WO2022194036A1 - Peripheral blood tcr marker for classical hodgkin lymphoma, detection kit therefor, and application thereof - Google Patents

Peripheral blood tcr marker for classical hodgkin lymphoma, detection kit therefor, and application thereof Download PDF

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WO2022194036A1
WO2022194036A1 PCT/CN2022/080301 CN2022080301W WO2022194036A1 WO 2022194036 A1 WO2022194036 A1 WO 2022194036A1 CN 2022080301 W CN2022080301 W CN 2022080301W WO 2022194036 A1 WO2022194036 A1 WO 2022194036A1
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张志新
卓越
杨鑫
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成都益安博生物技术有限公司
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    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/7051—T-cell receptor (TcR)-CD3 complex
    • 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
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    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
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    • G01N33/57488—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds identifable in body fluids
    • G—PHYSICS
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    • 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
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854—Immunoglobulins
    • G—PHYSICS
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    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G01N2333/7051—T-cell receptor (TcR)-CD3 complex

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  • the invention belongs to the technical field of genetic engineering, and in particular relates to a peripheral blood TCR marker of classic Hodgkin's lymphoma and a detection kit and application thereof.
  • HL Classical Hodgkin's Lymphoma
  • HL Hodgkin's disease
  • HL Classical Hodgkin's Lymphoma
  • Epidemiology shows that in European and American countries, Hodgkin lymphoma accounts for about 15% to 20%, while the average incidence of Hodgkin lymphoma in my country is low, with an average of 6 people per 1 million people each year, that is, About 8,000 people nationwide have been diagnosed with Hodgkin lymphoma.
  • Hodgkin's lymphoma accounts for only about 10% of all lymphomas, but the incidence of Hodgkin's lymphoma is high in children, accounting for 33%-55% of all malignant lymphomas, with a slightly higher prevalence in men Female (1.31-1.4:1).
  • lymphoma The cause of Hodgkin's lymphoma is still unclear. It is generally believed to be related to factors such as infection, immune dysfunction, genetics, occupational exposure and lifestyle. Under the action of various factors, immune cells in the body mutate, causing the activation of intracellular cancer genes and the inactivation of inhibitory genes. , lymphocytes proliferate malignantly and eventually form lymphoma.
  • Hodgkin lymphoma is a highly curable tumor. Internationally, the cure rate of newly treated Hodgkin's lymphoma can reach 75% to 85%. However, the overall cure rate of Hodgkin's lymphoma in China is still less than 75%, for several reasons. The first is related to the accuracy of the diagnosis.
  • Classical Hodgkin lymphoma is divided into four histological types: lymphocyte predominant, tuberous sclerosis, mixed cellularity, and lymphocyte depletion. In recent years, a nodular lymphocyte-predominant type has been added to the WHO classification. The most common type in my country is mixed cell type. Histological subtype is a major factor in determining the clinical presentation, prognosis and treatment of patients.
  • Anemia is more common in advanced patients, normochromic, normocytic anemia. Occasionally hemolytic anemia, 2% to 10% of patients with positive Coombs test. A small number of cases may appear neutropenia, eosinophilia. Peripheral blood lymphopenia ( ⁇ 1.0 ⁇ 10 9 /L), increased erythrocyte sedimentation rate, and increased serum lactate dehydrogenase can be used as disease detection indicators.
  • X-ray examination can observe the enlargement of the hilar and mediastinal lymph nodes, and at the same time observe whether the hilum is invaded by the tumor.
  • the anterior superior mediastinum and hilar lymph nodes are significantly enlarged, the possibility of lymphoma should be considered.
  • CT examination is still the most commonly used imaging method for lymphoma staging, restaging, efficacy evaluation and follow-up.
  • enhanced CT scans should be used as much as possible.
  • CT has obvious shortcomings, and cannot detect small lesions, especially small lesions in solid organs or at the edge; CT can only measure the size of the tumor, neither can it reliably distinguish between normal lymphocytes and tumor cells. Infiltrating lymph nodes, it is also impossible to determine whether the residual disease is fibrous necrotic scar tissue or always viable tumor cells.
  • B-ultrasound is also a commonly used method for staging. It is very important for the discovery and follow-up of superficial lymph nodes. According to whether the lymph node structure is damaged or not and blood flow, it can help to judge the benign and malignant of enlarged lymph nodes. The ability to detect lymph nodes larger than 2 cm in diameter is of great significance for assisting in the detection of hepatosplenomegaly lymph nodes.
  • MRI examination is the preferred examination method for the central nervous system, bone marrow and muscle; especially for those who are not suitable for enhanced CT scan, or as a further examination after CT findings can be used.
  • PET/CT Positron emission computed tomography
  • Tissue biopsy is the most reliable basis for diagnosis.
  • the lymph nodes of the diseased site are excised for pathological examination, and the key to the diagnosis of typical mirror cells (R-S cells) is found, and this is used as the basis for subtypes.
  • R-S cells typical mirror cells
  • Cytological examination has certain difficulties in determining Hodgkin's lymphoid subtypes, which are mainly auxiliary staging, and follow-up after treatment for recurrence.
  • Bone marrow aspirate can judge the state and function of the bone marrow. When the tumor invades the bone marrow, typical RS cells can be found in the bone marrow tissue.
  • Imaging detection also has its own advantages and disadvantages. Its detection requires the tumor to reach a certain size before it can be detected. There is a hysteresis, and it cannot be used as a means of real-time monitoring due to radiation and other reasons. Therefore, there is an urgent need for a more convenient and sensitive screening and testing method that can track the occurrence and development of Hodgkin lymphoma.
  • the present invention provides a peripheral blood TCR marker of classic Hodgkin's lymphoma and a detection kit and application thereof, which can accurately and quickly determine whether there is a higher classic Hodgkin in the sample to be tested. Gold lymphoma risk patients.
  • a peripheral blood TCR marker of classic Hodgkin's lymphoma includes at least one of the proteins whose sequences are shown in SEQ ID NO. 1 to 100, and the specific sequence is shown in Table 1.
  • serial number protein sequence SEQ ID NO.1 Ala Ser Ser Leu Asn Arg Val Ser Ser Ser Ser Glu Gln Phe SEQ ID NO.2 Ala Ser Ser Phe Gly Lys Gly Pro Asn Tyr Gly Tyr Thr
  • SEQ ID NO. 52 Ala Ser Ser Arg Glu Gly Ser Gly Ser Asn Gly Glu Leu Phe SEQ ID NO. 53 Ala Ser Arg Pro Ala Asp Trp Ser Pro Ser Thr Asp Thr Gln Tyr SEQ ID NO. 54 Ala Ser Ser Val Gly Arg Thr Ser Gly Leu Gly Arg Glu Thr Gln Tyr SEQ ID NO. 55 Ala Thr Ser Arg Asp Ser Arg Arg Thr Gly Ser Ser Tyr Glu Gln Tyr SEQ ID NO. 56 Ala Ser Ser Tyr Glu Thr Val Ser Val Ala Asn Thr Gly Glu Leu Phe SEQ ID NO.
  • protein sequence of the marker is that the sequence shown in SEQ ID NO. 1-100 can express a protein with the same function after one or more bases are substituted, deleted and/or replaced.
  • the marker is the peripheral blood TCR CDR3 sequence.
  • the preparation includes a T cell receptor containing the marker, or a plasmid, viral vector or nucleic acid fragment capable of expressing the T cell receptor producing the marker.
  • kits for the detection of classical Hodgkin's lymphoma comprising antibodies that can specifically bind to the above markers.
  • a preparation comprising an antibody capable of specifically binding to the above-mentioned marker; the preparation can be used for diagnosis, prediction, detection or screening of classical Hodgkin lymphoma.
  • a protein chip for detecting classical Hodgkin's lymphoma the protein chip comprises a substrate and a specific antibody spotted on the substrate, the specific antibody being an antibody that can specifically bind to the above marker.
  • B lymphocytes and T lymphocytes in the human body are two important types of cells in the acquired immune system.
  • B cells recognize antigens through the B cell receptor (BCR) on the cell surface. Later, when B cells differentiate into plasma cells, BCR is expressed as an antibody and secreted outside the cell.
  • T cells recognize antigens through T cell receptors (TCRs) on the cell surface.
  • BCR and TCR The diversity of BCR and TCR is the basis for the establishment of the adaptive immune system.
  • the theoretical value of BCR diversity is 10 18 and the theoretical value of TCR diversity is 10 14 .
  • antigenic determinant 3 CDR3
  • CDR3 antigenic determinant 3
  • BCR and TCR will change with different antigenic stimulation. Therefore, the occurrence and development of diseases can be tracked by using BCR or TCR high-throughput sequencing results.
  • MCHII histocompatibility antigen II
  • Antigen fragments presented by normal cells do not cause an immune response due to immune tolerance.
  • the abnormal protein expressed by the mutated gene and its fragments are presented on the cell surface, which will cause the human immune system to produce a targeted immune response. Therefore, analyzing the changes of BCR or TCR can detect the occurrence and development of tumors.
  • an artificial intelligence analysis model was first established by using 1300 non-classical Hodgkin lymphoma control samples and 70 classical Hodgkin lymphoma patients' TCR high-throughput sequencing data. Hodgkin's lymphoma-specific TCR sequence comparison can clearly determine whether there is a higher risk of classic Hodgkin's lymphoma in the sample to be tested.
  • TCR CDR3 sequence specific to classical Hodgkin lymphoma to analyze the effect of T cells in the human immune system on classical Hodgkin lymphoma.
  • the reaction is a new detection method.
  • the present invention simultaneously compares a huge number of specific TCR sequences, and has higher specificity and accuracy than detecting one or several markers alone.
  • the cost of the high-throughput sequencing instrument used in the present invention is lower than that of large-scale imaging equipment, and it can be outsourced to a third party.
  • the labor cost of sampling and processing is lower than the simultaneous detection of multiple markers, and it is also lower than that of a large number of cytology detection, therefore, the present invention greatly reduces the detection cost.
  • the present invention only needs to take a small amount of peripheral blood, and the sampling is simple and safe.
  • TCR CDR3 sequence described in the present invention can be used for immunotherapy of classical Hodgkin's lymphoma.
  • Fig. 1 is the characteristic TCR sequence of classical Hodgkin's lymphoma discovered by the immune-based big data analysis system in the present invention; wherein, the abscissa represents that the CDR3 sequence of a certain amino acid combination is added to the control sequence set or the classical Hodgkin's lymphoma
  • the order of the tumor characteristic sequence set, the ordinate represents the logarithm of the number of repeated occurrences of the sequence in a certain sample, C X ;
  • the immune map of patients with classic Hodgkin lymphoma has multiple types of classic Hodgkin with a high number of repetitions.
  • Golden lymphoma characteristic sequences healthy people rarely have classical Hodgkin lymphoma characteristic sequences, while unknown subjects have more obvious classical Hodgkin lymphoma characteristics, indicating a higher risk of classic Hodgkin lymphoma.
  • Fig. 2 is a comparative analysis of classical Hodgkin's lymphoma and other diseases using the characteristic index of classical Hodgkin's lymphoma in the present invention;
  • the lymphoma characteristic indices were significantly different from those of patients with classical Hodgkin lymphoma, proving the specificity of the characteristic sequence set of classical Hodgkin lymphoma, which can be used to determine whether an unknown subject suffers from classical Hodgkin lymphoma.
  • Example 1 Obtaining the classic Hodgkin's lymphoma TCR marker CDR3 sequence set by immunographic analysis
  • the peripheral blood (10 mL each) of 522 non-classical Hodgkin lymphoma tumor patients and 5 subjects with unknown health status was collected, and the TCR epitope 3 of the subjects and the control group was obtained by high-throughput sequencing (CDR3) amino acid sequence, ensure that the total number of CDR3 sequences of functional TCRs in each sample is not less than 25,000; the sequences of samples whose total number of CDR3 sequences of each functional TCR is more than 30,000 are randomly sampled without replacement, so that The total number of CDR3 sequences for this sample is 30,000.
  • the total number of CDR3 sequences of functional TCRs contained in all samples so far is 25,000-30,000.
  • the characteristic index of classical Hodgkin lymphoma is defined as: in a certain sample, the sum of the repeated occurrences C X of all CDR3 sequences belonging to the characteristic sequence set of classical Hodgkin lymphoma in the sample.
  • the analysis results are shown in Table 2 below and accompanying drawing 2.
  • the classical Hodgkin lymphoma TCR marker CDR3 sequence of the present invention does have significant classical Hodgkin lymphoma specificity, and can not only be used for classical Hodgkin lymphoma to predict subjects suffering from classical Hodgkin lymphoma.
  • the risk of Hodgkin lymphoma can also be used for biological immunotherapy of classical Hodgkin lymphoma in the future.

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Abstract

A peripheral blood TCR marker for classical Hodgkin lymphoma, a detection kit therefor, and an application thereof. The marker comprises at least one of proteins of which sequences are as shown in SEQ ID NOs. 1-100. On the basis of a high-throughput sequencing method, only a small amount of peripheral blood needs to be used, RNA is extracted, and an immune map library is established by treating a sample; then by means of high-throughput sequencing and TCR data analysis, a characteristic TCR sequence in the peripheral blood of classical Hodgkin lymphoma is determined first, and then a test result of a sample to be tested is compared with the characteristic TCR sequence, so as to determine whether a patient suffers from classical Hodgkin lymphoma. A huge number of specific TCR sequences of classical Hodgkin lymphoma can be compared at the same time, and compared with separate detection of one or more markers, the present invention has higher specificity and accuracy, and improves diagnosis efficiency.

Description

一种经典霍奇金淋巴瘤的外周血TCR标志物及其检测试剂盒和应用A peripheral blood TCR marker of classical Hodgkin's lymphoma and its detection kit and application 技术领域technical field
本发明属于基因工程技术领域，具体涉及一种经典霍奇金淋巴瘤的外周血TCR标志物及其检测试剂盒和应用。The invention belongs to the technical field of genetic engineering, and in particular relates to a peripheral blood TCR marker of classic Hodgkin's lymphoma and a detection kit and application thereof.
背景技术Background technique
经典霍奇金淋巴瘤(Hodgkin's Lymphoma，HL)，又称霍奇金病是淋巴瘤的一种独特类型，为青年人中最常见的恶性肿瘤之一。流行病学显示，在欧美国家，霍奇金淋巴瘤约占15％～20％，而在我国霍奇金淋巴瘤的平均发病率较低，平均每年每100万人群中有6人发病，即全国大约有8000人被诊断为霍奇金淋巴瘤。在所有淋巴瘤中，霍奇金淋巴瘤仅占到10％左右，但儿童群体中霍奇金淋巴瘤发病率高，占所有恶性淋巴瘤的33％-55％，男性患病率略高于女性(1.31～1.4:1)。Classical Hodgkin's Lymphoma (HL), also known as Hodgkin's disease, is a unique type of lymphoma and one of the most common malignant tumors in young adults. Epidemiology shows that in European and American countries, Hodgkin lymphoma accounts for about 15% to 20%, while the average incidence of Hodgkin lymphoma in my country is low, with an average of 6 people per 1 million people each year, that is, About 8,000 people nationwide have been diagnosed with Hodgkin lymphoma. Hodgkin's lymphoma accounts for only about 10% of all lymphomas, but the incidence of Hodgkin's lymphoma is high in children, accounting for 33%-55% of all malignant lymphomas, with a slightly higher prevalence in men Female (1.31-1.4:1).
霍奇金淋巴瘤的发病原因至今不是很清楚。一般认为是感染、免疫功能失调、遗传、职业暴露和生活方式等因素相关，在多种因素的作用下，机体内的免疫细胞发生突变，引起了细胞内癌症基因的激活和抑制基因的失活，淋巴细胞恶性增殖，最终形成淋巴瘤。The cause of Hodgkin's lymphoma is still unclear. It is generally believed to be related to factors such as infection, immune dysfunction, genetics, occupational exposure and lifestyle. Under the action of various factors, immune cells in the body mutate, causing the activation of intracellular cancer genes and the inactivation of inhibitory genes. , lymphocytes proliferate malignantly and eventually form lymphoma.
霍奇金淋巴瘤是一种治愈率较高的肿瘤。在国际上，初治霍奇金淋巴瘤的治愈率能达到75％～85％。但国内霍奇金淋巴瘤的整体治愈率目前还达不到75％，原因有好几个。首先是与诊断的准确性有关。经典霍奇金淋巴瘤分为4种组织学类型：淋巴细胞为主型、结节硬化型、混合细胞型和淋巴细胞耗竭型。近年来WHO分型中增加了一种结节性淋巴细胞为主型。我国最常见为混合细胞型。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。Hodgkin lymphoma is a highly curable tumor. Internationally, the cure rate of newly treated Hodgkin's lymphoma can reach 75% to 85%. However, the overall cure rate of Hodgkin's lymphoma in China is still less than 75%, for several reasons. The first is related to the accuracy of the diagnosis. Classical Hodgkin lymphoma is divided into four histological types: lymphocyte predominant, tuberous sclerosis, mixed cellularity, and lymphocyte depletion. In recent years, a nodular lymphocyte-predominant type has been added to the WHO classification. The most common type in my country is mixed cell type. Histological subtype is a major factor in determining the clinical presentation, prognosis and treatment of patients. According to the data of the 2019 Lymphoma White Paper, about 43% of the patients who participated in the survey had experienced misdiagnosis, and 51% of the patients were diagnosed after going to multiple hospitals. It took an average of 2.5 months for patients to go from initial diagnosis to final determination of their subtype, with some subtypes waiting longer for diagnosis. At the same time, the survey shows that most patients need to go to big cities with concentrated medical resources such as Beijing, Shanghai and Guangzhou in order to make a diagnosis; secondly, the patient's awareness of seeking medical treatment is low. Many patients are already in the late stage when they come to see the doctor, and the treatment effect is definitely not as good as the early stage. Therefore, early diagnosis and early treatment are urgent needs to improve the treatment effect of lymphoma, improve the prognosis of patients, and save social resources.
经典霍奇金淋巴瘤的常用检测诊断方法有：The commonly used detection and diagnosis methods for classical Hodgkin lymphoma are:
1、血常规：1. Blood routine:
贫血多见于晚期患者，为正色素、正细胞性贫血。偶见溶血贫血，2％～10％患者Coombs试验阳性。少数病例可出现中性粒细胞增多，嗜酸性粒细胞增多。外周血淋巴细胞减少(<1.0×10 9/L)、血沉增快、血清乳酸脱氢酶升高可作为病情检测指标。 Anemia is more common in advanced patients, normochromic, normocytic anemia. Occasionally hemolytic anemia, 2% to 10% of patients with positive Coombs test. A small number of cases may appear neutropenia, eosinophilia. Peripheral blood lymphopenia (<1.0×10 9 /L), increased erythrocyte sedimentation rate, and increased serum lactate dehydrogenase can be used as disease detection indicators.
2、影像学检查2. Imaging examination
a)X线检查可以观察肺门、纵膈淋巴结的肿大，同时观察肺门内是否被肿瘤侵犯。前上纵膈及肺门淋巴结明显肿大时，应考虑淋巴瘤的可能。a) X-ray examination can observe the enlargement of the hilar and mediastinal lymph nodes, and at the same time observe whether the hilum is invaded by the tumor. When the anterior superior mediastinum and hilar lymph nodes are significantly enlarged, the possibility of lymphoma should be considered.
b)CT检查仍作为淋巴瘤分期、再分期、疗效评价和随访的最为常用的影像学方法，于无碘对比剂禁忌证的患者，应尽可能采用增强CT扫描。但需要注意的是，CT有明显的缺点，不能发现小病灶，尤其是 实质脏器内或者边缘的小病灶；CT仅能测定肿瘤的大小，既不能可靠地区分正常的淋巴细胞结合被肿瘤细胞侵润的淋巴结，也不能判断残留病灶是纤维坏死瘢痕组织还是始终有活性的瘤细胞。b) CT examination is still the most commonly used imaging method for lymphoma staging, restaging, efficacy evaluation and follow-up. For patients without contraindications to iodine contrast agents, enhanced CT scans should be used as much as possible. However, it should be noted that CT has obvious shortcomings, and cannot detect small lesions, especially small lesions in solid organs or at the edge; CT can only measure the size of the tumor, neither can it reliably distinguish between normal lymphocytes and tumor cells. Infiltrating lymph nodes, it is also impossible to determine whether the residual disease is fibrous necrotic scar tissue or always viable tumor cells.
c)B超也是分期常用的手段，对于浅表淋巴结的发现及随诊十分重要，根据淋巴结结构有无破坏和血流情况可协助判断肿大淋巴结的良恶性。能够发现直径大于2cm的淋巴结，对于协助发现肝脾肿大淋巴结有重要意义。c) B-ultrasound is also a commonly used method for staging. It is very important for the discovery and follow-up of superficial lymph nodes. According to whether the lymph node structure is damaged or not and blood flow, it can help to judge the benign and malignant of enlarged lymph nodes. The ability to detect lymph nodes larger than 2 cm in diameter is of great significance for assisting in the detection of hepatosplenomegaly lymph nodes.
d)MRI检查是针对于中枢神经系统、骨髓和肌肉部位的首选检查手段；尤其是对于不宜行增强CT扫描，或者作为CT发现可以病变后的进一步检查。d) MRI examination is the preferred examination method for the central nervous system, bone marrow and muscle; especially for those who are not suitable for enhanced CT scan, or as a further examination after CT findings can be used.
e)正电子发射计算机体层现(PET/CT)随着影像技术的不断发展，18F-FDG PET/CT以其灵敏度高、特异性强、准确性好的优点越来越广泛用于恶性淋巴瘤的诊疗。目前首选PET/CT用于霍奇金淋巴患者治疗前分期、化疗前后疗效评估。e) Positron emission computed tomography (PET/CT) With the continuous development of imaging technology, 18F-FDG PET/CT is more and more widely used in malignant lymphoma due to its high sensitivity, high specificity and good accuracy. tumor diagnosis and treatment. At present, PET/CT is the first choice for pre-treatment staging of patients with Hodgkin's lymphoma and evaluation of efficacy before and after chemotherapy.
3、病理检查3. Pathological examination
a)组织活检是最可靠的诊断依据，切取病变部位的淋巴结进行病理检查，找到典型镜影细胞(R-S细胞)诊断的关键，并以此作为亚型的依据。a) Tissue biopsy is the most reliable basis for diagnosis. The lymph nodes of the diseased site are excised for pathological examination, and the key to the diagnosis of typical mirror cells (R-S cells) is found, and this is used as the basis for subtypes.
b)细胞学检查对确定霍奇金淋巴亚型有一定的困难其主要是辅助分期，治疗后随访有无复发。b) Cytological examination has certain difficulties in determining Hodgkin's lymphoid subtypes, which are mainly auxiliary staging, and follow-up after treatment for recurrence.
c)骨髓穿刺可以判断骨髓的状态和功能。当肿瘤侵犯骨髓时，在骨髓组织中可以找到典型的RS细胞。c) Bone marrow aspirate can judge the state and function of the bone marrow. When the tumor invades the bone marrow, typical RS cells can be found in the bone marrow tissue.
d)其他检查免疫组织化学、染色体分析和原位杂交等新技术对判断鉴别霍奇金淋巴瘤和非霍奇金淋巴瘤，判断恶性淋巴瘤和反应性增殖、恶性淋巴瘤和白血病等起到重要辅助作用。如果霍奇金淋巴瘤存在细胞免疫缺陷，CD15和CD30抗原识别RS细胞的重要免疫标志，抗原阳性提示肿瘤的可能。d) Other new technologies such as immunohistochemistry, chromosomal analysis and in situ hybridization play a role in judging and distinguishing Hodgkin's lymphoma and non-Hodgkin's lymphoma, and judging malignant lymphoma and reactive proliferation, malignant lymphoma and leukemia, etc. important auxiliary role. If there is cellular immune deficiency in Hodgkin lymphoma, CD15 and CD30 antigens recognize important immune markers of RS cells, and the positive antigens indicate the possibility of tumors.
综上所述，血液检测灵敏度不够，往往不能发现早期患者；而骨髓穿刺和活检，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。血液和骨髓穿刺往往需结合在一起，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，且可以追踪霍奇金淋巴瘤发生、发展状况的筛查、检验方法。To sum up, the sensitivity of blood testing is not enough, and early patients often cannot be detected; while bone marrow puncture and biopsy require puncture biopsy, sampling is difficult, cannot be used as a real-time monitoring method, and will also cause damage to the patient's body. Blood and bone marrow aspirate are often combined together, and the disease cannot be judged by one test, which also increases the burden on patients. Imaging detection also has its own advantages and disadvantages. Its detection requires the tumor to reach a certain size before it can be detected. There is a hysteresis, and it cannot be used as a means of real-time monitoring due to radiation and other reasons. Therefore, there is an urgent need for a more convenient and sensitive screening and testing method that can track the occurrence and development of Hodgkin lymphoma.
发明内容SUMMARY OF THE INVENTION
针对现有技术中的上述不足，本发明提供一种经典霍奇金淋巴瘤的外周血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经典霍奇金淋巴瘤风险患者。In view of the above deficiencies in the prior art, the present invention provides a peripheral blood TCR marker of classic Hodgkin's lymphoma and a detection kit and application thereof, which can accurately and quickly determine whether there is a higher classic Hodgkin in the sample to be tested. Gold lymphoma risk patients.
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：For realizing the above-mentioned purpose, the technical scheme that the present invention solves its technical problem adopts is:
一种经典霍奇金淋巴瘤的外周血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。A peripheral blood TCR marker of classic Hodgkin's lymphoma, the marker includes at least one of the proteins whose sequences are shown in SEQ ID NO. 1 to 100, and the specific sequence is shown in Table 1.
表1 标志物序列Table 1 Marker sequences
序列编号serial number 蛋白序列protein sequence
SEQ ID NO.1SEQ ID NO.1 Ala Ser Ser Leu Asn Arg Val Ser Ser Glu Gln PheAla Ser Ser Leu Asn Arg Val Ser Ser Ser Glu Gln Phe
SEQ ID NO.2SEQ ID NO.2 Ala Ser Ser Phe Gly Lys Gly Pro Asn Tyr Gly Tyr ThrAla Ser Ser Phe Gly Lys Gly Pro Asn Tyr Gly Tyr Thr
SEQ ID NO.3SEQ ID NO.3 Ala Ser Ser Ile Ala Gly Ile Ala Lys Pro Tyr Glu Gln TyrAla Ser Ser Ile Ala Gly Ile Ala Lys Pro Tyr Glu Gln Tyr
SEQ ID NO.4SEQ ID NO.4 Ala Ser Ser Phe Leu Leu Arg Glu Gly Asn Asn Glu Gln PheAla Ser Ser Phe Leu Leu Arg Glu Gly Asn Asn Glu Gln Phe
SEQ ID NO.5SEQ ID NO.5 Ala Ser Ser Leu Asn Thr Gly Thr Asn Gly His Thr Ile TyrAla Ser Ser Leu Asn Thr Gly Thr Asn Gly His Thr Ile Tyr
SEQ ID NO.6SEQ ID NO.6 Ala Ser Ser Glu Ile Gln Gly Met Tyr Thr Glu Ala PheAla Ser Ser Glu Ile Gln Gly Met Tyr Thr Glu Ala Phe
SEQ ID NO.7SEQ ID NO.7 Ala Ser Ser Phe Met Asp Val Ser Pro Asn Thr Gly Glu Leu PheAla Ser Ser Phe Met Asp Val Ser Pro Asn Thr Gly Glu Leu Phe
SEQ ID NO.8SEQ ID NO.8 Ala Ser Thr Pro Val Arg Leu Ala Ala Thr Ser Thr Asp Thr Gln TyrAla Ser Thr Pro Val Arg Leu Ala Ala Thr Ser Thr Asp Thr Gln Tyr
SEQ ID NO.9SEQ ID NO.9 Ala Ser Ile Arg Glu Leu Ala Ser Ser Tyr Glu Gln TyrAla Ser Ile Arg Glu Leu Ala Ser Ser Ser Tyr Glu Gln Tyr
SEQ ID NO.10SEQ ID NO.10 Ala Ser Ser Thr Arg Leu Ala Gly Ala Asn Glu Gln TyrAla Ser Ser Thr Arg Leu Ala Gly Ala Asn Glu Gln Tyr
SEQ ID NO.11SEQ ID NO.11 Ala Ser Glu Glu Phe Tyr Glu Gln TyrAla Ser Glu Glu Phe Tyr Glu Gln Tyr
SEQ ID NO.12SEQ ID NO.12 Ala Ser Ser Phe Ala Pro Gly Val Pro Asp Gly Tyr ThrAla Ser Ser Phe Ala Pro Gly Val Pro Asp Gly Tyr Thr
SEQ ID NO.13SEQ ID NO.13 Ala Ser Ser Lys Ser Val Lys Gly Asn Glu Gln PheAla Ser Ser Lys Ser Val Lys Gly Asn Glu Gln Phe
SEQ ID NO.14SEQ ID NO.14 Ala Ser Ser Leu Lys Arg Leu Ala Arg Tyr Asn Glu Gln PheAla Ser Ser Leu Lys Arg Leu Ala Arg Tyr Asn Glu Gln Phe
SEQ ID NO.15SEQ ID NO.15 Ala Gly Ser Leu Pro Gly Ser Gly Tyr Asn Glu Gln PheAla Gly Ser Leu Pro Gly Ser Gly Tyr Asn Glu Gln Phe
SEQ ID NO.16SEQ ID NO.16 Ala Ser Ser Pro Arg Thr Met Glu Ile Asp Thr Gln TyrAla Ser Ser Pro Arg Thr Met Glu Ile Asp Thr Gln Tyr
SEQ ID NO.17SEQ ID NO.17 Ala Ser Ser Ser Lys Thr Ser Val Tyr Asn Glu Gln PheAla Ser Ser Ser Ser Lys Thr Ser Val Tyr Asn Glu Gln Phe
SEQ ID NO.18SEQ ID NO.18 Ala Trp Thr Arg Gly Trp Trp Thr Glu Ala PheAla Trp Thr Arg Gly Trp Trp Thr Glu Ala Phe
SEQ ID NO.19SEQ ID NO.19 Ala Ser Ser Ser Thr Arg Thr Ser Gly Val Thr Asp Thr Gln TyrAla Ser Ser Ser Ser Thr Arg Thr Ser Gly Val Thr Asp Thr Gln Tyr
SEQ ID NO.20SEQ ID NO.20 Ala Ile Glu Thr Glu Asn Asn Glu Gln PheAla Ile Glu Thr Glu Asn Asn Glu Gln Phe
SEQ ID NO.21SEQ ID NO.21 Ala Ser Ser Glu Phe Lys Arg Ile Gly Val Ser His Asn Glu Gln PheAla Ser Ser Glu Phe Lys Arg Ile Gly Val Ser His Asn Glu Gln Phe
SEQ ID NO.22SEQ ID NO.22 Ala Trp Ser Thr Trp Val Ile Ser Ser Glu Glu Gln TyrAla Trp Ser Thr Trp Val Ile Ser Ser Ser Glu Glu Gln Tyr
SEQ ID NO.23SEQ ID NO.23 Ala Ser Ser Ser Gly Ala Ala Leu Gly Asn Glu Gln PheAla Ser Ser Ser Gly Ala Ala Leu Gly Asn Glu Gln Phe
SEQ ID NO.24SEQ ID NO.24 Ala Thr Arg Pro Asp Arg Gly Met Thr Pro Leu HisAla Thr Arg Pro Asp Arg Gly Met Thr Pro Leu His
SEQ ID NO.25SEQ ID NO.25 Ala Thr Pro Lys Arg Ala Gly Asp Tyr Glu Gln TyrAla Thr Pro Lys Arg Ala Gly Asp Tyr Glu Gln Tyr
SEQ ID NO.26SEQ ID NO.26 Ala Ser Pro Ser Gln Gly Asp Tyr Gly Tyr ThrAla Ser Pro Ser Gln Gly Asp Tyr Gly Tyr Thr
SEQ ID NO.27SEQ ID NO.27 Ser Ala Arg Ser Ser Gly Ile Trp Asp Thr Gln TyrSer Ala Arg Ser Ser Gly Ile Trp Asp Thr Gln Tyr
SEQ ID NO.28SEQ ID NO.28 Ala Ile Ser Val Pro Gly Gly Glu Thr Gln TyrAla Ile Ser Val Pro Gly Gly Glu Thr Gln Tyr
SEQ ID NO.29SEQ ID NO.29 Ala Ser Ser Ser Ser Gly Arg Gly Thr Arg TyrAla Ser Ser Ser Ser Ser Gly Arg Gly Thr Arg Tyr
SEQ ID NO.30SEQ ID NO.30 Ala Ser Ser Pro Pro Gly Ala Gln Gly Asn Glu Gln PheAla Ser Ser Pro Pro Gly Ala Gln Gly Asn Glu Gln Phe
SEQ ID NO.31SEQ ID NO.31 Ala Ser Thr Tyr Gly Ala Leu Asn Gln Pro Gln HisAla Ser Thr Tyr Gly Ala Leu Asn Gln Pro Gln His
SEQ ID NO.32SEQ ID NO.32 Ala Ser Ser Ala Asp Lys Gly Ser Gly Asn Thr Ile TyrAla Ser Ser Ala Asp Lys Gly Ser Gly Asn Thr Ile Tyr
SEQ ID NO.33SEQ ID NO.33 Ala Ser Ser Leu Leu Ala Asp Phe Tyr Glu Gln TyrAla Ser Ser Leu Leu Ala Asp Phe Tyr Glu Gln Tyr
SEQ ID NO.34SEQ ID NO.34 Ala Ser Asn Phe Gly Gln Ser Gly Met Asn Thr Glu Ala PheAla Ser Asn Phe Gly Gln Ser Gly Met Asn Thr Glu Ala Phe
SEQ ID NO.35SEQ ID NO.35 Ala Ser Ser Leu Leu Leu Glu Ala Pro Ala PheAla Ser Ser Leu Leu Leu Leu Glu Ala Pro Ala Phe
SEQ ID NO.36SEQ ID NO.36 Ala Ser Ser Leu Val Arg Trp Ala Arg Gly Gln Val His Ser Pro Leu HisAla Ser Ser Leu Val Arg Trp Ala Arg Gly Gln Val His Ser Pro Leu His
SEQ ID NO.37SEQ ID NO.37 Ala Ser Ser Pro Ala Asn Arg Asp Leu Tyr Gly Tyr ThrAla Ser Ser Pro Ala Asn Arg Asp Leu Tyr Gly Tyr Thr
SEQ ID NO.38SEQ ID NO.38 Ala Ser Ser Ala Glu Glu Thr Gly Gly Ser Glu Gln PheAla Ser Ser Ala Glu Glu Thr Gly Gly Ser Glu Gln Phe
SEQ ID NO.39SEQ ID NO.39 Ala Thr Ser Val Phe Gln Ser Gly Glu Leu PheAla Thr Ser Val Phe Gln Ser Gly Glu Leu Phe
SEQ ID NO.40SEQ ID NO.40 Ala Thr Asp Pro Gly Gly Gly Asn Gln Pro Gln HisAla Thr Asp Pro Gly Gly Gly Asn Gln Pro Gln His
SEQ ID NO.41SEQ ID NO.41 Ala Ser Gln Ser Gly Gly Asn Thr Asp Thr Gln TyrAla Ser Gln Ser Gly Gly Asn Thr Asp Thr Gln Tyr
SEQ ID NO.42SEQ ID NO.42 Ala Thr Ser Lys Asp Ser Gly Gly Gly Asn Glu Gln PheAla Thr Ser Lys Asp Ser Gly Gly Gly Asn Glu Gln Phe
SEQ ID NO.43SEQ ID NO.43 Ala Ser Ser Glu Asp Arg Gly Ser Ala Ala Lys Asn Ile Gln TyrAla Ser Ser Glu Asp Arg Gly Ser Ala Ala Lys Asn Ile Gln Tyr
SEQ ID NO.44SEQ ID NO.44 Ala Ser Ser Ile Gly Gln Gly Gly Leu Leu Thr Gln TyrAla Ser Ser Ile Gly Gln Gly Gly Leu Leu Thr Gln Tyr
SEQ ID NO.45SEQ ID NO.45 Ala Ser Ala Trp Gly Phe Gly Asp Thr Glu Ala PheAla Ser Ala Trp Gly Phe Gly Asp Thr Glu Ala Phe
SEQ ID NO.46SEQ ID NO.46 Ala Thr Ser Met Gly Gln Ile Asn Thr Glu Ala PheAla Thr Ser Met Gly Gln Ile Asn Thr Glu Ala Phe
SEQ ID NO.47SEQ ID NO.47 Ala Thr Arg Tyr Gly Asn Asn Val Leu ThrAla Thr Arg Tyr Gly Asn Asn Val Leu Thr
SEQ ID NO.48SEQ ID NO.48 Ala Asn Val Phe Gly Val Pro Asn Glu Gln PheAla Asn Val Phe Gly Val Pro Asn Glu Gln Phe
SEQ ID NO.49SEQ ID NO.49 Ala Thr Ser Arg Asn Thr Gly Ser Glu Thr Gln TyrAla Thr Ser Arg Asn Thr Gly Ser Glu Thr Gln Tyr
SEQ ID NO.50SEQ ID NO.50 Ala Ser Ser Leu Asn Ala Arg Gly Gln Thr Asp Gln Pro Gln HisAla Ser Ser Leu Asn Ala Arg Gly Gln Thr Asp Gln Pro Gln His
SEQ ID NO.51SEQ ID NO.51 Ser Val Glu Ala Ala Gly Pro Gln Asp Tyr Tyr Glu Gln TyrSer Val Glu Ala Ala Gly Pro Gln Asp Tyr Tyr Glu Gln Tyr
SEQ ID NO. 52SEQ ID NO. 52 Ala Ser Ser Arg Glu Gly Ser Gly Ser Asn Gly Glu Leu PheAla Ser Ser Arg Glu Gly Ser Gly Ser Asn Gly Glu Leu Phe
SEQ ID NO. 53SEQ ID NO. 53 Ala Ser Arg Pro Ala Asp Trp Ser Pro Ser Thr Asp Thr Gln TyrAla Ser Arg Pro Ala Asp Trp Ser Pro Ser Thr Asp Thr Gln Tyr
SEQ ID NO. 54SEQ ID NO. 54 Ala Ser Ser Val Gly Arg Thr Ser Gly Leu Gly Arg Glu Thr Gln TyrAla Ser Ser Val Gly Arg Thr Ser Gly Leu Gly Arg Glu Thr Gln Tyr
SEQ ID NO. 55SEQ ID NO. 55 Ala Thr Ser Arg Asp Ser Arg Arg Thr Gly Ser Ser Tyr Glu Gln TyrAla Thr Ser Arg Asp Ser Arg Arg Thr Gly Ser Ser Tyr Glu Gln Tyr
SEQ ID NO. 56SEQ ID NO. 56 Ala Ser Ser Tyr Glu Thr Val Ser Val Ala Asn Thr Gly Glu Leu PheAla Ser Ser Tyr Glu Thr Val Ser Val Ala Asn Thr Gly Glu Leu Phe
SEQ ID NO. 57SEQ ID NO. 57 Ala Ser Arg Thr Phe Ser Arg Asn Glu Gln PheAla Ser Arg Thr Phe Ser Arg Asn Glu Gln Phe
SEQ ID NO. 58SEQ ID NO. 58 Ala Ser Ser Trp Pro Ala Gln Gly Tyr Glu Gln TyrAla Ser Ser Trp Pro Ala Gln Gly Tyr Glu Gln Tyr
SEQ ID NO. 59SEQ ID NO. 59 Ala Ser Ala Arg Ser Leu Ser Thr Glu Ala PheAla Ser Ala Arg Ser Leu Ser Thr Glu Ala Phe
SEQ ID NO. 60SEQ ID NO. 60 Ala Ser Thr Gly Leu Ala Gly Gly Arg Ala Pro Tyr Glu Gln TyrAla Ser Thr Gly Leu Ala Gly Gly Arg Ala Pro Tyr Glu Gln Tyr
SEQ ID NO. 61SEQ ID NO. 61 Ala Ser Thr Val Thr Gly Glu Arg Arg Gln PheAla Ser Thr Val Thr Gly Glu Arg Arg Gln Phe
SEQ ID NO. 62SEQ ID NO. 62 Ala Thr Ser Arg Asp Leu Ala Gly Arg Gly Glu Asn Tyr Gly Tyr ThrAla Thr Ser Arg Asp Leu Ala Gly Arg Gly Glu Asn Tyr Gly Tyr Thr
SEQ ID NO. 63SEQ ID NO. 63 Pro Ala Val Thr Arg Asp Arg Glu Asp Thr Ser Ser ThrPro Ala Val Thr Arg Asp Arg Glu Asp Thr Ser Ser Ser Thr
SEQ ID NO. 64SEQ ID NO. 64 Ala Ser Thr Pro Ile Gly Val Glu Glu Glu Thr Gln TyrAla Ser Thr Pro Ile Gly Val Glu Glu Glu Thr Gln Tyr
SEQ ID NO. 65SEQ ID NO. 65 Ser Gly Leu Pro Tyr Ser Ala Asn Glu Gln PheSer Gly Leu Pro Tyr Ser Ala Asn Glu Gln Phe
SEQ ID NO. 66SEQ ID NO. 66 Ala Ser Ser Met Ser Gly Phe Thr Tyr Asn Glu Gln PheAla Ser Ser Met Ser Gly Phe Thr Tyr Asn Glu Gln Phe
SEQ ID NO. 67SEQ ID NO. 67 Ala Ser Ser Leu Thr Ala Gly Gly Cys Glu Gln PheAla Ser Ser Leu Thr Ala Gly Gly Cys Glu Gln Phe
SEQ ID NO. 68SEQ ID NO. 68 Ala Ser Ser Ile Asp Arg Asp Ser Tyr Gln Pro Gln HisAla Ser Ser Ile Asp Arg Asp Ser Tyr Gln Pro Gln His
SEQ ID NO. 69SEQ ID NO. 69 Ala Trp Ser Val Val Pro Glu Ile Thr Glu Ala PheAla Trp Ser Val Val Pro Glu Ile Thr Glu Ala Phe
SEQ ID NO. 70SEQ ID NO. 70 Ala Ser Ser Pro Leu Thr Tyr Arg Asp Thr Gln TyrAla Ser Ser Pro Leu Thr Tyr Arg Asp Thr Gln Tyr
SEQ ID NO. 71SEQ ID NO. 71 Ala Ser Ser Pro Pro Arg Thr Gly Gln Gly Tyr Tyr Glu Gln TyrAla Ser Ser Pro Pro Arg Thr Gly Gln Gly Tyr Tyr Glu Gln Tyr
SEQ ID NO. 72SEQ ID NO. 72 Ala Thr Ser Tyr Asp Gln Glu Thr Gln TyrAla Thr Ser Tyr Asp Gln Glu Thr Gln Tyr
SEQ ID NO. 73SEQ ID NO. 73 Ala Ile Ser Gly Ser Asp Pro Glu Asn Glu Gln PheAla Ile Ser Gly Ser Asp Pro Glu Asn Glu Gln Phe
SEQ ID NO. 74SEQ ID NO. 74 Ser Ala Lys Asn Thr Tyr Tyr Glu Gln TyrSer Ala Lys Asn Thr Tyr Tyr Glu Gln Tyr
SEQ ID NO. 75SEQ ID NO. 75 Ala Ser Ser Thr Thr Thr Gly Thr Thr Asn Glu Lys Leu PheAla Ser Ser Thr Thr Thr Gly Thr Thr Asn Glu Lys Leu Phe
SEQ ID NO. 76SEQ ID NO. 76 Ala Ser Asn Gln Gly Phe Gly Gly Ser Pro Leu HisAla Ser Asn Gln Gly Phe Gly Gly Ser Pro Leu His
SEQ ID NO. 77SEQ ID NO. 77 Ala Ser Ser Val Ala Leu Ser Val Thr Asp Thr Gln TyrAla Ser Ser Val Ala Leu Ser Val Thr Asp Thr Gln Tyr
SEQ ID NO. 78SEQ ID NO. 78 Ala Ser Ser Pro Ile Val Asn Arg Val Glu Thr Gln TyrAla Ser Ser Pro Ile Val Asn Arg Val Glu Thr Gln Tyr
SEQ ID NO. 79SEQ ID NO. 79 Ala Ser Ser Asp Arg Met Val Ala Gly Asp Tyr Glu Gln TyrAla Ser Ser Asp Arg Met Val Ala Gly Asp Tyr Glu Gln Tyr
SEQ ID NO. 80SEQ ID NO. 80 Ala Ser Lys Arg Trp Asn Gln Pro Gln HisAla Ser Lys Arg Trp Asn Gln Pro Gln His
SEQ ID NO. 81SEQ ID NO. 81 Ala Ser Ser Arg Val Val Ser Glu Gln PheAla Ser Ser Arg Val Val Val Ser Glu Gln Phe
SEQ ID NO. 82SEQ ID NO. 82 Ala Ser Met Gly Gly Gly Arg Arg Thr Asp Thr Gln TyrAla Ser Met Gly Gly Gly Arg Arg Thr Asp Thr Gln Tyr
SEQ ID NO. 83SEQ ID NO. 83 Ala Ser Ser Gly Thr Gly Gly Thr Arg Asn Gln Pro Gln HisAla Ser Ser Gly Thr Gly Gly Thr Arg Asn Gln Pro Gln His
SEQ ID NO. 84SEQ ID NO. 84 Ala Ser Ser Ser Arg Met Gly Gln Gly Arg Asn Asn Tyr Gly Tyr ThrAla Ser Ser Ser Arg Met Gly Gln Gly Arg Asn Asn Tyr Gly Tyr Thr
SEQ ID NO. 85SEQ ID NO. 85 Ala Ser Ser Ala Ala Gly Glu Gly Asn Tyr Gly Tyr ThrAla Ser Ser Ala Ala Gly Glu Gly Asn Tyr Gly Tyr Thr
SEQ ID NO. 86SEQ ID NO. 86 Ala Ser Thr Asn Gly Val Asn Trp Asn Thr Glu Ala PheAla Ser Thr Asn Gly Val Asn Trp Asn Thr Glu Ala Phe
SEQ ID NO. 87SEQ ID NO. 87 Ala Arg Gly Val Glu Asn Thr Asp Thr Gln TyrAla Arg Gly Val Glu Asn Thr Asp Thr Gln Tyr
SEQ ID NO. 88SEQ ID NO. 88 Ala Ser Thr Thr Pro Arg Gly Thr Gly Glu Leu PheAla Ser Thr Thr Pro Arg Gly Thr Gly Glu Leu Phe
SEQ ID NO. 89SEQ ID NO. 89 Ala Ser Ser Pro Gln Val Gln Thr Arg Tyr Gly Thr Glu Ala PheAla Ser Ser Pro Gln Val Gln Thr Arg Tyr Gly Thr Glu Ala Phe
SEQ ID NO. 90SEQ ID NO. 90 Ala Ser Ser Val Gly Gln Ser Ile Gln TyrAla Ser Ser Val Gly Gln Ser Ile Gln Tyr
SEQ ID NO. 91SEQ ID NO. 91 Ala Ser Ser Leu His Arg Met Asn Glu Ala PheAla Ser Ser Leu His Arg Met Asn Glu Ala Phe
SEQ ID NO. 92SEQ ID NO. 92 Ala Ser Arg Gly Gly Arg Val Met Glu Gly Tyr ThrAla Ser Arg Gly Gly Arg Val Met Glu Gly Tyr Thr
SEQ ID NO. 93SEQ ID NO. 93 Ala Ser Ser Ser Gly Tyr Arg Glu Gly His Glu Gln TyrAla Ser Ser Ser Gly Tyr Arg Glu Gly His Glu Gln Tyr
SEQ ID NO. 94SEQ ID NO. 94 Ala Ser Ser Ser Leu Arg Asp Val Asn Ser Asn Gln Pro Gln HisAla Ser Ser Ser Leu Arg Asp Val Asn Ser Asn Gln Pro Gln His
SEQ ID NO. 95SEQ ID NO. 95 Ala Thr Ser Arg Glu Gly Gly Ile Glu Thr Gln TyrAla Thr Ser Arg Glu Gly Gly Ile Glu Thr Gln Tyr
SEQ ID NO. 96SEQ ID NO. 96 Ala Ser Ser Arg Met Thr Gly Gly Pro Tyr Glu Gln TyrAla Ser Ser Arg Met Thr Gly Gly Pro Tyr Glu Gln Tyr
SEQ ID NO. 97SEQ ID NO. 97 Ala Ser Ser Pro Ser His Val Val Ser Ser Pro Leu HisAla Ser Ser Pro Ser His Val Val Val Ser Ser Ser Pro Leu His
SEQ ID NO. 98SEQ ID NO. 98 Ala Ser Ser Ile Glu Gly Thr Ser Gly Thr Glu Thr Gln TyrAla Ser Ser Ile Glu Gly Thr Ser Gly Thr Glu Thr Gln Tyr
SEQ ID NO. 99SEQ ID NO. 99 Ala Ser Ser Pro Arg Gly Gly Ala Arg Arg Glu Thr Gln TyrAla Ser Ser Pro Arg Gly Gly Ala Arg Arg Glu Thr Gln Tyr
SEQ ID NO. 100SEQ ID NO. 100 Ala Ser Ser Tyr Trp Ala Gly Lys Leu Tyr Gly Tyr ThrAla Ser Ser Tyr Trp Ala Gly Lys Leu Tyr Gly Tyr Thr
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。Further, the protein sequence of the marker is that the sequence shown in SEQ ID NO. 1-100 can express a protein with the same function after one or more bases are substituted, deleted and/or replaced.
进一步地，标志物为外周血TCR CDR3序列。Further, the marker is the peripheral blood TCR CDR3 sequence.
上述标志物在制备治疗经典霍奇金淋巴瘤的制剂中的应用。The application of the above markers in the preparation of preparations for the treatment of classic Hodgkin's lymphoma.
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。Further, the preparation includes a T cell receptor containing the marker, or a plasmid, viral vector or nucleic acid fragment capable of expressing the T cell receptor producing the marker.
一种用于经典霍奇金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。A kit for the detection of classical Hodgkin's lymphoma, comprising antibodies that can specifically bind to the above markers.
一种制剂，包括能与上述标志物发生特异性结合的抗体；所述制剂可用于对经典霍奇金淋巴瘤进行诊断、预测、检测或筛查。A preparation comprising an antibody capable of specifically binding to the above-mentioned marker; the preparation can be used for diagnosis, prediction, detection or screening of classical Hodgkin lymphoma.
一种检测经典霍奇金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。A protein chip for detecting classical Hodgkin's lymphoma, the protein chip comprises a substrate and a specific antibody spotted on the substrate, the specific antibody being an antibody that can specifically bind to the above marker.
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10 18，TCR多样性的理论值是10 14。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。 The principle of the present invention is as follows: B lymphocytes and T lymphocytes in the human body are two important types of cells in the acquired immune system. B cells recognize antigens through the B cell receptor (BCR) on the cell surface. Later, when B cells differentiate into plasma cells, BCR is expressed as an antibody and secreted outside the cell. T cells recognize antigens through T cell receptors (TCRs) on the cell surface. The diversity of BCR and TCR is the basis for the establishment of the adaptive immune system. The theoretical value of BCR diversity is 10 18 and the theoretical value of TCR diversity is 10 14 . Among BCR and TCR sequences, antigenic determinant 3 (CDR3) is the most important part in determining its antigenic specificity, so the sequence of CDR3 is considered to represent the properties of BCR and TCR sequences.
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。In various diseases, the diversity or expression levels of BCR and TCR will change with different antigenic stimulation. Therefore, the occurrence and development of diseases can be tracked by using BCR or TCR high-throughput sequencing results. In human cells, after degradation of senescent proteins, their fragments are transported to the cell surface and presented to T cells in the immune system through histocompatibility antigen II (MCHII). Antigen fragments presented by normal cells do not cause an immune response due to immune tolerance. Once normal cells become cancerous, the abnormal protein expressed by the mutated gene and its fragments are presented on the cell surface, which will cause the human immune system to produce a targeted immune response. Therefore, analyzing the changes of BCR or TCR can detect the occurrence and development of tumors.
本发明的有益效果为：The beneficial effects of the present invention are:
1、本发明中，首先利用1300个非经典霍奇金淋巴瘤的对照组样本、和70个经典霍奇金淋巴瘤病人的TCR高通量测序数据建立了人工智能分析模型，通过和这些经典霍奇金淋巴瘤特异性TCR序列对比，可以清楚的判断待测样本中是否有较高经典霍奇金淋巴瘤风险者。1. In the present invention, an artificial intelligence analysis model was first established by using 1300 non-classical Hodgkin lymphoma control samples and 70 classical Hodgkin lymphoma patients' TCR high-throughput sequencing data. Hodgkin's lymphoma-specific TCR sequence comparison can clearly determine whether there is a higher risk of classic Hodgkin's lymphoma in the sample to be tested.
2、通过高通量测序分析TCR变化可以发现很早期的经典霍奇金淋巴瘤，利用经典霍奇金淋巴瘤特有的TCR CDR3序列分析人的免疫系统中的T细胞对经典霍奇金淋巴瘤的反应，是一种新型的检测方法。2. Analysis of TCR changes by high-throughput sequencing can find very early classical Hodgkin lymphoma, and use the TCR CDR3 sequence specific to classical Hodgkin lymphoma to analyze the effect of T cells in the human immune system on classical Hodgkin lymphoma. The reaction is a new detection method.
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。3. By using high-throughput sequencing technology, the present invention simultaneously compares a huge number of specific TCR sequences, and has higher specificity and accuracy than detecting one or several markers alone.
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。4. The cost of the high-throughput sequencing instrument used in the present invention is lower than that of large-scale imaging equipment, and it can be outsourced to a third party. In addition, the labor cost of sampling and processing is lower than the simultaneous detection of multiple markers, and it is also lower than that of a large number of cytology detection, therefore, the present invention greatly reduces the detection cost.
5、本发明只需要采取少量外周血，采样简便、安全。5. The present invention only needs to take a small amount of peripheral blood, and the sampling is simple and safe.
6、本发明中所述TCR CDR3序列，可用于经典霍奇金淋巴瘤的免疫治疗。6. The TCR CDR3 sequence described in the present invention can be used for immunotherapy of classical Hodgkin's lymphoma.
附图说明Description of drawings
图1为本发明中利用基于免疫大数据分析系统发现的经典霍奇金淋巴瘤特征性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经典霍奇金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C X的对数值；经典霍奇金淋巴瘤患者的免疫图谱具有多个种类且重复次数较高的经典霍奇金淋巴瘤特征序列，健康人极少经典霍奇金淋巴瘤特征序列，而未知受试者的经典霍奇金淋巴瘤特征比较明显，说明罹患经典霍奇金淋巴瘤风险较高。 Fig. 1 is the characteristic TCR sequence of classical Hodgkin's lymphoma discovered by the immune-based big data analysis system in the present invention; wherein, the abscissa represents that the CDR3 sequence of a certain amino acid combination is added to the control sequence set or the classical Hodgkin's lymphoma The order of the tumor characteristic sequence set, the ordinate represents the logarithm of the number of repeated occurrences of the sequence in a certain sample, C X ; the immune map of patients with classic Hodgkin lymphoma has multiple types of classic Hodgkin with a high number of repetitions. Golden lymphoma characteristic sequences, healthy people rarely have classical Hodgkin lymphoma characteristic sequences, while unknown subjects have more obvious classical Hodgkin lymphoma characteristics, indicating a higher risk of classic Hodgkin lymphoma.
图2为本发明中利用经典霍奇金淋巴瘤特征特征性指数对比分析经典霍奇金淋巴瘤和其他疾病；健康人、非肿瘤病人、非经典霍奇金淋巴瘤肿瘤患者的经典霍奇金淋巴瘤特征性指数均与经典霍奇金淋巴瘤患者具有显著差异，证明了经典霍奇金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经典霍奇金淋巴瘤。Fig. 2 is a comparative analysis of classical Hodgkin's lymphoma and other diseases using the characteristic index of classical Hodgkin's lymphoma in the present invention; The lymphoma characteristic indices were significantly different from those of patients with classical Hodgkin lymphoma, proving the specificity of the characteristic sequence set of classical Hodgkin lymphoma, which can be used to determine whether an unknown subject suffers from classical Hodgkin lymphoma.
具体实施方式Detailed ways
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。The specific embodiments of the present invention are described below to facilitate those skilled in the art to understand the present invention, but it should be clear that the present invention is not limited to the scope of the specific embodiments. For those of ordinary skill in the art, as long as various changes Such changes are obvious within the spirit and scope of the present invention as defined and determined by the appended claims, and all inventions and creations utilizing the inventive concept are within the scope of protection.
实施例1 通过免疫图谱分析，获得经典霍奇金淋巴瘤TCR标志物CDR3序列集Example 1 Obtaining the classic Hodgkin's lymphoma TCR marker CDR3 sequence set by immunographic analysis
1、采样及免疫图谱分析(方法参考申请号为201910300069.9的专利)1. Sampling and immunochromatographic analysis (method reference patent application number 201910300069.9)
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)、71名经典霍奇金淋巴瘤患者(70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；Collected 1301 control groups (including healthy people and non-tumor disease patients, 1300 people were used for model establishment, 1 healthy person was used for validation), 71 classic Hodgkin lymphoma patients (70 people were used for model establishment, 1 person was used for validation) For verification) and peripheral blood of 1 subject with unknown health status (10mL per person), the antigenic determinant 3 (CDR3) amino acid sequences of TCR of the subjects and the control group were obtained by high-throughput sequencing to ensure that each The total number of CDR3 sequences of functional TCRs in the sample should not be less than 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C X； 2. Perform random non-replacement sampling on the sequences of the samples whose total number of CDR3 sequences of each functional TCR is higher than 30,000, so that the total number of CDR3 sequences of this sample is 30,000. The total number of CDR3 sequences of functional TCRs contained in all samples so far is 25,000-30,000. For any specific CDR3 sequence X, the number of repetitions in the single-sample sequencing result is counted as C X ;
3、通过分析TCR CDR3数据，确定经典霍奇金淋巴瘤TCR标志物CDR3序列：3. Determine the CDR3 sequence of classic Hodgkin lymphoma TCR marker by analyzing TCR CDR3 data:
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；a) All the CDR3 sequences of the 1300 control samples used to build the model were aggregated and deduplicated, and set as a control sequence set;
b)将70名用于建立模型的经典霍奇金淋巴瘤样本的所有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，设为经典霍奇金淋巴瘤特征序列集。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经典霍奇金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C X的对数值。 b) Summarize and deduplicate all CDR3 sequences of 70 classic Hodgkin lymphoma samples used to establish the model, and then remove all sequences containing sequence duplication with the control sequence set, and set it as the classic Hodgkin lymphoma characteristic sequence set . The drawing is shown in Figure 1A, in which the abscissa represents the sequence in which the CDR3 sequence of a specific amino acid combination is added to the control sequence set or the classic Hodgkin lymphoma characteristic sequence set, and the ordinate represents the sequence in a certain sample. The logarithm of the number of repetitions C X.
c)按照同样作图方法，将1名健康人、1名经典霍奇金淋巴瘤患者和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经典霍奇金淋巴瘤特征序列集合进行作图，见附图1B-D。从图中可见，经典霍奇金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经典霍奇金淋巴瘤特征序列(图1B)；健康人的免疫图谱中，只有极少量经典霍奇金淋巴瘤特征序列(图1C)；而未知健康状况受试者，有高于健康人的经典霍奇金淋巴瘤特征序列，说明此人有较高风险罹患经典霍奇金淋巴瘤(图1D)。c) According to the same mapping method, the immune profiles of 1 healthy person, 1 patient with classical Hodgkin lymphoma and 1 subject with unknown health status were compared with reference sequence set and classical Hodgkin lymphoma characteristic sequence set For mapping, see Figures 1B-D. It can be seen from the figure that the immune maps of patients with classical Hodgkin lymphoma contain more types and high repetitions of classical Hodgkin lymphoma characteristic sequences (Figure 1B); in the immune maps of healthy people, only extremely A small number of classic Hodgkin lymphoma signature sequences (Figure 1C); while subjects of unknown health status had higher classical Hodgkin lymphoma signature sequences than healthy individuals, indicating that this person was at higher risk for classic Hodgkin lymphoma tumor (Figure 1D).
d)将经典霍奇金淋巴瘤特征序列集中，将所有出现在两个及以上参与建模经典霍奇金淋巴瘤样本里的CDR3序列，按“所有参与建模经典霍奇金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经典霍奇金淋巴瘤样本数”从高到低排序，排名前100者即为经典霍奇金淋巴瘤TCR标志物CDR3序列，具体序列如SEQ ID NO.1～100所示。d) Collect the characteristic sequences of classical Hodgkin lymphoma, and put all the CDR3 sequences that appear in two or more samples of classical Hodgkin lymphoma participating in the modeling, and press the button of “All the samples participating in the modeling of classical Hodgkin lymphoma”. The sum of the number of repeated occurrences of CX in a single sample of this sequence × the number of samples of classical Hodgkin lymphoma that contains this sequence involved in modeling” is sorted from high to low, and the top 100 is the classic Hodgkin lymphoma TCR marker CDR3 Sequence, the specific sequence is shown in SEQ ID NO.1～100.
实施例2 验证经典霍奇金淋巴瘤TCR标志物CDR3序列集的特异性Example 2 Verification of the specificity of the classical Hodgkin lymphoma TCR marker CDR3 sequence set
1、采样及免疫图谱分析(方法参考申请号为201910300069.9的专利)1. Sampling and immunochromatographic analysis (method reference patent application number 201910300069.9)
采集522名非经典霍奇金淋巴瘤的肿瘤患者、5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。The peripheral blood (10 mL each) of 522 non-classical Hodgkin lymphoma tumor patients and 5 subjects with unknown health status was collected, and the TCR epitope 3 of the subjects and the control group was obtained by high-throughput sequencing (CDR3) amino acid sequence, ensure that the total number of CDR3 sequences of functional TCRs in each sample is not less than 25,000; the sequences of samples whose total number of CDR3 sequences of each functional TCR is more than 30,000 are randomly sampled without replacement, so that The total number of CDR3 sequences for this sample is 30,000. The total number of CDR3 sequences of functional TCRs contained in all samples so far is 25,000-30,000.
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。2. 100 healthy people and 43 non-tumor disease patients were randomly selected from the control group in Example 1.
3、根据来自实施例1的100名健康人、43名非肿瘤疾病病人、67名经典霍奇金淋巴瘤患者，以及实施例2新获取的522名非经典霍奇金淋巴瘤肿瘤患者、5名未知健康状况受试者的免疫图谱，分析其经典霍奇金淋巴瘤特征性指数。3. According to 100 healthy people, 43 non-tumor disease patients, 67 classic Hodgkin lymphoma patients from Example 1, and 522 non-classical Hodgkin lymphoma tumor patients newly obtained in Example 2, 5 Immune profiling of subjects with unknown health status and analysis of the classic Hodgkin lymphoma characteristic index.
其中经典霍奇金淋巴瘤特征性指数定义为：某个样本中，属于经典霍奇金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C X的总和。分析结果见下表2及附图2。经典霍奇金淋巴瘤组与健康人(p＝5.3E-76)、非肿瘤疾病(p＝7.7E-42)、其它肿瘤(p＝6.9E-295)都有显著差异，这证明了经典霍奇金淋巴瘤特征序列集的特异性。 The characteristic index of classical Hodgkin lymphoma is defined as: in a certain sample, the sum of the repeated occurrences C X of all CDR3 sequences belonging to the characteristic sequence set of classical Hodgkin lymphoma in the sample. The analysis results are shown in Table 2 below and accompanying drawing 2. The classic Hodgkin lymphoma group was significantly different from healthy people (p=5.3E-76), non-neoplastic disease (p=7.7E-42), and other tumors (p=6.9E-295), which proves the classic Specificity of the Hodgkin's lymphoma signature sequence set.
表2 不同样本组的经典霍奇金淋巴瘤特征性指数Table 2 Classical Hodgkin lymphoma characteristic indices in different sample groups
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Figure PCTCN2022080301-appb-000012
Figure PCTCN2022080301-appb-000012
4、分析各组的经典霍奇金淋巴瘤特征指数(表3)，5位未知健康状况受试者(检测样本)的经典霍奇金淋巴瘤特征指数显著高于“其它肿瘤”组的平均值+2×SD(134.0+2×476.2＝1086.4)，此5人具有较高风险罹患经典霍奇金淋巴瘤。与临床体检结果对照后，这5人确为早期经典霍奇金淋巴瘤患者。此实施例证明了利用经典霍奇金淋巴瘤特征序列集及经典霍奇金淋巴瘤特征性指数，预测受试者罹患经典霍奇金淋巴瘤风险的可行性。4. Analysis of the classic Hodgkin lymphoma characteristic index of each group (Table 3), the classic Hodgkin lymphoma characteristic index of 5 subjects with unknown health status (test samples) was significantly higher than the average of the "other tumors" group value+2×SD (134.0+2×476.2=1086.4), these 5 people have a higher risk of classic Hodgkin lymphoma. After comparing with the clinical physical examination results, these 5 people were indeed patients with early classic Hodgkin lymphoma. This example demonstrates the feasibility of using the classic Hodgkin lymphoma signature sequence set and the classic Hodgkin lymphoma signature index to predict the risk of a subject suffering from classic Hodgkin lymphoma.
表3 经典霍奇金淋巴瘤特征性指数分析Table 3 Characteristic index analysis of classical Hodgkin lymphoma
Figure PCTCN2022080301-appb-000013
Figure PCTCN2022080301-appb-000013
综上所述，本发明所述经典霍奇金淋巴瘤TCR标志物CDR3序列，确实具有显著的经典霍奇金淋巴瘤特异性，不仅可以用于经典霍奇金淋巴瘤预测受试者罹患经典霍奇金淋巴瘤风险，未来还可用于经典霍奇金淋巴瘤的生物免疫治疗。To sum up, the classical Hodgkin lymphoma TCR marker CDR3 sequence of the present invention does have significant classical Hodgkin lymphoma specificity, and can not only be used for classical Hodgkin lymphoma to predict subjects suffering from classical Hodgkin lymphoma. The risk of Hodgkin lymphoma can also be used for biological immunotherapy of classical Hodgkin lymphoma in the future.

Claims (7)

  1. 一种经典霍奇金淋巴瘤的外周血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。A peripheral blood TCR marker for classic Hodgkin's lymphoma, characterized in that the marker comprises at least one of the proteins whose sequences are shown in SEQ ID NO. 1-100.
  2. 根据权利要求1所述的经典霍奇金淋巴瘤的外周血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。The peripheral blood TCR marker for classic Hodgkin's lymphoma according to claim 1, wherein the protein sequence of the marker is the sequence shown in SEQ ID NO. 1-100 with substitution, deletion and/or After replacing one or more bases, the protein with the same function can be expressed.
  3. 权利要求1所述的标志物在制备治疗经典霍奇金淋巴瘤的制剂中的应用。The application of the marker of claim 1 in the preparation of a preparation for the treatment of classic Hodgkin's lymphoma.
  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。The use according to claim 3, wherein the preparation comprises a T cell receptor containing the marker, or a plasmid, viral vector or nucleic acid fragment capable of expressing the T cell receptor producing the marker.
  5. 一种用于经典霍奇金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。A kit for detection of classical Hodgkin's lymphoma, characterized by comprising an antibody that can specifically bind to the marker of claim 1.
  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；所述制剂可用于对经典霍奇金淋巴瘤进行诊断、预测、检测或筛查。A preparation is characterized by comprising an antibody that can specifically bind to the marker of claim 1; the preparation can be used for diagnosis, prediction, detection or screening of classical Hodgkin lymphoma.
  7. 一种检测经典霍奇金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。A protein chip for detecting classical Hodgkin's lymphoma, characterized in that the protein chip comprises a substrate and a specific antibody spotted on the substrate, and the specific antibody is capable of producing the marker described in claim 1. specific binding antibodies.
PCT/CN2022/080301 2021-03-15 2022-03-11 Peripheral blood tcr marker for classical hodgkin lymphoma, detection kit therefor, and application thereof WO2022194036A1 (en)

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