CN113030473A - 一种急性b淋巴细胞白血病的外周血tcr标志物及其检测试剂盒和应用 - Google Patents
一种急性b淋巴细胞白血病的外周血tcr标志物及其检测试剂盒和应用 Download PDFInfo
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Abstract
本发明公开了一种急性B淋巴细胞白血病的外周血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，首先确定急性B淋巴细胞白血病外周血中特征性TCR序列，然后将待测样本测试结果与该特征性TCR序列比对，从而确定是否患有急性B淋巴细胞白血病。本发明能够同时比较巨大数量的急性B淋巴细胞白血病特异性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，具体涉及一种急性B淋巴细胞白血病的外周血TCR标志物及其检测试剂盒和应用。
背景技术
急性淋巴细胞白血病(Acute Lymphoblastic Leukemia，ALL)是一种起源于B系或T系淋巴祖细胞在骨髓内异常增生的恶性肿瘤性疾病。异常增生的原始细胞可在骨髓聚集并抑制正常造血功能，同时也可侵及骨髓外的组织，如脑膜、淋巴结、性腺、肝等。我国曾进行过白血病发病情况调查，ALL发病率约为0.67/10万。在油田、污染区发病率明显高于全国发病率。ALL儿童期(0～9岁)为发病高峰，可占儿童白血病的70％以上。ALL在成人中占成人白血病的20％左右。
目前针对急性淋巴细胞白血病的诊断手段主要有血常规检测、细胞化学检查、免疫分型、PCR检测、骨髓象检测、腰椎穿刺、骨髓活检。
在血常规检测中超过90％以上患者在诊断时有明显血液学异常，异常的严重程度反映了白血病细胞侵及的程度。贫血为正细胞、正色素性。约有50％以上ALL白细胞总数增高，3/4患者有血小板减少。大部分患者在末梢血涂片中可以见到数量不等的幼稚淋巴细胞。该种检测只能作为急性淋巴细胞白血病的常规检测之一，无法单独通过该种检测判定是否患有急性淋巴细胞白血病，而且不同的患者检测结果不具有一致性的白细胞总数增高或血小板减少或均可见幼稚淋巴细胞。
AL细胞在细胞化学检查糖原染色时多数细胞中有特异的阳性颗粒，以粗块状为典型表现，过氧化物酶和苏丹黑染色呈阴性。借助细胞化学染色来认识ALL各亚型有一定的一定的鉴别意义，但该种检测不能作为早期诊断定性急性淋巴细胞白血病的检测。
免疫分型应用系列单克隆抗体对白血病细胞进行标记，常用多参数流式细胞仪进行分析，确定白血病类型。该种检测测量速度快,可多参数测量，但灵敏度不够且ALL患者微小残留病可出现抗原漂移导致流式结果呈假阴性。
PCR包括实时荧光定量RCR(qPCR)、逆转录PCR(RT-PCR)两种,检测微小残留病的灵敏度均较高,但结果易出现假阴性。
骨髓象检测多数患者骨髓细胞增生活跃或明显活跃，骨髓细胞分类可见到≥20％原始淋巴细胞，并有部分幼稚淋巴细胞。腰椎穿刺涉及从脊柱取一小部分脑脊液并检查是否存在异常的癌细胞。对于癌症已经扩散到中枢神经系统的患者，这可能是积极的。如果患者血液检查异常，则开具骨髓活检。活组织检查包括使用注射器取骨髓样本。通常将针刺入髋骨并抽取样本。该过程是痛苦的并且在局部麻醉下进行。该过程大约需要15分钟才能完成，并在门诊进行。
除以上检测之外，目前可用于诊断急性淋巴细胞白血病的标志物非常少。
发明内容
针对现有技术中的上述不足，本发明提供一种急性B淋巴细胞白血病的外周血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急性B淋巴细胞白血病风险患者。
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：
一种急性B淋巴细胞白血病的外周血TCR标志物，该标志物包括序列为SEQ IDNO.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急性B淋巴细胞白血病的制剂中的应用。
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。
一种用于急性B淋巴细胞白血病检测的试剂盒，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；所述制剂可用于对急性B淋巴细胞白血病进行诊断、预测、检测或筛查。
一种检测急性B淋巴细胞白血病的蛋白质芯片，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个非急性B淋巴细胞白血病的对照组样本、和55个急性B淋巴细胞白血病病人的TCR高通量测序数据建立了人工智能分析模型，通过和这些急性B淋巴细胞白血病特异性TCR序列对比，可以清楚的判断待测样本中是否有较高急性B淋巴细胞白血病风险者；
2、通过高通量测序分析TCR变化可以发现很早期的急性B淋巴细胞白血病，利用急性B淋巴细胞白血病特有的TCR CDR3序列分析人的免疫系统中的T细胞对急性B淋巴细胞白血病的反应，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，可用于急性B淋巴细胞白血病的免疫治疗。
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急性B淋巴细胞白血病特征性TCR序列。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急性B淋巴细胞白血病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；急性B淋巴细胞白血病患者的免疫图谱具有多个种类且重复次数较高的急性B淋巴细胞白血病特征序列，健康人极少急性B淋巴细胞白血病特征序列，而未知受试者的急性B淋巴细胞白血病特征比较明显，说明罹患急性B淋巴细胞白血病风险较高。
图2为本发明中，利用急性B淋巴细胞白血病特征特征性指数对比分析急性B淋巴细胞白血病和其他疾病。健康人、非肿瘤病人、非急性B淋巴细胞白血病肿瘤患者的急性B淋巴细胞白血病特征性指数均与急性B淋巴细胞白血病患者具有显著差异，证明了急性B淋巴细胞白血病特征序列集的特异性。据此可以判断未知受试者是否罹患急性B淋巴细胞白血病。
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，获得急性B淋巴细胞白血病TCR标志物CDR3序列集
1、采样及免疫图谱分析
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)、56名急性B淋巴细胞白血病患者(55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，确定急性B淋巴细胞白血病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55名用于建立模型的急性B淋巴细胞白血病样本的所有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，设为急性B淋巴细胞白血病特征序列集。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急性B淋巴细胞白血病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名急性B淋巴细胞白血病患者和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急性B淋巴细胞白血病特征序列集合进行作图，见附图1B-D。从图中可见，急性B淋巴细胞白血病患者的免疫图谱中，含有较多种类且较高重复出现次数的急性B淋巴细胞白血病特征序列(图1B)；健康人的免疫图谱中，只有极少量急性B淋巴细胞白血病特征序列(图1C)；而未知健康状况受试者，有高于健康人的急性B淋巴细胞白血病特征序列，说明此人有较高风险罹患急性B淋巴细胞白血病(图1D)。
d)将急性B淋巴细胞白血病特征序列集中，将所有出现在两个及以上参与建模急性B淋巴细胞白血病样本里的CDR3序列，按“所有参与建模急性B淋巴细胞白血病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急性B淋巴细胞白血病样本数”从高到低排序，排名前100者即为急性B淋巴细胞白血病TCR标志物CDR3序列，具体序列如SEQ ID NO.1～100所示。
实施例2验证急性B淋巴细胞白血病TCR标志物CDR3序列集的特异性
1、采样及免疫图谱分析
采集626名非急性B淋巴细胞白血病的肿瘤患者、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97名健康人及41名非肿瘤疾病病人。
3、根据来自实施例1的97名健康人、41名非肿瘤疾病病人、47名急性B淋巴细胞白血病患者，以及实施例2新获取的626名非急性B淋巴细胞白血病肿瘤患者、7名未知健康状况受试者的免疫图谱，分析其急性B淋巴细胞白血病特征性指数。
其中急性B淋巴细胞白血病特征性指数定义为：某个样本中，属于急性B淋巴细胞白血病特征序列集的所有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。急性B淋巴细胞白血病组与健康人(p＝2.4E-80)、非肿瘤疾病(p＝5.4E-43)、其它肿瘤(p趋于0)都有显著差异，这证明了急性B淋巴细胞白血病特征序列集的特异性。
表2不同样本组的急性B淋巴细胞白血病特征性指数
4、分析各组的急性B淋巴细胞白血病特征指数(表3)，7位未知健康状况受试者(检测样本)的急性B淋巴细胞白血病特征指数显著高于“其它肿瘤”组的平均值+2×SD(141.4+2×359.4＝800.2)，此7人具有较高风险罹患急性B淋巴细胞白血病。与临床体检结果对照后，这7人确为早期急性B淋巴细胞白血病患者。此实施例证明了利用急性B淋巴细胞白血病特征序列集及急性B淋巴细胞白血病特征性指数，预测受试者罹患急性B淋巴细胞白血病风险的可行性。
表3急性B淋巴细胞白血病特征性指数分析
综上所述，本发明所述急性B淋巴细胞白血病TCR标志物CDR3序列，确实具有显著的急性B淋巴细胞白血病特异性，不仅可以用于急性B淋巴细胞白血病预测受试者罹患急性B淋巴细胞白血病风险，未来还可用于急性B淋巴细胞白血病的生物免疫治疗。
序列表
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<400> 11
Ala Ser Ser Lys Glu Gly Arg Ala Ser Tyr Glu Gln Tyr
1 5 10
<210> 12
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Ala Ser Ser Leu Ile Asp Arg Gly Ser Gly Ser Pro Leu His
1 5 10
<210> 13
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 13
Ala Ser Ser Trp Gly Leu Gly Thr Gly Glu Gly Tyr Thr
1 5 10
<210> 14
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
Ala Ser Ser Leu Lys Gly Val His Glu Gln Phe
1 5 10
<210> 15
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
Ala Ser Ser His Leu Thr Ser Gly Phe Ser Asn Glu Gln Phe
1 5 10
<210> 16
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
Ser Val Glu Tyr Ala Thr Arg Gly Leu Ala Asp Asn Glu Gln Phe
1 5 10 15
<210> 17
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 17
Ala Ser Asn His Arg Gly Arg Ser Glu Pro Gln His
1 5 10
<210> 18
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
Ala Ser Ser Leu Ser Ser Ser His Gly Ala Phe
1 5 10
<210> 19
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Ala Ser Arg Pro Arg Gly Thr Gly His Asn Glu Gln Phe
1 5 10
<210> 20
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Ala Ser Ser Gln Asp Asn Thr Pro Tyr Gly Gly Tyr Thr
1 5 10
<210> 21
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 21
Ala Ser Ser Lys Ala Pro Gly Lys Gly Cys Gln Pro Gln His
1 5 10
<210> 22
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
Gln Gln Pro Arg Arg Ala Ser Gly Leu Trp Gly Gln Arg Pro Asp
1 5 10 15
<210> 23
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 23
Ala Ser Ser Tyr Glu Thr Gly Ile Phe Asp Tyr Gly Tyr Thr
1 5 10
<210> 24
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 24
Ala Ser Ser Asn Leu Trp Asp Arg Gly Gly Gln Asn Glu Lys Leu Phe
1 5 10 15
<210> 25
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 25
Ser Val Glu Ile Pro Arg Gly Leu Glu Asp Ser Tyr Gly Tyr Thr
1 5 10 15
<210> 26
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Ser Val Glu Tyr Gly Ser Ser Ala Gly Glu Leu Phe
1 5 10
<210> 27
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 27
Ala Ser Ser Pro Leu Ala Gly Leu Ala Asn Asn Glu Gln Phe
1 5 10
<210> 28
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
Ala Ser Ser Ser Thr Pro Leu Ala Gly Glu Gln Tyr
1 5 10
<210> 29
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 29
Ala Ser Ser Leu Asp Gly Gly Met Tyr Glu Gln Phe
1 5 10
<210> 30
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 30
Ala Thr Pro Ser Glu Arg Thr Leu Thr Asn Thr Gly Glu Leu Phe
1 5 10 15
<210> 31
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 31
Ala Ser Ser Pro Glu Pro Gln Arg Ser Ala Gly Asp Gln Pro Gln His
1 5 10 15
<210> 32
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
Ala Ser Ser Val Arg Gln Gly Phe Ile Tyr
1 5 10
<210> 33
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 33
Ala Ser Ser Pro Val Asp Ala Gly Gly Gly Tyr Thr
1 5 10
<210> 34
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 34
Ala Trp Ser Asp Arg Gly Ala Phe Ser Asp Thr Gln Tyr
1 5 10
<210> 35
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 35
Ala Ile Gln Glu Gly Arg Glu Gly Asn Asn Glu Gln Phe
1 5 10
<210> 36
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 36
Ala Ser Ser Leu Glu Gln Ser Pro Tyr Gly Tyr Thr
1 5 10
<210> 37
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 37
Ala Ser Ser Leu Gly Gly Arg Ser Asn Gly Ala Asn Val Leu Thr
1 5 10 15
<210> 38
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 38
Ala Ser Ser Phe Gly Ile Ser Glu Val Ala Phe
1 5 10
<210> 39
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 39
Ala Ser Ser Leu Val Arg Asp Ser Leu Asn Glu Gln Tyr
1 5 10
<210> 40
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 40
Ala Ser Ser Tyr Gly Gln His Ser Thr Gln Glu Thr Gln Tyr
1 5 10
<210> 41
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 41
Gln Gln Leu Ser Arg Glu Ile Leu Trp Gly Gln Arg Pro Asp
1 5 10
<210> 42
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 42
Ala Ser Asp His Pro Thr Ser Gly Ser Phe Glu Thr Gln Tyr
1 5 10
<210> 43
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 43
Ala Ser Thr Pro Pro Ile Thr Asn Thr Gly Glu Leu Phe
1 5 10
<210> 44
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 44
Ala Ser Ser Gly Glu Arg Asp Gly Tyr Asn Glu Gln Phe
1 5 10
<210> 45
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 45
Ala Trp Gln Gly Ala Gly Tyr Gly Tyr Thr
1 5 10
<210> 46
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 46
Thr Ser Ser Gln Glu Ser Ala Gly Leu Leu Gln Glu Thr Gln Tyr
1 5 10 15
<210> 47
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 47
Ala Ile Ser Gly Gly Ser Lys Asn Thr Glu Ala Phe
1 5 10
<210> 48
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 48
Ala Thr Ser Thr Gly Ser Arg Ser Tyr Glu Gln Tyr
1 5 10
<210> 49
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 49
Ala Ser Ser Leu Arg Gly Gln Ser Glu Gly Gly Thr Gln Tyr
1 5 10
<210> 50
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 50
Ala Ser Ser Leu Ala Gly Gly Thr Ser Val Glu Asn Thr
1 5 10
<210> 51
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 51
Gln Gln Gln Arg Gly Gln Gly Leu Leu Arg Ala Val
1 5 10
<210> 52
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 52
Ala Ser Ser Arg Arg Gly Gly Arg Arg Ser Tyr Asn Glu Gln Phe
1 5 10 15
<210> 53
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 53
Gln Gln Arg Arg Lys Gly Ile His Arg Tyr Ala Val
1 5 10
<210> 54
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 54
Ala Ser Arg Met Pro Ala Gly Ala Leu Gln Glu Thr Gln Tyr
1 5 10
<210> 55
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 55
Ser Ala Arg Asp Asp Gln Phe Tyr Ser Asn Gln Pro Gln His
1 5 10
<210> 56
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 56
Ala Ser Ser Pro Arg Asp Thr Ser Asn Asn Glu Gln Phe
1 5 10
<210> 57
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 57
Ala Ser Ser Gln Gly Gly Pro Leu Glu Ala Phe
1 5 10
<210> 58
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 58
Ala Ser Ser Pro Met Asp Arg Val Gly Ile Trp
1 5 10
<210> 59
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 59
Pro Ala Tyr Arg Ser Leu Leu Thr Ser Ser Thr
1 5 10
<210> 60
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 60
Ala Ser Ser Ile Gly Pro Thr Ser Gly Gly Ser Asn Glu Gln Phe
1 5 10 15
<210> 61
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 61
Ala Ser Ser Pro Ser Gly Gly Arg Ser Gly Ala Asn Val Leu Thr
1 5 10 15
<210> 62
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 62
Ala Thr Ser Asp Ser Phe Gly Gly Arg Ala Asn Glu Gln Phe
1 5 10
<210> 63
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 63
Ala Ser Ser Gln Ala Ser Asn Gly Asp Glu Gln Phe
1 5 10
<210> 64
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 64
Gln Leu Ile Ala Ala Phe Leu Arg Ala Val
1 5 10
<210> 65
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 65
Ala Ser Ser Ala Asp Leu Gly Ser Ser Gly Asn Thr Ile Tyr
1 5 10
<210> 66
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 66
Ala Ser Asn Asp Arg Ser Ser Ser Gly Asn Thr Ile Tyr
1 5 10
<210> 67
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 67
Ala Trp Ser Val Val Ala Gly Gly His Ser Tyr Asn Glu Gln Phe
1 5 10 15
<210> 68
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 68
Ala Ser Thr Pro Lys Arg Gln Trp Asn Thr Glu Ala Phe
1 5 10
<210> 69
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 69
Ala Ser Ser Tyr Ser Met Gly Lys Gly Ser His Arg Asp Glu Gln Phe
1 5 10 15
<210> 70
<211> 18
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 70
Ala Ser Ser Leu Glu Gly Ser Arg Gly Ile Thr Tyr Ser Tyr Asn Glu
1 5 10 15
Gln Phe
<210> 71
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 71
Ser Val Ile Asp Pro Gly Thr Arg Ser Tyr Glu Gln Tyr
1 5 10
<210> 72
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 72
Ala Ile Ser Val Gly Tyr Gly Thr Glu Ala Phe
1 5 10
<210> 73
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 73
Ala Ser Ser Ser Thr Glu Arg Glu Trp Gly Asn Glu Gln Phe
1 5 10
<210> 74
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 74
Ala Ser Ser Pro Gln Lys Asp Ala Gly Thr Ser Val Asn Glu Gln Phe
1 5 10 15
<210> 75
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 75
Ala Ser Thr Glu Trp Gly Leu Gly Tyr Glu Gln Tyr
1 5 10
<210> 76
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 76
Ala Ser Ser Thr Gly Leu Glu Leu Tyr Asn Glu Gln Phe
1 5 10
<210> 77
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 77
Ala Ser Ser Pro Thr Gly Thr Gly Lys Asn Asn Glu Gln Phe
1 5 10
<210> 78
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 78
Ala Ser Ser His His Leu Thr Gly Pro Glu Ala Phe
1 5 10
<210> 79
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 79
Ala Ser Ser Leu Ser Gly Gln Gly Val Phe Thr Glu Ala Phe
1 5 10
<210> 80
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 80
Ala Ser Arg His Ser Gly Tyr Gly Tyr Thr
1 5 10
<210> 81
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 81
Pro Ala Arg Arg Gly Gln Arg Pro Ser Thr
1 5 10
<210> 82
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 82
Ala Ser Ser Pro Arg Gly Ile Phe Gly Asn Thr Ile Tyr
1 5 10
<210> 83
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 83
Ala Ser Ser Lys Glu Gly Thr Ser Gly Glu Thr Gln Tyr
1 5 10
<210> 84
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 84
Ala Ser Leu Asp Ser Gly Arg Tyr Asn Glu Gln Phe
1 5 10
<210> 85
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 85
Ala Trp Ser Ala Gln Pro Gly Asn Glu Gln Phe
1 5 10
<210> 86
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 86
Ala Ser Ser His Gly Val Leu Arg Glu Asn Glu Gln Tyr
1 5 10
<210> 87
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 87
Ala Ser Ser Pro Asn Arg Trp Thr Gly Pro His Glu Lys Leu Phe
1 5 10 15
<210> 88
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 88
Ala Ser Ser Arg Gly Asp Gln Asn Glu Gln Phe
1 5 10
<210> 89
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 89
Ala Ser Ser Phe Gly Thr Asp Phe Ser Ser Thr Asp Thr Gln Tyr
1 5 10 15
<210> 90
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 90
Ala Ser Ser Leu Asp Pro Ala Asp Pro Tyr Glu Gln Tyr
1 5 10
<210> 91
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 91
Ala Ser Ser Glu Thr Gly Gly Gly Ser Ala Asp Thr Gln Tyr
1 5 10
<210> 92
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 92
Ala Ser Arg Val Ala Glu Lys Ser Ala Tyr Asn Glu Gln Phe
1 5 10
<210> 93
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 93
Ala Thr Arg Gly Gln Gly Ala Ser Pro Leu His
1 5 10
<210> 94
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 94
Ala Ser Ser Gln Ser Ser Ser Gly Glu Gln Phe
1 5 10
<210> 95
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 95
Ala Ile Ser Glu Ser Ser Gly Leu Ala Gly Leu Ile Ile Gly Gln Phe
1 5 10 15
<210> 96
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 96
Ala Trp Thr Pro Leu Ala Gly Gly Thr Gly Glu Leu Phe
1 5 10
<210> 97
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 97
Ala Ser Ser Leu Ala Leu Gly Arg Val Gly Val Tyr Glu Gln Tyr
1 5 10 15
<210> 98
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 98
Ala Ser Ser Leu Gln Ser Gly Gln Ala Tyr Glu Gln Tyr
1 5 10
<210> 99
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 99
Ala Thr Ser Arg Asp Leu Gly Gly Thr Gly Tyr Thr
1 5 10
<210> 100
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 100
Ala Ile Ser Gln Ile Asp Asn Glu Gln Tyr
1 5 10
Claims (7)
1.一种急性B淋巴细胞白血病的外周血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所述的急性B淋巴细胞白血病的外周血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急性B淋巴细胞白血病的制剂中的应用。
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。
5.一种用于急性B淋巴细胞白血病检测的试剂盒，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；所述制剂可用于对急性B淋巴细胞白血病进行诊断、预测、检测或筛查。
7.一种检测急性B淋巴细胞白血病的蛋白质芯片，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/CN2022/080317 WO2022194039A1 (zh) | 2021-03-15 | 2022-03-11 | 一种急性b淋巴细胞白血病的外周血tcr标志物及其检测试剂盒和应用 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
| CN113563454A (zh) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | 一种IgA肾病的外周血TCR标志物及其检测试剂盒和应用 |
| WO2022194039A1 (zh) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | 一种急性b淋巴细胞白血病的外周血tcr标志物及其检测试剂盒和应用 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2022194039A1 (zh) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | 一种急性b淋巴细胞白血病的外周血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 |
| CN113563454A (zh) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | 一种IgA肾病的外周血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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