高亲和力结合。
进一步的,本发明中,序列通过以下步骤制备:
1.1通过GeneBank数据库或UniProt全球蛋白资源数据库检索获得乙肝病毒蛋白氨基酸序列;
1.2利用六种表位预测数据库SYFPEITHI、BIMAS、SVMHC、IEDB、NETMHC和EPIJEN分别预测步骤1.1所得乙型肝炎病毒抗原中能分别与HLA-A*0101、A*0301、A*3001、A*3101、A*3201、A*3303、A*6801分子呈高亲和力结合的多肽序列;
1.3对步骤1.2所得六种在线表位预测网站的预测结果,选择同一HLA-A分子至少有两种及以上方法都预测获得高亲和力结果的某一多肽序列作为乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列。
本发明的一种将上述的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列在制备多肽疫苗或基因疫苗中的应用,是将与HLA-A分子呈高亲和力结合的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽制备成多肽疫苗或基因疫苗。
本发明的一种将上述乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列在检测乙型肝炎病毒抗原特异性T细胞中的应用,是以与HLA-A分子呈高亲和力结合的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽作为试剂,检测乙型肝炎病毒抗原表位肽特异性胸腺依赖性淋巴细胞。
本发明的上述乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列在检测乙型肝炎病毒抗原特异性T细胞中的应用,所述试剂为酶联免疫斑点法试剂、酶联免疫吸附试验试剂、人类白细胞抗原多聚体荧光染色或流式细胞分析法试剂。
本发明利用六种在线表位预测数据库预测6种HLA-A等位基因型限制性的HBV特异性抗原表位肽序列,获得一组能分别与HLA-A*0101、A*0301、A*3001、A*3101、A*3201、A*3303和A*6801等HLA-A分子呈高亲和力结合的HBV特异性抗原表位肽序列,为制备HBV感染的治疗和预防性疫苗以及研制HBV抗原特异性T细胞的检测试剂等提供了候选抗原表位肽。
1.选择乙肝病毒核心抗原(HBcAg)、表面抗原(HBsAg)、聚合酶蛋白(HBpol)和X蛋白(HBx)氨基酸序列为靶向序列;
2.选择预测结果获得研究者的公认、具有较高准确性的、常用的六种表位预测数据库:SYFPEITHI、BIMAS、SVMHC、IEDB、NETMHC和EPIJEN预测上述6种HLA-A分子限制性的HBV特异性抗原表位肽序列;
3.根据一定的预测标准,对六个在线表位预测网站的预测结果进行整合分析,获得六个网站预测结果较一致的候选抗原肽序列。
本发明为乙肝病毒核心抗原(HBcAg)、表面抗原(HBsAg)、聚合酶蛋白(HBpol)和X蛋白(HBx)中能分别与HLA-A*0101、A*0301、A*3001、A*3101、A*3201、A*3303和A*6801等蛋白分子呈高亲和力结合的抗原肽序列;还涉及到以上述抗原肽为基础的乙型肝炎多肽疫苗、基因疫苗、乙型肝炎的治疗与预防方法,以及以上述抗原肽为基础的检测乙型肝炎病毒抗原特异性T细胞的试剂和方法。
有益效果:本发明与现有技术相比,具有以下优点:
预测获得的HLA-A*0101、A*0301、A*3001、A*3101、A*3201、A*3303和A*6801等HLA-A分子限制性的HBV特异性抗原表位肽以前也没有被报道过。这些HLA-A分子以前也没有被报道有限制性的HBV抗原肽。因此这些新的抗原表位肽序列将为研制针对乙型肝炎治疗性和预防性多肽疫苗和基因疫苗、设计检测乙型肝炎病毒抗原特异性T细胞的试剂和方法等提供所需的关键抗原组分,即抗原表位肽序列;同时这些抗原表位肽也为针对这些特定HLA-A等位基因的乙型肝炎患者进行个体化检测和精准医疗提供了关键的抗原组分。
具体实施方式
本发明的HLA-A分子限制性HBV特异性抗原肽,包括以下步骤:
1.通过GeneBank数据库和UniProt全球蛋白资源数据库检索获得HBV蛋白氨基酸序列,选择研究最多的蛋白序列,最终选定的蛋白序列为HBsAg(P03138)、HBcAg(P03146)、HBpol(P03156)和HBx(P03165)。
2.利用六种常用表位预测数据库SYFPEITHI、BIMAS、SVMHC、IEDB、NETMHC和EPIJEN(http://www.syfpeithi.de/、http://www-bimas.cit.nih.gov/molbio/hla_bind/、ttp://www.sbc.su.se/~pierre/svmhc/、http://www.immuneepitope.org/tools.do/、http://www.cbs.dtu.dk/services/NetMHC/、http://www.ddg-pharmfac.net/epijen/EpiJen/EpiJen.htm/)分别将上述乙肝病毒抗原HBsAg(P03138)、HBcAg(P03146)、HBpol(P03156)和HBx(P03165)的氨基酸序列与HLA-A*0101、A*0301、A*3001、A*3101、A*3201、A*3303和A*6801等HLA-A分子中的每一种进行亲和力和空间构象的吻合度分析,根据一定的标准预测筛选每种HLA-A分子限制性的抗原表位肽序列,分别选择长度为9和10个氨基酸的结合肽作为候选抗原表位肽。具体的筛选标准如下:SYFPEITHI预测选取评分>20或者排名在前10的肽序列;BIMAS预测选取评分>10或者排名在前10的肽序列;SVMHC预测选取评分>0.5或者排名在前10的肽序列;IEDB预测选取评IC50<500nM或者排名在前10的肽序列;NETMHC预测选取Rank Threshold<2.0Rank Threshold<2.0或者排名在前10的肽序列;EPIJEN预测选取IC50<500nM或者排名在前10的肽序列。
3.针对每一种HLA-A分子,经不同数据库预测出的表位肽再根据亲和力从高到低进行排列,选取高亲和力的肽序列作为该种HLA-A分子限制性的HBV候选抗原表位肽。同时对六种在线表位预测网站的预测结果进行整合分析,对每一种HLA-A分子,筛选至少达到两种及以上方法筛选标准的某一表位肽作为该HLA-A分子限制性的候选抗原表位肽。
表1是通过上述六个在线表位预测网站预测并最终筛选出的HLA-A限制性HBV特异性抗原表位肽序列。
四种被用来预测抗原表位的乙型肝炎病毒抗原的氨基酸序列如下:
SEQ ID NO1:
HBsAg(P03138):
SEQ ID NO2:
HBcAg(P03146):
表1 HLA-A限制性HBV相关抗原表位的预测结果
---:该预测网站没有针对该MHC分子的预测方法;
NS:没有获得该表位的计算数据。
SEQ ID NO3:
HBpol(P03156):
SEQ ID NO4:
HBx(P03165):
上述实施例仅是本发明的优选实施方式,应当指出:对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和等同替换,这些对本发明权利要求进行改进和等同替换后的技术方案,均落入本发明的保护范围。
SEQUENCE LISTING
<110> 东南大学
<120> 乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列
<130> 2017
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 389
<212> PRT
<213> 人工合成
<400> 1
Met Gly Gln Asn Leu Ser Thr Ser Asn Pro Leu Gly Phe Phe Pro Asp
1 5 10 15
His Gln Leu Asp Pro Ala Phe Arg Ala Asn Thr Ala Asn Pro Asp Trp
20 25 30
Asp Phe Asn Pro Asn Lys Asp Thr Trp Pro Asp Ala Asn Lys Val Gly
35 40 45
Ala Gly Ala Phe Gly Leu Gly Phe Thr Pro Pro His Gly Gly Leu Leu
50 55 60
Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Gln Thr Leu Pro Ala Asn
65 70 75 80
Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr Pro
85 90 95
Leu Ser Pro Pro Leu Arg Asn Thr His Pro Gln Ala Met Gln Trp Asn
100 105 110
Ser Thr Thr Phe His Gln Thr Leu Gln Asp Pro Arg Val Arg Gly Leu
115 120 125
Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Asn Pro Val Leu
130 135 140
Thr Thr Ala Ser Pro Leu Ser Ser Ile Phe Ser Arg Ile Gly Asp Pro
145 150 155 160
Ala Leu Asn Met Glu Asn Ile Thr Ser Gly Phe Leu Gly Pro Leu Leu
165 170 175
Val Leu Gln Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile Pro
180 185 190
Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly Thr
195 200 205
Thr Val Cys Leu Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His Ser
210 215 220
Pro Thr Ser Cys Pro Pro Thr Cys Pro Gly Tyr Arg Trp Met Cys Leu
225 230 235 240
Arg Arg Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile Phe
245 250 255
Leu Leu Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro Leu
260 265 270
Ile Pro Gly Ser Ser Thr Thr Ser Thr Gly Pro Cys Arg Thr Cys Met
275 280 285
Thr Thr Ala Gln Gly Thr Ser Met Tyr Pro Ser Cys Cys Cys Thr Lys
290 295 300
Pro Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp Ala
305 310 315 320
Phe Gly Lys Phe Leu Trp Glu Trp Ala Ser Ala Arg Phe Ser Trp Leu
325 330 335
Ser Leu Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro Thr
340 345 350
Val Trp Leu Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser Leu
355 360 365
Tyr Ser Ile Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe Phe Cys
370 375 380
Leu Trp Val Tyr Ile
385
<210> 2
<211> 183
<212> PRT
<213> 人工合成
<400> 2
Met Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Thr Val Glu Leu Leu
1 5 10 15
Ser Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp
20 25 30
Thr Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys
35 40 45
Ser Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu
50 55 60
Leu Met Thr Leu Ala Thr Trp Val Gly Val Asn Leu Glu Asp Pro Ala
65 70 75 80
Ser Arg Asp Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys
85 90 95
Phe Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg
100 105 110
Glu Thr Val Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr
115 120 125
Pro Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro
130 135 140
Glu Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr
145 150 155 160
Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser
165 170 175
Gln Ser Arg Glu Ser Gln Cys
180
<210> 3
<211> 832
<212> PRT
<213> 人工合成
<400> 3
Met Pro Leu Ser Tyr Gln His Phe Arg Arg Leu Leu Leu Leu Asp Asp
1 5 10 15
Glu Ala Gly Pro Leu Glu Glu Glu Leu Pro Arg Leu Ala Asp Glu Gly
20 25 30
Leu Asn Arg Arg Val Ala Glu Asp Leu Asn Leu Gly Asn Leu Asn Val
35 40 45
Ser Ile Pro Trp Thr His Lys Val Gly Asn Phe Thr Gly Leu Tyr Ser
50 55 60
Ser Thr Val Pro Val Phe Asn Pro His Trp Lys Thr Pro Ser Phe Pro
65 70 75 80
Asn Ile His Leu His Gln Asp Ile Ile Lys Lys Cys Glu Gln Phe Val
85 90 95
Gly Pro Leu Thr Val Asn Glu Lys Arg Arg Leu Gln Leu Ile Met Pro
100 105 110
Ala Arg Phe Tyr Pro Lys Val Thr Lys Tyr Leu Pro Leu Asp Lys Gly
115 120 125
Ile Lys Pro Tyr Tyr Pro Glu His Leu Val Asn His Tyr Phe Gln Thr
130 135 140
Arg His Tyr Leu His Thr Leu Trp Lys Ala Gly Ile Leu Tyr Lys Arg
145 150 155 160
Glu Thr Thr His Ser Ala Ser Phe Cys Gly Ser Pro Tyr Ser Trp Glu
165 170 175
Gln Asp Leu Gln His Gly Ala Glu Ser Phe His Gln Gln Ser Ser Gly
180 185 190
Ile Leu Ser Arg Pro Pro Val Gly Ser Ser Leu Gln Ser Lys His Arg
195 200 205
Lys Ser Arg Leu Gly Leu Gln Ser Gln Gln Gly His Leu Ala Arg Arg
210 215 220
Gln Gln Gly Arg Ser Trp Ser Ile Arg Ala Gly Phe His Pro Thr Ala
225 230 235 240
Arg Arg Pro Phe Gly Val Glu Pro Ser Gly Ser Gly His Thr Thr Asn
245 250 255
Phe Ala Ser Lys Ser Ala Ser Cys Leu His Gln Ser Pro Val Arg Lys
260 265 270
Ala Ala Tyr Pro Ala Val Ser Thr Phe Glu Lys His Ser Ser Ser Gly
275 280 285
His Ala Val Glu Phe His Asn Leu Pro Pro Asn Ser Ala Arg Ser Gln
290 295 300
Ser Glu Arg Pro Val Phe Pro Cys Trp Trp Leu Gln Phe Arg Asn Ser
305 310 315 320
Lys Pro Cys Ser Asp Tyr Cys Leu Ser Leu Ile Val Asn Leu Leu Glu
325 330 335
Asp Trp Gly Pro Cys Ala Glu His Gly Glu His His Ile Arg Ile Pro
340 345 350
Arg Thr Pro Ser Arg Val Thr Gly Gly Val Phe Leu Val Asp Lys Asn
355 360 365
Pro His Asn Thr Ala Glu Ser Arg Leu Val Val Asp Phe Ser Gln Phe
370 375 380
Ser Arg Gly Asn Tyr Arg Val Ser Trp Pro Lys Phe Ala Val Pro Asn
385 390 395 400
Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu Ser
405 410 415
Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro Ala Ala
420 425 430
Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr Val Ala
435 440 445
Arg Leu Ser Ser Asn Ser Arg Ile Leu Asn Asn Gln His Gly Thr Met
450 455 460
Pro Asp Leu His Asp Tyr Cys Ser Arg Asn Leu Tyr Val Ser Leu Leu
465 470 475 480
Leu Leu Tyr Gln Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His Pro
485 490 495
Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser Pro
500 505 510
Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg Arg
515 520 525
Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val Leu
530 535 540
Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Phe Thr Ala Val Thr
545 550 555 560
Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr Lys
565 570 575
Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val Ile Gly Cys Tyr
580 585 590
Gly Ser Leu Pro Gln Glu His Ile Ile Gln Lys Ile Lys Glu Cys Phe
595 600 605
Arg Lys Leu Pro Ile Asn Arg Pro Ile Asp Trp Lys Val Cys Gln Arg
610 615 620
Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly Tyr
625 630 635 640
Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala Phe
645 650 655
Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu Asn
660 665 670
Leu Tyr Pro Val Ala Arg Gln Arg Pro Gly Leu Cys Gln Val Phe Ala
675 680 685
Asp Ala Thr Pro Thr Gly Trp Gly Leu Val Met Gly His Gln Arg Met
690 695 700
Arg Gly Thr Phe Ser Ala Pro Leu Pro Ile His Thr Ala Glu Leu Leu
705 710 715 720
Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Asn Ile Ile Gly Thr
725 730 735
Asp Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp Leu
740 745 750
Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val Tyr
755 760 765
Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg Leu
770 775 780
Gly Leu Ser Arg Pro Leu Leu Arg Leu Pro Phe Arg Pro Thr Thr Gly
785 790 795 800
Arg Thr Ser Leu Tyr Ala Asp Ser Pro Ser Val Pro Ser His Leu Pro
805 810 815
Asp Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro Pro
820 825 830
<210> 4
<211> 154
<212> PRT
<213> 人工合成
<400> 4
Met Ala Ala Arg Leu Cys Cys Gln Leu Asp Pro Ala Arg Asp Val Leu
1 5 10 15
Cys Leu Arg Pro Val Gly Ala Glu Ser Arg Gly Arg Pro Phe Ser Gly
20 25 30
Ser Leu Gly Thr Leu Ser Ser Pro Ser Pro Ser Ala Val Pro Thr Asp
35 40 45
His Gly Ala His Leu Ser Leu Arg Gly Leu Pro Val Cys Ala Phe Ser
50 55 60
Ser Ala Gly Pro Cys Ala Leu Arg Phe Thr Ser Ala Arg Arg Met Glu
65 70 75 80
Thr Thr Val Asn Ala His Gln Ile Leu Pro Lys Val Leu His Lys Arg
85 90 95
Thr Leu Gly Leu Ser Ala Met Ser Thr Thr Asp Leu Glu Ala Tyr Phe
100 105 110
Lys Asp Cys Leu Phe Lys Asp Trp Glu Glu Leu Gly Glu Glu Ile Arg
115 120 125
Leu Lys Val Phe Val Leu Gly Gly Cys Arg His Lys Leu Val Cys Ala
130 135 140
Pro Ala Pro Cys Asn Phe Phe Thr Ser Ala
145 150
Claims (7)
1.一种乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列,其特征在于,该序列为如下所示序列中的任一种:
2.根据权利要求1所述的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列,其特征在于,该序列为乙肝病毒核心抗原HBcAg、表面抗原HBsAg、聚合酶蛋白HBpol或X蛋白HBx的抗原肽序列。
3.根据权利要求1所述的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列,其特征在于,该序列能分别与HLA-A*0101、A*0301、A*3001、A*3101、A*3201、A*3303、A*6801分子呈高亲和力结合。
4.根据权利要求1、2或3所述的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列,其特征在于,所述序列通过以下步骤制备:
1.1通过GeneBank数据库或UniProt全球蛋白资源数据库检索获得乙肝病毒蛋白氨基酸序列;
1.2利用六种表位预测数据库SYFPEITHI、BIMAS、SVMHC、IEDB、NETMHC和EPIJEN分别预测步骤1.1所得乙型肝炎病毒抗原中能分别与HLA-A*0101、A*0301、A*3001、A*3101、A*3201、A*3303、A*6801分子呈高亲和力结合的多肽序列;
1.3对步骤1.2所得六种在线表位预测网站的预测结果,选择同一HLA-A分子至少有两种及以上方法都预测获得高亲和力结果的某一多肽序列作为乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列。
5.一种将权利要求1、2、3或4所述的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列在制备多肽疫苗或基因疫苗中的应用,其特征在于,该应用是将与HLA-A分子呈高亲和力结合的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽制备成多肽疫苗或基因疫苗。
6.一种将权利要求1、2、3或4所述的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列在检测乙型肝炎病毒抗原特异性T细胞中的应用,其特征在于,该应用是以与HLA-A分子呈高亲和力结合的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽作为试剂,检测乙型肝炎病毒抗原表位肽特异性胸腺依赖性淋巴细胞。
7.根据权利要求6所述的乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列在检测乙型肝炎病毒抗原特异性T细胞中的应用,其特征在于,所述试剂为酶联免疫斑点法试剂、酶联免疫吸附试验试剂、人类白细胞抗原多聚体荧光染色或流式细胞分析法试剂。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710191364.6A CN107022006A (zh) | 2017-03-28 | 2017-03-28 | 乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710191364.6A CN107022006A (zh) | 2017-03-28 | 2017-03-28 | 乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN107022006A true CN107022006A (zh) | 2017-08-08 |
Family
ID=59526019
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201710191364.6A Pending CN107022006A (zh) | 2017-03-28 | 2017-03-28 | 乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN107022006A (zh) |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110172080A (zh) * | 2019-05-16 | 2019-08-27 | 南京大户生物科技有限公司 | 乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽及其应用 |
CN111393504A (zh) * | 2020-03-18 | 2020-07-10 | 北京鼎成肽源生物技术有限公司 | 一种肝癌抗原组合及其应用、细胞毒性t淋巴细胞 |
CN111848787A (zh) * | 2020-08-21 | 2020-10-30 | 北京安必奇生物科技有限公司 | preS1抗体、乙型肝炎病毒前S1抗体酶免检测试剂盒、制备方法、使用方法 |
CN114478711A (zh) * | 2022-01-05 | 2022-05-13 | 成都朗谷生物科技股份有限公司 | 一种针对乙型肝炎病毒的抗原肽及其应用 |
CN114573705A (zh) * | 2022-03-17 | 2022-06-03 | 杭州师范大学 | 特异性启动抗乙型肝炎病毒t细胞免疫的双特异性抗体及其应用 |
CN114591404A (zh) * | 2022-03-23 | 2022-06-07 | 成都朗谷生物科技股份有限公司 | 适用于白细胞抗原单倍型为hla-a2个体的乙型肝炎病毒抗原肽及其应用 |
CN116751262A (zh) * | 2023-07-31 | 2023-09-15 | 重庆医科大学国际体外诊断研究院 | 靶向乙型肝炎病毒核心蛋白的多肽及其应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1454082A (zh) * | 2000-09-08 | 2003-11-05 | 艾普免疫公司 | 使用肽和核酸组合物诱导针对乙型肝炎病毒的细胞免疫应答 |
US20050063983A1 (en) * | 1993-03-05 | 2005-03-24 | Epimmune Inc. | Inducing cellular immune responses to hepatitis B virus using peptide and nucleic acid compositions |
CN101979405A (zh) * | 2010-08-27 | 2011-02-23 | 中国人民解放军第三军医大学 | 乙肝病毒核心抗原的免疫显性hla-a3超型限制性ctl表位及其鉴定方法和应用 |
-
2017
- 2017-03-28 CN CN201710191364.6A patent/CN107022006A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050063983A1 (en) * | 1993-03-05 | 2005-03-24 | Epimmune Inc. | Inducing cellular immune responses to hepatitis B virus using peptide and nucleic acid compositions |
CN1454082A (zh) * | 2000-09-08 | 2003-11-05 | 艾普免疫公司 | 使用肽和核酸组合物诱导针对乙型肝炎病毒的细胞免疫应答 |
CN101979405A (zh) * | 2010-08-27 | 2011-02-23 | 中国人民解放军第三军医大学 | 乙肝病毒核心抗原的免疫显性hla-a3超型限制性ctl表位及其鉴定方法和应用 |
Non-Patent Citations (2)
Title |
---|
BRINCK-JENSEN ET AL.: "Immunogenicity of twenty peptides representing epitopes of the hepatitis B core and surface antigens by IFN-γ response in chronic and resolved HBV", 《BMC IMMUNOLOGY》 * |
王文博: "HBV相关抗原HLA-A3超型CTL表位的预测与鉴定", 《中国优秀硕士学位论文全文数据库(电子期刊)医药卫生科技辑》 * |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110172080A (zh) * | 2019-05-16 | 2019-08-27 | 南京大户生物科技有限公司 | 乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽及其应用 |
CN111393504A (zh) * | 2020-03-18 | 2020-07-10 | 北京鼎成肽源生物技术有限公司 | 一种肝癌抗原组合及其应用、细胞毒性t淋巴细胞 |
CN111393504B (zh) * | 2020-03-18 | 2023-03-31 | 北京鼎成肽源生物技术有限公司 | 一种肝癌抗原组合及其应用、细胞毒性t淋巴细胞 |
CN111848787A (zh) * | 2020-08-21 | 2020-10-30 | 北京安必奇生物科技有限公司 | preS1抗体、乙型肝炎病毒前S1抗体酶免检测试剂盒、制备方法、使用方法 |
CN111848787B (zh) * | 2020-08-21 | 2024-05-17 | 北京安必奇生物科技有限公司 | preS1抗体、乙型肝炎病毒前S1抗体酶免检测试剂盒、制备方法、使用方法 |
CN114478711A (zh) * | 2022-01-05 | 2022-05-13 | 成都朗谷生物科技股份有限公司 | 一种针对乙型肝炎病毒的抗原肽及其应用 |
CN114573705A (zh) * | 2022-03-17 | 2022-06-03 | 杭州师范大学 | 特异性启动抗乙型肝炎病毒t细胞免疫的双特异性抗体及其应用 |
CN114573705B (zh) * | 2022-03-17 | 2024-05-14 | 杭州师范大学 | 特异性启动抗乙型肝炎病毒t细胞免疫的双特异性抗体及其应用 |
CN114591404A (zh) * | 2022-03-23 | 2022-06-07 | 成都朗谷生物科技股份有限公司 | 适用于白细胞抗原单倍型为hla-a2个体的乙型肝炎病毒抗原肽及其应用 |
CN116751262A (zh) * | 2023-07-31 | 2023-09-15 | 重庆医科大学国际体外诊断研究院 | 靶向乙型肝炎病毒核心蛋白的多肽及其应用 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107022006A (zh) | 乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽序列 | |
Iesa et al. | SARS-CoV-2 and Plasmodium falciparum common immunodominant regions may explain low COVID-19 incidence in the malaria-endemic belt | |
Sanchez-Mazas | HLA studies in the context of coronavirus outbreaks | |
Pan et al. | Sixty years (1957–2017) of research on toxoplasmosis in China—an overview | |
Brien et al. | Protective capacity and epitope specificity of CD8+ T cells responding to lethal West Nile virus infection | |
Gottstein et al. | Echinococcus metacestode: in search of viability markers | |
Wang et al. | CTL epitopes for influenza A including the H5N1 bird flu; genome-, pathogen-, and HLA-wide screening | |
Taborda et al. | Mapping of the T-cell epitope in the major 43-kilodalton glycoprotein of Paracoccidioides brasiliensis which induces a Th-1 response protective against fungal infection in BALB/c mice | |
Brown et al. | Designing blood-stage vaccines against Babesia bovis and B. bigemina | |
US20130064843A1 (en) | Identification of conserved peptide blocks in homologous polypeptides | |
Long | Q fever vaccine development: current strategies and future considerations | |
CN110172080A (zh) | 乙型肝炎病毒抗原的胸腺依赖性淋巴细胞抗原表位肽及其应用 | |
Amit et al. | Immunomodulation mediated through Leishmania donovani protein disulfide isomerase by eliciting CD8+ T-cell in cured visceral leishmaniasis subjects and identification of its possible HLA class-1 restricted T-cell epitopes | |
CN101935345A (zh) | Hsp60的肽和apl型衍生物及药物组合物 | |
Zobayer | In silico characterization and homology modeling of histamine receptors | |
Qiao et al. | Personalized workflow to identify optimal T-cell epitopes for peptide-based vaccines against COVID-19 | |
Prakash et al. | Genome-wide asymptomatic B-cell, CD4+ and CD8+ T-cell epitopes, that are highly conserved between human and animal coronaviruses, identified from SARS-CoV-2 as immune targets for pre-emptive pan-Coronavirus vaccines | |
Bishop et al. | More than three decades of Bm86: what we know and where to go | |
Ito et al. | Vaccine progress | |
Madan et al. | Use of a synthetic peptide epitope of Asp f 1, a major allergen or antigen of Aspergillus fumigatus, for improved immunodiagnosis of allergic bronchopulmonary aspergillosis | |
WO2015025165A1 (en) | T cell epitopes of classical swine fever virus | |
CN102219858A (zh) | 微小隐孢子虫ctl多表位基因和融合蛋白及其应用 | |
Guerfali et al. | An in silico immunological approach for prediction of CD8+ T cell epitopes of Leishmania major proteins in susceptible BALB/c and resistant C57BL/6 murine models of infection | |
Chauhan et al. | Designing peptide-based vaccine candidates for Plasmodium falciparum erythrocyte binding antigen 175 | |
Fonseca et al. | Human T cell epitope mapping of the Schistosoma mansoni 14-kDa fatty acid-binding protein using cells from patients living in areas endemic for schistosomiasis |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170808 |
|
RJ01 | Rejection of invention patent application after publication |