CN113092761A - 一种弥漫大b细胞淋巴瘤的外周血tcr标志物及其检测试剂盒和应用 - Google Patents
一种弥漫大b细胞淋巴瘤的外周血tcr标志物及其检测试剂盒和应用 Download PDFInfo
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Abstract
本发明公开了一种弥漫大B细胞淋巴瘤的外周血TCR标志物及其检测试剂盒和应用。该标志物包括序列为SEQ ID NO.1～100所示的蛋白中的至少一种。本发明基于高通量测序方法，只需要采取少量外周血，提取RNA，通过对样本的处理建立免疫图谱文库，再经过高通量测序和TCR数据分析，首先确定弥漫大B细胞淋巴瘤外周血中特征性TCR序列，然后将待测样本测试结果与该特征性TCR序列比对，从而确定是否患有弥漫大B细胞淋巴瘤。本发明能够同时比较巨大数量的弥漫大B细胞淋巴瘤特异性TCR序列，相比单独检测一种或几种标记物，具有更高的特异性和准确性，提高了诊断效率。
Description
技术领域
本发明属于基因工程技术领域，具体涉及一种弥漫大B细胞淋巴瘤的外周血TCR标志物及其检测试剂盒和应用。
背景技术
非霍奇金淋巴瘤(Non-Hodgkin Lymphoma，NHL)是一组起源于淋巴结和淋巴组织的恶性肿瘤的总称，NHL病变是主要发生在淋巴结、脾脏、胸腺等淋巴器官，也可发生在淋巴结外的淋巴组织和器官的淋巴造血系统的恶性肿瘤。依据细胞来源将其分为三种基本类型：B细胞、T细胞和NK/T细胞NHL。
流行病学显示，我国非霍奇金淋巴瘤患者相对较少，但近年来发病率逐渐上升。最近的一项调查显示，非霍奇金淋巴瘤占每年新发肿瘤的4％，占所有恶性淋巴瘤的90％，全国每年约有54000人被诊断为非霍奇金淋巴瘤，每年死亡人数约19000人。该病发病率随年龄的增长而显著上升，男性患者比女性多见。
弥漫大B细胞淋巴瘤(Diffuse Large B-Cell Lymphoma，DLBCL)是NHL中最常见的类型，在临床上最常见的病理学类型、最具代表性的侵袭性淋巴瘤。DLBCL是肿瘤性大B淋巴细胞呈弥漫性生长，肿瘤细胞的核与正常组织细胞的核大小相近或大于组织细胞的核，通常大于正常淋巴细胞的2倍。该癌种可发生于任何年龄段，但常见于老年人，几乎全身任何部位均可发生弥漫性大B细胞淋巴瘤。其发病率占非霍奇金淋巴瘤(NHL)的31％～34％，在亚洲国家一般大于40％。我国2011年一项由24个中心联合进行、共收集10 002例病例样本的分析报告指出，在中国DLBCL占所有NHL的45.8％，占所有淋巴瘤的40.1％。
非霍奇金淋巴瘤的病因和发病机制目前尚未完全清楚，一般认为其与感染、免疫状态、环境和饮食等危险因素相关。在这些危险因素影响下，可引起B细胞和T细胞内癌症基因的激活及肿瘤抑制基因的失活，使淋巴细胞增殖失控，最终形成非霍奇金淋巴瘤。
淋巴瘤生存率相对较高，在规范化综合治疗的情况下，淋巴瘤治愈率可超过60％，其中霍奇金氏淋巴瘤治愈率为70％-90％，50％的非霍奇金氏淋巴瘤患者可获得长期生存。但根据《柳叶刀》2018统计数据，我国淋巴瘤患者的5年生存率约为38.3％，相比日本和美国的57.3％，68.1％，仍存在明显差距。原因有好几个。首先是与诊断的准确性有关，不同亚型的确定治疗方案不同。组织学亚型是决定患者临床表现、预后和治疗的主要因素。2019淋巴瘤白皮书数据显示，参与调研的患者中，约43％患者曾有过误诊经历，51％的患者辗转多家医院后方得以确诊。患者从初次诊断到最终确定所患亚型，平均耗费时长为2.5个月，而一些亚型患者确诊所需等待时间更为漫长。同时，调研表明，大部分患者为了确诊，需奔赴北上广等医疗资源集中的大城市；其次是患者的就诊意识较低。很多患者前来就诊时就已经是晚期，治疗效果肯定不如早期。所以早诊断、早治疗，是提高淋巴瘤治疗效果，改善患者预后，节约社会资源的迫切需求。
弥漫大B细胞淋巴瘤的诊断检查可分为创伤性和非创伤性检查。创伤性检查多是取组织细胞学标本为诊断而进行，非创伤性检查多是为寻找病变位置、评估病变范围和评估治疗疗效而进行。
1、创伤性检查
主要根据病变的大小、生长的部位和创伤检查可能带来的风险而决定操作的方式。
浅表肿大淋巴结直接穿刺；或在超声、CT定位引导下经皮进行深部淋巴结及占位病变穿刺针吸细胞获取标本。此种方法价值存在一定的争议。
根据病变部位可在支气管镜、纵隔镜、胃镜、结肠镜等内镜辅助下取组织细胞标本。各种手术方式获取淋巴结或病变部位细胞标本。
骨髓穿刺和活检。
腰椎穿刺进行脑脊液细胞学检查。
2、非创伤性检查
实验室检查包括全血细胞计数、血沉、乳酸脱氢酶、β2微球蛋白和肝肾功能等。
影像学检查包括全身CT、磁共振、正电子发射型断层扫描技术(PET/CT)等。
综上所述，非创伤性检查中的血液检测只能大致判断有疾病存在，无法分辨针对具体疾病；创伤性检查中的骨髓穿刺和活检，需要穿刺活检，取样困难，不能作为实时监测手段，对患者身体也会造成损伤。往往需将血液和骨髓穿刺检查结果结合在一起，不能通过检测一项而判断疾病，这也增加了患者负担。影像学检测也都有各自优缺点，其检测都需要肿瘤达到一定体积大小才能检测到，存在滞后性，因辐射等原因不能作为实时监控的手段。因此，亟需一种能更便捷灵敏，且可以追踪弥漫大B细胞淋巴瘤发生、发展状况的筛查、检验方法。
发明内容
针对现有技术中的上述不足，本发明提供一种弥漫大B细胞淋巴瘤的外周血TCR标志物及其检测试剂盒和应用，能准确快速的判断待测样本中是否有较高弥漫大B细胞淋巴瘤风险患者。
为实现上述目的，本发明解决其技术问题所采用的技术方案是：
一种弥漫大B细胞淋巴瘤的外周血TCR标志物，该标志物包括序列为SEQ ID NO.1～100所示的蛋白中的至少一种，具体序列见表1。
表1标志物序列
进一步地，标志物的蛋白序列为SEQ ID NO.1～100所示的序列经取代、缺失和/或替换一个或多个碱基后，能表达相同功能的蛋白。
进一步地，标志物为外周血TCR CDR3序列。
上述标志物在制备治疗弥漫大B细胞淋巴瘤的制剂中的应用。
进一步地，制剂中包括含有该标志物的T细胞受体，或能表达产生该标志物的T细胞受体的质粒、病毒载体或核酸片段。
一种用于弥漫大B细胞淋巴瘤检测的试剂盒，包括能与上述标志物发生特异性结合的抗体。
一种制剂，包括能与上述标志物发生特异性结合的抗体；所述制剂可用于对弥漫大B细胞淋巴瘤进行诊断、预测、检测或筛查。
一种检测弥漫大B细胞淋巴瘤的蛋白质芯片，该蛋白质芯片包括基片和点样在基片上特异性抗体，该特异性抗体为能与上述标志物发生特异性结合的抗体。
本发明的原理为：人体内的B淋巴细胞和T淋巴细胞是获得性免疫系统中重要的两类细胞。B细胞通过细胞表面的B细胞受体(BCR)识别抗原，后期BCR在B细胞分化成浆细胞时，表达成抗体，分泌到细胞外。T细胞通过细胞表面的T细胞受体(TCR)识别抗原。BCR和TCR的多样性是建立获得性免疫系统的基础。BCR多样性的理论值是1018，TCR多样性的理论值是1014。BCR与TCR序列中，抗原决定簇3(CDR3)是决定其抗原特异性最重要的部分，因此CDR3的序列被认为可以代表BCR、TCR序列的特性。
在各种疾病中，随着不同的抗原刺激，BCR和TCR的多样性或者表达水平都会发生改变。因此，利用BCR或者TCR高通量测序结果可以追踪疾病的发生、发展。人体内细胞中，衰老蛋白质降解后，其片段会被运输到细胞表面，通过组织相容性抗原II(MCHII)呈递给免疫系统中的T细胞。正常细胞呈递的抗原片段，由于免疫耐受的关系，不会引起免疫反应。一旦当正常细胞出现癌变后，突变的基因表达的异常蛋白，其片段被呈递到细胞表面后，就会引起人体免疫系统产生针对性的免疫反应。因此，分析BCR或TCR的变化，能够检测出肿瘤的发生和发展。
本发明的有益效果为：
1、本发明中，首先利用1300个非弥漫大B细胞淋巴瘤的对照组样本、和40个弥漫大B细胞淋巴瘤病人的TCR高通量测序数据建立了人工智能分析模型，通过和这些弥漫大B细胞淋巴瘤特异性TCR序列对比，可以清楚的判断待测样本中是否有较高弥漫大B细胞淋巴瘤风险者。
2、通过高通量测序分析TCR变化可以发现很早期的弥漫大B细胞淋巴瘤，利用弥漫大B细胞淋巴瘤特有的TCR CDR3序列分析人的免疫系统中的T细胞对弥漫大B细胞淋巴瘤的反应，是一种新型的检测方法。
3、本发明通过采用高通量测序技术同时比较数量巨大的特异性TCR序列，比起单独检测一种或几种标记物，具有更高的特异性和准确性。
4、本发明中使用的高通量测序仪器成本低于大型影像学设备，且可向第三方外包，此外，采样、处理的人力成本低于同时检测多种标记物，也低于大量细胞学检测，因此，本发明大大降低了检测成本。
5、本发明只需要采取少量外周血，采样简便、安全。
6、本发明中所述TCR CDR3序列，可用于弥漫大B细胞淋巴瘤的免疫治疗。
附图说明
图1为本发明中，利用基于免疫大数据分析系统发现弥漫大B细胞淋巴瘤特征性TCR序列；其中，横坐标代表某一特定氨基酸组合的CDR3序列被加入对照序列集合或弥漫大B细胞淋巴瘤特征序列集合的先后顺序，纵坐标代表该序列在某一样本中重复出现次数CX的对数值；弥漫大B细胞淋巴瘤患者的免疫图谱具有多个种类且重复次数较高的弥漫大B细胞淋巴瘤特征序列，健康人极少弥漫大B细胞淋巴瘤特征序列，而未知受试者的弥漫大B细胞淋巴瘤特征比较明显，说明罹患弥漫大B细胞淋巴瘤风险较高；
图2为本发明中，利用弥漫大B细胞淋巴瘤特征特征性指数对比分析弥漫大B细胞淋巴瘤和其他疾病；健康人、非肿瘤病人、非弥漫大B细胞淋巴瘤肿瘤患者的弥漫大B细胞淋巴瘤特征性指数均与弥漫大B细胞淋巴瘤患者具有显著差异，证明了弥漫大B细胞淋巴瘤特征序列集的特异性，据此可以判断未知受试者是否罹患弥漫大B细胞淋巴瘤。
具体实施方式
下面对本发明的具体实施方式进行描述，以便于本技术领域的技术人员理解本发明，但应该清楚，本发明不限于具体实施方式的范围，对本技术领域的普通技术人员来讲，只要各种变化在所附的权利要求限定和确定的本发明的精神和范围内，这些变化是显而易见的，一切利用本发明构思的发明创造均在保护之列。
实施例1通过免疫图谱分析，获得弥漫大B细胞淋巴瘤TCR标志物CDR3序列集
1、采样及免疫图谱分析(方法参考申请号为201910300069.9的专利)
采集1301名对照组(包括健康人和非肿瘤疾病病人，1300人用于建立模型，1个健康人用于验证)、41名弥漫大B细胞淋巴瘤患者(40人用于建立模型，1人用于验证)及1名未知健康状况受试者的外周血(每人10mL)，通过高通量测序得到受试者和对照组的TCR的抗原决定簇3(CDR3)氨基酸序列，保证每个样本的功能性TCR的CDR3序列总数综合不低于25000；
2、对每个功能性TCR的CDR3序列总数综合高于30000的样本的序列进行随机不放回抽样，使该样本的CDR3序列数量总和为30000。至此所有样本包含的功能性TCR的CDR3序列总数为25000-30000。对任一特定CDR3序列X，在单样本测序结果中重复出现次数计为CX；
3、通过分析TCR CDR3数据，确定弥漫大B细胞淋巴瘤TCR标志物CDR3序列：
a)将1300名用于建立模型的对照组样本的所有CDR3序列归总去重，设为对照序列集；
b)将40名用于建立模型的弥漫大B细胞淋巴瘤样本的所有CDR3序列归总去重，再去除所有与对照序列集中包含序列重复的序列，设为弥漫大B细胞淋巴瘤特征序列集。作图如附图1A所示，其中横坐标代表某一特定氨基酸组合的CDR3序列被加入对照序列集合或弥漫大B细胞淋巴瘤特征序列集合的先后顺序，纵坐标代表该序列在某一样本中重复出现次数CX的对数值。
c)按照同样作图方法，将1名健康人、1名弥漫大B细胞淋巴瘤患者和1名健康状况未知受试者的免疫图谱，参照对照序列集合和弥漫大B细胞淋巴瘤特征序列集合进行作图，见附图1B-D。从图中可见，弥漫大B细胞淋巴瘤患者的免疫图谱中，含有较多种类且较高重复出现次数的弥漫大B细胞淋巴瘤特征序列(图1B)；健康人的免疫图谱中，只有极少量弥漫大B细胞淋巴瘤特征序列(图1C)；而未知健康状况受试者，有高于健康人的弥漫大B细胞淋巴瘤特征序列，说明此人有较高风险罹患弥漫大B细胞淋巴瘤(图1D)。
d)将弥漫大B细胞淋巴瘤特征序列集中，将所有出现在两个及以上参与建模弥漫大B细胞淋巴瘤样本里的CDR3序列，按“所有参与建模弥漫大B细胞淋巴瘤样本里该序列单样本中重复出现次数CX的总和×包含该序列的参与建模弥漫大B细胞淋巴瘤样本数”从高到低排序，排名前100者即为弥漫大B细胞淋巴瘤TCR标志物CDR3序列，具体序列如SEQ IDNO.1～100所示。
实施例2验证弥漫大B细胞淋巴瘤TCR标志物CDR3序列集的特异性
1、采样及免疫图谱分析(方法参考申请号为201910300069.9的专利)
采集577名非弥漫大B细胞淋巴瘤的肿瘤患者、7名未知健康状况受试者的外周血(每人10mL)，通过高通量测序得到受试者和对照组的TCR的抗原决定簇3(CDR3)氨基酸序列，保证每个样本的功能性TCR的CDR3序列总数综合不低于25000；对每个功能性TCR的CDR3序列总数综合高于30000的样本的序列进行随机不放回抽样，使该样本的CDR3序列数量总和为30000。至此所有样本包含的功能性TCR的CDR3序列总数为25000-30000。
2、从实施例1的对照组中随机挑选100名健康人及43名非肿瘤疾病病人。
3、根据来自实施例1的100名健康人、43名非肿瘤疾病病人、38名弥漫大B细胞淋巴瘤患者，以及实施例2新获取的577名非弥漫大B细胞淋巴瘤肿瘤患者、7名未知健康状况受试者的免疫图谱，分析其弥漫大B细胞淋巴瘤特征性指数。
其中弥漫大B细胞淋巴瘤特征性指数定义为：某个样本中，属于弥漫大B细胞淋巴瘤特征序列集的所有CDR3序列在该样本内重复出现次数CX的总和。分析结果见下表2及附图2。弥漫大B细胞淋巴瘤组与健康人(p＝3.1E-93)、非肿瘤疾病(p＝8.6E-50)、其它肿瘤(p趋近于0)都有显著差异，这证明了弥漫大B细胞淋巴瘤特征序列集的特异性。
表2不同样本组的弥漫大B细胞淋巴瘤特征性指数
4、分析各组的弥漫大B细胞淋巴瘤特征指数(表3)，7位未知健康状况受试者(检测样本)的弥漫大B细胞淋巴瘤特征指数显著高于“其它肿瘤”组的平均值+2×SD(152.7+2×587.4＝1327.5)，此7人具有较高风险罹患弥漫大B细胞淋巴瘤。与临床体检结果对照后，这7人确为早期弥漫大B细胞淋巴瘤患者。此实施例证明了利用弥漫大B细胞淋巴瘤特征序列集及弥漫大B细胞淋巴瘤特征性指数，预测受试者罹患弥漫大B细胞淋巴瘤风险的可行性。
表3弥漫大B细胞淋巴瘤特征性指数分析
综上所述，本发明所述弥漫大B细胞淋巴瘤TCR标志物CDR3序列，确实具有显著的弥漫大B细胞淋巴瘤特异性，不仅可以用于弥漫大B细胞淋巴瘤预测受试者罹患弥漫大B细胞淋巴瘤风险，未来还可用于弥漫大B细胞淋巴瘤的生物免疫治疗。
序列表
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<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 6
Ala Ser Ser Leu Tyr Gly Pro Gly Ala Asn Thr Glu Ala Phe
1 5 10
<210> 7
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 7
Ala Ser Ser Ile Gly Gly Ser Met Val Gln Pro Gln His
1 5 10
<210> 8
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 8
Ala Thr Gln Asp Pro Ser Arg Pro Gln His
1 5 10
<210> 9
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 9
Ser Ala Leu Gly Gln Thr Val Lys Thr Tyr Glu Gln Tyr
1 5 10
<210> 10
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 10
Pro Pro Ala Val Pro Pro Glu Arg Pro Ser Thr
1 5 10
<210> 11
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 11
Ala Ser Ser Ser Ala Thr Ser Gly Val Tyr Glu Gln Tyr
1 5 10
<210> 12
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Ala Ser Ser Gln Glu Pro Ser Gly Ser Ser Gln Glu Thr Gln Tyr
1 5 10 15
<210> 13
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 13
Ala Ser Ser Asn Ser Arg Val Asp Gln Pro Gln His
1 5 10
<210> 14
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
Ala Ser Thr Leu Leu His Arg Asp Asn Glu Gln Phe
1 5 10
<210> 15
<211> 19
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
Ala Ser Ser Tyr Ser Leu Pro Gly Leu Ala Gly Lys Glu Gly Gln Glu
1 5 10 15
Thr Gln Tyr
<210> 16
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
Ala Ser Arg His Arg Glu Ser Phe Thr Asp Thr Gln Tyr
1 5 10
<210> 17
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 17
Ala Ser Ser Asp Asp Gly Ala Trp Glu Asp Glu Gln Phe
1 5 10
<210> 18
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
Ala Asn Thr Leu Pro Glu Gly Gly Ser Glu Gln Phe
1 5 10
<210> 19
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Ala Ser Ser His Arg Thr Ser Gly Arg Ala Leu Tyr Glu Gln Tyr
1 5 10 15
<210> 20
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Ala Ser Ser Gln Gln Val Pro Pro Thr Asn Thr Gly Glu Leu Phe
1 5 10 15
<210> 21
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 21
Ser Val Glu Pro His Ser Gly Arg Ser Glu Thr Gln Tyr
1 5 10
<210> 22
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
Ala Ser Ser Ser Gly Thr Gly Pro Gly Leu Pro Gln His
1 5 10
<210> 23
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 23
Ser Val Glu Glu Val Arg Glu Gln Tyr
1 5
<210> 24
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 24
Ala Ser Ile Ser Gly His Ser Tyr Glu Gln Tyr
1 5 10
<210> 25
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 25
Ala Ser Ser Thr Arg Asp Arg Val Glu Met Asn Thr Glu Ala Phe
1 5 10 15
<210> 26
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Ala Ser Ser Leu Gly Thr Thr Ala Thr Asp Thr Gly Glu Leu Phe
1 5 10 15
<210> 27
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 27
Ala Ser Lys Gly Trp Thr Thr Ser Pro Leu His
1 5 10
<210> 28
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
Ala Ile Arg Arg Asp Ile Val Lys Thr Glu Ala Phe
1 5 10
<210> 29
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 29
Ala Ser Ser Glu Pro Ala Ser Leu Asp Thr Gln Tyr
1 5 10
<210> 30
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 30
Ala Ser Gly Leu Ser Gly Gly Gly Gln Tyr
1 5 10
<210> 31
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 31
Ala Ser Thr Arg Gln Ala Gln Met Ser Asn Gln Pro Gln His
1 5 10
<210> 32
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
Gln Gln Gln Arg Thr Gly Gly Ala His Arg Tyr Ala Val
1 5 10
<210> 33
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 33
Ala Ser Thr Ser Ser Arg Gly Gly Ser Gly Ala Asn Val Leu Thr
1 5 10 15
<210> 34
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 34
Ala Ser Ser Leu Met Arg Val Lys Ser His Glu Gln Tyr
1 5 10
<210> 35
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 35
Ala Ser Ser Pro Ala Gln Arg Gly Val Glu Gln Phe
1 5 10
<210> 36
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 36
Ala Thr Ser Gly Asp Ser Leu Ala Glu Gln Phe
1 5 10
<210> 37
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 37
Ser Asp Leu Leu Leu Gly Thr Gln Tyr
1 5
<210> 38
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 38
Ala Ile Ser Glu Pro Thr Tyr Ala Ala Ser Ala Ser
1 5 10
<210> 39
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 39
Ala Ser Ser Ala Pro Arg Leu Ala Asp Asn Glu Gln Phe
1 5 10
<210> 40
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 40
Ser Ala Thr Gln Thr Asp Ile Tyr Asn Glu Gln Phe
1 5 10
<210> 41
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 41
Ala Ser Thr Thr Gly Gly Gly Pro Ile Ala Gln Tyr
1 5 10
<210> 42
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 42
Ala Ser Arg Asp Gly Ser Gln Gly Asn Ser Pro Leu His
1 5 10
<210> 43
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 43
Ala Ser Ser Gly Gly Thr Gly Ala Leu Asn Glu Gln Phe
1 5 10
<210> 44
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 44
Ala Ser Thr Gln Ala Gly Ala Gln Ala Asp Thr Gln Tyr
1 5 10
<210> 45
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 45
Ala Ser Ser Ile Ser Gly Gly Gly Ala Val Thr Glu Gln Phe
1 5 10
<210> 46
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 46
Ala Ser Arg Lys Gly Ser Gln Glu Gly Thr Gln Tyr
1 5 10
<210> 47
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 47
Ala Ser Ser Leu Gly Gly Arg Gly Val Ala Gly Glu Leu Phe
1 5 10
<210> 48
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 48
Ala Ser Ser Asp Thr Gly Thr Ser Tyr His Glu Gln Tyr
1 5 10
<210> 49
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 49
Ala Ser Ser Leu Asp Gly Asp Tyr Gln Glu Thr Gln Tyr
1 5 10
<210> 50
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 50
Ala Thr Gly Leu Arg Gln Gly Asn Thr Gly Glu Leu Phe
1 5 10
<210> 51
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 51
Ala Ser His Ser Gly Arg Ile Asn Thr Glu Ala Phe
1 5 10
<210> 52
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 52
Ala Ser Ser Lys Ala Lys Gly Thr Gly Ser Tyr Asn Glu Gln Phe
1 5 10 15
<210> 53
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 53
Ala Ser Ser Pro Leu Thr Gly Thr Ala Gly Ser Tyr Thr
1 5 10
<210> 54
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 54
Ala Thr Ser Glu Glu Ala Gln Glu Thr Gln Tyr
1 5 10
<210> 55
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 55
His Gln Gln Tyr Arg Arg Arg Asp Pro Val
1 5 10
<210> 56
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 56
Ala Thr Ser Arg Gly Glu Thr Asp Tyr Gln Glu Thr Gln Tyr
1 5 10
<210> 57
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 57
Ala Trp Asn Leu Lys Asp Thr Arg Gln Glu Thr Gln Tyr
1 5 10
<210> 58
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 58
Ala Ser Ser Leu Ala Gly Gln Gly Thr Gly Glu Gln Tyr
1 5 10
<210> 59
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 59
Ala Thr Ser Leu Ile Gln Gly Arg Thr Glu Ala Phe
1 5 10
<210> 60
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 60
Pro Ala Asp His Arg Phe Pro Thr Ser Ser Thr
1 5 10
<210> 61
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 61
Ala Ser Ser Ser Ile Leu Gly Thr Asn Glu Gln Phe
1 5 10
<210> 62
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 62
Ala Ser Arg Ala Ala Asn Gly Gln Lys Glu Thr Gln Tyr
1 5 10
<210> 63
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 63
Ala Ser Ser Leu Leu Gln Met Asn Asn Glu Gln Phe
1 5 10
<210> 64
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 64
Ser Ala Glu Thr Gly Phe Ser Asn Gln Pro Gln His
1 5 10
<210> 65
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 65
Ala Ser Ser Leu Gly Glu Thr Gly Thr His Asn Ser Pro Leu His
1 5 10 15
<210> 66
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 66
Ala Trp Arg Asp Thr Tyr Arg Gly Ala Glu Ala Phe
1 5 10
<210> 67
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 67
Ala Ser Ser Ser Ile Arg Gly Gln Gly Asn Ser Asn Glu Gln Phe
1 5 10 15
<210> 68
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 68
Ala Ser Ser Pro Asp Gly Thr Ser Gly Met Glu Thr Gln Tyr
1 5 10
<210> 69
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 69
Ala Ser Ser Thr Gln Gly Ser Val Ser Tyr Glu Gln Tyr
1 5 10
<210> 70
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 70
Ala Ser Ser Gln Met Thr Ser Gly Glu Gln Phe
1 5 10
<210> 71
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 71
Ala Ser Ser Leu Ser Trp Gly Ser Ser Gly Ala Asn Val Leu Thr
1 5 10 15
<210> 72
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 72
Ala Ser Ser Gln Thr Val Gly Pro Asp Glu Gln Tyr
1 5 10
<210> 73
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 73
Ala Ser Leu Met Gly His Arg Pro Gly Asn Glu Gln Phe
1 5 10
<210> 74
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 74
Ala Cys Gly Arg Ala Asp Asn Glu Gln Phe
1 5 10
<210> 75
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 75
Ala Ser Ser Phe Leu Ser Asn Pro Gly Asn Thr Ile Tyr
1 5 10
<210> 76
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 76
Ala Ser Ser Ile Leu Gly Gly Ala Ala Asp Thr Gln Tyr
1 5 10
<210> 77
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 77
Ala Asp Pro Phe Asp Arg Gly Leu Glu Ala Phe
1 5 10
<210> 78
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 78
Ala Ser Ser Leu Gly Thr Ile Ile Gly Ser Gly Asn Thr Ile Tyr
1 5 10 15
<210> 79
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 79
Ala Ser Asp Trp Gly Pro Ala Leu His Glu Gln Tyr
1 5 10
<210> 80
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 80
Ala Ser Ser Ser Ser Thr Gly Leu Ala Gly Gly Pro Tyr Glu Gln Tyr
1 5 10 15
<210> 81
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 81
Ala Ser Ser Glu Leu Gly Gly Val Trp Ser Glu Ala Phe
1 5 10
<210> 82
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 82
Ala Ser Ser Gly Pro Thr Thr Gly Val Arg Asn Asn Glu Gln Phe
1 5 10 15
<210> 83
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 83
Ala Thr Ser Ser Tyr Trp Pro Gln His
1 5
<210> 84
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 84
Ala Ser Ser Ile Asp Val Val Ser Gly Ser Phe Pro Tyr Glu Gln Tyr
1 5 10 15
<210> 85
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 85
Ala Ser Ser Ser Thr Gly Gly Thr Gly Ile Tyr Asn Glu Gln Phe
1 5 10 15
<210> 86
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 86
Ala Ser Ser Ser Ser Phe Asp Thr Gly Glu Leu Phe
1 5 10
<210> 87
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 87
Ala Ser Ser Pro Val Ala Glu Gly Glu Glu Thr Gln Tyr
1 5 10
<210> 88
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 88
Ala Ser Ser Leu Ala Asp Phe Leu Ser Tyr Asn Glu Gln Phe
1 5 10
<210> 89
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 89
Ala Ile Ser Glu Pro Met Val Gly Gly Thr Glu Ala Phe
1 5 10
<210> 90
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 90
Ala Ser Ser Pro Gly Gln Gly Glu Pro Thr Ile Tyr
1 5 10
<210> 91
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 91
Ala Ser Thr Ser Glu Thr Ser Phe Tyr Glu Gln Phe
1 5 10
<210> 92
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 92
Pro Ala Val Thr Arg Gly Val Ser Thr Ser Ser Thr
1 5 10
<210> 93
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 93
Ala Ser Ser Gln Glu Tyr Ser Gly Ser Ala Tyr Glu Gln Tyr
1 5 10
<210> 94
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 94
Ala Thr Lys Arg Arg Asp Arg Gly Asn Glu Gln Phe
1 5 10
<210> 95
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 95
Ala Ser Thr Pro His Leu Ser Ser Tyr Glu Gln Tyr
1 5 10
<210> 96
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 96
Ala Ser Ser Thr Asp Arg Phe Ala Gln His
1 5 10
<210> 97
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 97
Ala Ile Ser Glu Pro Thr Thr Ser Gly Gly Gly Leu Leu Glu Thr Gln
1 5 10 15
Tyr
<210> 98
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 98
Ala Ser Ser Lys Gly Thr Ser Trp Phe Leu Ala Tyr Glu Gln Tyr
1 5 10 15
<210> 99
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 99
Ala Ser Ser Leu Val Val Thr Gln Gly Leu Thr Asp Thr Gln Tyr
1 5 10 15
<210> 100
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 100
Ala Thr Thr Ser Gly Leu Ala Gly Gly Tyr Leu Ser Gly Glu Leu Phe
1 5 10 15
Claims (7)
1.一种弥漫大B细胞淋巴瘤的外周血TCR标志物，其特征在于，所述标志物包括序列为SEQ ID NO.1～100所示的蛋白中的至少一种。
2.根据权利要求1所述的弥漫大B细胞淋巴瘤的外周血TCR标志物，其特征在于，所述标志物的蛋白序列为SEQ ID NO.1～100所示的序列经取代、缺失和/或替换一个或多个碱基后，能表达相同功能的蛋白。
3.权利要求1所述的标志物在制备治疗弥漫大B细胞淋巴瘤的制剂中的应用。
4.根据权利要求3所述的应用，其特征在于，所述制剂包括含有该标志物的T细胞受体，或能表达产生该标志物的T细胞受体的质粒、病毒载体或核酸片段。
5.一种用于弥漫大B细胞淋巴瘤检测的试剂盒，其特征在于，包括能与权利要求1所述标志物发生特异性结合的抗体。
6.一种制剂，其特征在于，包括能与权利要求1所述标志物发生特异性结合的抗体；所述制剂可用于对弥漫大B细胞淋巴瘤进行诊断、预测、检测或筛查。
7.一种检测弥漫大B细胞淋巴瘤的蛋白质芯片，其特征在于，所述蛋白质芯片包括基片和点样在基片上特异性抗体，所述特异性抗体为能与权利要求1所述标志物发生特异性结合的抗体。
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| PCT/CN2022/080282 WO2022194033A1 (zh) | 2021-03-15 | 2022-03-11 | 一种弥漫大b细胞淋巴瘤的外周血tcr标志物及其检测试剂盒和应用 |
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| WO2022194033A1 (zh) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | 一种弥漫大b细胞淋巴瘤的外周血tcr标志物及其检测试剂盒和应用 |
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| KR101773690B1 (ko) * | 2007-03-05 | 2017-08-31 | 인터내셔널 인스티튜트 오브 캔서 이무놀로지 인코퍼레이티드 | 암 항원 특이적 ｔ 세포의 수용체 유전자 및 그것에 따라 코드되는 펩티드 및 이들의 사용 |
| CN101225388A (zh) * | 2008-01-24 | 2008-07-23 | 暨南大学 | 弥漫性大B细胞淋巴瘤相关抗原特异TCR Vα6亚家族的MR基因序列及应用 |
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| CN113092761A (zh) * | 2021-03-15 | 2021-07-09 | 成都益安博生物技术有限公司 | 一种弥漫大b细胞淋巴瘤的外周血tcr标志物及其检测试剂盒和应用 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2022194033A1 (zh) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | 一种弥漫大b细胞淋巴瘤的外周血tcr标志物及其检测试剂盒和应用 |
| CN113567682A (zh) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | 一种阿尔茨海默病的外周血tcr标志物及其检测试剂盒和应用 |
| WO2023000688A1 (zh) * | 2021-07-23 | 2023-01-26 | 成都益安博生物技术有限公司 | 一种阿尔茨海默病的外周血tcr标志物及其检测试剂盒和应用 |
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