101 results about "Human leukocyte antigen" patented technology

The invention discloses a human leukocyte antigen HLA-A and HLA-B gene full-length sequencing method and an HLA gene sequencing and typing method. The HLA-A and HLA-B gene full-length sequencing method comprises the following steps of: a, performing PCR amplification on about 4kb full-length sequences of HLA-A and HLA-B genes by using a pair of primers respectively; and b, cloning the amplification products to a pGEM-Tea sy vector, sequencing the full-length sequences by using ten walking sequencing primers in positive and negative directions respectively, and totally obtaining 38 allele 4.3kb full-length sequences of the HLA-A and 30 allele 3.7kb full-length sequences of the HLA-B. The HLA-A and HLA-B sequencing and typing method comprises the following steps of: performing PCR amplification on typing target areas of the HLA-A and HLA-B by using two pairs of primers respectively; and performing two directional sequencing on products by using fourteen sequencing primers respectively, wherein the HLA-DRB1 and HLA-DQB1 sequencing and typing method comprises the following steps of: amplifying sequences of second and third exons of DRB1 and DQB1 by adopting group specificity primers respectively; performing two directional sequencing on the second and third exons of the DRB1 by adopting eight group specificity primers and three sequencing primers; and performing two directional sequencing on the second and third exons of the DQB1 by adopting four sequencing primers respectively.

The invention relates to nucleic acid molecules which code for the tumor rejection antigen precursor MAGE-3. Also disclosed are vectors, cell lines, and so forth, which utilize the nucleic acid molecule, and optionally, molecules coding for human leukocyte antigen HLA-A1. Uses of these materials in therapeutic and diagnostic contexts are also a part of the invention.

The invention discloses an HLA (human leukocyte antigen) gene specific PCR (polymerase chain reaction) amplification primer, an HLA typing method and an HLA typing kit. A sequence of the PCR amplification primer comprises a primer sequences shown as SEQ ID No. (sequence identification number) 1-2, 3-4, 5-6, 7-8 and 9-10. The HLA gene typing method comprises the steps that the HLA gene specific PCR amplification primer is used for conducting PCR amplification and purification on sample DNA (deoxyribonucleic acid); a gene exon sequencing primer is used for conducting sequencing PCR amplification, purification and sequencing on an HLA gene amplification product, the sequence of the HLA gene amplification product is compared with a standard sequence; and the type of the HLA gene is determined. The HLA gene typing kit comprises the PCR amplification primer and the sequencing primer. A qualitative leap in aspects of detection flux, data quality, cost control and the like is achieved; data is more reliable and realer; and the problem that typing cannot be conducted when nucleotide of certain alleles is positioned outside an amplification area is solved.

The invention provides an HLA (Human Leukocyte Antigen)-A,B genotyping PCR (Polymerase Chain Reaction) primer, and a method of applying the PCR primer to HLA gene analysis based on a sequencing method.

The invention provides a human mucin-1 tumor antigenic polypeptide with an amino acid sequence shown in SEQ ID NO: 1 or variants thereof, wherein the polypeptide can be combined with an HLA (Human Leukocyte Antigen) I and can be recognized by a cell CD8<+>T. The invention further provides nucleic acids coding the polypeptide. The invention further provides an antigen presenting cell capable of presenting the polypeptide on the surface of the cell and an immune effector cell capable of recognizing the polypeptide or antigen presenting cell. The invention further provides application of the polypeptide or variants thereof, the nucleic acids, the antigen presenting cell or the immune effector cell in the preparation of vaccines or pharmaceutical compositions for treating or preventing cancer. Tumor vaccines provided by the invention have a good treatment effect on relatively large crowds and are particularly applicable to the Asian, e.g. Chinese.

The invention discloses a primer probe assembly for high-specificity amplification HLA (Human Leukocyte Antigen)-B*5801 alleles on the basis of a TaqMan probe detection method. The primer probe assembly comprises an upstream primer Fp: 5'-AGGGGCCGGAGTATTGGGATG-3', a downstream primer Rp: 5'-TTGGCCTCAACTGAAAATGAAAC-3' and a probe: 5'-HEX-TCAGGGAGGCGGATCTCGGAC-BHQ2-3'. Primers and probes of reference genes ACTB are used simultaneously, target genes and the reference genes are added into a tube for dual-channel fluorescent quantitative PCR reaction, and a result is analyzed through an amplification curve. The method has the characteristics of simplicity, convenience, flexibility, quickness, high specificity, high flux, zero pollution, high resolution and the like, and is applicable to detection of the HLA-B*5801 alleles of a whole-genome DNA (Deoxyribonucleic Acid) sample in the peripheral blood and the saliva of a human body.

The invention discloses an HLA (Human Leukocyte Antigen)-based Adams simulation model integrated platform, to solve the defect in the prior art that an Adams simulation model is hard to be subjected to HLA integration. The HLA-based Adams simulation model integrated platform comprises a simulation model unit, an adaptor unit and an HLA system, wherein the simulation model unit comprises at least one Adams simulation module; the adaptor unit comprises a user defined module, a shared memory space module and a main body module; and the user defined module comprises an input sub-module and an output sub-module. The invention discloses an HLA-based Adams simulation model integrated method. With the adoption of the HLA-based Adams simulation model integrated method, the advantages of a commercial simulation software solver are maintained, the packaging of an Adams model is realized under the condition that the simulation model needs not to be modified or is slightly configured, and the reusability of the Adams model is greatly improved.

The invention provides a TCR (T Cell Receptor) which is capable of specifically binding oligopeptides KVLEYVIKV derived from an MAGE-A1 (Melanoma antigen A1). A compound can be formed by the oligopeptides KVLEYVIKV derived from the MAGE-A1 and an HLA (Human Leukocyte Antigen) A0201, and the oligopeptides KVLEYVIKV and the HLA A0201 can be transferred to cell surfaces together. The invention also provides a nucleic acid molecule for coding the TCR and a carrier comprising the nucleic acid molecule. In addition, the invention also provides a cell for transducing the TCR.

The invention discloses joint application of three types of monoclonal antibody-coupled immunomagnetic beads to sorting tumor cells and a method for sorting breast cancer tumor cells. The three types of monoclonal antibody-coupled immunomagnetic beads include anti-HLA-G (human leukocyte antigen-G) monoclonal antibody-coupled immunomagnetic beads, anti-EpCAM (epithelial cell adhesion molecule) monoclonal antibody-coupled immunomagnetic beads and anti-CK8 / 18 (casein kinase 8 / 18) monoclonal antibody-coupled immunomagnetic beads. The joint application and the method have the advantages that the three types of jointly applied antibody-coupled immunomagnetic beads are high in breast cancer tumor cell enrichment ratio and good in sensitivity and repeatability, and the relevant tumor cells can be detected; the three types of immunomagnetic beads (the anti-EpCAM immunomagnetic beads, the anti-HLA-G immunomagnetic beads and the anti-CK8 / 18 immunomagnetic beads) are jointly applied to enriching the tumor cells, so that the circulating tumor cell coverage rate can be increased to the greatest extent, the enrichment ratio can reach 80%, and a technology is international pioneer; the tumor cell enrichment specificity and sensitivity can be particularly guaranteed owing to usage of the anti-HLA-G monoclonal antibody-coupled immunomagnetic beads.

The invention provides a preparation method of a breast cancer-specific epitope polypeptide-loaded dendritic cell vaccine, and the method comprises the following steps: loading dendritic cells with identified human leukocyte antigen A201 positive epitope polypeptide of human epidermal growth factor receptor 2, breast cancer drug resistance protein and IQ motif and Sec7 structural domain 1 protein which specifically express in breast cancer (namely polypeptide comprising the amino acid sequence shown in any of SEQ ID No.1-6) to prepare into the dendritic cell vaccine which can induce strong breast cancer-targeting cytotoxic T lymphocyte response. The invention also provides a kit comprising the breast cancer-specific epitope polypeptide-loaded dendritic cell vaccine.

The invention relates to a detection method of HLA (Human Leukocyte Antigen)-B*58:01 allele. The detection method comprises the following steps: providing a mixture of a sample to be tested, a nucleic acid amplification system and a fluorescence detection system; circularly amplifying target polynucleotide through an amplified reaction; indirectly combining a fluorescence generating group and an amplified target polynucleotide sequence; and detecting the fluorescent amount generated by the fluorescence generating group so as to determine existence of the target polynucleotide and the relative amount thereof. The invention further discloses a kit for detecting the allele. The kit comprises a plurality of sealed centrifuge tubes which are respectively filled with a reaction liquid 1, a reaction liquid 2, a reaction liquid 3, an EZ Taq enzyme mixed liquid, a standard liquid 1 and a standard liquid 2 as well as the kit which separates and packages the centrifuge tubes in a centralized manner. The kit and the detection method provided by the invention are simple and convenient to operate, short in time consumed, strong in specificity and high in sensitivity. The kit and the detection device can be widely applied to detecting the HLA-B*58:01 allele and clinically avoiding severe untoward effects of skins of the HLA-B*58:01 allele patients caused by using allopurinol.

The invention discloses a chimeric antibody and an immunocyte. The chimeric antibody provided by the invention has an amino acid sequence containing a target antibody, a transmembrane domain segment of a HLA-A2 (Human Leukocyte Antigen-A2) molecule and an intracellular domain segment of a CD3 compound protein zeta chain in sequence from a terminal N to a terminal C. The chimeric antibody can be expressed on the surfaces of any cells, when the chimeric antibody is expressed on the surface of an immunocyte, the immunocyte is endowed with a specific immunity by virtue of the target antibody on the chimeric antibody, and thus, the chimeric antibody provided by the invention can be specifically combined with a plurality of antigens along with different target antibodies in the chimeric antibody. The chimeric antibody, a single-domain antibody and an immunocyte for expressing the chimeric antibody, provided by the invention, are about to have broad application prospects in the cellular immunotherapy.

The invention discloses an HLA (human leucocyte antigen) typing method based on a PacBio RS II sequencing platform. A collected sample is subjected to DNA extraction and then to PCR (polymerase chain reaction) amplification, PCR products are mixed to establish a 10k library, and PacBio RS II sequencing is performed; then, original data obtained by sequencing are corrected, and HLA typing is performed with software programs. Compared with existing HLA typing methods, the HLA typing method has super-high resolution and is of important value to applications such as clinical graft tissue matching, population genetics, anthropology and evolutiology, as well as basic research work.

Hematopoietic stem cells of healthy volunteers are separated and purified in a 100 grade GMP (Good Manufacturing Practice) plant, and are induced to divide to NK cells under a special condition of culture. Qualified NK cells are put in storage or KIR-matched with HLA (Human Leukocyte Antigen) and KN cells of the patients. The qualified NK cells are intravenously infused to tumor patients to prevent recurrence and metastasis of cancer cells of the tumor patients. The hematopoietic stem cells used by the method have the advantages of abundant source, material object storage and the like, are free from toxic and side effects and adverse reaction for treating tumor, and remarkably improve the survival quality of the tumor patients.

The invention discloses a genetic testing method for risk of causing serious adverse reactions of carbamazepine. The genetic testing method comprises the following steps: collecting oral mucosa cells of a subject, extracting genome DNA (deoxyribonucleic acid) of the oral mucosa cells, testing HLA (human leukocyte antigen)-B*1502 allelotype of the genome DNA and evaluating the risk of causing the serious adverse reactions of the skin of an anti-epileptic medicament-carbamazepine, thereby providing reference basis for clinical individual medication.

A method for altering the composition of tumor infiltrating mononuclear cells, increasing CD4+ / CD8+ ratio, increasing tumor stroma / epithelial ratio and modulating HLA (Human Leukocyte Antigen) class II expression on a tumor cell surface with a serum-free and mitogen-free mixture having specific cytokine ratios from the group of IL-1β, TNF-α, IFN-γ, GM-CSF, and Interleukin-2 (IL-2) with specific ratios of IL-1β, TNF-α, IFN-γ, GM-CSF to IL-2, respectively. The serum-free and mitogen-free mixtures comprised of cytokine ratios include Leukocyte Interleukin Injection (LI) or Multikine®, which can be further used alone or in combination with other drugs for the treatment of cancer thereby increasing the success of cancer treatment and the disease free survival of cancer patients.

The invention provides a method and a system for detecting HLA (human leukocyte antigen) heterozygosity deficiency. The method comprises the following steps: data acquisition: acquiring sequencing data of a tumor sample and a control sample; hLA typing detection: detecting HLA molecular types of the tumor sample and the contrast sample; HLA allele imbalance detection: comparing the sequencing data with an HLA typing result to obtain an HLA allele imbalance detection result; copy number variation detection: performing copy number variation detection on all the target areas to obtain a copy number variation detection result of the HLA gene loci; and HLA heterozygosity deletion judgment: judging whether HLA heterozygosity deletion exists or not according to an HLA allele imbalance detection result and a copy number variation detection result. According to the invention, the sequence information on the HLA gene is used independently, and the sequence information near the HLA gene is also combined, so that an accurate HLA LOH result is obtained.

The invention discloses a preparation method of an HLA-A0201 (Human Leukocyte Antigen-A0201) restrictive anti-MAGE (Micro Array and Gene Expression) antigenic specificity CTL (Cytotoxic T Lymphocyte). The method comprises the following steps of: carrying out hemapheresis to collect peripheral blood mononuclear cells; enriching and purifying CD8+T lymphocytes; utilizing a mature dendritic cell loaded with an HLA-A0201 restrictive anti-MAGE antigen polypeptide to stimulate the CD8+T lymphocytes; utilizing rhIL-2 and rhIL-7 to accelerate the T cells to grow; utilizing a Tetramer marking method and a flow type cell separation technology to purify the target CTL; utilizing an anti-human CD3 monoclonal antibody covered by a solid phase and an IL-2 to stimulate the target CTL to grow; adding an autologous PBMC (Peripheral Blood Mononuclear Cell) radiated by gamma rays to enhance the activation of the target CTL; adding rhIL-15 to cultivate and reproduce; and collecting and identifying. The prepared target CTL has high purity, high reproduction capability, high killing activity and high ratio CTL-CM, and can be used for immune treatment of tumors and the like.

The invention discloses a method of detecting common ambiguous types of HLA (human leukocyte antigen) typing locus A. The method includes: (1) acquiring ambiguous types of locus A through HLA typing results, and selecting specific primers necessary to distinguish the ambiguous types, wherein the specific primers are shown as SEQ ID NO. 1 to 9; (2) using the selected specific primers to perform sequencing PCR (polymerase chain reaction) on the purified HLA-A gene amplification product; (3) purifying the post-sequencing product, acquiring a product sequence through a sequencer, comparing the product sequence with the ambiguous types so as to distinguish the ambiguous types, and finally determining HLA-A type information. Compared with traditional PCR-SSP (PCR-sequence specific primer) methods to determine gene ambiguous types, the method herein can accurately and precisely judge the types, has greatly reduced cost, is simple to perform, can easily provide high throughput, and is a time-saving economical option for large-scale donor HLA typing.

The invention provides a PCR (polymerase chain reaction) amplification primer group and Taqm probes used for detecting HLA (human leukocyte antigen)-B*1502 allelic genes. The PCR amplification primer group and Taqm probes include two pairs of primers designed according to specific sequence of HLA-B*1502 and two corresponding probes, further include one pair of probes used for filtering out false positive results of the HLA-B*1502 and one corresponding probe, and further include one pair of internal-contract primers and one corresponding probe. The primer group has the technical advantages that the genes of the HLA-B*1502 are positive or not are determined through two reaction tubes, the false positive result of the HLA-B*1502 is eliminated through one reaction tube, and the false positive results of the genes of the sampled HLA-B*1502 are eliminated by adding the internal-control primers and the probes in reaction holes.

The invention discloses tumor-associated antigen XAGE-1b nonapeptide and application thereof. The sequence of XAGE-1b oligopeptide is RQKKIRIQL. CTLs (cytotoxic T lymphocytes) induced by the XAGE-1b antigen peptide do not produce immune responses to testicular cells, but only kill tumor cells. Induced CTL clones have a good tumor cell specificity lethal effect. In the process of establishing the CTLs, the XAGE-1b antigen peptide screened out by the inventor has proper affinity with HLAs (human leukocyte antigens) on DC cells and can effectively stimulate and induce the specific CTLs, thereby having a good potential of serving as a polypeptide vaccine and a DC vaccine. Virus carriers are created by cloning specific TCR (T cell receptor) genes of the XAGE-1b antigen peptide, and peripheral circulation CD8+T cells are transduced to obtain clones of TCR gene modified T cells (TCR-T); the clones also have the tumor cell specificity lethal effect same as the parental CTLs, thereby having a good prospect in clinical transformation and practical application.

The invention discloses an HLA (human leukocyte antigen) gene amplification primer. The HLA gene amplification primer comprises a mixture consisting of any one or more of the following nine primer groups of SEQ ID NO: 1 and SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4; SEQ ID NO: 5 and SEQ ID NO: 6; SEQ ID NO: 7 and SEQ ID NO: 8; SEQ ID NO: 9 and SEQ ID NO: 10; SEQ ID NO: 11 and SEQ ID NO: 12; SEQ ID NO: 12 and SEQ ID NO: 13; SEQ ID NO: 14 and SEQ ID NO: 15; SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO: 18. The invention also discloses a kit for sequencing the HLA gene, and the kit comprises the HLA gene amplification primer. The invention also discloses a construction method of a gene sequencing library based on the HLAHLA amplification primer. The invention also discloses an HLA gene sequencing method based on the construction method of the HLA gene sequencing library.

The invention discloses an assay kit for detecting human leukocyte antigen-B (HLA-B)*57:01 and HLA complex P5 (HCP5) alleles. The assay kit comprises a polymerase chain reaction (PCR) reaction liquid containing primers for detecting HLA-B*57:01 and HCP5, wherein the primers for detecting HLA-B*57:01 are primers B57F, B57R1 and B57R2; the primer B57F has a sequence shown in SEQIDNO:1, the primer B57R1 has a sequence shown in SEQIDNO:2, and the primer B57R2 has a sequence shown in SEQIDNO:3; and the primers for detecting the HCP5 are primers 631R and 568GF, wherein the primer 631R has a sequence shown in SEQIDNO:4, and the primer 568GF has a sequence shown in SEQIDNO:5. The assay kit has high detection sensitivity and good specificity, and can be used for accurately separating genotypes without sequencing.

The invention discloses a primer group, a kit and a method for amplification of a human leukocyte antigen-B (HLA)-B gene, and the primer group, the kit and the method for genotyping of the HLA-B gene,belonging to the field of gene detection. According to the primer group, provided by the invention, for the amplification of the HLA-B gene, at least two sets of primers are designed according to theeight exon regions of the HLA-B gene; the primers are used for performing polymerase chain reaction (PCR) amplification on the HLA-B gene, so that a product having a gene fragment length of less than1.5 kb is obtained; after the HLA-B gene is subjected to the PCR amplification by the primers, the gene fragment length of the amplified product is less than 1.5 kb, so that the requirement for template integrity is reduced, the amplification efficiency is high, amplification reaction time is short, and the cost is low. The primer groups, the kits and the methods can meet the needs of amplification or genotyping of the HLA-B gene in mass samples, provide a more accurate basis for clinical HLA matching, provide suitable transplant donors for patients, reduce the rejection in a transplantationprocess and improve the success rate of organ transplantation and the patient survival rate.

The invention belongs to the technical field of gene engineering, and discloses a primer combination for HLA (human leukocyte antigen) genotyping, a kit containing the primer combination and a method for HLA genotyping. Thus, the sequence-specific HLA genotyping (GSA-SBT) technique is quick, simple, accurate and visual. The primer combination can be synchronously used for amplification and sequencing with the commercialized kit, thereby lowering the experimentation cost. Besides, the specific design of mismatched bases are introduced to the 3' terminal of the primer, thereby enhancing the specificity of the primer amplification and ensuring the accuracy of the typing result.

The invention relates to a gene detecting kit that is used in clinical detection, in particular to a DNA microarray chip detecting kit, which can realize subtype detection of human leukocyte antigen DQB1(HLA-DQB1) gene in high flux, high efficiency and high specificity.

The invention discloses a method and a system for extracting an immunotherapy neoantigen. The method comprises the following steps: S1, acquiring conventional proteomes of a tumor tissue and a normaltissue of a sample; S2, obtaining a base monomer unit sequence library of a sample tumor tissue and a sample normal tissue and a specific proteome of the sample tumor tissue; S3, based on the conventional proteome of the sample tumor tissue, the specific proteome of the sample tumor tissue and human leukocyte antigen (HLA) molecular typing, obtaining a plurality of candidate tumor specific neoantigens; S4, based on the plurality of obtained candidate tumor specific neoantigens, respectively calculating the existence conditions in a sample tumor tissue conventional proteome, a sample normal tissue conventional proteome, a sample tumor tissue basic group monomer unit sequence library and a sample normal tissue basic group monomer unit sequence library, and taking the existence conditions andgene expression change multiples as filtering rules to obtain the tumor specific neoantigen. From the aspect of source, the tumor specific neoantigen part discovered by the scheme of the invention comes from a genome non-coding region and is not limited to a coding region, so that more neoantigens can be discovered.

A method for altering the composition of tumor infiltrating mononuclear cells, increasing CD4+ / CD8+ ratio, increasing tumor stroma / epithelial ratio and modulating HLA (Human Leukocyte Antigen) class II expression on a tumor cell surface with a serum-free and mitogen-free mixture having specific cytokine ratios from the group of IL-1β, TNF-α, IFN-γ, GM-CSF, and Interleukin-2 (IL-2) with specific ratios of IL-1β, TNF-α, IFN-γ, GM-CSF to IL-2, respectively. The serum-free and mitogen-free mixtures comprised of cytokine ratios include Leukocyte Interleukin Injection (LI) or Multikine®, which can be further used alone or in combination with other drugs for the treatment of cancer thereby increasing the success of cancer treatment and the disease free survival of cancer patients.