CN111568930A - Application of MSC (mesenchymal stem cell) in adjusting number of NK (natural killer) cells - Google Patents
Application of MSC (mesenchymal stem cell) in adjusting number of NK (natural killer) cells Download PDFInfo
- Publication number
- CN111568930A CN111568930A CN202010444831.3A CN202010444831A CN111568930A CN 111568930 A CN111568930 A CN 111568930A CN 202010444831 A CN202010444831 A CN 202010444831A CN 111568930 A CN111568930 A CN 111568930A
- Authority
- CN
- China
- Prior art keywords
- msc
- cells
- cell
- preparation
- patients
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 210000004027 cell Anatomy 0.000 title claims description 50
- 210000002901 mesenchymal stem cell Anatomy 0.000 title abstract description 84
- 210000000822 natural killer cell Anatomy 0.000 claims abstract description 46
- 238000002360 preparation method Methods 0.000 claims abstract description 39
- 238000011282 treatment Methods 0.000 claims abstract description 39
- 230000009385 viral infection Effects 0.000 claims abstract description 26
- 208000036142 Viral infection Diseases 0.000 claims abstract description 22
- 238000011084 recovery Methods 0.000 claims abstract description 16
- 239000003814 drug Substances 0.000 claims abstract description 8
- 239000006285 cell suspension Substances 0.000 claims description 15
- 230000029087 digestion Effects 0.000 claims description 15
- 210000003954 umbilical cord Anatomy 0.000 claims description 15
- 230000001105 regulatory effect Effects 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 10
- 238000011284 combination treatment Methods 0.000 claims description 8
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 claims description 7
- 108010019160 Pancreatin Proteins 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 6
- 229940055695 pancreatin Drugs 0.000 claims description 6
- 239000012089 stop solution Substances 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 239000000725 suspension Substances 0.000 claims description 6
- 238000012546 transfer Methods 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 5
- 238000012797 qualification Methods 0.000 claims description 5
- 238000002648 combination therapy Methods 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 238000012163 sequencing technique Methods 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 3
- 238000001514 detection method Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 210000005259 peripheral blood Anatomy 0.000 claims description 3
- 239000011886 peripheral blood Substances 0.000 claims description 3
- 239000002504 physiological saline solution Substances 0.000 claims description 3
- 238000005070 sampling Methods 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 238000009472 formulation Methods 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 210000004271 bone marrow stromal cell Anatomy 0.000 claims 20
- 210000005087 mononuclear cell Anatomy 0.000 claims 1
- 241000700605 Viruses Species 0.000 abstract description 16
- 230000001965 increasing effect Effects 0.000 abstract description 5
- 229940079593 drug Drugs 0.000 abstract description 3
- 230000028709 inflammatory response Effects 0.000 abstract 1
- 230000008092 positive effect Effects 0.000 abstract 1
- 210000001519 tissue Anatomy 0.000 description 11
- 208000015181 infectious disease Diseases 0.000 description 9
- 201000010099 disease Diseases 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 241001678559 COVID-19 virus Species 0.000 description 5
- 206010061218 Inflammation Diseases 0.000 description 5
- 230000036541 health Effects 0.000 description 5
- 230000004054 inflammatory process Effects 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 241000725303 Human immunodeficiency virus Species 0.000 description 4
- 210000001744 T-lymphocyte Anatomy 0.000 description 4
- 206010069351 acute lung injury Diseases 0.000 description 4
- 230000000840 anti-viral effect Effects 0.000 description 4
- 230000006907 apoptotic process Effects 0.000 description 4
- 210000000130 stem cell Anatomy 0.000 description 4
- 208000024891 symptom Diseases 0.000 description 4
- 241000711573 Coronaviridae Species 0.000 description 3
- 102000001398 Granzyme Human genes 0.000 description 3
- 108060005986 Granzyme Proteins 0.000 description 3
- KHGNFPUMBJSZSM-UHFFFAOYSA-N Perforine Natural products COC1=C2CCC(O)C(CCC(C)(C)O)(OC)C2=NC2=C1C=CO2 KHGNFPUMBJSZSM-UHFFFAOYSA-N 0.000 description 3
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 3
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 3
- 229930192851 perforin Natural products 0.000 description 3
- 102000003390 tumor necrosis factor Human genes 0.000 description 3
- 208000025721 COVID-19 Diseases 0.000 description 2
- 206010008909 Chronic Hepatitis Diseases 0.000 description 2
- 241000700721 Hepatitis B virus Species 0.000 description 2
- 206010037660 Pyrexia Diseases 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 230000035876 healing Effects 0.000 description 2
- 208000006454 hepatitis Diseases 0.000 description 2
- 230000009390 immune abnormality Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 241000712461 unidentified influenza virus Species 0.000 description 2
- 208000010444 Acidosis Diseases 0.000 description 1
- 206010001052 Acute respiratory distress syndrome Diseases 0.000 description 1
- 101150013553 CD40 gene Proteins 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 206010050685 Cytokine storm Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 241000701022 Cytomegalovirus Species 0.000 description 1
- 201000011001 Ebola Hemorrhagic Fever Diseases 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 101150089023 FASLG gene Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 208000029433 Herpesviridae infectious disease Diseases 0.000 description 1
- 102000018713 Histocompatibility Antigens Class II Human genes 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 1
- 208000032420 Latent Infection Diseases 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 108091054438 MHC class II family Proteins 0.000 description 1
- 206010027417 Metabolic acidosis Diseases 0.000 description 1
- 208000025370 Middle East respiratory syndrome Diseases 0.000 description 1
- 102000004503 Perforin Human genes 0.000 description 1
- 108010056995 Perforin Proteins 0.000 description 1
- 208000037581 Persistent Infection Diseases 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 208000013616 Respiratory Distress Syndrome Diseases 0.000 description 1
- 206010040070 Septic Shock Diseases 0.000 description 1
- 201000003176 Severe Acute Respiratory Syndrome Diseases 0.000 description 1
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 1
- 206010070863 Toxicity to various agents Diseases 0.000 description 1
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 201000000028 adult respiratory distress syndrome Diseases 0.000 description 1
- 230000000735 allogeneic effect Effects 0.000 description 1
- -1 and CD86 Proteins 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 230000022534 cell killing Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 210000001612 chondrocyte Anatomy 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 206010052015 cytokine release syndrome Diseases 0.000 description 1
- 230000007402 cytotoxic response Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000005860 defense response to virus Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 208000017574 dry cough Diseases 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 230000008143 early embryonic development Effects 0.000 description 1
- 210000003981 ectoderm Anatomy 0.000 description 1
- 210000003038 endothelium Anatomy 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 208000002672 hepatitis B Diseases 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 208000021601 lentivirus infection Diseases 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 210000003041 ligament Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 210000003716 mesoderm Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 210000004287 null lymphocyte Anatomy 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 210000001778 pluripotent stem cell Anatomy 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000012372 quality testing Methods 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 230000036303 septic shock Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000002636 symptomatic treatment Methods 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0665—Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
- C12N2509/10—Mechanical dissociation
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Developmental Biology & Embryology (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Animal Behavior & Ethology (AREA)
- Cell Biology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Hematology (AREA)
- Virology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Rheumatology (AREA)
- Oncology (AREA)
- Microbiology (AREA)
- Communicable Diseases (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention specifically discloses an application of MSC (mesenchymal stem cell) in adjusting the number of NK cells, in particular to an application of MSC in adjusting the number of NK cells of a virus infected patient, relating to the technical field of biomedical engineering. The prepared MSC preparation is added into an MSC treatment group to carry out combined medication treatment on patients with viral infection, so that the number of NK cells of the patients after recovery is effectively increased, and the MSC preparation has a positive effect on the inflammatory response of the patients after recovery.
Description
Technical Field
The invention relates to the technical field of biomedical engineering, in particular to application of MSC (mesenchymal stem cell) in regulating the number of NK (natural killer) cells.
Background
Natural killer cells (NK cells), which are a third class of lymphocytes that lack specific markers for T and B cells, such as TCR and SmIg, on their surface, were previously referred to as null cells. These cells, independent of antigen stimulation, can spontaneously lyse a variety of tumor cells and virus-infected cells, called natural killer cells, which are mainly present in peripheral blood and spleen, accounting for 5% to 10% of lymphocytes in human peripheral blood. NK cell surface receptors (NKRs) can recognize polysaccharide molecules expressed on the surface of cells infected with viruses. The killing effect is mediated by toxic molecules released after activation, such as perforin, granzyme and TNFa (tumor necrosis factor). The target cells for NK cell killing are mainly tumor cells, virus infected cells, larger pathogens (such as fungi and parasites), allograft organs, tissues and the like. It has wide spectrum antitumor effect, especially on lymphoma and leukemia cells. NK can release perforin and granzyme, and the perforin perforates the surface of a target cell so that granzyme b enters the target cell to induce apoptosis of the target cell. At the same time, a large amount of cytokines such as ifn-v, tnf-x, gm-csf, il-3, m-csf and the like are secreted, and directly act on target cells or attack the target cells by further activating other kinds of immune cells. And can express protein (fasl) capable of inducing apoptosis and TRAIL (TRAIL) for inducing apoptosis of tumor necrosis factor, so as to make target cell enter into programmed apoptosis state.
Viral infection refers to infectious diseases caused by viruses that can parasitize and propagate in the human body and cause diseases. Mainly shows general toxic symptoms such as fever, headache, general malaise and the like and local symptoms caused by inflammatory injury caused by virus hosts and invading tissues and organs. Viral infections of the human body are classified into recessive infections, dominant infections, and lentivirus infections. In most cases, the infection is recessive (after the infection of the human body with the virus, no symptoms appear, but specific antibodies can be produced). A few are dominant infections (meaning that symptoms appear after the human body is infected with viruses). The majority of the dominant infections are acute infections, the onset of diseases is acute, the course of diseases is short, and the minority of the dominant infections are latent infections (such as herpes virus infections) and chronic infections (such as hepatitis B virus infections). At present, the virus infection diseases lack specific treatment, mainly support and symptomatic treatment, and antiviral drugs and hormone treatment cannot play an effective treatment role. And viral infections (such as influenza, SARS, MERS and ebola) spread rapidly in humans, posing a significant threat to human health. Especially, the novel coronavirus which is outbreaked worldwide in 2020 causes serious loss to human health and economy.
The etiology of COVID-19 has been shown to be a novel coronavirus, now known as Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). At present, no specific medicine or vaccine aiming at the virus is successfully developed, and the appearance of SARS-CoV-2 brings a difficult treatment dilemma to clinicians. Most infected patients exhibit non-specific symptoms such as fever, dry cough, and fatigue. The prognosis of most patients is good, and some patients with severe disease can rapidly develop acute respiratory distress syndrome, septic shock, metabolic acidosis, blood coagulation dysfunction and even die. The deterioration of the patient's condition may be due to a cytokine storm in the body, and since SARS-CoV-2 is a novel strain that causes a global pandemic, there is an urgent need for effective targeted therapies, including antiviral and immunotherapy. Currently, although many clinical trials have begun, there is currently no effective antiviral or immunomodulatory therapy for treating COVID-19.
Clinical studies have found that MSC is an effective tool for treating diseases associated with Human Immunodeficiency Virus (HIV) immune abnormality, Hepatitis B Virus (HBV) chronic hepatitis, influenza virus Acute Lung Injury (ALI), and the like. Mesenchymal Stem Cells (MSCs) refer to a group of pluripotent stem cells with diverse differentiation potential that have differentiated into osteoblasts, chondroblasts and adipoblasts. They are derived from immature embryonic connective tissues of mesoderm and ectoderm in early embryonic development stage, and can differentiate into various tissue cells such as fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, endothelium and even blood under specific induction conditions in vitro and in vivo. Clinical studies have found that MSC is an effective tool for treating diseases associated with Human Immunodeficiency Virus (HIV) immune abnormality, Hepatitis B Virus (HBV) chronic hepatitis, influenza virus Acute Lung Injury (ALI), and the like. MSCs have low immunogenicity as well as significant and extensive immune-regulatory functions. MSCs do not express co-stimulatory molecules such as CD40, CD80, and CD86, as well as MHC-class II molecules. The T cell inhibitory effect of MSCs is not restricted by MHC, i.e. the effect on autologous and allogeneic T cells is similar. The antiviral response is critical to eradicate the virus and prevent the development of virus-related diseases. If antiviral specific T cells are allowed to function in the presence of MSCs, they have the ability to maintain the integrity of the host's defense against infection. EB virus-specific cytotoxic T Cells (CTL) or cytomegalovirus CTL cultured together with MSC have been found to maintain the ability to proliferate and produce interferon-in vitro and have a killing effect on virus-infected cells. MSC-derived interferon-is thought to counteract the immunosuppressive effects of MSCs by mediating a partial cytotoxic response during viral infection. Therefore, when using MSCs as regenerative medicine, it is very important to recognize the dual role of MSCs in fighting viral infections of the immune system.
Because the cured virus infected patient still has a certain degree of inflammation after the virus turns negative, how to participate in the inflammatory reaction in the recovery period after the cure by increasing the number of NK cells of the cured virus infected patient is a technical problem to be solved urgently by the technical staff in the technical field.
Disclosure of Invention
The invention aims to provide application of MSC (mesenchymal stem cell) in regulating the number of NK cells, in particular application of MSC in regulating the number of NK cells of a patient with viral infection. By adding the MSC preparation into the MSC treatment group for combined treatment, the number of NK cells of the healed patient is effectively increased.
The invention provides an application of MSC (mesenchymal stem cell) in regulating the number of NK (natural killer) cells of a virus infection patient, which comprises the following specific steps:
s1, preparing an MSC preparation;
s2, screening patients with viral infection and bringing the screened patients with viral infection into an MSC treatment group for combination treatment;
and S3, sampling and analyzing the peripheral blood mononuclear cell sample of the healed patient, thereby obtaining the number of NK cells of the healed patient.
Preferably, the preparation method of the MSC preparation comprises:
s11, collecting an umbilical cord, placing the umbilical cord in a culture dish, and then cleaning an umbilical cord tissue through physiological saline;
s12, cutting the cleaned umbilical cord tissue into small tissue blocks and planting the small tissue blocks in a culture dish for culture;
s13, removing the culture solution in the culture dish, cleaning the culture dish by using normal saline, adding pancreatin for digestion, adding a stop solution to stop digestion until the cells in the culture dish are digested, transferring the cell suspension into a centrifuge tube for centrifugation, discarding supernatant, resuspending the cells by using a proper amount of culture solution, counting, and finally planting the cell suspension into a new culture dish for culture according to a counting result;
s14, removing the culture solution in the new culture dish in the step S13, then washing with normal saline, adding pancreatin for digestion, adding a stop solution to stop digestion after cell digestion in the culture dish is finished, filtering with a cell sieve, transferring the filtered cell suspension into a centrifuge tube, counting, centrifuging, discarding supernatant, preparing cell preparation suspension, adding cell preparation suspension for cell suspension resuspension, transferring the cell suspension into a transfer bag, and putting the transfer bag into a low-temperature environment for taking, thereby completing preparation of the MSC preparation;
s15, carrying out qualification detection on the MSC preparation prepared in the step S14.
Preferably, the combination therapy in step S2 is: the MSC preparation prepared in step S1 is incorporated into an MSC combination treatment regimen for combination administration to a patient suffering from viral infection.
Preferably, the combination in the combination therapy refers to: MSC preparation is added in the original treatment scheme as a unique interference factor, and the condition of the combined medicine is tracked and recorded in the process of combined treatment.
Preferably, the post-cure viral infection patient includes a post-cure viral infection patient who is included in the MSC combination treatment regimen and a post-cure viral infection patient who is not included in the MSC combination treatment regimen.
Preferably, the step S3 is to analyze the sample of peripheral blood mononuclear cells by single cell sequencing, so as to obtain the number of NK cells of the patient after healing.
Compared with the prior art, the prepared MSC preparation is incorporated into the MSC treatment group to carry out combined treatment on the virus infection patients, thereby effectively increasing the number of NK cells of the healed patients, participating in the post-healing inflammatory reaction and the antiviral process in the recovery period after healing and playing a positive role.
Drawings
FIG. 1 is a flow chart of an application of MSC for regulating NK cell number of a patient with viral infection in the present invention,
figure 2 is a flow chart of a method of preparing the MSC formulation of the present invention,
FIG. 3 is a graph showing the ratio of the number of NK cells in the total number of grouped cells in patients after MSC-treated group recovery, patients without MSC-treated group recovery and healthy group persons in the present invention,
FIG. 4 is a multiple of the ratio of NK cell numbers of each group in the present invention with respect to the healthy group, wherein the ratio of NK cell numbers of healthy group persons was set to 1.
Detailed Description
In order to make the technical solutions of the present invention better understood, the present invention is further described in detail below with reference to the accompanying drawings.
It should be noted that the original treatment plan in the present invention refers to a treatment plan adopted in a novel coronavirus pneumonia diagnosis and treatment plan issued by the office of the national Health and care committee and the office of the national traditional Chinese medicine administration, MSC + represents an MSC treatment group, MSC-represents a non-MSC treatment group, and Health represents a healthy group.
As shown in figure 1, the application of MSC for regulating the number of NK cells of a virus infection patient comprises the following specific steps:
s1, preparing an MSC preparation;
s2, screening patients with viral infection and bringing the screened patients with viral infection into an MSC treatment group for combination treatment;
and S3, sampling and analyzing the peripheral blood mononuclear cells of the healed patient, thereby obtaining the number of NK cells of the healed patient.
As shown in fig. 2, the preparation method of the MSC preparation comprises:
s11, collecting an umbilical cord, placing the umbilical cord in a culture dish, and then cleaning an umbilical cord tissue through physiological saline;
s12, cutting the cleaned umbilical cord tissue into small tissue blocks and planting the small tissue blocks in a culture dish for culture;
s13, removing the culture solution in the culture dish, cleaning the culture dish by using normal saline, adding pancreatin for digestion, adding a stop solution to stop digestion until the cells in the culture dish are digested, transferring the cell suspension into a centrifuge tube for centrifugation, discarding supernatant, resuspending the cells by using a proper amount of culture solution, counting, and finally planting the cell suspension into a new culture dish for culture according to a counting result;
s14, removing the culture solution in the new culture dish in the step S13, then washing with normal saline, adding pancreatin for digestion, adding a stop solution to stop digestion after cell digestion in the culture dish is finished, filtering with a cell sieve, transferring the filtered cell suspension into a centrifuge tube, counting, centrifuging, discarding supernatant, preparing cell preparation suspension, adding cell preparation suspension for cell suspension resuspension, transferring the cell suspension into a transfer bag, and putting the transfer bag into a low-temperature environment for taking, thereby completing preparation of the MSC preparation;
s15, carrying out qualification detection on the MSC preparation prepared in the step S14.
In this embodiment, the MSC preparation is prepared from Human umbilical cord mesenchymal stem cells (hiuc-MSC), the preparation process is performed in a sterilized clean bench, instruments and consumables and the like required in the preparation process are sterilized with 75% alcohol, and the collection or cells in the culture dish and the specific operation process need to be labeled in detail in the preparation process; meanwhile, the stem cells can be frozen between the step S13 and the step S14, and if the frozen stem cells need to be revived before the step S14 is performed on the stem cell preparation, the stem cell freezing and reviving operations both belong to the prior art, and are not described in detail herein.
In this embodiment, the qualification testing of the MSC preparation further includes quality testing of the collected material, warehousing testing of the primary cell bank of the umbilical cord mesenchymal stem cells, and warehousing testing of the main cell bank of the umbilical cord mesenchymal stem cells, and the step and standard of the qualification testing are consistent with those of the conventional passing testing, which is not repeated here.
The combined treatment in the MSC treatment group refers to that the MSC preparation prepared in the step S1 is brought into the MSC treatment group and is used as a unique interference factor in the original treatment scheme to carry out combined medication on patients with viral infection, and meanwhile, the situation of the combined medication is tracked and recorded in the combined treatment scheme5-5×108Cell number/ml, MSC preparation was administered by peripheral intravenous infusion throughout the MSC treatment group and was used several times. Wherein the number of times of use can be controlled between 1 and 10 times according to actual conditions.
In this embodiment, the selected common new coronary positive patients (i.e., SARS-CoV-2 virus infected patients) are equally divided into two groups and used as MSC treatment group samples and non-MSC treatment group (i.e., original treatment scheme) samples, after the virus infected patients in the two groups of samples are cured, the peripheral blood mononuclear cells of the cured patients are sampled, in order to better explain the working principle and technical effect of the present invention, the peripheral blood mononuclear cells of corresponding number of healthy group personnel are simultaneously selected and sampled, then the collected peripheral blood mononuclear cell samples are respectively subjected to group analysis by single cell sequencing and the analysis results are processed, and the NK cell number relationship between the MSC treatment group cured patients, the non-MSC treatment group cured patients and the healthy group personnel is obtained (see table 1). In other embodiments, other virally infected patients may be selected as samples for the MSC-treated group and the non-MSC-treated group.
TABLE 1 relationship of NK cell number between patients after MSC-treated group recovery, patients after MSC-untreated group recovery and healthy group personnel
Meanwhile, drawing a relationship schematic diagram of the number of NK cells between the healed patients and healthy group personnel in two groups of samples according to the processing result data after single cell sequencing analysis (as shown in fig. 3 and fig. 4, wherein the first represents Health, the second represents MSC +, and the third represents MSC-), the abscissa in fig. 3 represents the NK cells, the ordinate represents the proportion of the number of the NK cells in each group in the total number of the corresponding cells, the abscissa in fig. 4 represents the NK cells, and the ordinate represents the change multiple of the number ratio of the NK cells in each group relative to the healthy group.
As can be seen from Table 1, FIG. 3 and FIG. 4, the number of NK cells in the patients after the MSC-treated group was cured accounted for 20.57% of the total number of cells, the number of cells in the patients without the MSC-treated group was accounted for 15.93% of the total number of cells, the number of cells in the healthy group accounted for 9.103% of the total number of cells, compared with the healthy group, the number of NK cells of the patients after the MSC treatment group is cured and the patients without the MSC treatment group are increased, moreover, the number of NK cells of the patients after the MSC treatment group is 2.259 times that of the healthy group, the number of NK cells of the patients after the MSC treatment group is 1.292 times that of the patients without the MSC treatment group, and in view of the fact that the patients still have certain inflammation in the recovery period after the virus turns negative, while NK cells are involved in inflammatory reactions in the recovery stage of the patient after recovery, the patient after recovery after MSC treatment group combination treatment is more beneficial to the recovery of the recovery stage after recovery.
The use of the MSC of the present invention for regulating NK cell number is described in detail above. The principles and embodiments of the present invention are explained herein using specific examples, which are presented only to assist in understanding the core concepts of the present invention. It should be noted that, for those skilled in the art, it is possible to make various improvements and modifications to the present invention without departing from the principle of the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention.
Claims (8)
1. Use of MSC for modulating NK cell number.
2. The use of the MSCs of claim 1 for modulating NK cell number, wherein said MSCs are used for modulating NK cell number in a virally infected patient.
3. The use of the MSCs for modulating NK cell number as claimed in claim 2, wherein the specific steps of use of the MSCs for modulating NK cell number in a virally infected patient comprise:
s1, preparing an MSC preparation;
s2, screening patients with viral infection and bringing the screened patients with viral infection into an MSC treatment group for combination treatment;
and S3, sampling and analyzing the peripheral blood mononuclear cell sample of the healed patient, thereby obtaining the number of NK cells of the healed patient.
4. The use of the MSCs as claimed in claim 3 for modulating NK cell number, wherein the preparation of the MSC formulation comprises:
s11, collecting an umbilical cord, placing the umbilical cord in a culture dish, and then cleaning an umbilical cord tissue through physiological saline;
s12, cutting the cleaned umbilical cord tissue into small tissue blocks and planting the small tissue blocks in a culture dish for culture;
s13, removing the culture solution in the culture dish, cleaning the culture dish by using normal saline, adding pancreatin for digestion, adding a stop solution to stop digestion until the cells in the culture dish are digested, transferring the cell suspension into a centrifuge tube for centrifugation, discarding supernatant, resuspending the cells by using a proper amount of culture solution, counting, and finally planting the cell suspension into a new culture dish for culture according to a counting result;
s14, removing the culture solution in the new culture dish in the step S13, then washing with normal saline, adding pancreatin for digestion, adding a stop solution to stop digestion after cell digestion in the culture dish is finished, filtering with a cell sieve, transferring the filtered cell suspension into a centrifuge tube, counting, centrifuging, discarding supernatant, preparing cell preparation suspension, adding cell preparation suspension for cell suspension resuspension, transferring the cell suspension into a transfer bag, and putting the transfer bag into a low-temperature environment for taking, thereby completing preparation of the MSC preparation;
s15, carrying out qualification detection on the MSC preparation prepared in the step S14.
5. The use of the MSC of claim 4 for regulating NK cell number, wherein the combination therapy of step S2 is: the MSC preparation prepared in step S1 is incorporated into an MSC combination treatment regimen for combination administration to a patient suffering from viral infection.
6. The use of the MSC of claim 5 for regulating NK cell number wherein the combination in combination therapy is: MSC preparation is added in the original treatment scheme as a unique interference factor, and the condition of the combined medicine is tracked and recorded in the process of combined treatment.
7. The use of the MSC of claim 6 for regulating the number of NK cells, wherein the patients after recovery comprise patients with viral infection cured in the MSC-treated group and patients with viral infection cured in the non-MSC-treated group.
8. The use of the MSC in claim 7 for regulating the number of NK cells, wherein the step S3 is performed by analyzing the mononuclear cell sample of peripheral blood by single cell sequencing to obtain the number of NK cells of the healed patient.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010444831.3A CN111568930A (en) | 2020-05-23 | 2020-05-23 | Application of MSC (mesenchymal stem cell) in adjusting number of NK (natural killer) cells |
| PCT/CN2020/105751 WO2021237930A1 (en) | 2020-05-23 | 2020-07-30 | Application of mesenchymal stem cells in treatment of patients with viral infection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010444831.3A CN111568930A (en) | 2020-05-23 | 2020-05-23 | Application of MSC (mesenchymal stem cell) in adjusting number of NK (natural killer) cells |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111568930A true CN111568930A (en) | 2020-08-25 |
Family
ID=72121311
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010444831.3A Pending CN111568930A (en) | 2020-05-23 | 2020-05-23 | Application of MSC (mesenchymal stem cell) in adjusting number of NK (natural killer) cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111568930A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101626772A (en) * | 2007-03-22 | 2010-01-13 | 奥西里斯治疗公司 | Mesenchymal stem cells and their uses |
| CN110540959A (en) * | 2019-10-08 | 2019-12-06 | 孟明耀 | Umbilical cord mesenchymal stem cell isolation culture amplification method |
| CN111514169A (en) * | 2020-05-23 | 2020-08-11 | 湖南源品细胞生物科技有限公司 | Application of MSC (Mobile switching center) in inhibiting virus replication |
-
2020
- 2020-05-23 CN CN202010444831.3A patent/CN111568930A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101626772A (en) * | 2007-03-22 | 2010-01-13 | 奥西里斯治疗公司 | Mesenchymal stem cells and their uses |
| CN110540959A (en) * | 2019-10-08 | 2019-12-06 | 孟明耀 | Umbilical cord mesenchymal stem cell isolation culture amplification method |
| CN111514169A (en) * | 2020-05-23 | 2020-08-11 | 湖南源品细胞生物科技有限公司 | Application of MSC (Mobile switching center) in inhibiting virus replication |
Non-Patent Citations (3)
| Title |
|---|
| BING LIANG等: "Clinical remission of a critically ill COVID-19 patient treated by human umbilical cord mesenchymal stem cells" * |
| WEN WEN等: ""Immune cell profiling of COVID-19 patients in the recovery stage by single-cell sequencing" * |
| 古利明等: "人脐带间充质干细胞联合抗病毒等方法治疗新型冠状病毒肺炎" * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111557951A (en) | Novel method for treating virus infection patient by mesenchymal stem cells | |
| US20240197848A1 (en) | Therapeutic vaccine for treatment of diabetes type 1 in children, application of the cell sorter and the method of multiplying treg cells to produce therapeutic vaccine for treatment of diabetes type 1 | |
| CN111514169A (en) | Application of MSC (Mobile switching center) in inhibiting virus replication | |
| JP2025072455A (en) | Method for producing highly pure and highly efficient naturally killed cells and its use | |
| CN109731002B (en) | Application of sapogenin R in preparation of hematopoietic stem cell mobilizing agent | |
| CN111568928A (en) | Application of MSC (mesenchymal stem cell) in regulating and controlling gene expression of effector CD8+ T cells | |
| CN111568931A (en) | Application of MSC (mesenchymal stem cell) in regulating and controlling human body effect CD4+ T memory cell gene expression | |
| CN113109570B (en) | Method for evaluating anti-cytomegalovirus effect of NK (Natural killer) cells | |
| CN111568930A (en) | Application of MSC (mesenchymal stem cell) in adjusting number of NK (natural killer) cells | |
| WO1989005657A1 (en) | Lymphokine activation of cells for adoptive immunotherapy, e.g. of hiv infection | |
| CN111529551A (en) | Application of MSC (mesenchymal stem cell) in adjusting number of plasma cells | |
| CN111557950A (en) | Application of MSC (mesenchymal stem cell) in regulating and controlling NK (natural killer) cell gene expression | |
| CN112852731A (en) | Method for inducing hematopoietic stem cells to differentiate into regulatory T cells in vitro | |
| CN112057474A (en) | Application of mesenchymal stem cells in regulating and controlling mononuclear cells of virus infection patient | |
| CN102512448B (en) | Application of DC-CIK (Dendritic Cell-Cytokine-Induced Killer) cells in preparation of medicine used for resisting HIV (Human Immunodeficiency Virus) infection | |
| CN111568927A (en) | Application of MSC (mesenchymal stem cell) in regulating number of memory B cells | |
| CN111544452A (en) | Application of MSC (mesenchymal stem cell) in regulating number of CD4+ T cells | |
| CN111568929A (en) | Application of a MSC for regulating the number of effector CD8+ T cells | |
| US20220081678A1 (en) | Use of mesenchymal stem cells in the treatment of viral infections and/or complications caused by viral infections | |
| CN108853129B (en) | Application of cimicin in preparation of myelogenous cytostatic drug | |
| US20210308190A1 (en) | Human umbilical cord blood mesenchymal stem cell transfusion immunotherapy for treatment of cytokine storm associated with coronavirus infection | |
| CN102488678B (en) | Application of embelin to preparation of medicament for treating autoimmune disease | |
| WO2021237930A1 (en) | Application of mesenchymal stem cells in treatment of patients with viral infection | |
| CN119842608B (en) | A method for improving the killing power of NK cells derived from umbilical cord blood and its application | |
| CN110656084B (en) | BAK cell and preparation kit and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200825 |
|
| RJ01 | Rejection of invention patent application after publication |