CN117987297B - 一株肠道菌及其在抗肿瘤免疫治疗中的应用 - Google Patents
一株肠道菌及其在抗肿瘤免疫治疗中的应用 Download PDFInfo
- Publication number
- CN117987297B CN117987297B CN202311603539.1A CN202311603539A CN117987297B CN 117987297 B CN117987297 B CN 117987297B CN 202311603539 A CN202311603539 A CN 202311603539A CN 117987297 B CN117987297 B CN 117987297B
- Authority
- CN
- China
- Prior art keywords
- tumor
- mice
- anaerobic
- ibiome007
- cells
- 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.)
- Active
Links
- 230000000968 intestinal effect Effects 0.000 title abstract description 21
- 241000894006 Bacteria Species 0.000 title description 19
- 238000011398 antitumor immunotherapy Methods 0.000 title description 3
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 101
- 241000186216 Corynebacterium Species 0.000 claims abstract description 64
- 241001505572 Anaerostipes caccae Species 0.000 claims abstract description 35
- 210000004027 cell Anatomy 0.000 claims description 70
- 238000011282 treatment Methods 0.000 claims description 30
- 230000028327 secretion Effects 0.000 claims description 27
- 239000003814 drug Substances 0.000 claims description 20
- 239000012636 effector Substances 0.000 claims description 14
- 238000004321 preservation Methods 0.000 claims description 13
- 230000004614 tumor growth Effects 0.000 claims description 13
- 239000008194 pharmaceutical composition Substances 0.000 claims description 12
- 206010009944 Colon cancer Diseases 0.000 claims description 10
- 238000000338 in vitro Methods 0.000 claims description 9
- 201000001441 melanoma Diseases 0.000 claims description 9
- 206010006187 Breast cancer Diseases 0.000 claims description 8
- 208000026310 Breast neoplasm Diseases 0.000 claims description 8
- 210000004698 lymphocyte Anatomy 0.000 claims description 8
- 229940124597 therapeutic agent Drugs 0.000 claims description 8
- 208000029742 colonic neoplasm Diseases 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 230000001737 promoting effect Effects 0.000 claims description 7
- 239000012228 culture supernatant Substances 0.000 claims description 6
- 210000001035 gastrointestinal tract Anatomy 0.000 claims description 6
- 238000001727 in vivo Methods 0.000 claims description 6
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 claims description 5
- 239000012271 PD-L1 inhibitor Substances 0.000 claims description 5
- 239000002246 antineoplastic agent Substances 0.000 claims description 5
- 229940121656 pd-l1 inhibitor Drugs 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 4
- 230000002265 prevention Effects 0.000 claims description 4
- 239000002775 capsule Substances 0.000 claims description 3
- 239000002552 dosage form Substances 0.000 claims description 3
- 239000008187 granular material Substances 0.000 claims description 3
- 210000004400 mucous membrane Anatomy 0.000 claims description 3
- 239000000725 suspension Substances 0.000 claims description 3
- 239000003826 tablet Substances 0.000 claims description 3
- 230000001093 anti-cancer Effects 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 210000003171 tumor-infiltrating lymphocyte Anatomy 0.000 claims description 2
- 102000008096 B7-H1 Antigen Human genes 0.000 claims 2
- 108010074708 B7-H1 Antigen Proteins 0.000 claims 2
- 230000000694 effects Effects 0.000 abstract description 29
- 238000000034 method Methods 0.000 abstract description 10
- 230000004044 response Effects 0.000 abstract description 6
- 238000009169 immunotherapy Methods 0.000 abstract description 5
- 230000005809 anti-tumor immunity Effects 0.000 abstract description 3
- 201000011510 cancer Diseases 0.000 abstract description 3
- 241000699670 Mus sp. Species 0.000 description 80
- 101100519207 Mus musculus Pdcd1 gene Proteins 0.000 description 32
- 241000699666 Mus <mouse, genus> Species 0.000 description 28
- 102100037850 Interferon gamma Human genes 0.000 description 20
- 108010074328 Interferon-gamma Proteins 0.000 description 20
- 230000002496 gastric effect Effects 0.000 description 19
- 230000002550 fecal effect Effects 0.000 description 18
- 210000003608 fece Anatomy 0.000 description 15
- 230000012010 growth Effects 0.000 description 15
- 230000001580 bacterial effect Effects 0.000 description 14
- 238000003304 gavage Methods 0.000 description 14
- 239000003242 anti bacterial agent Substances 0.000 description 11
- 239000002207 metabolite Substances 0.000 description 11
- 229940088710 antibiotic agent Drugs 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 10
- 230000002401 inhibitory effect Effects 0.000 description 10
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 9
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 9
- 238000002347 injection Methods 0.000 description 9
- 239000007924 injection Substances 0.000 description 9
- 208000001382 Experimental Melanoma Diseases 0.000 description 8
- 238000011068 loading method Methods 0.000 description 8
- 230000003115 biocidal effect Effects 0.000 description 7
- 238000007920 subcutaneous administration Methods 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 108020004465 16S ribosomal RNA Proteins 0.000 description 6
- 241000252852 Campylobacter faecalis Species 0.000 description 6
- 229930193140 Neomycin Natural products 0.000 description 6
- 210000001744 T-lymphocyte Anatomy 0.000 description 6
- 108010059993 Vancomycin Proteins 0.000 description 6
- 229960000723 ampicillin Drugs 0.000 description 6
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 229960000282 metronidazole Drugs 0.000 description 6
- VAOCPAMSLUNLGC-UHFFFAOYSA-N metronidazole Chemical compound CC1=NC=C([N+]([O-])=O)N1CCO VAOCPAMSLUNLGC-UHFFFAOYSA-N 0.000 description 6
- 229960004927 neomycin Drugs 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 229960001572 vancomycin hydrochloride Drugs 0.000 description 6
- LCTORFDMHNKUSG-XTTLPDOESA-N vancomycin monohydrochloride Chemical compound Cl.O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 LCTORFDMHNKUSG-XTTLPDOESA-N 0.000 description 6
- 238000000684 flow cytometry Methods 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 230000001965 increasing effect Effects 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 210000002784 stomach Anatomy 0.000 description 5
- 238000002560 therapeutic procedure Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 230000000259 anti-tumor effect Effects 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 210000005206 intestinal lamina propria Anatomy 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 150000003384 small molecules Chemical class 0.000 description 4
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 241001529936 Murinae Species 0.000 description 3
- 230000000903 blocking effect Effects 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 230000002708 enhancing effect Effects 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000002808 molecular sieve Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 3
- 230000000638 stimulation Effects 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 2
- 208000034309 Bacterial disease carrier Diseases 0.000 description 2
- 238000009631 Broth culture Methods 0.000 description 2
- 238000011740 C57BL/6 mouse Methods 0.000 description 2
- 102000008203 CTLA-4 Antigen Human genes 0.000 description 2
- 108010021064 CTLA-4 Antigen Proteins 0.000 description 2
- 229940045513 CTLA4 antagonist Drugs 0.000 description 2
- 108010022366 Carcinoembryonic Antigen Proteins 0.000 description 2
- 102100025475 Carcinoembryonic antigen-related cell adhesion molecule 5 Human genes 0.000 description 2
- 102000029816 Collagenase Human genes 0.000 description 2
- 108060005980 Collagenase Proteins 0.000 description 2
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 2
- 241000305071 Enterobacterales Species 0.000 description 2
- 229940076838 Immune checkpoint inhibitor Drugs 0.000 description 2
- 108091008036 Immune checkpoint proteins Proteins 0.000 description 2
- 102000037982 Immune checkpoint proteins Human genes 0.000 description 2
- 238000012404 In vitro experiment Methods 0.000 description 2
- 108091008026 Inhibitory immune checkpoint proteins Proteins 0.000 description 2
- 102000037984 Inhibitory immune checkpoint proteins Human genes 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 206010027480 Metastatic malignant melanoma Diseases 0.000 description 2
- 241000288906 Primates Species 0.000 description 2
- 241000283984 Rodentia Species 0.000 description 2
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 2
- 229910000831 Steel Inorganic materials 0.000 description 2
- 229940125644 antibody drug Drugs 0.000 description 2
- 238000011394 anticancer treatment Methods 0.000 description 2
- 229940041181 antineoplastic drug Drugs 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 229940044683 chemotherapy drug Drugs 0.000 description 2
- 229960002424 collagenase Drugs 0.000 description 2
- 230000009849 deactivation Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000005746 immune checkpoint blockade Effects 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 239000012274 immune-checkpoint protein inhibitor Substances 0.000 description 2
- 230000001024 immunotherapeutic effect Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 208000021039 metastatic melanoma Diseases 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 210000000822 natural killer cell Anatomy 0.000 description 2
- 239000003504 photosensitizing agent Substances 0.000 description 2
- 239000012221 photothermal agent Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000010959 steel Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 239000001974 tryptic soy broth Substances 0.000 description 2
- 238000000108 ultra-filtration Methods 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- WLDRFJDGKMTPPV-UHFFFAOYSA-N 1,2-dimethylhydrazine dihydrochloride Chemical compound Cl.Cl.CNNC WLDRFJDGKMTPPV-UHFFFAOYSA-N 0.000 description 1
- 235000011470 Adenanthera pavonina Nutrition 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 0.000 description 1
- 108020000946 Bacterial DNA Proteins 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 241001608472 Bifidobacterium longum Species 0.000 description 1
- WNBCMONIPIJTSB-BGNCJLHMSA-N Cichoriin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1)c1c(O)cc2c(OC(=O)C=C2)c1 WNBCMONIPIJTSB-BGNCJLHMSA-N 0.000 description 1
- 241000193403 Clostridium Species 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 1
- 201000008808 Fibrosarcoma Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000605909 Fusobacterium Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 1
- 102100033096 Interleukin-17D Human genes 0.000 description 1
- 108010066979 Interleukin-27 Proteins 0.000 description 1
- 208000008839 Kidney Neoplasms Diseases 0.000 description 1
- 241000428378 Lopa Species 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 1
- 208000015634 Rectal Neoplasms Diseases 0.000 description 1
- 206010038389 Renal cancer Diseases 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- AWUCVROLDVIAJX-UHFFFAOYSA-N alpha-glycerophosphate Natural products OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 description 1
- 229960001931 ampicillin sodium Drugs 0.000 description 1
- KLOHDWPABZXLGI-YWUHCJSESA-M ampicillin sodium Chemical compound [Na+].C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C([O-])=O)(C)C)=CC=CC=C1 KLOHDWPABZXLGI-YWUHCJSESA-M 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 238000002869 basic local alignment search tool Methods 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940009291 bifidobacterium longum Drugs 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000002619 cancer immunotherapy Methods 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- YTRQFSDWAXHJCC-UHFFFAOYSA-N chloroform;phenol Chemical compound ClC(Cl)Cl.OC1=CC=CC=C1 YTRQFSDWAXHJCC-UHFFFAOYSA-N 0.000 description 1
- 208000006990 cholangiocarcinoma Diseases 0.000 description 1
- 230000007012 clinical effect Effects 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 201000010897 colon adenocarcinoma Diseases 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 238000007877 drug screening Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- XHCADAYNFIFUHF-TVKJYDDYSA-N esculin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1)O)=CC2=C1OC(=O)C=C2 XHCADAYNFIFUHF-TVKJYDDYSA-N 0.000 description 1
- 229940093496 esculin Drugs 0.000 description 1
- AWRMZKLXZLNBBK-UHFFFAOYSA-N esculin Natural products OC1OC(COc2cc3C=CC(=O)Oc3cc2O)C(O)C(O)C1O AWRMZKLXZLNBBK-UHFFFAOYSA-N 0.000 description 1
- 201000004101 esophageal cancer Diseases 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000017188 evasion or tolerance of host immune response Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 102000034287 fluorescent proteins Human genes 0.000 description 1
- 108091006047 fluorescent proteins Proteins 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 210000003736 gastrointestinal content Anatomy 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 230000005934 immune activation Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000006058 immune tolerance Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000008944 intestinal immunity Effects 0.000 description 1
- 210000005025 intestinal intraepithelial lymphocyte Anatomy 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 201000010982 kidney cancer Diseases 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 201000004792 malaria Diseases 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000000581 natural killer T-cell Anatomy 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 231100000989 no adverse effect Toxicity 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 206010038038 rectal cancer Diseases 0.000 description 1
- 201000001275 rectum cancer Diseases 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- AWUCVROLDVIAJX-GSVOUGTGSA-N sn-glycerol 3-phosphate Chemical compound OC[C@@H](O)COP(O)(O)=O AWUCVROLDVIAJX-GSVOUGTGSA-N 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 108010050327 trypticase-soy broth Proteins 0.000 description 1
Classifications
-
- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- 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
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Genetics & Genomics (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Mycology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Endocrinology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明涉及癌症的免疫治疗领域。具体的,本发明涉及肠道特定菌株粪厌氧棒状菌(Anaerostipes caccae)ibiome007对抗肿瘤免疫的作用,并提供提高患者肿瘤免疫治疗响应率的方法。
Description
技术领域
本发明涉及癌症的免疫治疗领域。具体的,本发明涉及肠道特定菌株对抗肿瘤免疫的作用,并提供提高患者肿瘤免疫治疗响应率的方法。
背景技术
程序性死亡受体1(PD-1),是一种主要表达于活化的CD4和CD8 T细胞、B细胞、单核细胞、NK细胞和树突状细胞表面的免疫抑制分子,通过抑制T细胞炎症活动促进免疫耐受。肿瘤细胞通过上调PD-1的配体PD-L1,促进T细胞耗竭,进而实现免疫逃逸。近年来,针对程序性死亡1 (PD-1)及其配体(PD-L1)的抑制剂,由于在晚期恶性肿瘤中具有显著的临床效果,受到了广泛关注,并获得2018年生理学及医学奖。然而,免疫检查点阻断(immunecheckpoint blockade,ICB)的治疗效果目前仅局限于15-30%的癌症患者,仍有很大比例的患者对ICB治疗不响应。
小鼠的临床前研究表明,相比无特定病原体(SPF)小鼠,无菌(GF)荷瘤小鼠使用ICB对肿瘤无显著疗效。与此类似,对SPF小鼠进行抗生素处理能显著抑制ICB的疗效。此外,Matson等人通过对42名转移性黑色素瘤患者的ICB前粪便16s测序分析,发现长双歧杆菌在PD-1响应患者体内富集。在小鼠体内定植双歧杆菌协同PD-1阻断剂能够显著减小肿瘤体积(Vyara Matson, et al., The commensal microbiome is associated with anti-PD-1efficacy in metastatic melanoma patients. Science. 2018 Jan 5; 359(6371):104-108. doi: 10.1126/science.aao3290.)。这些结果表明,某些特定的肠道微生物类群可以通过与宿主免疫系统的相互作用调节抗PD1治疗的反应。因此,寻找特定的能增强PD1治疗疗效的菌株对肿瘤的ICB治疗至关重要,并在肿瘤的治疗领域具有广泛的应用前景。
发明内容
本发明提供一种肠道菌,能够通过增强CD8T细胞效应因子分泌能力,从而增加免疫检查点阻断疗法的疗效,达到改善肿瘤患者生存期的目的。
因此,发明人首先建立包含了人肠道微生物的主要类群的肠道菌库,通过培养上清液的IFN-γ+CD8T细胞刺激试验的体外筛选,筛选到了能提高机体免疫应答的粪厌氧棒状菌(Anaerostipes caccae)ibiome007,保藏于中国典型培养物保藏中心保藏,地址为湖北省武汉市武昌区八一路武汉大学中国典型培养物保藏中心,保藏日期2023年8月11日,保藏号为CCTCC M 20231456。
该菌株能在GAM肉汤中厌氧培养长出灰白色大菌落。其16s rRNA序列示于SEQ IDNo:1中。
在体外筛选中,该菌株来源的代谢物能够显著增强CD8T细胞IFN-γ分泌情况。在体外杀伤实验中,该菌株来源代谢物能提高CD8T细胞IFN-γ分泌能力以及IFN-γ阳性的CD8T细胞中,PD-1阳性和TNF-α阳性细胞占的比例。
所述粪厌氧棒状菌(Anaerostipes caccae)ibiome007能够减少荷瘤小鼠的肿瘤体积,提高抗肿瘤免疫治疗响应率,在受试者体内定植,有望增强基于抗-PD-1的肿瘤治疗效果。
本发明是通过以下技术方案实现的:
粪厌氧棒状菌(Anaerostipes caccae)ibiome007,在中国典型培养物保藏中心保藏,地址为湖北省武汉市武昌区八一路武汉大学中国典型培养物保藏中心,保藏日期2023年8月11日,保藏号为CCTCC M 20231456。
本发明还保护上述的粪厌氧棒状菌(Anaerostipes caccae)ibiome007在制备用于预防肿瘤产生或治疗肿瘤的药物中的用途,其中,所述粪厌氧棒状菌(Anaerostipes caccae)ibiome007优选为活菌或其代谢物,更优选为分子量小于3kDa的代谢物;当所述药物用于治疗肿瘤时可以单用或与其他抗癌治疗联合施用而增强抗癌效果。
优选地,所述预防肿瘤产生或治疗肿瘤包括但不限于如下至少一种表型:
(a)在体外促进CD8T细胞效应因子(如TNF-α和/或IFN-γ)的分泌;
(b)在体内促进肠道固有层淋巴细胞中CD8T细胞的效应因子TNF-α和/或IFN-γ的分泌;
(c)在体内促进肿瘤浸润淋巴细胞中CD8T细胞的效应因子TNF-α和/或IFN-γ的分泌;
(d)减缓肿瘤生长曲线;
(e)减小肿瘤体积;
(f)减轻肿瘤重量。
优选地,所述其他抗癌治疗包括抗PD-1治疗剂(优选为抗PD-1的抗体、CTLA-4抗体、PD-L1抗体和/或PD-L1抑制剂),或除了抗PD-1抗体以外的其他抗体药物(优选为化疗药物、光敏剂、光热剂、免疫治疗药物中的至少一种)。
进一步地,所述肿瘤为实体瘤(优选为选自由胃癌、食管癌、结肠癌、直肠癌、肝癌、胰腺癌、乳腺癌、肾癌、纤维肉瘤、肺癌、胆管癌、黑色素瘤组成的组中的一种或多种,更优选为结肠癌、黑色素瘤或乳腺癌)。
本发明还保护上述的粪厌氧棒状菌(Anaerostipes caccae)ibiome007在制备提高生物体(优选为哺乳动物,更优选为人)的免疫应答(优选为提高T细胞中CD8阳性T细胞的比率)的药物中的用途。
本发明还保护药物组合物,包含上述的粪厌氧棒状菌(Anaerostipes caccae)ibiome007、或所述粪厌氧棒状菌(Anaerostipes caccae)ibiome007的培养物或其加工物(优选为活菌、活菌代谢物,更优选为分子量小于3kDa的代谢物),和药学上可接受的载体。
优选地,所述药物组合物还包括抗PD-1治疗剂(优选为抗PD-1的抗体、CTLA-4抗体、PD-L1抗体或PD-L1抑制剂),或除了抗PD-1抗体以外的其他抗体药物(优选为化疗药物、光敏剂、光热剂、免疫治疗药物中的至少一种)。
优选地,所述药物组合物的剂型选自由片剂、颗粒剂、胶囊剂、悬液、冻干制剂组成的组。
本发明还保护提高生物体免疫应答的方法,包括对生物体施用上述的粪厌氧棒状菌(Anaerostipes caccae)ibiome007,或所述粪厌氧棒状菌(Anaerostipes caccae)ibiome007的培养物或其加工物,或上述的药物组合物。
本发明还保护检查上述的粪厌氧棒状菌(Anaerostipes caccae)ibiome007是否在生物体中定植的方法,包括从生物体的粪便或肠道提取物中提取细菌16s rRNA,并以其为模板,使用具有如SEQ ID No:2所示的序列的上游引物和具有如SEQ ID No:3所示的序列的下游引物进行PCR。
本发明所述抗PD-1治疗剂指通过阻止或抑制PD-1工作而发挥作用,使PD-1下游的信号阻止或抑制的物质,可以为抗PD-1的抗体或PD-L1抑制剂等。
在本发明的药物组合物中,可以包含本领域通常使用的药学上可接受的溶剂、乳化剂、悬浮剂、防腐剂、润滑剂等,但不限于此。
本发明的药物组合物可以制备为常用的剂型,例如片剂、颗粒剂、胶囊剂、悬液、冻干制剂等。
“包含上述的粪厌氧棒状菌(Anaerostipes caccae)ibiome007”不仅指包含所述粪厌氧棒状菌(Anaerostipes caccae)ibiome007的活菌形式,也包括菌体分解物、培养上清液,以及分离的所述菌的代谢产物。
受试者为哺乳动物,可以为啮齿类例如大鼠、小鼠等,也可以为灵长类,优选为灵长类动物,例如人。
本发明的菌株或含有其的药物组合物的用量没有特别限定,只要粪厌氧棒状菌(Anaerostipes caccae)ibiome007能够在受试者的肠道定植即可。当受试者为人时,以每次给予0.2mL计,可以以例如5×106~5×1012CFU/mL,优选为5×108~5×1010CFU/mL的浓度进行施用。接种频率或施用频率可以为每1~3天一次,优选2~3天一次。为了成功地在受试者肠道定植,连续施用时间例如为两周,一个月,需要时可以进行延长。施用方式可以为胃肠道方式给予,例如口服。可以通过在施用后检查受试者的粪便确认菌株的定植情况。
本发明的有益效果在于:
1.本菌株粪厌氧棒状菌ibiome007从健康志愿者体内分离,易于在体外培养,能够在小鼠肠道中定殖。
2.在体外实验中,该菌株能够增强CD8T细胞的效应因子IFN-γ分泌能力,提高CD8T细胞效应功能。
3.该菌株提高CD8T细胞效应功能主要依赖其产生的小分子代谢物。
4.在小鼠体内实验中,通过检测定殖后小鼠肠道CD8T细胞效应因子分泌情况,发现该菌株能促进小鼠LPL(固有淋巴细胞)细胞CD8T细胞分泌IFN-γ,结果与体外实验相吻合。
5.对SPF小鼠皮下荷瘤MC38,并灌胃粪厌氧棒状菌ibiome007,无论是否进行抗生素预处理,该菌株均能发挥肿瘤抑制功能,且能够联合PD-1抗体,发挥更优的肿瘤抑制功能,扩大了临床的用药范围,不受患者治疗之前使用抗生素的个体差异而影响药物疗效。
6.对无菌小鼠皮下荷瘤MC38,并灌胃粪厌氧棒状菌ibiome007,该菌株能够联合PD-1抗体,发挥肿瘤抑制功能,表明粪厌氧棒状菌单一菌株就能达到联合PD-1的功能。
7. 对SPF小鼠皮下荷瘤B16黑色素瘤,并灌胃粪厌氧棒状菌ibiome007,该菌株能发挥肿瘤抑制功能,且能够显著抑制B16黑色素瘤中浸润的CD8 T细胞表面程序性死亡受体1(PD-1),并显著增加LPL中的CD8 T细胞分泌肿瘤坏死因子-α(TNF-α)。
8.该菌株具有协同肿瘤免疫治疗、提高免疫检查点阻断疗法疗效、提高肿瘤患者anti-PD-1疗法响应率、提高患者生存期的潜力。
生物保藏
粪厌氧棒状菌(Anaerostipes caccae)ibiome007,保藏日期2023年8月11日,保藏地点为中国典型培养物保藏中心,地址为湖北省武汉市武昌区八一路武汉大学中国典型培养物保藏中心,保藏号为CCTCC M 20231456。
附图说明
图1为实施例3采用培养基对照及其小分子或粪厌氧棒状菌ibiome007培养上清液及其小分子刺激CD8T细胞后,使用流式细胞术检测CD8T细胞效应因子IFN-γ的分泌情况;其中,***代表p<0.001。
图2为实施例4中粪厌氧棒状菌ibiome007在SPF小鼠体内定殖实验结果。
图3为实施例5粪厌氧棒状菌ibiome007定殖对肠道免疫的影响;其中,3A为流式细胞术划门策略,3B为流式细胞术的统计结果图。
图4为实施例6粪厌氧棒状菌ibiome007对SPF小鼠MC38肿瘤生长的影响;其中,4A为肿瘤注射后各组的肿瘤生长曲线,4B为肿瘤注射后14天对小鼠肿瘤组织进行的剥离拍照图,4C为4B的肿瘤称重统计结果。
图5为实施例7粪厌氧棒状菌ibiome007对SPF小鼠MC38肿瘤生长的影响;其中,5A为肿瘤注射后各组的肿瘤生长曲线,5B为肿瘤注射后28天对小鼠肿瘤组织进行的剥离拍照图。
图6为实施例8粪厌氧棒状菌ibiome007对SPF小鼠皮下B16黑色素瘤生长的影响;其中,6A为肿瘤注射后各组的肿瘤生长曲线,6B为肿瘤注射后34天对小鼠肿瘤组织进行的剥离拍照图,6C显示粪厌氧棒状菌ibiome007能够显著抑制B16黑色素瘤中浸润的CD8 T细胞表面程序性死亡受体1(PD-1),6D显示粪厌氧棒状菌ibiome007能够显著增加LPL中的CD8T细胞分泌肿瘤坏死因子-α(TNF-α)。
图7为实施例9中粪厌氧棒状菌ibiome007在无菌小鼠体内定殖实验结果;其中,7A为无菌鼠粪便的涂布平板生长情况,7B为灌胃了粪厌氧棒状菌ibiome007的无菌鼠粪便的涂布平板生长情况,7C为7B的菌落鉴定结果。
图8为实施例10粪厌氧棒状菌ibiome007及联合PD-1对无菌(GF)小鼠肿瘤生长的影响。
图9为实施例11粪厌氧棒状菌ibiome007灌胃小鼠后的体重监测图。
图10为实施例12粪厌氧棒状菌ibiome007对SPF小鼠E0771乳腺癌肿瘤生长的影响;其中,10A为肿瘤注射后各组的肿瘤生长曲线,10B为肿瘤称重统计结果,10C为肿瘤注射后35天对小鼠肿瘤组织进行的剥离拍照图。
实施方式
为更好理解本发明,下面结合实施例及附图对本发明作进一步描述,以下实施例仅是对本发明进行说明而非对其加以限定。
实施例1:菌库建立
从10名健康成人、青少年、婴儿及1名PD-1阻断治疗响应患者粪便中分离培养单菌落:志愿者粪便样品用20%的甘油磷酸盐缓冲液保存,分别梯度稀释至10-5、10-6、10-7倍,将各浓度分别涂布于GAM肉汤(Solarbio LA4450)、BBE(拟胆汁七叶苷固体培养基,SolarbioLA7310),RCM(梭菌增菌培养基,青岛海博HB0316),MRS肉汤(青岛海博HB0384-1)、TSB(胰酪大豆胨液体培养基,青岛海博HB4114-19)、BHI(脑心浸出液培养基,OXOID,CM1136)、哥伦比亚血平板(比克曼生物)。挑单克隆至相应的液体培养基,通过16s rRNA通用引物(上游引物27F:AGAGTTTGATCCTGGCTCAG,下游引物1492R:TACGGCTACCTTGTTACGACTT)测序确定种属(扩增条件:预变性:95℃ 3min;[变性 95℃ 15s、退火 58℃ 15s、延伸 72℃ 30s] 35个循环;彻底延伸 72℃ 5min),用甘油于-80℃保存。将细菌利用37℃摇床培养至OD600=0.8,3000rpm离心10min,收集细菌培养上清液,-40℃冻存备用。共建立包含5个门、12个纲、15个目、30个科、1000株菌的菌库,包含了人肠道微生物的主要类群。
实施例2:IFN-γ+ CD8T细胞体外筛选
IFN-γ是一种通常由NK细胞、NKT细胞、CD4以及CD8T细胞表达的细胞因子,具有广泛的刺激和调节免疫系统的能力。IFN-γ+ CD8T细胞在抗肿瘤免疫中也起着至关重要的作用,并被报道能够影响免疫检查点抑制剂(ICI)疗法的疗效。Honda实验室的研究发现,从健康人体内分离的11种肠道菌的混合物能够通过上调CD8T细胞分泌IFN-γ的能力,在小鼠实验中,达到显著抑制肿瘤生长的作用。因此,能够刺激CD8T细胞IFN-γ分泌的肠道菌,可作为筛选潜在的抗肿瘤功能菌的指标。
然而,先前的研究采用的是用抗生素富集的方法,11种肠道菌的混合物具有氨苄青霉素抗性,有在服用抗生素情况下大量增殖的可能,在安全性方面具有风险。此外,由于先前的研究尚未确定具体的菌株,并且混合物中的梭杆菌对肿瘤的影响仍有争议,因此,寻找具体的、单一的作用菌株,具有重要意义。
从6-8周的IFN-γ荧光报告小鼠(在IFN-γ蛋白转录序列后插入IRES序列和katushka荧光蛋白序列,使IFN-γ表达的同时细胞表达katushka荧光,构建方法可参考Villegas-Mendez, A. et al. Parasite-Specific CD4+ IFN-γ+ IL-10+ T CellsDistribute within Both Lymphoid and Nonlymphoid Compartments and AreControlled Systemically by Interleukin-27 and ICOS during Blood-Stage MalariaInfection. Infection and immunity84, 34-46, doi:10.1128/iai.01100-15 (2016).)脾脏中分离CD8T细胞:脾脏组织去除表面脂肪,加入磷酸盐缓冲溶液,研磨成细胞悬液,用钢网过滤残余组织。通过500g、4℃离心5min,将细胞收集于管底,加入2mL红细胞裂解液去除红细胞。剩余细胞通过CD8T细胞磁珠分选试剂盒(Biolegend 480035)分离CD8T细胞。
对细胞进行计算,取3×105 的CD8T细胞加入anti-CD3抗体(终浓度5μg/mL)、anti-CD28抗体(终浓度2μg/mL)(分别为Biolegend 100223、Biolegend 102112)体外活化CD8T细胞,按3×105/孔分于96孔板。向孔中分别加入实施例1冻存的细菌上清液各20μL进行刺激,培养(即刺激)48h。
收集上述经刺激后的细胞(约1×106/管),每孔加入60 µL含有特异性表面标记荧光抗体的混合液(FITC-CD3 Biolegend 100204 0.2µL/孔,APC-Cy7 CD8 Biolegend100714 0.2µg/孔),4℃避光放置15-20min。加满1×PBS 洗一遍(6000rpm×2min)。
用200~300µL 1×PBS 重悬细胞,上样浓度在3~5×106/mL。细胞悬液经200目尼龙网过滤去除杂质,转至Ep管中,加DAPI瞬染(200倍稀释,博士德AR1177)上机检测,用Flowjo软件分析。以IFN-γ为例,先以FSC-H与FSC-A为横纵轴圈出单细胞,在单细胞门中圈出DAPI阴性的活细胞,之后圈出CD3阳性、CD8阳性的CD8T细胞,检测CD8T细胞IFN-γ分泌情况。
分析检测结果,发现一株健康志愿者来源、GAM肉汤培养基分离、并能在GAM肉汤厌氧培养、长出灰白色大菌落的菌株,该菌株来源的代谢物能够显著增强CD8T细胞IFN-γ分泌情况,以及提高IFN-γ阳性的CD8T细胞中TNF-α阳性细胞的比例。
经实施例1的方法测序得到该菌株的16s rRNA基因序列(SEQ ID NO.1)提交到NCBI Basic Local Alignment Search Tool,进行菌株16s rRNA基因分析。比对结果表明,与其相似性最高的菌株是Anaerostipes caccae strain L1-92 (相似度99.93%),因此该菌株属于粪厌氧棒状菌(Anaerostipes caccae)种,将其命名为粪厌氧棒状菌(Anaerostipes caccae) ibiome007,保藏至中国典型培养物保藏中心,保藏地址为:湖北省武汉市武昌区八一路珞珈山;保藏日期为:2023年8月11日;保藏编号为:CCTCC M20231456。
实施例3:粪厌氧棒状菌小分子代谢物增加体外培养的CD8 T细胞的IFN-γ分泌能力
收集粪厌氧棒状菌(Anaerostipes caccae)ibiome007上清液和GAM肉汤培养基,分别用0.2µm的滤膜过滤除菌,用孔径<3kDa的分子筛超滤,除去上清中的蛋白组分待用。准备4组样品:(1)GAM肉汤培养基,(2)经分子筛超滤后的GAM肉汤培养基,(3)粪厌氧棒状菌(Anaerostipes caccae)ibiome007上清液,(4)经分子筛超滤后的粪厌氧棒状菌(Anaerostipes caccae)ibiome007上清液。提前一天用4µg/mL的CD3抗体60µL铺板96孔板。
按照实施例2的方法,仅在刺激培养时替换加入上述4组样品,分别检测4组样品存在情况下CD8T细胞IFN-γ分泌情况。
结果如图1所示,粪厌氧棒状菌培养上清液具有促进CD8T细胞效应因子分泌的能力,且发挥功能的有效成分为细菌来源的小分子代谢物(<3kDa)。
实施例4:粪厌氧棒状菌(Anaerostipes caccae) ibiome007在SPF小鼠模型中的定殖效果检测
对购买自集萃药康C57/B6J小鼠喂ABX(1g/L氨苄青霉素、新霉素、甲硝唑,以及0.5g/L万古霉素盐酸盐)清菌2周(以ABX替换饮用水),随后分为两组:对照组灌胃PBS,定殖组灌胃5×106CFU/mL的菌液,各200μL,两天一次,一周后收集小鼠粪便,检测粪便中粪厌氧棒状菌含量。用20%(w/v)的SDS裂菌,用酚氯仿抽提细菌DNA,用菌株特异性引物SEQ ID No:2和SEQ ID No:3(F:TAGAGGCTTCGGTGCC,R:CCTGTCTCCAATGTCTCG)进行PCR,以16s rRNA通用引物(qPCR-27F:AGAGTTTGATCMTGGCTCAG,qPCR-342R:CTGCTGCSYCCCGTAG)为内参,检测菌株定殖效果。
结果如图2所示:与对照组相比,定殖组小鼠粪便中粪厌氧棒状菌ibiome007含量明显增高。表明灌胃后,小鼠肠道内粪厌氧棒状菌ibiome007水平增加,粪厌氧棒状菌ibiome007能在小鼠体内定殖。
实施例5:粪厌氧棒状菌(Anaerostipes caccae) ibiome007增加肠道固有层CD8T细胞的IFN-γ分泌能力
6-8周的集萃药康C57/B6J小鼠提前用四联抗生素(ABX)清除肠道菌:每250mL水分别加入0.25g氨苄青霉素、新霉素、甲硝唑,以及0.125g万古霉素盐酸盐,饲喂2周。之后换为普通水饮用,将小鼠随机分为2组:实验组灌胃5×108CFU/mL粪厌氧棒状菌(Anaerostipes caccae) ibiome007菌液,对照组灌胃PBS,各200μL。ibiome007定殖2周后,牺牲小鼠,用PBS冲洗肠道内容物并剖开肠管,肠子剪成1-2cm小段投入25mL预消化液中(RPMI 65mL,FBS1.2mL,EDTA 120μL),37℃,220rpm摇20min。将所得的培养基通过200目钢网过滤,肠段加入预消化液供后续处理,过滤后的培养基离心可得肠上皮细胞及上皮内淋巴细胞。重复上述步骤2次,剩余组织加入含胶原酶和DNA酶的消化液(RPMI 10mL,DNASE I STOCK 20μL,TypeII Collagenase 5mg (sigma),FBS 0.2mL),过滤后所得为肠道固有层淋巴细胞。对固有层淋巴细胞用特异性荧光抗体标记,流式上机检测IFN-γ分泌情况。
如图3所示,上图(3A)为流式细胞术的划门策略:分离小鼠肠道上皮细胞后,先圈出活细胞,进一步圈出CD45+的淋巴细胞,之后圈出CD3+的T细胞,进一步区分CD4和CD8,最后检测CD8 IFN-γ分泌情况。下图(3B)为流式细胞术的结果统计图,图中表明,与空白对照组相比,粪厌氧棒状菌灌胃组的小肠固有层内淋巴细胞中,CD8T细胞的效应因子IFN-γ和TNF-α分泌增多。
实施例6:粪厌氧棒状菌(Anaerostipes caccae) ibiome007对SPF小鼠的抗MC38肿瘤效应
MC38,小鼠结肠癌细胞,又名MCA38。来源于长期暴露于致癌物DMH (1,2-二甲基肼二盐酸盐)而患有结肠腺癌的C57BL6小鼠结肠肿瘤组织,并可高水平表达人癌胚抗原(CEA)。结肠直肠癌(CRC)是全球癌症死亡的第二大原因。而啮齿动物结肠癌模型研究是新药物临床前评估的有效方法。MC38细胞对免疫调节抗体具有良好的应答特性,证实了肿瘤微环境可修饰免疫激活。因此,MC38被定位为功能强大的免疫肿瘤学模型,在药物研发中具有重要的用途,常用于抗肿瘤药物、特别是抗结肠癌肿瘤药物在体内的免疫功能研究。
6-8周的集萃药康C57/B6J小鼠提前用四联抗生素(ABX)清除肠道菌,每250mL水分别加入0.25g氨苄青霉素、新霉素、甲硝唑,以及0.125g万古霉素盐酸盐,饲喂2周。对小鼠进行随机分组,分别为空白对照组、粪厌氧棒状菌组、PD-1抗体治疗组和粪厌氧棒状菌联合PD-1抗体治疗组共4组,粪厌氧棒状菌组和粪厌氧棒状菌联合PD-1抗体治疗组每天灌胃200μL 2.5×107CFU/mL的粪厌氧棒状菌活菌ibiome007菌液,对照组和PD-1抗体治疗组每天灌胃PBS。定殖2周后对所有组小鼠剃毛,皮下荷瘤MC38,3*105/只。荷瘤后隔天测量肿瘤体积,采用0.5*长*宽*宽近似计算肿瘤体积。荷瘤后7天、9天、11天,PD-1抗体治疗组和粪厌氧棒状菌联合PD-1抗体治疗组对小鼠腹腔注射anti-PD-1(BioXcell BE0146),注射剂量为100μg/只。荷瘤后第14天牺牲小鼠,对小鼠肿瘤进行拍照称重。如图4所示,粪厌氧棒状菌ibiome007能够联合PD-1抗体,达到抑制肿瘤生长的效果。具体来说,单独灌胃粪厌氧棒状菌ibiome007即可使肿瘤生长曲线明显变缓(图4A)、体积明显变小(图4B)、肿瘤重量明显减轻(图4C),且效果优于PD-1;当灌胃粪厌氧棒状菌ibiome007的同时还注射PD-1抗体,效果更佳,生长曲线(图4A)、体积(图4B)、重量(图4C)的效果均明显优于PD-1抗体治疗组。
实施例7:粪厌氧棒状菌(Anaerostipes caccae) ibiome007对SPF小鼠的抗MC38肿瘤效应不受原肠道菌群影响
将集萃药康C57/B6J雌性鼠适应性饲养一周后随机分为两组:灌胃PBS对照组和灌胃粪厌氧棒状菌治疗组。对灌胃粪厌氧棒状菌治疗组隔天灌胃粪厌氧棒状菌ibiome007菌粉,菌粉使用无菌PBS溶解,每只鼠灌胃200μL(5×106CFU)。灌胃PBS对照组每只鼠隔天灌胃200μL PBS。灌胃4次后对所有组小鼠剃毛,皮下荷瘤MC38,5*105/只。荷瘤第7天后开始测量肿瘤体积,此后每隔天测量一次,采用0.5*长*宽*宽近似计算肿瘤体积。在荷瘤后28天左右牺牲小鼠,对小鼠肿瘤进行拍照。结果如图5所示,粪厌氧棒状菌ibiome007能够抑制MC38肿瘤生长的效果。结合实施例6,无论是否使用抗生素对小鼠原来的肠道菌进行处理,ibiome007均体现出了良好的抗肿瘤效应。且不进行抗生素处理更倾向于临床实际治疗场景,大大提高了临床用药范围,即ibiome007的治疗疗效不受患者肠道菌的个体差异和抗生素等用药情况的影响。
实施例8:粪厌氧棒状菌(Anaerostipes caccae) ibiome007对SPF小鼠皮下B16黑色素瘤的抗肿瘤效应
B16,小鼠黑色素瘤细胞,是来源于C57BL/6小鼠皮肤的自发性肿瘤,此瘤株主要发生肺转移。B16细胞在小鼠体内的生长及转移特性与人黑色素瘤非常相似,为理想的黑色素瘤动物模型,已广泛地用于研究黑色素瘤发生发展、转移和药物筛选等研究。
将集萃药康C57/B6J雄性鼠适应性饲养一周后,用四联抗生素(ABX)清除肠道菌,每250mL水分别加入0.25g氨苄青霉素、新霉素、甲硝唑,以及0.125g万古霉素盐酸盐,饲喂3周。停用四联抗生素(ABX)后四天对小鼠进行随机分组:灌胃PBS对照组和灌胃粪厌氧棒状菌治疗组。对灌胃粪厌氧棒状菌治疗组隔天灌胃粪厌氧棒状菌ibiome007菌粉,菌粉使用无菌PBS溶解,每只鼠灌胃200μL(5×106CFU)。灌胃PBS对照组每只鼠隔天灌胃200μL PBS。灌胃8次后对所有组小鼠剃毛,皮下荷瘤B16,5*105/只。荷瘤第7天后开始测量肿瘤体积并记录,此后每隔天测量一次,采用0.5*长*宽*宽近似计算肿瘤体积。在荷瘤后34天左右牺牲小鼠,对小鼠肿瘤进行拍照。结果如图6A和6B所示,粪厌氧棒状菌ibiome007能够显著抑制B16黑色素瘤的生长效果。
拍照后将小鼠的肿瘤制成单细胞悬液,用于流式染色分析,结果如图6C所示,粪厌氧棒状菌ibiome007能够显著抑制B16黑色素瘤中浸润的CD8 T细胞表面程序性死亡受体1(PD-1)。同时取小鼠的结肠肠道黏膜固有层淋巴细胞(LPL),使用eBioscience™ CellStimulation Cocktail刺激4小时后,做流式染色分析,结果如图6D所示,粪厌氧棒状菌ibiome007能够显著增加LPL中的CD8 T细胞分泌肿瘤坏死因子-α(TNF-α)。表明粪厌氧棒状菌ibiome007发挥抑制肿瘤生长作用是由于其减少B16黑色素瘤中浸润的CD8 T细胞表面PD-1,从而使该组别的肿瘤细胞无法逃逸自身的免疫,同时粪厌氧棒状菌ibiome007使肠道固有层CD8 T细胞分泌 TNF-α,进一步杀伤肿瘤细胞。总体实验表明粪厌氧棒状菌ibiome007不仅对小鼠结肠癌瘤MC38的生长有抑制作用,也可以对小鼠黑色素瘤B16的生长有显著抑制作用。
实施例9:粪厌氧棒状菌(Anaerostipes caccae) ibiome007在无菌(GF)小鼠上的定殖
将集萃药康C57/B6J雄性GF鼠随机分为对照组和粪厌氧棒状菌定殖组,对照组不处理,粪厌氧棒状菌定殖组灌胃粪厌氧棒状菌ibiome007菌液,每只鼠200μL,5×106CFU/mL,一次。定殖后第五天对粪厌氧棒状菌定殖组小鼠进行细菌定殖检测(qPCR,同实施例4),对对照组小鼠进行无菌检测(取无菌鼠粪便至无菌水里,使用上清液在GAM肉汤培养基上涂平板)。
结果如图7所示,无菌鼠粪便没有细菌生长(图7A),粪厌氧棒状菌ibiome007定殖后,小鼠粪便可涂布出单一形态的菌落(图7B)。图7C的菌落鉴定结果表明,定殖后小鼠粪便中仅有粪厌氧棒状菌定殖。
实施例10:粪厌氧棒状菌(Anaerostipes caccae) ibiome007及联合PD-1抗体增强无菌(GF)小鼠抗MC38肿瘤效应
将集萃药康C57/B6J雄性GF鼠随机分为两组:PD-1抗体治疗组和粪厌氧棒状菌联合PD-1抗体治疗组。对粪厌氧棒状菌联合PD-1抗体治疗组灌胃粪厌氧棒状菌ibiome007,每只鼠200μL,5×106CFU/mL。定殖1周后对所有组小鼠剃毛,皮下荷瘤MC38,3*105/只。荷瘤后隔天测量肿瘤体积,采用0.5*长*宽*宽近似计算肿瘤体积。荷瘤后的7天、9天、11天、13天,对小鼠腹腔注射anti-PD-1,注射剂量为200 μg/只。在荷瘤后16天左右牺牲小鼠,对小鼠肿瘤进行拍照。结果如图8所示,粪厌氧棒状菌ibiome007能够联合PD-1抗体,达到进一步抑制肿瘤生长的效果,表明粪厌氧棒状菌ibiome007单一菌株就能达到联合PD-1的功能。
实施例11:粪厌氧棒状菌(Anaerostipes caccae) ibiome007具有良好的安全性
将集萃药康C57/B6J雄性鼠适应性饲养一周后,用四联抗生素(ABX)清除肠道菌,每250mL水分别加入0.25g氨苄青霉素、新霉素、甲硝唑,以及0.125g万古霉素盐酸盐,小鼠自由饮用24天。停用四联抗生素(ABX)后4天对小鼠进行随机分组:灌胃PBS对照组和灌胃粪厌氧棒状菌治疗组。对灌胃粪厌氧棒状菌治疗组隔天灌胃粪厌氧棒状菌ibiome007菌粉,菌粉使用无菌PBS溶解,每只鼠灌胃200μL(1×1012CFU)。灌胃PBS对照组每只鼠隔天灌胃200μLPBS。抗生素处理开始为第0天,第42天对所有组小鼠剃毛,皮下荷瘤B16,5*105/只,小鼠全程体重监测直至实验的第73天(图9)。监测期间随机间隔测量小鼠体重(计小数点后一位)。实验全程小鼠未出现任何不良反应,且两组小鼠的体重无显著性差异(P=0.55),初步说明该菌给与小鼠灌胃对小鼠无不良影响,具有较好的安全性。
实施例12:粪厌氧棒状菌(Anaerostipes caccae) ibiome007对SPF小鼠皮下E0771乳腺癌的抗肿瘤效应
将集萃药康C57/B6J雄性鼠适应性饲养一周后,用四联抗生素(ABX)清除肠道菌,每250mL水分别加入0.75g氨苄青霉素钠、新霉素、甲硝唑,以及0.375g万古霉素盐酸盐,灌胃1周。停用四联抗生素(ABX)十天后对小鼠进行随机分组:灌胃PBS对照组和灌胃粪厌氧棒状菌治疗组。对灌胃粪厌氧棒状菌治疗组以1周6次的频次灌胃粪厌氧棒状菌ibiome007菌粉,菌粉使用无菌PBS溶解,每只鼠灌胃200μL(106CFU)。灌胃PBS对照组每只鼠以1周6次的频次灌胃200μL PBS。灌胃7次后对所有组小鼠剃毛,皮下荷瘤E0771乳腺癌细胞,5*105/只。荷瘤第11天后开始测量肿瘤体积并记录,此后每2-3天测量一次肿瘤体积,采用0.5*长*宽*宽近似计算肿瘤体积。在荷瘤后第35天牺牲小鼠,对小鼠肿瘤进行拍照和称重。结果如图10所示,无论是从肿瘤整体的生长曲线(图10A)、体积(图10C)来看,还是从肿瘤最后的称重情况来看(图10B),实验均能证明粪厌氧棒状菌ibiome007能够显著抑制E0771乳腺癌生长的效果。总体实验表明粪厌氧棒状菌ibiome007不仅对小鼠结肠癌瘤MC38和小鼠黑色素瘤B16的生长有抑制作用,对小鼠乳腺癌E0771的生长也具有显著的抑制作用。
以上所述实施方式仅仅是对本发明的优选实施方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通技术人员对本发明的技术方案作出的各种变形和改进,均应落入本发明的权利要求书确定的保护范围内。
Claims (10)
1.粪厌氧棒状菌(Anaerostipes caccae)ibiome007,其特征在于:所述的粪厌氧棒状菌菌株在中国典型培养物保藏中心保藏,地址为湖北省武汉市武昌区八一路武汉大学中国典型培养物保藏中心,保藏日期2023年8月11日,保藏号为CCTCC NO:M 20231456。
2. 权利要求1所述的粪厌氧棒状菌(Anaerostipes caccae)ibiome007在制备用于预防肿瘤产生或治疗肿瘤的药物中的用途,其特征在于:所述肿瘤为结肠癌瘤、黑色素瘤或乳腺癌瘤。
3.根据权利要求2所述的用途,其特征在于,所述预防肿瘤产生或治疗肿瘤包括但不限于如下至少一种表型:
(a)在体外促进CD8T细胞效应因子的分泌;
(b)在体内促进肠道固有层淋巴细胞中CD8T细胞的效应因子TNF-α和/或IFN-γ的分泌;
(c)在体内促进肿瘤浸润淋巴细胞中CD8T细胞的效应因子TNF-α和/或IFN-γ的分泌;
(d)减缓肿瘤生长;
(e)减小肿瘤体积;
(f)减轻肿瘤重量。
4.根据权利要求2所述的用途,其特征在于:所述药物与其他抗癌治疗剂联合施用而增强抗癌效果,所述其他抗癌治疗剂为抗PD-1治疗剂。
5.根据权利要求4所述的用途,其特征在于:所述抗PD-1治疗剂为抗PD-1的抗体、PD-L1抗体和/或PD-L1抑制剂。
6. 权利要求1所述的粪厌氧棒状菌(Anaerostipes caccae)ibiome007或其培养上清液在制备促进CD8 T细胞分泌IFN-γ药物中的用途。
7. 药物组合物,其特征在于:包含权利要求1所述的粪厌氧棒状菌(Anaerostipes caccae)ibiome007,和药学上可接受的载体。
8.根据权利要求7所述的药物组合物,其特征在于:还包括抗PD-1治疗剂。
9.根据权利要求8所述的药物组合物,其特征在于:所述抗PD-1治疗剂为抗PD-1的抗体、PD-L1抗体和/或PD-L1抑制剂。
10.根据权利要求7或8或9所述的药物组合物,其特征在于:所述药物组合物的剂型选自片剂、颗粒剂、胶囊剂、悬液、冻干制剂。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311010511 | 2023-08-11 | ||
CN2023110105117 | 2023-08-11 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN117987297A CN117987297A (zh) | 2024-05-07 |
CN117987297B true CN117987297B (zh) | 2024-06-14 |
Family
ID=90887827
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202311603539.1A Active CN117987297B (zh) | 2023-08-11 | 2023-11-28 | 一株肠道菌及其在抗肿瘤免疫治疗中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN117987297B (zh) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114540229A (zh) * | 2021-04-22 | 2022-05-27 | 上海耀旦生物科技有限公司 | 一种增强免疫检查点抑制剂治疗效应的副干酪乳杆菌株及其应用 |
CN115806893A (zh) * | 2021-09-13 | 2023-03-17 | 中国科学技术大学 | 普通拟杆菌及其组合物在辅助癌症免疫治疗中的应用 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020136879A1 (ja) * | 2018-12-28 | 2020-07-02 | 国立大学法人大阪大学 | 悪性腫瘍を治療するための免疫療法剤の有用性の提示方法及び悪性腫瘍を治療するための免疫療法剤 |
-
2023
- 2023-11-28 CN CN202311603539.1A patent/CN117987297B/zh active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114540229A (zh) * | 2021-04-22 | 2022-05-27 | 上海耀旦生物科技有限公司 | 一种增强免疫检查点抑制剂治疗效应的副干酪乳杆菌株及其应用 |
CN115806893A (zh) * | 2021-09-13 | 2023-03-17 | 中国科学技术大学 | 普通拟杆菌及其组合物在辅助癌症免疫治疗中的应用 |
Also Published As
Publication number | Publication date |
---|---|
CN117987297A (zh) | 2024-05-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN110496140B (zh) | 脆弱拟杆菌或阿克曼粘细菌在制备用于预防或治疗肿瘤的药物中的应用 | |
MX2012008817A (es) | Composicion probiotica para uso en el tratamiento de la inflamacion del intestino. | |
CN115806893B (zh) | 普通拟杆菌及其组合物在辅助癌症免疫治疗中的应用 | |
JP2021524751A (ja) | 組成物及びその使用 | |
CN114540229A (zh) | 一种增强免疫检查点抑制剂治疗效应的副干酪乳杆菌株及其应用 | |
CN115969888B (zh) | 一株唾液联合乳杆菌及其在制备治疗癌症的药物中的应用 | |
EP3436065B1 (en) | Anti-cancer oncolytic virus combination therapies and elite responder selection platforms | |
JP2023539910A (ja) | 免疫チェックポイント阻害剤の増強剤の調製における細菌の使用 | |
Xu et al. | Lactobacillus casei JY300-8 generated by 12C6+ beams mutagenesis inhibits tumor progression by modulating the gut microbiota in mice | |
JP2024050573A (ja) | 腫瘍浸潤リンパ球(tils)を活性化する方法 | |
CN117987297B (zh) | 一株肠道菌及其在抗肿瘤免疫治疗中的应用 | |
CN117503802A (zh) | Ac菌在抗肿瘤免疫治疗中的应用 | |
CN116855418B (zh) | 一种具有预防和/或治疗炎症性肠病作用的产气柯林斯菌及其应用 | |
CN116103196B (zh) | 一种活泼瘤胃球菌及其应用 | |
CN117089490B (zh) | 青春双歧杆菌basj001及应用 | |
WO2024140860A1 (zh) | 克里斯滕森菌或包含其的组合物在预防或治疗肿瘤中的用途以及包含其的药物 | |
CN117025479B (zh) | 格氏乳杆菌shmb 0001及其在预防和缓解急性结肠炎中的应用 | |
JP2024025452A (ja) | 機能的免疫賦活乳酸菌 | |
Sobol | ANTINEOPLASTIC AND ANTIMUTAGENIC EFFECTS OF A NEW PROBIOTIC PRODUCT AT THE CELLULAR LEVEL AND IN RATS WITH TRANSPLANTED FAST-GROWING PLISS'LYMPHOSARCOMA | |
CN115747097A (zh) | 一种Bifidobacterium bifidum菌株及其在肿瘤辅助治疗中的应用 | |
KR20240123764A (ko) | 락토바실러스 플란타룸 균주 및 면역세포의 병용 요법을 이용한 암 예방 또는 치료용 조성물 | |
KR20230123429A (ko) | 암 예방 또는 치료용 약학적 조성물 | |
CN118185812A (zh) | 一种约氏乳杆菌及其应用 | |
CN115838662A (zh) | 一株具有缓解克罗恩病潜力的两歧双歧杆菌 | |
CN118440861A (zh) | 嗜黏蛋白阿克曼氏菌及其在抗肿瘤免疫治疗中的用途 |
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 | ||
GR01 | Patent grant | ||
GR01 | Patent grant |