CN113563454A - IgA nephropathy peripheral blood TCR marker and detection kit and application thereof - Google Patents
IgA nephropathy peripheral blood TCR marker and detection kit and application thereof Download PDFInfo
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- CN113563454A CN113563454A CN202110834998.5A CN202110834998A CN113563454A CN 113563454 A CN113563454 A CN 113563454A CN 202110834998 A CN202110834998 A CN 202110834998A CN 113563454 A CN113563454 A CN 113563454A
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a TCR marker of peripheral blood of IgA nephropathy and a detection kit and application thereof. The marker comprises at least one of proteins with sequences shown as SEQ ID NO. 1-100. Based on a high-throughput sequencing method, only a small amount of peripheral blood is needed to be taken, RNA is extracted, an immune big data library is established through sample processing, and through high-throughput sequencing and TCR data analysis, a characteristic TCR sequence in IgA nephropathy peripheral blood is determined firstly, and then a test result of a sample to be tested is compared with the characteristic TCR sequence, so that whether IgA nephropathy exists or not is determined. The invention can simultaneously compare a great number of IgA nephropathy specific TCR sequences, has higher specificity and accuracy compared with the single detection of one or more markers, and improves the diagnosis efficiency.
Description
Technical Field
The invention belongs to the technical field of genetic engineering, and particularly relates to a T Cell Receptor (TCR) marker in peripheral blood of IgA nephropathy and a detection kit and application thereof.
Background
One) brief introduction to IgA nephropathy
IgA nephropathy, also known as Berger's disease, is the most common primary glomerular disease in clinical settings and is characterized by the presence of IgA antibodies or IgA-based immune complex deposition in the glomerular mesangial region. The incidence of IgA nephropathy has obvious regional difference, accounts for 15% -40% of primary glomerular diseases in Europe and Asia, and accounts for 26% -34% of primary glomerular diseases in China, and shows a gradually rising trend in recent years.
IgA nephropathy can occur in all ages, and male young and old with the age of 20-30 are the most common. IgA nephropathy is often chronic in course and the early symptoms of the patient are mild, but if not treated in time, statistically up to 50% of affected patients will progress slowly to different degrees of renal failure, and therefore need to be diagnosed as early as possible and treated in time.
II) etiology and pathogenesis
The etiology and pathogenesis of IgA nephropathy are studied, and a plurality of evidences show that the IgA nephropathy is a polygenic and multifactorial complex character disease. The etiology may arise from familial genetic or autoimmune dysregulation, but also may be caused by other diseases, particularly infectious and autoimmune diseases, such as HIV infection, viral hepatitis, leprosy, allergic purpura, systemic lupus erythematosus, rheumatoid arthritis, psoriasis, seronegative spondyloarthritis, tumors, and the like.
IgA nephropathy is caused by immune injury, and IgA nephropathy is caused by stimulating the body to produce excessive abnormally glycosylated IgA and inducing autoantibodies to form immune complexes and deposit on the mesangial region of glomeruli, so that mesangial cell hyperplasia, mesangial stroma increase, focal segmental hyperplasia or sclerosis are caused, and a small number of patients can have more crescent bodies and the like.
Third) existing diagnostic methods
1. Common clinical manifestations
The patient usually has visual hematuria which is easy to repeatedly attack after 1-3 days of upper respiratory tract infection, can turn into microscopic hematuria after lasting for hours to days, and can be accompanied by abdominal pain, lumbago, myalgia or low fever, urine abnormity is found in part of patients during physical examination, and the patients are asymptomatic albuminuria and/or microscopic hematuria, and a few patients have continuous visual hematuria and proteinuria with different degrees and can be accompanied by edema and hypertension. However, the clinical manifestations of IgA nephropathy may be confused with urinary system infection, acute glomerulonephritis, hereditary glomerulopathy, etc., and need to be differentiated by the experience of a doctor, and cannot be used as a basis for accurate diagnosis.
2. Laboratory examination
1) Immunological examination of serum
Serum IgA levels are elevated in 50% of patients. A specific circulating immune complex containing IgA was detected in 37% to 75% of patients. However, there is also a large proportion of patients with serum IgA in the normal range in routine immunological examinations.
2) Examination of urine
The quantification and typing of proteinuria are important for the judgment and prognosis estimation of IgA nephropathy. Proteinuria of <1g/24h is usually mild and mainly caused by focal mesangial hyperplasia. Moderate-to-severe proteinuria is usually diffuse mesenteric hyperplasia, often accompanied by crescentic and glomerular sclerosis. Hematuria: urine RBC morphologies are polymorphic, suggesting that the source of hematuria is glomerular. However, many patients with renal disease are accompanied by abnormal urine index, and urine examination can be used as a basis for judging the condition of patients with confirmed IgA nephropathy, but cannot be used as a diagnosis basis.
3) Examination of renal function
The increase of creatinine to 1.5mg/dl (132.6umol/L) is usually the disease progression. When GFR is less than 20ml/min, the pathological change is above grade III. Similar to urine examination, renal function examination can be used as a basis for judging the condition of IgA nephropathy patients, but cannot be used as a diagnosis basis.
3. Biopsy pathology diagnosis
The definitive diagnosis of IgA nephropathy requires renal biopsy pathology, supported by immunofluorescence or immunohistochemistry. The diagnosis characteristics are as follows: diffuse mesangial hyperplasia or focal segmental proliferative glomerulonephritis common under the light mirror; immunofluorescence visible mesangial region IgA or IgA-predominant immune complex deposition is a diagnostic marker for IgA nephropathy. However, the renal biopsy is painful to the patient, and has a considerable risk of missed diagnosis and misdiagnosis.
In view of the above, there is a need for a convenient, accurate and convenient diagnostic method for IgA nephropathy in clinical practice.
Disclosure of Invention
Aiming at the urgent need of IgA nephropathy clinical diagnosis at present and the defects in the prior art, the invention provides a detection marker for rapidly and accurately judging whether a person to be detected has the T cell reaction specific to IgA nephropathy by using a TCR marker which is characteristic in peripheral blood of an IgA nephropathy patient, a detection kit and application thereof, and the detection marker can be used for non-invasively, accurately and rapidly judging whether a patient with higher IgA nephropathy risk exists in a sample to be detected.
In order to achieve the purpose, the technical scheme adopted by the invention for solving the technical problems is as follows:
a peripheral blood TCR marker of IgA nephropathy comprises at least one of proteins with sequences shown in SEQ ID No. 1-100, and the specific sequences are shown in Table 1.
TABLE 1 marker sequences
Furthermore, the protein sequence of the marker is a protein which can realize the same function after one or more amino acids are substituted, deleted and/or replaced by the sequence shown in SEQ ID NO. 1-100.
Further, the marker is a protein sequence of peripheral blood TCR epitope 3(CDR 3).
The use of the above markers in the preparation of a formulation for the treatment of IgA nephropathy.
Further, the preparation includes a T cell receptor gene sequence containing the marker, or a plasmid, viral vector or nucleic acid fragment capable of expressing the T cell receptor producing the marker.
A kit for IgA nephropathy detection comprising an antibody capable of specifically binding to the above marker.
An agent comprising an antibody that specifically binds to the marker; the formulations are useful for diagnosis, prognosis, detection or screening of IgA nephropathy.
A protein chip for detecting IgA nephropathy comprises a substrate and a specific antibody spotted on the substrate, wherein the specific antibody is an antibody capable of specifically binding with the marker.
The principle of the invention is as follows: b-lymphocytes and T-lymphocytes in the human body are two important types of cells in the adaptive immune system. B cells recognize antigens through a B Cell Receptor (BCR) on the cell surface, and later, BCR expresses antibodies and is secreted extracellularly when B cells differentiate into plasma cells. T cells recognize antigens via T Cell Receptors (TCRs) on the cell surface. The diversity of BCRs and TCRs is the basis for establishing an adaptive immune system. The theoretical value of the diversity of BCR is 1018Theoretical value of TCR diversity is 1014. Among the BCR and TCR sequences, epitope 3(CDR3) is the most important part in determining the antigenic specificity, and therefore the sequence of CDR3 is considered to represent the properties of the BCR and TCR sequences.
In various diseases, the diversity or expression level of both BCR and TCR changes with different antigenic stimuli. 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 the senescent protein, fragments thereof are transported to the cell surface and presented to T cells in the immune system by histocompatibility antigen II (MCHII). Antigen fragments presented by normal cells, due to immune tolerance, do not elicit an immune response. Once normal cells are diseased and the mutated gene expresses an abnormal protein, a fragment of which is presented on the cell surface, which causes a targeted immune response in the human immune system. Thus, analysis of changes in BCR or TCR can detect the occurrence and progression of tumors or other diseases.
In the case of IgA nephropathy, a disease-specific T cell immune response is developed in the patient. We can use these IgA nephropathy patient-specific immune response characteristics to determine disease.
The invention has the beneficial effects that:
1. in the invention, 1627 samples of a control group of non-IgA nephropathy and TCR high-throughput sequencing data of 47 IgA nephropathy patients are firstly utilized to establish a big data analysis model, and whether a higher IgA nephropathy risk person exists in a sample to be detected can be clearly judged by comparing with the IgA nephropathy specific TCR sequences;
2. the early IgA nephropathy can be found by analyzing TCR change through high-throughput sequencing, and the T cell response to the IgA nephropathy in the human immune system is analyzed by utilizing the specific TCR CDR3 sequence of the IgA nephropathy, so that the method is a novel detection method;
3. the invention adopts high-throughput sequencing technology to simultaneously compare a great number of specific TCR sequences, and has higher specificity and accuracy compared with the method of singly detecting one or more markers;
4. the high-throughput sequencing instrument used in the invention has lower cost than large-scale imaging equipment, can be outsourced to a third party, and in addition, the labor cost for sampling and processing is lower than the labor cost for simultaneously detecting various markers and is also lower than the labor cost for a large number of cytological detections, so the detection cost is greatly reduced;
5. the invention only needs to adopt a small amount of peripheral blood, is simple and safe to sample, and is a non-invasive test method;
6. the TCR CDR3 sequence disclosed by the invention can be used for immunotherapy of IgA nephropathy.
Drawings
FIG. 1 shows the discovery of TCR sequences characteristic of IgA nephropathy using an immune-based big data analysis system in accordance with the present invention. The abscissa represents the sequence in which a CDR3 sequence of a specific amino acid combination is added to the IgA nephropathy-specific sequence set, and the ordinate represents the number of repetitions of the sequence in a sample CXA logarithmic value of; immunization summary of IgA nephropathy patientsThe IgA nephropathy characteristic sequences with multiple types and high repetition times are obtained, healthy people have few IgA nephropathy characteristic sequences, and the IgA nephropathy characteristic of unknown subjects is obvious, which indicates that the risk of suffering from IgA nephropathy is high.
FIG. 2 is a graph showing comparative analysis of IgA nephropathy and other diseases using characteristic indexes of IgA nephropathy in the present invention. The characteristic indexes of IgA nephropathy of healthy people and other non-tumor disease patients are obviously different from those of IgA nephropathy patients, and the specificity of the characteristic sequence set of IgA nephropathy is proved. Therefore, whether the unknown subject suffers from IgA nephropathy can be judged.
Detailed Description
The following description of the embodiments of the present invention is provided to facilitate the understanding of the present invention by those skilled in the art, but it should be understood that the present invention is not limited to the scope of the embodiments, and it will be apparent to those skilled in the art that various changes may be made without departing from the spirit and scope of the invention as defined and defined in the appended claims, and all matters produced by the invention using the inventive concept are protected.
Example 1 IgA nephropathy TCR marker CDR3 sequence set obtained by immunoblot analysis
1. Sampling and immune big data analysis
Collecting 1628 control groups (including healthy people and other non-tumor non-nephropathy patients, 1627 people for model establishment, 1 healthy person for verification, 48 IgA nephropathy patients (47 people for model establishment and 1 person for verification) and peripheral blood (10 mL per person) of 1 unknown health condition subject, obtaining the antigenic determinant 3(CDR3) amino acid sequences of TCR of the subject and the control groups by high-throughput sequencing, and ensuring that the total number of CDR3 sequences of functional TCR of each sample is not less than 25000 comprehensively;
2. the total number of CDR3 sequences for each functional TCR summed over the sequence of the sample 30000 was randomly sampled without back sampling to give a total of 30000 CDR3 sequences for that sample. All samples thus far contained a total number of CDR3 sequences for functional TCRs of 25000-30000. For any particular CDR3 sequence X, the number of repeats in a single sample sequencing result was counted as CX;
3. By analyzing the TCR CDR3 data, IgA nephropathy TCR marker CDR3 sequences were determined:
a) summarizing and de-duplicating all CDR3 sequences of 1627 control group samples for establishing the model, and setting the CDR3 sequences as a control sequence set;
b) all CDR3 sequences of 47 IgA nephropathy samples used for modeling were grouped together and de-duplicated, and all sequences including sequence repeats in the control sequence set were removed to prepare IgA nephropathy characteristic sequence set. The graph is shown in FIG. 1A, in which the CDR3 sequence with the abscissa representing a specific amino acid combination is added to the sequence order of the IgA nephropathy characteristic sequence set and the ordinate represents the number of repeats C of the sequence in a sampleXThe logarithmic value of (c).
c) The immunodata of 1 healthy person, 1 IgA nephropathy patient and 1 subject with unknown health status were plotted against the IgA nephropathy signature sequence set according to the same plotting method, as shown in FIGS. 1B-D. As can be seen from the figure, IgA nephropathy characteristic sequences having a large number of types and a high number of repeats are contained in the immune data of IgA nephropathy patients (FIG. 1B); among the immune data of healthy persons, there were only a very small number of IgA nephropathy signature sequences (FIG. 1C); whereas a subject of unknown health status had a higher signature sequence for IgA nephropathy than a healthy person, indicating that this person is at higher risk for IgA nephropathy (FIG. 1D).
d) Centralizing IgA nephropathy characteristic sequences, and repeating the occurrence times C of all CDR3 sequences in two or more samples participating in modeling IgA nephropathyXThe sum of the sequences is multiplied by the number of IgA nephropathy samples participating in modeling, wherein the samples are ranked from high to low, the top 100 are IgA nephropathy TCR marker CDR3 sequences, and the specific sequences are shown in SEQ ID NO. 1-100.
Example 2 validation of the specificity of the IgA nephropathy TCR marker CDR3 sequence set
1. Sampling and immune big data analysis
Collecting peripheral blood (10 mL per person) of 95 healthy persons, 44 other non-renal disease non-tumor disease patients and 3 subjects with unknown health conditions, and obtaining the amino acid sequences of antigenic determinant 3(CDR3) of TCR of the subjects and the control group by high-throughput sequencing to ensure that the total number of CDR3 sequences of functional TCR of each sample is comprehensively not less than 25000; the total number of CDR3 sequences for each functional TCR summed over the sequence of the sample 30000 was randomly sampled without back sampling to give a total of 30000 CDR3 sequences for that sample. All samples thus far contained a total number of CDR3 sequences for functional TCRs of 25000-30000.
3. IgA nephropathy characteristic indices were analyzed based on the immunological data of 47 IgA nephropathy patients from example 1, and 95 healthy persons newly obtained in example 2, 44 other non-renal disease non-tumor patients, and 3 unknown health condition subjects.
Wherein the IgA nephropathy characteristic index is defined as: in a sample, all CDR3 sequences belonging to the IgA nephropathy signature sequence set are repeated within the sample by the number of times CXThe sum of (a) and (b). The results of the analysis are shown in Table 2 below and FIG. 2. The specificity of the characteristic sequence set of IgA nephropathy is proved by the remarkable difference between the IgA nephropathy group and healthy people (p is 6.96E-138) and non-renal non-tumor diseases (p is 4.89E-82).
TABLE 2 IgA nephropathy characteristic index for different sample groups
4. The IgA nephropathy characteristic index of each group (table 3) was analyzed, and the IgA nephropathy characteristic index of 3 subjects with unknown health conditions (test samples) was significantly higher than the average value of "healthy person" group +2 × SD (45.1+2 × 15.8 ═ 76.6) and the average value of "non-renal disease non-tumor" group +2 × SD (49.8+2 × 32.5 ═ 114.8), and these 3 persons had a higher risk of acquiring IgA nephropathy. After comparison with the results of clinical examinations, these 3 patients were identified as early-stage IgA nephropathy patients. This example demonstrates the feasibility of using the IgA nephropathy signature sets and IgA nephropathy signature indices to predict a subject's risk of contracting IgA nephropathy.
TABLE 3 IgA nephropathy characteristic index analysis
In conclusion, the IgA nephropathy TCR marker CDR3 sequence provided by the invention has obvious IgA nephropathy specificity, can be used for early diagnosis of IgA nephropathy and evaluation of curative effect of medicines for treating IgA nephropathy, and can be used for designing a new biological immunotherapy scheme according to the immune characteristics in the future.
Sequence listing
<110> Chengdu Yianbo Biotech Ltd
<120> IgA nephropathy peripheral blood TCR marker, detection kit and application thereof
<160> 100
<170> SIPOSequenceListing 1.0
<210> 1
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 1
Ala Ser Ser Gln Glu Asp Gly Pro Thr Tyr Glu Gln Phe
1 5 10
<210> 2
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 2
Ala Ser Ser Pro Thr Pro Gly Asp Arg Tyr Tyr Gly Tyr Thr
1 5 10
<210> 3
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 3
Ala Ser Ser Ala Glu Gln Arg Val Arg Ser Asp Glu Gln Tyr
1 5 10
<210> 4
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 4
Ser Val Val Phe Gln Gly Pro Asp Ile Gln Tyr
1 5 10
<210> 5
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 5
Ala Ser Ser Leu Glu Gln Ala Ala Arg Ala Glu Gln Phe
1 5 10
<210> 6
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 6
Ser Tyr Arg Asp Gly Asp Ser Ser Tyr Glu Gln Tyr
1 5 10
<210> 7
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 7
Ala Ser Ser Leu Ser Val Tyr Trp Ala Tyr Glu Gln Tyr
1 5 10
<210> 8
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 8
Ala Ser Ser Gln Val Tyr Gly Asp Gly Gly Tyr Glu Gln Tyr
1 5 10
<210> 9
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 9
Ala Thr Ser Leu Gly Thr Gly Glu Glu Lys Leu Phe
1 5 10
<210> 10
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 10
Ser Ala Ile Arg Gly His Gln Pro Gln His
1 5 10
<210> 11
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 11
Ser Ala Leu Pro Leu Ala Ile Asp Glu Gln Phe
1 5 10
<210> 12
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 12
Ala Ser Ser Gly Gly Asp Gly Ile Asn Tyr Glu Gln Tyr
1 5 10
<210> 13
<211> 16
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 13
Ala Ser Ser Pro Pro Pro Gly Leu Ala Lys Ala Lys Asn Ile Gln Tyr
1 5 10 15
<210> 14
<211> 17
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 14
Ala Ser Ser Pro Ser His Val Ala Ala Ala Arg Ala Lys Asn Ile Gln
1 5 10 15
Tyr
<210> 15
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 15
Ala Ser Ser Pro Leu Asp Arg Ile Asp Tyr Glu Gln Tyr
1 5 10
<210> 16
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 16
Ala Ser Ser Gln Val Tyr Gly Thr Ala Phe Gly Pro Leu His
1 5 10
<210> 17
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 17
Ala Ser Ser Tyr Ser Pro Leu Asp Thr Gly Tyr Gly Tyr Thr
1 5 10
<210> 18
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 18
Ala Ile Ser Glu Arg Gly Gln Gly Glu Asn Tyr Gly Tyr Thr
1 5 10
<210> 19
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 19
Ala Ser Thr Lys Ser Gly Thr Thr Asn Glu Lys Leu Phe
1 5 10
<210> 20
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 20
Ala Ile Ser Glu Val Ala Gly Arg Asp Glu Gln Phe
1 5 10
<210> 21
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 21
Ala Ile Ser Tyr Thr Thr Gly Thr Gly Gly Asn Glu Gln Phe
1 5 10
<210> 22
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 22
Ala Ser Gly Ser Gly Gly Phe Asp Asn Glu Gln Phe
1 5 10
<210> 23
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 23
Ser Ala Arg Ser Tyr Phe Pro Pro Ser Thr Tyr Asn Glu Gln Phe
1 5 10 15
<210> 24
<211> 9
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 24
Ala Thr Ser Asp Asn Ala Glu Gln Phe
1 5
<210> 25
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 25
Ala Ser Ser Glu Leu Arg Gly Asp Asn Ser Pro Leu His
1 5 10
<210> 26
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 26
Ala Ser Ser Phe Ile Leu Ala Ser Thr Asn Glu Gln Phe
1 5 10
<210> 27
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 27
Ser Val Glu Leu Ala Asn Arg Gly Asn Tyr Gly Tyr Thr
1 5 10
<210> 28
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 28
Ala Ser Ser Glu Glu Gly Ser Val Ser Thr Asp Thr Gln Tyr
1 5 10
<210> 29
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 29
Ala Ser Ser Leu Phe Pro Gly Val His Asn Glu Gln Phe
1 5 10
<210> 30
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 30
Ala Ser Lys Leu Gly Gly Gly Thr Gly Glu Leu Phe
1 5 10
<210> 31
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 31
Ser Val Ala Pro Ser Ala Gly Gly Glu Leu Phe
1 5 10
<210> 32
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 32
Ala Trp Ser Val Arg Arg Gly His Gln Asn Thr Glu Ala Phe
1 5 10
<210> 33
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 33
Ala Ser Ser Thr Ala Gly Gly Leu Asn Glu Gln Phe
1 5 10
<210> 34
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 34
Ala Ser Ser Tyr Asp Ala Leu Ala Gly Tyr Asn Glu Gln Phe
1 5 10
<210> 35
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 35
Ala Ser Asn Gly Gly Arg His Asp Glu Gln Phe
1 5 10
<210> 36
<211> 18
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 36
Ala Ser Ser Gln Thr Ile Thr Gly Gln Thr Leu Trp Ser Phe Tyr Gly
1 5 10 15
Tyr Thr
<210> 37
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 37
Ala Ser Ser Val Asp Arg Asn Arg Lys Asn Thr Glu Ala Phe
1 5 10
<210> 38
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 38
Ala Ser Ser Pro Lys Gln Gly Arg Ile Thr Glu Ala Phe
1 5 10
<210> 39
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 39
Ala Thr Ser Ser Tyr Leu Phe Asp Glu Gln Phe
1 5 10
<210> 40
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 40
Ala Ser Arg Pro Trp Leu Ala Ala Asp Thr Gln Tyr
1 5 10
<210> 41
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 41
Ala Trp Ser Pro Asp Pro Leu Phe Thr Glu Ala Phe
1 5 10
<210> 42
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 42
Ala Ile Ser Asp Tyr Gln Thr Ser Arg Glu Thr Gln Tyr
1 5 10
<210> 43
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 43
Ser Val Ser Leu Asp Arg Gly Tyr Glu Gln Tyr
1 5 10
<210> 44
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 44
Ala Ile Ser Glu Ser Arg Thr Gly Ala Ser Glu Thr Gln Tyr
1 5 10
<210> 45
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 45
Ser Ala Arg Gly Glu Gly Glu Val Asn Glu Gln Phe
1 5 10
<210> 46
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 46
Ala Ser Ser Ser Ile Ala Gly Val Val Tyr Asp Glu Gln Phe
1 5 10
<210> 47
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 47
Ser Ala Thr Gly Arg Ala Pro Gly Glu Leu Phe
1 5 10
<210> 48
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 48
Ala Thr Ser Arg Asp Arg Tyr Arg Asn Tyr Glu Gln Tyr
1 5 10
<210> 49
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 49
Ala Ser Ser Asp Pro Gly Gly Ser Asp Val Asn Glu Gln Phe
1 5 10
<210> 50
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 50
Ala Ser Ser Tyr Gly Ala Gly Glu Gly Tyr Tyr Gly Tyr Thr
1 5 10
<210> 51
<211> 17
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 51
Ala Ser Ser Pro Val Ala Leu Ala Gly Phe Lys Phe Ser Asp Thr Gln
1 5 10 15
Tyr
<210> 52
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 52
Ala Thr Ser Arg Ala Leu Ala Glu Thr Ser Thr Asp Thr Gln Tyr
1 5 10 15
<210> 53
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 53
Ala Thr Ser Arg Asp Arg Arg Ala Arg Glu Glu Ser Pro Leu His
1 5 10 15
<210> 54
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 54
Ala Thr Ser Ile Pro Arg Asp Gln Glu Thr Gln Tyr
1 5 10
<210> 55
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 55
Ser Ala Arg Gln Ser Asp Ser Tyr Asn Glu Gln Phe
1 5 10
<210> 56
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 56
Ala Ser Ser Pro Leu Thr Ser Gly Ser Asn Asp Glu Gln Phe
1 5 10
<210> 57
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 57
Ala Ser Ser Pro Leu Pro Gly Thr Ser Ser Phe Asp Glu Gln Phe
1 5 10 15
<210> 58
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 58
Ala Ser Ser Pro Pro Tyr Arg Gly His Gly Glu Pro Gln His
1 5 10
<210> 59
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 59
Ser Ala Leu Gln Thr Ser Gly Ser Asp Glu Gln Phe
1 5 10
<210> 60
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 60
Ser Ala Thr Pro Leu Gly Gly Arg Leu Lys Thr Gly Glu Leu Phe
1 5 10 15
<210> 61
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 61
Ala Ser Ser Pro Pro Gly Thr Asp Tyr Glu Lys Leu Phe
1 5 10
<210> 62
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 62
Ala Ser Ser Leu Pro Trp Pro Tyr Asn Glu Gln Phe
1 5 10
<210> 63
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 63
Ser Ala Glu Gly Asp Asp Thr Ala Ser Thr Asp Thr Gln Tyr
1 5 10
<210> 64
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 64
Ala Trp Ser Asn Arg Leu Lys Thr Glu Ala Phe
1 5 10
<210> 65
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 65
Ala Ser Ser Gly Ser Gly Thr Ile His Glu Gln Tyr
1 5 10
<210> 66
<211> 9
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 66
Ala Trp Ser Gln Ser Asn Glu Gln Tyr
1 5
<210> 67
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 67
Ala Asn Pro Thr Ile Ala Gly Gly Asn Glu Gln Phe
1 5 10
<210> 68
<211> 16
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 68
Ala Ser Ser Leu Glu Val Tyr Arg Gly Phe Met Asp Gln Pro Gln His
1 5 10 15
<210> 69
<211> 17
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 69
Ala Ser Ser Leu Arg Gly Pro Ala Gly Ala His Gly Thr Asp Thr Gln
1 5 10 15
Tyr
<210> 70
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 70
Ala Ile Ser Glu Arg Thr Ser Gly Ala Ser Tyr Asn Glu Gln Phe
1 5 10 15
<210> 71
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 71
Ala Ser Ser Pro His Gly Val Gly Val His Ser Tyr Glu Gln Tyr
1 5 10 15
<210> 72
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 72
Ala Thr Ser Ser Tyr Gly Ala Gln Ser Ser Tyr Asn Glu Gln Phe
1 5 10 15
<210> 73
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 73
Ala Trp Asp Arg Gly Ala Gly Lys Thr Gln Tyr
1 5 10
<210> 74
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 74
Ala Ser Ser Thr Gly Arg Arg Asp Glu Gln Tyr
1 5 10
<210> 75
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 75
Ser Thr Lys Gly His Ser Met Asn Thr Glu Ala Phe
1 5 10
<210> 76
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 76
Ala Ser Ser Pro Gln Arg Gln Arg Asn Thr Glu Ala Phe
1 5 10
<210> 77
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 77
Ala Ser Ser Gly Pro Gly Arg Ile Ala Lys Asn Ile Gln Tyr
1 5 10
<210> 78
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 78
Ala Ser Ser Gln Thr Ser Gly Gly Glu Asn Glu Gln Tyr
1 5 10
<210> 79
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 79
Ala Ile Ser Asp Pro Gly Arg Gly Gln Asp Glu Asp Thr Gln Tyr
1 5 10 15
<210> 80
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 80
Ser Ala Arg Arg Gly Leu Thr Tyr Ser Gly Asn Thr Ile Tyr
1 5 10
<210> 81
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 81
Ala Ser Ser Leu Ser Ser Val Gly Gln Gln Phe
1 5 10
<210> 82
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 82
Ala Thr Ser Val Val Phe Ser Gly Ala Asn Val Leu Thr
1 5 10
<210> 83
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 83
Pro Pro Pro Glu Leu Gln Gly Arg Pro Ser Thr
1 5 10
<210> 84
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 84
Ala Ser Ser Ser His Trp Thr Thr Gln Tyr
1 5 10
<210> 85
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 85
Ala Ser Ser Pro Ser Thr Gly Leu Asp Ser Pro Leu His
1 5 10
<210> 86
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 86
Ala Ser Ser Asp Glu Pro Gly Gln Gly Ser Asn Glu Gln Phe
1 5 10
<210> 87
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 87
Ala Ser Ser Ser Thr Gly Leu Ala Gly Pro Ser Gly Glu Leu Phe
1 5 10 15
<210> 88
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 88
Ala Ser Ser Ala Gly Ala Gly Thr Ser Asn Glu Gln Phe
1 5 10
<210> 89
<211> 17
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 89
Ala Ser Ser Leu Glu Gly Arg Gly Gly Glu His Leu Tyr Asn Glu Gln
1 5 10 15
Phe
<210> 90
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 90
Ala Ser Thr Glu Pro Trp Lys Gln Glu Thr Gln Tyr
1 5 10
<210> 91
<211> 9
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 91
Ala Leu Arg Asp Pro Tyr Glu Gln Tyr
1 5
<210> 92
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 92
Ala Ser Lys Gly Thr Arg Gly Ser Asn Asn Glu Gln Phe
1 5 10
<210> 93
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 93
Ala Ser Ser Ser Gly Gln Gly Leu Leu Asn Glu Gln Phe
1 5 10
<210> 94
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 94
Ala Thr Ser Asp Met Val Arg Glu Thr Gln Tyr
1 5 10
<210> 95
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 95
Ala Ser Ser Ser Gly Arg Tyr Tyr Tyr Glu Gln Tyr
1 5 10
<210> 96
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 96
Ala Ile Lys Ala Gly Ala Gly Thr Asp Thr Gln Tyr
1 5 10
<210> 97
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 97
Ala Ser Ser Pro Pro Leu Arg Pro Gly Glu Ala Phe
1 5 10
<210> 98
<211> 19
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 98
Ala Thr Ser Arg Glu Arg Thr Gly Ala Ile Asn Pro Glu Asn Thr Gly
1 5 10 15
Glu Leu Phe
<210> 99
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 99
Ala Ser Thr Phe Pro Gly Leu Asn Thr Glu Ala Phe
1 5 10
<210> 100
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 100
Ala Ser Ser Ser Arg Tyr Trp Glu Gly Asn Glu Gln Phe
1 5 10
Claims (7)
1. A TCR marker for peripheral blood of IgA nephropathy, which comprises at least one TCR protein sequence shown in SEQ ID NO. 1-100.
2. The IgA nephropathy peripheral blood TCR marker according to claim 1, wherein the protein sequence of the marker is a protein which can achieve the same function by substituting, deleting and/or substituting one or more amino acids in the sequences represented by SEQ ID No.1 to 100.
3. Use of a TCR marker as claimed in claim 1 in the manufacture of a formulation for the treatment of IgA nephropathy.
4. The use of claim 3, wherein the preparation comprises a T cell receptor comprising the marker, or a plasmid, viral vector or nucleic acid fragment capable of expressing the T cell receptor producing the marker.
5. A kit for IgA nephropathy detection comprising an antibody that specifically binds to the TCR marker of claim 1.
6. An agent comprising an antibody that specifically binds to a TCR marker of claim 1; the formulations are useful for diagnosis, prognosis, detection or screening of IgA nephropathy.
7. A protein chip for detecting IgA nephropathy, comprising a substrate and a specific antibody spotted on the substrate, wherein the specific antibody is an antibody capable of specifically binding to the TCR marker of claim 1.
Priority Applications (2)
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CN202110834998.5A CN113563454A (en) | 2021-07-23 | 2021-07-23 | IgA nephropathy peripheral blood TCR marker and detection kit and application thereof |
PCT/CN2022/080395 WO2023000689A1 (en) | 2021-07-23 | 2022-03-11 | Peripheral blood tcr marker for iga nephropathy, detection kit thereof, and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
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CN202110834998.5A CN113563454A (en) | 2021-07-23 | 2021-07-23 | IgA nephropathy peripheral blood TCR marker and detection kit and application thereof |
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CN113563454A true CN113563454A (en) | 2021-10-29 |
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CN202110834998.5A Pending CN113563454A (en) | 2021-07-23 | 2021-07-23 | IgA nephropathy peripheral blood TCR marker and detection kit and application thereof |
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CN (1) | CN113563454A (en) |
WO (1) | WO2023000689A1 (en) |
Cited By (1)
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---|---|---|---|---|
WO2023000689A1 (en) * | 2021-07-23 | 2023-01-26 | 成都益安博生物技术有限公司 | Peripheral blood tcr marker for iga nephropathy, detection kit thereof, and application thereof |
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CN111665359A (en) * | 2020-07-11 | 2020-09-15 | 成都益安博生物技术有限公司 | Lung cancer peripheral blood TCR marker and detection kit and application thereof |
CN113030473A (en) * | 2021-03-15 | 2021-06-25 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of acute B lymphocyte leukemia and detection kit and application thereof |
CN113109564A (en) * | 2021-03-15 | 2021-07-13 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of acute myelocytic leukemia and detection kit and application thereof |
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CN102124340B (en) * | 2008-06-02 | 2014-12-17 | 卫材R&D管理有限公司 | IgA nephropathy detection method and detection kit |
CN109161592A (en) * | 2018-08-02 | 2019-01-08 | 深圳市人民医院 | A kind of kit and the application in IgA nephrosis |
CN109182487A (en) * | 2018-08-15 | 2019-01-11 | 深圳慈海医院 | A kind of kit found for monitoring the biomarker of IgA nephrosis |
CN110045130B (en) * | 2019-04-17 | 2022-03-29 | 中山大学附属第一医院 | Immunoassay detection kit for IgA nephropathy-related polypeptide |
CN113563454A (en) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | IgA nephropathy peripheral blood TCR marker and detection kit and application thereof |
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2021
- 2021-07-23 CN CN202110834998.5A patent/CN113563454A/en active Pending
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2022
- 2022-03-11 WO PCT/CN2022/080395 patent/WO2023000689A1/en unknown
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CN111665359A (en) * | 2020-07-11 | 2020-09-15 | 成都益安博生物技术有限公司 | Lung cancer peripheral blood TCR marker and detection kit and application thereof |
CN113030473A (en) * | 2021-03-15 | 2021-06-25 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of acute B lymphocyte leukemia and detection kit and application thereof |
CN113109564A (en) * | 2021-03-15 | 2021-07-13 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of acute myelocytic leukemia and detection kit and application thereof |
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WO2023000689A1 (en) * | 2021-07-23 | 2023-01-26 | 成都益安博生物技术有限公司 | Peripheral blood tcr marker for iga nephropathy, detection kit thereof, and application thereof |
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