61 results about "Peripheral blood lymphocyte" patented technology

The invention relates to the field of cells, in particular to a cytokine combination, a culture medium, a tumor-specific T cell population and application thereof. The preparation method comprises the following steps: firstly, activating a solidified CD3 monoclonal antibody, enabling T cells to automatically secrete part of cell factors, and meanwhile, enabling mononuclear cells in peripheral blood lymphocytes to be mature into dendritic cells under the action of adding exogenous cell factors; adding non-activated peripheral blood lymphocytes again, and activating and proliferating T cells under the action of dendritic cells and cell factors; then, activation and proliferation of T cells are further promoted, so that a tumor specific T cell population is obtained, and the T cell population can directly kill tumors and inhibit the growth of the tumors.

The invention discloses a nano antibody for resisting a listeria monocytogenes MurA protein and application of the nano antibody, and relates to the technical field of biology. The amino acid sequence of the nano antibody is as shown in SEQ ID NO.3. MurA protein of prokaryotically expressed listeria monocytogenes is used as an immunogen to immunize alpaca, total RNA of peripheral blood lymphocytes is extracted, VHH fragments are amplified through reverse transcription and two rounds of nested PCR, the VHH fragments and a pComb3Xss vector are converted into TG1 competent cells after enzyme digestion and connection, and a phage display technology is used for three rounds of elutriation, so that the total RNA of the listeria monocytogenes is obtained. The nanometer antibody for resisting the listeria monocytogenes MurA protein is successfully screened out. The nano antibody can specifically recognize MurA protein and whole listeria monocytogenes, and the immunocolorimetric sensor based on thiolated bacteriophage induced colloidal gold aggregation, which is established by utilizing the nano antibody, is strong in specificity, high in sensitivity and low in detection limit, and can greatly shorten the detection time.

The application discloses a use of Ejiao Qianggu oral liquid in regulating immunity, and particularly relates to an improving effect of the Ejiao Qianggu oral liquid on an immune system, which is realized by the following aspects. On one hand, the Ejiao Qianggu oral liquid promotes proliferation and transformation (mainly early T lymphocytes and part of B lymphocytes) and active functions of spleen lymphocytes; on the other hand, the Ejiao Qianggu oral liquid improves phagocytosis of macrophages. The Ejiao Qianggu oral liquid has the following characteristics: firstly, the Ejiao Qianggu oral liquid acts on early lymphocyte maturation, and has no significant effect on proliferation of peripheral blood lymphocytes; secondly, the Ejiao Qianggu oral liquid can significantly act on immune target organs, thymus and spleen tissues, and does not excessively stimulate the spleen, unlike positive drugs such as colony stimulating factor which can cause significant edema of the spleen; and thirdly, the Ejiao Qianggu oral liquid has a significant recovery effect on body weight, white blood cells, lymphocytes, red blood cells and hemoglobin in blood of patients with low immunity.

The application discloses an application of baicalin in treating coronavirus infectious diseases. The new application of baicalin is verified through in-vitro cell tests and mouse tests infected with human coronavirus HCoV-229E, the technical scheme is novel, and a curative effect test of baicalin in treating pneumonia caused by HCoV-229E virus infection is carried out, and the results show that baicalin has a certain inhibitory effect on HCoV-229E infected cells in vitro; baicalin shows good drug efficacy in lung index and inhibition rate of pneumonia mice infected with HCoV-229E virus, can reduce the virus load of lung tissue of pneumonia mice infected with HCoV-229E virus, increase the proportion of peripheral blood lymphocytes of mice, and reduce the content of IL-2, IL-10, TNF-alpha and IFN-beta in lung tissue of mice.

The invention provides a method for detecting insulin sensitivity, which comprises the following steps: (1) transfecting a cell to be detected by using a GLUT4myc cDNA plasmid: transfecting the cell to be detected by using the GLUT4myc cDNA plasmid, so that the cell stably expresses GLUT4myc; (2) measuring the endocytosis rate of the GLUT4myc: performing endocytosis stimulation to enable the GLUT4myc combined with the anti-myc antibody to be endocytosed, and regularly detecting the amount of the GLUT4myc combined with the anti-myc antibody remained on the surface of the cell; (3) measuring the GLUT4myc exocytosis rate: performing exocytosis stimulation to enable the GLUT4myc combined with the anti-myc antibody in the cell to exocytosis, and regularly detecting the translocation amount of the GLUT4myc combined with the anti-myc antibody in the cell to the surface of the cell; (4) calculating the difference value between the GLUT4myc exocytosis rate and the GLUT4myc endocytosis rate, and calculating the insulin sensitivity according to the difference value; the insulin sensitivity evaluation method has the beneficial effects that the insulin sensitivity is evaluated by detecting the exocytosis and endocytosis rates of fat cells, muscle cells or peripheral blood lymphocytes GLUT4, so that early-stage diabetic patients can be identified, and the pathogenesis of diabetes mellitus can be researched.

The invention provides an anti-CD30 rabbit Fab antibody as well as a preparation method and application thereof. Human CD30 protein is expressed through in-vitro recombination, a rabbit is immunized for three times by using the CD30 protein, peripheral blood lymphocytes are separated out, total RNA of the peripheral blood lymphocytes is extracted, the peripheral blood lymphocytes are reversely transcribed into cDNA, the cDNA is taken as a template, and a Fab sequence is amplified by using a specific primer; according to the invention, a rabbit Fc structural domain is used as an antigen, a Fab antibody capable of being combined with CD30 protein is obtained by screening through a yeast display technology, a Fab sequence is combined with the rabbit Fc structural domain to obtain high-purity fusion protein, and the fact that the Fab antibody has relatively strong activity with CD30 and has specific binding capacity with tumor cells is verified at the cellular level.

This invention discloses a T-cell training method based on tumor organ-on-a-chip. The method includes: preparing an organ-on-a-chip with a micropillar array and a biomimetic fishbone structure; introducing primary tumor cells from a patient into the chip, causing them to spontaneously form uniform three-dimensional tumor spheroids; dynamically pumping pre-activated peripheral blood lymphocytes from the same patient into the chip and co-culturing them with the tumor spheroids; collecting the co-cultured lymphocytes and repeating the co-culture process multiple times to obtain a lymphocyte population with enhanced anti-tumor activity. This invention utilizes organ-on-a-chip technology to highly simulate the dynamic tumor immune microenvironment in vivo. Through multiple cycles of "training," it effectively activates and expands tumor-specific T cells, avoiding immune exhaustion, and providing a highly efficient and controllable new platform for personalized immunotherapy of solid tumors.

The invention discloses a peripheral blood lymphocyte culture medium and a preparation method of a synchronization kit, and relates to the technical field of cell culture media, the peripheral blood lymphocyte culture medium comprises an RPMI-1640 culture solution, fetal calf serum, PHA phytolectin, HEPES free acid, penicillin, streptomycin, interleukin-2 and L-glutamic acid; basic nutrition and growth factors are provided through an RPMI-1640 culture solution and fetal calf serum, PHA phytolectin is used for stimulating cell proliferation, HEPES free acid maintains pH stability of the culture medium, penicillin and streptomycin prevent bacterial infection, interleukin-2 promotes proliferation and activation of T cells, L-glutamic acid serves as important amino acid to support cell metabolism, and the culture medium has the advantages that the culture medium can be used for promoting T cell proliferation and activation; the composition of the components is used for culturing peripheral blood lymphocytes and is beneficial to providing an optimal culture environment.

According to the method and device for predicting recurrence after HCC local treatment, multi-time-point peripheral blood lymphocyte subgroups are utilized, an integrated lymphocyte subgroup index is established through related factor screening and model optimization, and clinical information is assisted to establish an immune-clinical combined model to evaluate microwave ablation prognosis of liver cancer patients, so that liver cancer MWA postoperative recurrence and death risks are predicted in the early stage; the dynamic change of the lymphatic subgroup of the patient is longitudinally evaluated for multiple times through the verification queue to serve as input information to optimize the model, and guidance is provided for MWA preoperative planning and prognosis dynamic monitoring. The method comprises the following steps: (1) screening and constructing an accurate prediction model based on a circulating lymphatic subgroup; (2) explaining immune level change characteristics of the patient after ablation through the model, and performing prognosis risk assessment; and (3) verifying the efficacy of the post-treatment lymphatic subgroup dynamic monitoring and prediction model and re-optimizing the model.

Provided herein are methods of making tumor reactive peripheral blood lymphocytes (trPBLs) using PBMCs obtained from a patient. The method comprises enriching trPBLs from PBMCs using markers that are expressed or secreted by the trPBLs, such as 4- IBB, CD103, CXCL13, IFN-y, CXCR6, CD200, HLA-DR, CD69, PD-1, CD39, CD38, CD49a, etc. Further provided are use of the trPBLs or pharmaceutical compositions thereof for the treatment of cancers.

The invention discloses a screening method and application of a shark-derived anti-idiotypic nano antibody, and belongs to the technical field of biological pharmacy and vaccines. The screening method comprises the following steps: immunizing adult shark by using a target pathogen specific antigen, collecting peripheral blood, separating lymphocytes, extracting RNA (Ribonucleic Acid), carrying out reverse transcription to synthesize cDNA (Complementary Deoxyribonucleic Acid), amplifying a nano antibody variable region fragment of a shark IgNAR (Immunoglobulin Nucleic Acid Receptor) gene, and constructing a phage display gene bank of which the storage capacity is greater than or equal to 1 * 10 < 8 > CFU and the recombination rate is greater than or equal to 85%; a variable region of a target pathogen specific antibody is taken as a target, specific bacteriophage is enriched through 3-4 rounds of gradient washing and elutriation (Tween-20 concentration and washing time are gradually improved), and the shark source anti-idiotypic nano antibody with high affinity and high purity (greater than or equal to 95%) is obtained by combining ELISA screening positive clone, gene sequencing de-weighting and SPR detection (combining dissociation constant KDlt, 1 * 10 <-7 > mol / L).

The invention relates to the technical field of biological pharmacy, in particular to a human serum albumin targeted nano antibody and application thereof. An alpaca is immunized by using an HSA protein, cDNA is obtained through reverse transcription of total RNA of peripheral blood lymphocytes to serve as a template to amplify a nano antibody sequence, finally, five nano antibodies are obtained through separation and are named as antiHSA-A6, antiHSA-A7, antiHSA-B1, antiHSA-E8 and antiHSA-H7 respectively, the five nano antibodies have different antigen complementarity determining regions, and an ELISA result shows that the five nano antibodies have different antigen complementarity determining regions. And the five nano antibodies all show high-affinity binding with human HSA (Human Serum Albumin) protein. The nano antibody provided by the invention is expected to be used for preparing vaccines for treating and / or diagnosing diseases related to HSA, or for prolonging the half-life period of macromolecular drugs in vivo, and has important significance.

The invention discloses a peripheral blood lymphocyte culture standardization process, and relates to the technical field of cell culture, and the peripheral blood lymphocyte culture standardization process comprises the following steps: S1, blood sample planting; s2, cell culture; s3, harvesting the cells; s4, wafer dropping; s5, carrying out banding treatment; and S6, scanning and analyzing. The standardized process intends to standardize the whole peripheral blood chromosome preparation process, manual errors are reduced through instrument operation, experience judgment is replaced with specific numerical values, the standard operation process can avoid the slide preparation gap caused by human factors, then the peripheral blood chromosome karyotype analysis resolution is improved, a guarantee is provided for the reliability of clinical results, and the method is suitable for large-scale popularization and application. The step is the key of cytogenetics research, and detailed cell and chromosome data are provided; the standardized process aims to ensure that high-quality data output can be achieved in the culture, treatment and analysis process of the lymphocytes, so that the final analysis result is accurate and reliable.

Methods of expanding peripheral blood lymphocytes (PBLs) from blood of patients with hematological malignancies, including lymphomas and leukemias, genetic modifications of expanded PBLs to incorporate chimeric antigen receptors, genetically modified T cell receptors, and other genetic modifications, and uses of such expanded and / or modified PBLs in the treatment of diseases such as cancers and hematological malignancies are disclosed herein.

The present invention provides novel processes, compositions, and methods for analyzing or assaying the potency and / or functionality of tumor infiltrating lymphocyte (TIL) products for use in therapy, including human cancer therapy, and analyzing or assaying the potency and / or functionality of other polyclonal products, such as marrow infiltrating lymphocyte (MIL) and peripheral blood lymphocyte (PBL) products. Compositions, methods, and kits for preparing and treating cancer using TIL, MIL, and PBL products are also provided.

The present invention provides novel processes, compositions, and methods for analyzing or assaying the potency and / or functionality of tumor infiltrating lymphocyte (TIL) products for use in therapy, including human cancer therapy, and analyzing or assaying the potency and / or functionality of other polyclonal products, such as marrow infiltrating lymphocyte (MIL) and peripheral blood lymphocyte (PBL) products. Compositions, methods, and kits for preparing and treating cancer using TIL, MIL, and PBL products are also provided.

The invention discloses application of a detection reagent in malignant differentiation detection of Carbosi sarcoma, and belongs to the technical field of biological medicine. Aiming at the problem of insufficient specificity of a malignant differentiation marker of Carbosi sarcoma in the prior art, the invention provides application of a detection reagent in detection of the malignant differentiation of Carbosi sarcoma, and the detection reagent can simultaneously detect two biomarkers, namely CXCL12 and VWF, and is used for identifying endothelial cell malignant differentiation subgroups in tissue focuses of Carbosi sarcoma. By detecting the expression of CXCL12 and VWF in a tested sample, the endothelial cell malignant differentiation subgroup can be stably identified, and meanwhile, the expression level of CXCL12 can be used as an indicator of disease progression: the higher the expression level of CXCL12 is, the higher the lesion progression degree is, the higher the risk is, and the expression level is also positively correlated with the counting of peripheral blood lymphocytes. According to the invention, high-specificity recognition and standardized reading of the endothelial cell malignant differentiation subgroups are realized.

According to the present disclosure, optimization of treatment for a rejection reaction by immune tolerance induction is provided.According to the present disclosure, there is provided a method for evaluating quality of a medicament for achieving immune tolerance, which is administered to a patient who has undergone living-donor liver transplantation, the method including: (a) a step of mixing and culturing peripheral blood lymphocytes collected from a patient before the living-donor liver transplantation and peripheral blood lymphocytes collected from a donor of the living-donor liver transplantation; (b) a step of mixing and culturing peripheral blood lymphocytes collected from the patient before the living-donor liver transplantation, peripheral blood lymphocytes collected from the donor of the living-donor liver transplantation, and the medicament for achieving immune tolerance in the patient; and (c) a step of measuring cytokine production in steps (a) and (b), in which in a case where the cytokine production in step (b) has changed compared to step (a), the medicament is considered to be a medicament for immune tolerance therapy.

The invention relates to the technical field of biological pharmacy, in particular to a nano antibody with high affinity with a human DR3 extracellular domain. The method comprises the following steps: immunizing alpaca by using a DR3 extracellular domain of an insect expression system, separating peripheral blood lymphocytes, performing total RNA extraction, performing reverse transcription, amplifying a nano-antibody sequence, and finally separating to obtain a nano-antibody named as H10. The SPR (Surface Plasmon Resonance) result shows that the nano antibody can be combined with the extracellular domain of human DR3 with high affinity, and the nano antibody provided by the invention is expected to provide an experimental factual basis for the treatment thought of DR3-related diseases.

The invention discloses a marker, a primer combination and a kit for epilepsy diagnosis / detection and application, and belongs to the technical field of minute biology. Wherein the marker is one or a combination of two of a QSOX1 gene and an ABCC1 gene. The primer sequences contained in the primer combination are as shown in SEQ ID NO.1-6. The invention finds that the mRNA expression level of QSOX1 and ABCC1 in peripheral blood lymphocytes is obviously higher than that of a normal group in epilepsy patients, and verifies that the change of the expression level can reflect epilepsy conditions. Based on the detection kit or the primer combination developed by the invention, the mRNA level change of the peripheral blood lymphocytes QSOX1 and ABCC1 is detected, so that the epilepsy illness condition can be reflected, and the detection kit or the primer combination can also be used for early warning of epilepsy diagnosis. The detection result is more accurate and reliable, and the stability and repeatability are good.

The present invention provides a method for expanding tumor-infiltrating lymphocytes (TILs) (including peripheral blood lymphocytes and bone marrow-infiltrating lymphocytes) from the blood and / or bone marrow of patients with hematological malignancies (e.g., liquid tumors, including lymphomas and leukemias), and the use of the TILs so expanded in treating diseases (e.g., cancer and hematological malignancies).

The present application relates to the technical field of biopharmaceuticals, and particularly relates to a kind of nanobody with high affinity to human DR3 extracellular domain.The present application immunizes alpaca with the extracellular domain of DR3 of insect expression system, separates peripheral blood lymphocyte to carry out total RNA extraction, reverses transcription and amplifies nanobody sequence, finally separates and obtains nanobody, and is named H10.SPR result shows that the nanobody shows high affinity binding to human DR3 extracellular domain, and the nanobody provided by the present application has the potential to provide experimental fact basis for the treatment idea of DR3 related diseases.

The application provides an HE chromatin control product, a preparation method and application thereof, including a control slide, the control slide includes a first smear area and a second smear area, the first smear area and the second smear area are respectively arranged in different areas of the control slide, the first smear area is uniformly coated with mammalian peripheral blood mature red blood cells, and the second smear area is uniformly coated with mammalian peripheral blood lymphocytes. The HE chromatin control product provided by the application includes the combination of the mature red blood cell first smear area and the lymphocyte second smear area, can respectively and specifically reflect the staining effect of cytoplasm and nucleus in HE staining, realizes independent quality control of nucleus and plasma, the blood cell source is uniform, the difference of human body structure can be eliminated, batch uniformity and stability can be ensured, quantitative detection can be realized through a microspectrophotometer, objective and quantitative staining quality can be realized, the subjective and standardized difficulty problem of traditional quality control can be solved, and the accuracy and consistency of pathological quality control are improved.

The invention belongs to the field of biological medicine, and discloses a culture method and application of an organoid. The culture method is easy to operate and high in success rate. The organoid constructed by the method can be used for constructing an in-vitro organoid and peripheral blood lymphocyte co-culture system to evaluate the curative effect of tumor immunotherapy, the co-culture system predicts that AUC which is possibly invalid in immunotherapy is 0.82 (95% CI, 0.71-0.93), the sensitivity can reach 90.90%, the specificity is 73.50%, the positive predictive value is 77.42%, and the negative predictive value is 88.98%, which are obviously higher than PD-L1 (the sensitivity is 63.60%; the specificity is 37.20%), a new screening index is provided for determination of a precise immunotherapy scheme, and the treatment effect of a patient is improved.

This invention discloses a shark-derived nanobody targeting the SARS-CoV-2 RBD protein and its applications. The nanobody's nucleotide sequence is shown in SEQ ID NO: 1, and its amino acid sequence is shown in SEQ ID NO: 5. Preparation involves immunizing a striped bamboo shark with the RBD protein, extracting total RNA from peripheral blood lymphocytes and reverse transcribing it into cDNA. The amplified VNAR fragment is then seamlessly ligated with amplified pR2 phage particles, transformed into TG1 competent cells, and a phage antibody library is constructed. SARS-CoV-2 RBD protein-specific nanobody sequences are screened from the library and recombinantly expressed to obtain the targeted shark-derived nanobody. The nanobody of this invention has a small molecular weight, allowing for the detection of the SARS-CoV-2 RBD antigen using paired nanobodies. It can be applied to immunoblotting, enzyme-linked immunosorbent assays (ELISA), and COVID-19 pharmaceutical applications related to the SARS-CoV-2 RBD protein.

The invention discloses application of radix trichosanthis polysaccharide to preparation of a medicine for regulating immunity and inhibiting postoperative recurrence of colorectal cancer. The radix trichosanthis polysaccharide is found to be capable of promoting M2 phenotype macrophages to be polarized to M1 phenotype, and has an immunoregulation function; meanwhile, proliferation of peripheral blood lymphocytes and spleen lymphocytes is promoted, the proportion of CD4 +, CD8 + and NK cells is increased, and the effect of enhancing immunity is achieved. Furthermore, the hydrogel CMOT prepared by taking the radix trichosanthis polysaccharide as the raw material has good biocompatibility in vitro and in vivo and has the qualified characteristic of hydrogel, and the CMOT hydrogel is found to have the effect of inhibiting the recurrence of the colorectal cancer when being applied to the resection part of the mouse colorectal cancer operation. Therefore, the radix trichosanthis polysaccharide hydrogel has the potential of becoming a postoperative wound dressing after cancer resection.

The invention discloses an application of a reagent for detecting B cell surface markers in peripheral blood in preparation of a diagnostic kit, which is characterized in that on the basis of spectral flow data of B lymphocytes in peripheral blood, a chronic graft versus host disease patient characteristic subgroup is found, and eight surface markers of the subgroup are clearly identified; wherein three surface markers can be used for accurately defining a B cell characteristic subgroup of the chronic graft versus host disease patient; establishing a monitoring system for distinguishing a chronic graft versus host disease patient and a non-chronic graft versus host disease patient: cGPS; the cGPS system can be used for dynamically monitoring the condition of a chronic graft versus host disease patient and accurately judging whether the patient is in the active period of the disease or not; meanwhile, the three surface markers can also be used for diagnosing the health state of a human body and effectively distinguishing health from sub-health.

The invention relates to the technical field of biological pharmacy, in particular to a DR3 extracellular domain targeting nano antibody. The method comprises the following steps: immunizing alpaca by using a DR3 extracellular domain of an insect expression system, separating peripheral blood lymphocytes, carrying out total RNA extraction, carrying out reverse transcription, amplifying a nano-antibody sequence, and finally separating to obtain the nano-antibody named as A6. The SPR (Surface Plasmon Resonance) result shows that the nano antibody can be combined with the extracellular domain of human DR3 with high affinity, and the nano antibody provided by the invention is expected to provide an experimental factual basis for the treatment thought of DR3-related diseases.

The present application relates to the technical field of biopharmaceuticals, and particularly relates to a nano-antibody targeting DR3 extracellular domain. The present application immunizes a llama with DR3 extracellular domain of an insect expression system, extracts total RNA from peripheral blood lymphocytes, reversely transcribes and amplifies nano-antibody sequences, and finally separates and obtains a nano-antibody, named A6. SPR results show that the nano-antibody exhibits high affinity binding with human DR3 extracellular domain, and the nano-antibody provided by the present application is expected to provide experimental fact basis for the treatment idea of DR3 related diseases.

The invention belongs to the field of medicine, and particularly relates to a marker for evaluating the severity and clinical outcome of systemic lupus erythematosus disease, and the systemic lupus erythematosus disease is lupus nephritis, namely LN; the marker comprises a first combined marker, and the first combined marker is the number of CD3 + TIM3 + T lymphocytes or the proportion of CD3 + TIM3 + T lymphocytes in the total amount of peripheral blood T lymphocytes of a patient; the marker can also be one or more of a first combined marker CD3 + TIM3 + T lymphocyte, a second combined marker CD4 + CD25 + CD127-T lymphocyte, a third combined marker CD3 + LAG3 + T lymphocyte and a fourth combined marker CD3 + CTLA4 + T lymphocyte. The marker for evaluating the severity of the lupus nephritis disease is more accurate than an SLE traditional index, and the marker can be used for evaluating the clinical outcome of the lupus nephritis.