CN1767847A - 用胸腺素α1治疗曲霉菌感染 - Google Patents
用胸腺素α1治疗曲霉菌感染 Download PDFInfo
- Publication number
- CN1767847A CN1767847A CN 200480008490 CN200480008490A CN1767847A CN 1767847 A CN1767847 A CN 1767847A CN 200480008490 CN200480008490 CN 200480008490 CN 200480008490 A CN200480008490 A CN 200480008490A CN 1767847 A CN1767847 A CN 1767847A
- Authority
- CN
- China
- Prior art keywords
- cell
- treatment
- dcs
- mice
- infection
- 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.)
- Granted
Links
- 108010078233 Thymalfasin Proteins 0.000 title abstract description 3
- 102400000800 Thymosin alpha-1 Human genes 0.000 title abstract description 3
- NZVYCXVTEHPMHE-ZSUJOUNUSA-N thymalfasin Chemical compound CC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O NZVYCXVTEHPMHE-ZSUJOUNUSA-N 0.000 title abstract description 3
- 229960004231 thymalfasin Drugs 0.000 title abstract description 3
- 201000002909 Aspergillosis Diseases 0.000 title description 14
- 208000036641 Aspergillus infections Diseases 0.000 title description 14
- 238000000034 method Methods 0.000 claims abstract description 24
- 208000015181 infectious disease Diseases 0.000 claims abstract description 18
- 238000010322 bone marrow transplantation Methods 0.000 claims abstract description 17
- 210000004443 dendritic cell Anatomy 0.000 claims abstract description 9
- 201000009085 invasive aspergillosis Diseases 0.000 claims description 16
- 230000000843 anti-fungal effect Effects 0.000 claims description 14
- 229940121375 antifungal agent Drugs 0.000 claims description 12
- 108010046075 Thymosin Proteins 0.000 claims description 11
- 102000007501 Thymosin Human genes 0.000 claims description 11
- LCJVIYPJPCBWKS-NXPQJCNCSA-N thymosin Chemical compound SC[C@@H](N)C(=O)N[C@H](CO)C(=O)N[C@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@H](C(C)C)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](C(C)C)C(=O)N[C@H](CO)C(=O)N[C@H](CO)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@H]([C@@H](C)CC)C(=O)N[C@H]([C@H](C)O)C(=O)N[C@H](C(C)C)C(=O)N[C@H](CCCCN)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@H](CCCCN)C(=O)N[C@H](CCCCN)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@H](C(C)C)C(=O)N[C@H](C(C)C)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@H](CCC(O)=O)C(O)=O LCJVIYPJPCBWKS-NXPQJCNCSA-N 0.000 claims description 11
- 102000004127 Cytokines Human genes 0.000 claims description 10
- 108090000695 Cytokines Proteins 0.000 claims description 10
- FIVPIPIDMRVLAY-UHFFFAOYSA-N aspergillin Natural products C1C2=CC=CC(O)C2N2C1(SS1)C(=O)N(C)C1(CO)C2=O FIVPIPIDMRVLAY-UHFFFAOYSA-N 0.000 claims description 8
- FIVPIPIDMRVLAY-RBJBARPLSA-N gliotoxin Chemical compound C1C2=CC=C[C@H](O)[C@H]2N2[C@]1(SS1)C(=O)N(C)[C@@]1(CO)C2=O FIVPIPIDMRVLAY-RBJBARPLSA-N 0.000 claims description 8
- 210000000447 Th1 cell Anatomy 0.000 claims description 7
- APKFDSVGJQXUKY-KKGHZKTASA-N Amphotericin-B Natural products O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1C=CC=CC=CC=CC=CC=CC=C[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-KKGHZKTASA-N 0.000 claims description 6
- 241000124008 Mammalia Species 0.000 claims description 6
- APKFDSVGJQXUKY-INPOYWNPSA-N amphotericin B Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/C=C/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-INPOYWNPSA-N 0.000 claims description 6
- 229960003942 amphotericin b Drugs 0.000 claims description 6
- 208000029462 Immunodeficiency disease Diseases 0.000 claims description 5
- 230000037396 body weight Effects 0.000 claims description 5
- 230000002265 prevention Effects 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 2
- 241000228212 Aspergillus Species 0.000 abstract description 9
- 206010061598 Immunodeficiency Diseases 0.000 abstract description 2
- 230000003213 activating effect Effects 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 30
- 241000699670 Mus sp. Species 0.000 description 29
- 230000000694 effects Effects 0.000 description 28
- 102000002689 Toll-like receptor Human genes 0.000 description 20
- 108020000411 Toll-like receptor Proteins 0.000 description 20
- 102000008235 Toll-Like Receptor 9 Human genes 0.000 description 14
- 108010060818 Toll-Like Receptor 9 Proteins 0.000 description 14
- 210000004072 lung Anatomy 0.000 description 14
- 108010065805 Interleukin-12 Proteins 0.000 description 12
- 102000013462 Interleukin-12 Human genes 0.000 description 12
- 210000000440 neutrophil Anatomy 0.000 description 12
- 102000003814 Interleukin-10 Human genes 0.000 description 10
- 108090000174 Interleukin-10 Proteins 0.000 description 10
- 241000233866 Fungi Species 0.000 description 9
- 230000001419 dependent effect Effects 0.000 description 9
- 108090000765 processed proteins & peptides Proteins 0.000 description 9
- 108090001007 Interleukin-8 Proteins 0.000 description 8
- 230000014509 gene expression Effects 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 7
- 102000010168 Myeloid Differentiation Factor 88 Human genes 0.000 description 7
- 108010077432 Myeloid Differentiation Factor 88 Proteins 0.000 description 7
- 230000037361 pathway Effects 0.000 description 7
- 102100022297 Integrin alpha-X Human genes 0.000 description 6
- 102000006992 Interferon-alpha Human genes 0.000 description 6
- 108010047761 Interferon-alpha Proteins 0.000 description 6
- 108010057466 NF-kappa B Proteins 0.000 description 6
- 102000003945 NF-kappa B Human genes 0.000 description 6
- 210000001744 T-lymphocyte Anatomy 0.000 description 6
- 239000002158 endotoxin Substances 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 102000002574 p38 Mitogen-Activated Protein Kinases Human genes 0.000 description 6
- 108010068338 p38 Mitogen-Activated Protein Kinases Proteins 0.000 description 6
- 230000000638 stimulation Effects 0.000 description 6
- 206010017533 Fungal infection Diseases 0.000 description 5
- 208000031888 Mycoses Diseases 0.000 description 5
- 239000000427 antigen Substances 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 238000007912 intraperitoneal administration Methods 0.000 description 5
- OHDXDNUPVVYWOV-UHFFFAOYSA-N n-methyl-1-(2-naphthalen-1-ylsulfanylphenyl)methanamine Chemical compound CNCC1=CC=CC=C1SC1=CC=CC2=CC=CC=C12 OHDXDNUPVVYWOV-UHFFFAOYSA-N 0.000 description 5
- 238000001890 transfection Methods 0.000 description 5
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 4
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 4
- 102100037850 Interferon gamma Human genes 0.000 description 4
- 108010074328 Interferon-gamma Proteins 0.000 description 4
- 108090000978 Interleukin-4 Proteins 0.000 description 4
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 4
- 208000036142 Viral infection Diseases 0.000 description 4
- 229920000392 Zymosan Polymers 0.000 description 4
- 239000002671 adjuvant Substances 0.000 description 4
- 210000001185 bone marrow Anatomy 0.000 description 4
- 229960004397 cyclophosphamide Drugs 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 230000003203 everyday effect Effects 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000036039 immunity Effects 0.000 description 4
- 229920006008 lipopolysaccharide Polymers 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 230000001225 therapeutic effect Effects 0.000 description 4
- 230000009385 viral infection Effects 0.000 description 4
- 238000011740 C57BL/6 mouse Methods 0.000 description 3
- 102000043136 MAP kinase family Human genes 0.000 description 3
- 108091054455 MAP kinase family Proteins 0.000 description 3
- 206010057249 Phagocytosis Diseases 0.000 description 3
- 241000053227 Themus Species 0.000 description 3
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 description 3
- 238000013459 approach Methods 0.000 description 3
- 230000006399 behavior Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 150000002337 glycosamines Chemical class 0.000 description 3
- 210000005174 lung dendritic cell Anatomy 0.000 description 3
- 210000002540 macrophage Anatomy 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 230000008782 phagocytosis Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 2
- 238000011725 BALB/c mouse Methods 0.000 description 2
- 101150013553 CD40 gene Proteins 0.000 description 2
- 208000027219 Deficiency disease Diseases 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 229930182566 Gentamicin Natural products 0.000 description 2
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 2
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 description 2
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 description 2
- 239000007995 HEPES buffer Substances 0.000 description 2
- 101000669460 Homo sapiens Toll-like receptor 5 Proteins 0.000 description 2
- 102100034343 Integrase Human genes 0.000 description 2
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 2
- 229930182816 L-glutamine Natural products 0.000 description 2
- 102000043131 MHC class II family Human genes 0.000 description 2
- 108091054438 MHC class II family Proteins 0.000 description 2
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 2
- 210000004241 Th2 cell Anatomy 0.000 description 2
- 102100039357 Toll-like receptor 5 Human genes 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 210000005006 adaptive immune system Anatomy 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000012636 effector Substances 0.000 description 2
- 238000003114 enzyme-linked immunosorbent spot assay Methods 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 230000002538 fungal effect Effects 0.000 description 2
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 238000003119 immunoblot Methods 0.000 description 2
- 230000007365 immunoregulation Effects 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 210000004969 inflammatory cell Anatomy 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 239000008176 lyophilized powder Substances 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 208000004235 neutropenia Diseases 0.000 description 2
- 230000017111 nuclear migration Effects 0.000 description 2
- 210000001539 phagocyte Anatomy 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 230000002685 pulmonary effect Effects 0.000 description 2
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 2
- 238000005829 trimerization reaction Methods 0.000 description 2
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 2
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- -1 5(6)-Carboxyfluorescein oxalic acid succinimide ester Chemical class 0.000 description 1
- 208000030507 AIDS Diseases 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000758250 Aspergillus fumigatus A1163 Species 0.000 description 1
- 108010021800 B7-2 Antigen Proteins 0.000 description 1
- 102000007697 B7-2 Antigen Human genes 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 206010010356 Congenital anomaly Diseases 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- QTANTQQOYSUMLC-UHFFFAOYSA-O Ethidium cation Chemical compound C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 QTANTQQOYSUMLC-UHFFFAOYSA-O 0.000 description 1
- 238000012413 Fluorescence activated cell sorting analysis Methods 0.000 description 1
- 206010017711 Gangrene Diseases 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 101001046686 Homo sapiens Integrin alpha-M Proteins 0.000 description 1
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 108010091358 Hypoxanthine Phosphoribosyltransferase Proteins 0.000 description 1
- 102000018251 Hypoxanthine Phosphoribosyltransferase Human genes 0.000 description 1
- 102100022338 Integrin alpha-M Human genes 0.000 description 1
- 108010002386 Interleukin-3 Proteins 0.000 description 1
- 102000004890 Interleukin-8 Human genes 0.000 description 1
- 238000011050 LAL assay Methods 0.000 description 1
- 241000239218 Limulus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 108010071384 Peptide T Proteins 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 108010093965 Polymyxin B Proteins 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 1
- 241001222774 Salmonella enterica subsp. enterica serovar Minnesota Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000000516 activation analysis Methods 0.000 description 1
- 210000002821 alveolar epithelial cell Anatomy 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000006909 anti-apoptosis Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000005208 blood dendritic cell Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 210000000621 bronchi Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 210000004718 centriole Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000002301 combined effect Effects 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000000890 drug combination Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 231100000284 endotoxic Toxicity 0.000 description 1
- 230000002346 endotoxic effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 230000007274 generation of a signal involved in cell-cell signaling Effects 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 238000010231 histologic analysis Methods 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000007813 immunodeficiency Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 238000007852 inverse PCR Methods 0.000 description 1
- 239000002085 irritant Substances 0.000 description 1
- 231100000021 irritant Toxicity 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 231100000636 lethal dose Toxicity 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000000527 lymphocytic effect Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 238000007898 magnetic cell sorting Methods 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- NBNNDUZYMXBCOX-UHFFFAOYSA-N n-[4-[4-(4-methoxyphenoxy)-2,6-dimethylphenyl]-1,3-thiazol-2-yl]pyridine-4-carboxamide Chemical compound C1=CC(OC)=CC=C1OC(C=C1C)=CC(C)=C1C1=CSC(NC(=O)C=2C=CN=CC=2)=N1 NBNNDUZYMXBCOX-UHFFFAOYSA-N 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000000242 pagocytic effect Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 229940115272 polyinosinic:polycytidylic acid Drugs 0.000 description 1
- 229920000024 polymyxin B Polymers 0.000 description 1
- 229960005266 polymyxin b Drugs 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229950010131 puromycin Drugs 0.000 description 1
- 208000019539 pyogenic bacterial infections due to MyD88 deficiency Diseases 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000002787 reinforcement Effects 0.000 description 1
- 230000008521 reorganization Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 229940054269 sodium pyruvate Drugs 0.000 description 1
- 210000004988 splenocyte Anatomy 0.000 description 1
- 238000003153 stable transfection Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Endocrinology (AREA)
- Zoology (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
一种利用胸腺素α1为刺激树突状细胞的免疫刺激剂来治疗人类曲霉菌感染的方法。该方法尤其适用于预防骨髓移植后免疫缺乏症宿主的曲霉菌感染。
Description
相关文献
本申请要求2003年3月28日提交的申请号为60/457,911的临时申请为优先权申请。
技术领域
本发明涉及真菌感染的治疗。尤其是涉及曲霉菌感染如与骨髓移植相关的侵入性曲霉病感染的治疗和预防。
背景技术
侵入性曲霉病(IA)的特征是菌丝侵入，它破坏肺部组织并扩散到其他器官中，这是导致医院内肺炎和异源性骨髓移植(BMT)死亡的主要原因，估计感染率为5至10％，随之的死亡率为90至100％。引起IA的最重要的风险因素一直被认为是嗜中性白血球减少症，因此，在异源性骨髓移植鼠模型中，骨髓祖细胞的重建为防止IA发挥了作用。但是，近来在BMT接受者中进行的IA流行病学研究表明，与嗜中性白血球减少症相关的感染减少了，而伴随着针对宿主疾病的移植出现的“之后开始”的感染增加了。
本领域仍需要治疗曲霉菌感染的方法。
发明概述
根据本发明，治疗或预防哺乳动物中曲霉菌感染的方法包括给哺乳动物施用含有抗真菌有效量的胸腺素α1(TA1)的药物组合物。
发明详述
临床和实验证据表明Th1细胞反应性在控制IA中的作用。树枝状细胞(DCs)在体内和体外使Th1导向真菌。有证据表明肺部的DCs使适宜的T细胞对真菌抗原产生应答的能力可能受局部免疫调节行为的影响，这些局部免疫调节行为包括通过Toll样受体(TLRs)传导信号。DCs可能是介入免疫治疗和疫苗开发的有前景的靶点，从而从药物介入的研究重心转向“佐剂”。佐剂既能刺激机体产生适宜应答，进行最佳调节从而抑制感染，同时又能有效抑制生物体的免疫反应，对治疗是有利的。
胸腺素α1(TA1)是一种天然产生的胸腺肽。合成的有28个氨基酸残基的的TA1在全球范围内进行临床试验用于治疗一些病毒感染，这种治疗可以采用单独治疗或者与α-干扰素进行联合治疗。治疗一些免疫缺陷性疾病、恶性肿瘤以及HIV/AIDS病毒感染，这些是TA1的其它适应症。合成的多肽TA1的作用机理还不完全清楚，目前认为与免疫调节行为相关，主要作用在于增大了T细胞的功能。由于对先天的免疫系统的免疫细胞具有免疫调节作用，包括激活具有促进细胞分裂能力的蛋白激酶(MAPKs)以及巨噬细胞基因的表达，我们认为TA1能作为佐剂来刺激DCs引导Th1定向于曲霉菌。本发明提供了一种治疗曲霉菌感染的方法，其中TA1可通过TLRs信号传导刺激DCs引导抗真菌的Th1。
本发明提供了一种治疗曲霉菌感染的哺乳动物的方法，其包括给哺乳动物施用抗真菌有效量的TA1。在一个优选实施例中，TA1有效抑制侵入性曲霉病(IA)。有效剂量的TA1足以能刺激树突状细胞产生促Th1细胞的细胞因子。用于治疗真菌感染的优选剂量为每日200至400微克/公斤体重。在一个优选实施例中，哺乳动物是免疫缺乏症宿主，尤其是人类。该方法尤其适用于治疗免疫缺乏症的患者，特别是那些接受了骨髓移植的患者。
本发明还提供了一种预防哺乳动物感染曲霉菌的方法，其包括给哺乳动物施用抗真菌有效剂量的TA1。本发明尤其适用于预防免疫缺乏症宿主中的IA。在一个优选实施例中，该方法可预防免疫缺乏症患者，尤其是接受骨髓移植的患者的所述感染。TA1的有效剂量能足以刺激树突状细胞产生促Th1细胞的细胞因子。用于治疗真菌感染的优选剂量为每日200至400微克/公斤体重。
没有结合任何具体理论，本发明是基于TA1用于治疗或防止曲霉菌感染的新的免疫调节活性的发现。在各种类型的DCs中，通过MyD88-依赖性途径，TA1促进了促Th1生成的细胞因子IL-12 p70，IL-10和IFN-α的生成。
在转染了TLR的细胞中，TA1似乎直接刺激TLR9而不是TLR2传导信号，后者是通过对相关配基的应答得到了加强。因此，TA1直接或间接地刺激了TLR传导信号。数据表明TA1可利用TLR2-依赖性途径使得骨髓树突状细胞(MDCs)生成IL-12 p70，利用TLR9-依赖性途径使得血液树突状细胞(PDCs)来生成IFN-α和IL-10。
由于通过DCs生成IL-10可能是记忆保护性的抗真菌免疫性的组成之一，平衡DCs和/或不同DC亚群上生成的IL-12/IL-10可能是TA1在曲霉病中的辅助作用的本质原因。
用TA1治疗感染曲霉菌后的BMT小鼠模型，TA1导致CD4+和CD8+细胞的增加，以及嗜中心粒细胞总数的增加。TA1治疗后，Th1细胞(生成IFN-γ)数量增加了，而Th2细胞(生成IL-4)数减少了。
重要的是，用TA1治疗感染了曲霉菌的BMT小鼠使得治疗肺中真菌生长所需药物剂量减少，并且采用高剂量药物能将感染完全治愈。TA1还能增加两性霉素B的治疗效果。
TA1对DCs的作用与它的抗凋亡活性相一致。由于DCs在免疫病理学、免疫学和自身免疫学之间的平衡中起着主要作用，且在胸腺中PDCs通过TLR9传导信号，因此TA1调节DC功能的能力表明TA1是通过TLR的作用对先天的、适应性的免疫系统进行调节的内源调节因子。这就为TA1在一些病毒感染的治疗处方中的作用提供了理论基础，在这些病毒感染病例中PDCs生成α-IFN被认为起着关键作用。为了在PDCs中生成IFNα，TLR9是必需的。并且，PDCs还参与造血细胞移植后的免疫应答，这可以解释TA1在BMT哺乳动物中进行免疫重建的有益作用。
TLRs刺激先天免疫系统不仅有助于适应性免疫系统而且还具有直接的抗菌效应子活性。由于TA1刺激DCs使Th1定位于曲霉菌，还使具有相关效应的嗜中性粒细胞达到抗真菌程度，这进一步表明TA1在治疗真菌感染中的有益效果。
曲霉菌具有独特性质，因为它是一种在免疫缺乏症的宿主中繁殖的腐生真菌。本发明提供了精选的细胞靶点和细胞介导的免疫途径，并提高了对曲霉菌的抵抗力，其中TA1是通过利用TLR途径使Th1对真菌产生适宜反应性的佐剂。
通过下述实施例对本发明进行举例说明，不应理解为对本发明的限制。
实施例1
动物
雌性，8周至10周，BALB/c和C57BL6小鼠来自查尔斯河(CharlesRiver)，NOD/SCID来自杰克逊。成对饲养纯合的TLR2-缺陷、TLR9-缺陷和MyD88-缺陷型C57BL6小鼠；纯合的IFN-γ-缺陷和IL-4-缺陷型BALB/c小鼠，在没有特定病菌的条件下进行饲养。
微生物的感染和治疗
为了感染上A.fumigatus，连续三天通过小鼠鼻内注射悬浮液2×107分生孢子/20微升盐水。通过检测几丁质观察肺中真菌生长数量。几丁质的含量以每个器官中葡糖胺的微克量表示。将未经感染的小鼠的肺中的葡糖胺含量作为阴性对照，对照含量为0.80至2.25微克葡糖胺/器官。将肺切除并立即保存在福尔马林中以进行组织学分析。用过碘酸希夫染色法对嵌入石蜡的组织部分(3至4微米)进行染色。将胸腺素α1(TA1)和杂乱的多肽纯化为无菌乙酰化多肽冻干粉，通过标准的内毒素鲎试剂测定法(limulus lysate assay)，其内毒素水平低于0.03pg/ml。序列如下：Ac-Ser-Asp-Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu-Lys-Glu-Lys-Lys-Glu-Val-Glu-Glu-Ala-Glu-Asn-O(胸腺素α1)以及Ac-Ala-Lys-Ser-Asp-Val-Lys-Ala-Glu-Thr-Ser-Ser-Glu-Ile-Asp-Thr-Thr-Glu-Leu-Asp-Glu-Lys-Val-Glu-Val-Lys-Ala-Asn-Glu-OH(s胸腺素α1)。在无菌水中制备冻干粉。
治疗如下：在BMT-小鼠中，以不同剂量腹膜内给药TA1，腹膜内给予胸腺素α1400μg/kg或静脉给予人重组G-CSF 250μg/kg，在开始灌输BM的那天起每天给药，随着感染的进行，继续给药3天。每天腹膜内给予两性霉素B 4000μg/kg，随着感染的进行，给药3天。该剂量能治愈环磷酰胺处理后的小鼠的IA感染。感染前一天腹膜内给药环磷酰胺150mg/kg。在环磷酰胺处理后的小鼠中，在开始感染的那天起连续5天腹膜内给予剂量400μg/kg的TA1。
感染前一天和感染后一天，通过静脉内注射1mg的Gr-1中和的RB6-8C5抗体，使得小鼠嗜中性粒细胞减少。该处理使肺的嗜中性粒细胞急剧减少，但没有减少DCs数量(在处理前后，CD11c+，MHC Class II+，F480-细胞为5至6×105)。对照小鼠接受等量的纯化的鼠IgG2b。用环磷酰胺处理后一天进行FACS肺细胞分析，可见白血球明显减少且持久(达5天)。F480+细胞的百分比(约20％)以及CD11c+，MHC Class II+，F480-DCs(＜3％)的百分比都没有受到处理的影响。
TLR配体
酵母聚糖来自Saccharomyces cerevisiae，脂磷壁酸质(LTA)来自Staphylococcus aureus，以及脂多糖(LPS)来自Salmonella Minnesota Re 595。CpG寡核苷酸1826和2006被证明是具有免疫刺激活性的序列。
BMT小鼠的形成
给予C57BL6小鼠致命剂量的9Gy并灌注取自BALB/c小鼠的不含T细胞的供体细胞。从脾细胞上表达的供体型MHC I抗原可以看出，95％以上的小鼠存活并表现出稳定的状态，供体型的造血细胞嵌合体。
树突状细胞的分离和培养
通过磁性细胞的分选，从CD14+单核细胞中得到血液CD11c+骨髓DCs(MDCs)，并在Iscove′s修饰的培养基中培养5天，此培养基含有10％胎牛血清、50μmol的2-巯基乙醇、丙酮酸钠(1mM)、2mM的L-谷胺酰胺、HEPES(10mM)以及50μg/ml的庆大霉素，同时还含有50ng/ml的r-人GM-CSF和200U/ml的r-人IL-4，。未成熟MDCs与1000ng/ml的三聚人CD40配体-亮氨酸-拉链融合蛋白共同培养24个小时得到成熟的DCs。利用BDCA-4分离试剂盒分离CD123+类浆树突状细胞(PDCs)。CD123+细胞的纯度＞96％。
将未成熟的DCs与如上的三聚人CD40配体和IL-3(10ng/ml)一起培养得到成熟的PDCs。FACS分析显示PDCs的特征表面受体为CD123bright，CD4+，CD45RA+和CD11c-，而MDCs则为CD1a+，CD11c+，CD11b+，CD4+，CD14low以及CD8-。在未成熟和成熟DCs中HLA II型、CD80和CD86的表达都很高。通过磁性细胞分选分离出鼠肺中的CD11c+DCs(5-7％为CD8α阳性，30-35％为Gr-1阳性)。
在检测吞噬作用中，将DCs预先置于100ng/ml TA1中60分钟，随后在37℃下再与曲霉菌分生孢子孵育60分钟。计算吞噬的百分数并拍摄照片。在评价功能成熟化和细胞因子的检测中，将纯化的DCs再次悬浮在Iscove′s培养基中(不含血清，含有多粘菌素B，以避免血清成分和内毒素的非特异性刺激)，然后只与100ng/ml TA1或同时包括TLR配体或未被吞噬的曲霉菌分生孢子一同进行震动混合24个小时。
表型分析
将样品与结合了FITC或PE的大鼠的抗小鼠抗体进行反应来评价细胞表面表型。将无关的hisotype匹配的抗体作为对照。
抗真菌效应子活性
在吞噬作用检测中，支气管小泡巨噬细胞和外周嗜中性粒细胞预先置于100ng/ml TA1中60分钟，在37℃下与未吞噬(unosponized)的曲霉菌分生孢子孵育60分钟。此外，通过确定集落形成单元的数量和集落形成单元的抑制百分数(平均±SE)来评价分生孢子的活性，称为分生孢子活性。
转染的HEK293细胞的测定
野生型或稳定转染了人TLR2、TLR9和TLR4/CD1427的人胚肾细胞系HEK293在含有10％FCS、HEPES(10nM)、L-谷胺酰胺(2μg/ml)以及庆大霉素(50μg/ml)的低糖DMEM培养基(Dulbecco′s modified Eagle′s medium)中培养。转染细胞中又加有嘌呤霉素(100μg/ml)。为进行刺激试验，将细胞在12孔组织培养板中以细胞密度为3-5×105个细胞/孔培养过夜。清洗细胞，在检测上清液中IL-8前，用100ng/ml TA1单独或与TLR配基一起刺激细胞5个小时。
细胞因子和酶联免疫吸附斑点(ELISPOT)的检测
用试剂盒ELISAs检测培养物上清液中TNF-α、IL-10、IL-12 p70、IFN-α和IL-8的浓度。检测限(pg/ml)分别为：TNF-α的检测限＜3(人)、＜32(鼠)；IL-10的检测限＜12(鼠)、＜5(人)，IL-12 p70的检测限＜16(鼠)、＜3(人)；IL-8的检测限＜25(人)。人的IFN-α＜3ng/ml。为了计算产生细胞因子的细胞数，对来自于肺中的纯化的CD4+T细胞和DCs进行ELISPOT检测。
利用流式细胞分析进行增殖检测
在肺DCs存在的情况下，通过10μg/ml ConA刺激或加热的非活化分生孢子刺激肺CD4+T淋巴细胞的增殖，然后用荧光染料CFSE(5(6)-羧基荧光素二乙酸琥珀酰亚胺酯)标记后进行检测。
逆转录酶(RT)-PCR
从未成熟的DCs中提取总RNA，先用100ng/ml的TA1预处理未成熟的DCs 60分钟，随后与未吞噬的曲霉菌分生孢子接触60分钟(如上述试验所述)。合成cDNA，并采用正向和反向PCR引物以及在鼠和人TLRs和HPRT上所采用的循环数对cDNA进行PCR扩增。在2％琼脂凝胶上电泳分离合成的PCR产物，并用溴乙非啶染色显示。
p38和NF-KB的活化分析
通过与曲霉菌分生孢子和/或100ng/ml的TA1混合20分钟使肺DCs上的P38和NF-kB活化。细胞溶解物印迹与识别未磷酸化的p38 MAPK或识别双磷酸化的p38 MAPK(Thr-180/Tyr-182)的兔多克隆抗体共孵育，或者是与RelA特异性抗体共孵育，其中该RelA是人NF-kB的65kDa DNA的结合亚单位，随后加入联有辣根过氧化物酶的山羊抗兔IgG(按照厂商指示)。用增强型化学发光检测试剂盒显示印迹。免疫印迹在柯达RX胶卷上显示呈条带。为确保每条泳道含有相似的蛋白，将有磷光的免疫印迹剥离，用针对p38和NF-kB的抗体对膜进行重新标记。
胸腺素α1(TA1)刺激树突状细胞(DCs)
前述表明鼠DCs在被感染部位体外吞噬曲霉菌。用TA1，而非杂乱的肽，来刺激肺DCs吞噬未被吞噬的分生孢子(不仅是菌丝)，协同刺激抗原表达以及产生细胞因子。相比较，单独的曲霉菌分生孢子并没有表现出能诱导DCs活性的足够刺激，而与TA1结合使用则明显促进了MHCII型抗原、CD86和CD40分子的表达，并使IL-12 p70产生的DCs增多。有趣的是，仅胸腺素也能使IL-12 p70产生的DCs增多。TA1还能刺激人MDC和PDC亚群(subsets)。未成熟和成熟的DC亚群都能吞噬分生孢子。TA1增加了未成熟DCs的吞噬能力，影响了DC形态学特征(在未成熟的MDCs中检测到更多的细胞质突起)并正向调节了HLA II型抗原以及协同刺激了对分生孢子应答的分子表达。TA1显著增加了未成熟MDCs应答分生孢子和酵母聚糖时IL-12 p70的释放量，以及未成熟PDCs应答分生孢子时IL-10的释放量。IFN-α可以通过PDCs对TLR9配基CpG产生应答来生成，该生成过程能被TA1显著增强。相比较，杂乱的肽就不能正向调节II型抗原，也不能协同刺激分子表达，不能通过对分生孢子产生应答诱导DCs产生细胞因子。
这些数据表明了TA1在DCs被激活和发挥功能中的全新的、从未披露过的免疫调节作用。
TA1通过TLR信号传导刺激MvD88依赖性途径
在应答曲霉菌分生孢子时产生了TLR信号传导，它能介导对真菌的功能应答。TA1有力刺激了鼠DCs上的TLR2、TLR5和TLR9的表达。当与分生孢子和TA1混合物进行接触时，TLR2和TLR9的表达仍受到了刺激，而TLR5的表达却受到了抑制。再一次发现，杂乱的肽不能刺激TLR2和TLR9的表达，无论是仅对肽应答还是对分生孢子的应答。
TA1具有刺激TLR依赖性信号传导的能力通过如下研究得到支持，即对转染了TLR2、TLR9和TLR4/CD14的HEK293细胞进行研究，测定该细胞仅对TA1产生应答或对TA1与相关的TLR配基共同产生应答而生成的IL-8。。在这些HEK293细胞中，TA1能通过TLR9转染的细胞单独产生的应答或细胞对TA1与TLR9配基CpG共同产生的应答而显著增加IL-8。但是，TA1不能在转染TLR2的细胞中单独刺激产生IL-8，但是在这些细胞对酵母聚糖应答时能轻微促进IL-8的生成。无论是单独作用还是与TLR4配基LPS的共同作用，TA1都不能诱导转染TLR4/CD14的细胞产生IL-8。TA1还能影响鼠DCs对这些病菌的TLR配基产生应答而产生IL-12 p70和IL-10的能力。TA1并不会影响细胞对Poly(I：C)或LPS(TLR4)的应答而产生细胞因子的能力。在用酵母聚糖和LTA(TLR2)和CpG(TLR9)刺激后，TA1使IL-12 p70的生成显著增加，而使IL-10的生成显著减少。因此，TA1能直接通过TLR9传导信号并通过相关配基增强TLR2的信号。
NF-和p38 MAPK的激活在促发TLR诱导的基因表达中发挥作用，TA1在前面也显示了刺激MAPK转换的途径。为了支持TA1在TLR诱导的途径中的关系，TA1诱导了NF-kB的核迁移以及p38的磷酸化(不被分生孢子、杂乱的肽或杂乱的肽和分生孢子所刺激)。且，NF-kB核迁移抑制剂(SN50)或p38MAPK(SB202190)消除了TA1对DCs的作用。
骨髓分化因子88(MyD88)是激活NF-kB和MAPK以及通过TLRs传导信号生成IL-12 p70的必要连接蛋白之一。TA1和分生孢子对生成IL-12 p70的作用以及TAI1对产生IL-10的作用在MyD88-缺陷型小鼠中明显消除。因此，MyD88依赖性途径似乎对TA1的体外作用机理起着主要作用。为了确定是否MyD88依赖性途径在TA1体内作用中仍发挥主要作用，对感染曲霉菌的野生型、TLR2-缺陷型、TRL9-缺陷型或MyD88-缺陷型小鼠的局部真菌生长进行评价。在TLR2-缺陷型和TRL9-缺陷型的小鼠中，真菌的生长与野生型小鼠相当，在接受胸腺素治疗后，真菌的生长同样受到减弱。MyD88-缺陷的小鼠中真菌生长也相当，但这些小鼠在接受TA1治疗后真菌生长并未受到减弱。因此，无论TLR的使用程度如何，MyD88-依赖性信号传导途径对TA1体外和体内的活性都是必需的。
TA1能防止BMT小鼠患IA
正如存活率得到提高、同时肺中真菌生长相应减少所揭示的，用TA1而不是杂乱的肽进行治疗，似乎能治愈BMT小鼠的IA。治疗效果是剂量依赖性的，用200和400μg/kg TA1治疗小鼠可达到完全治疗(＞60天的存活)，并且这样的效果超出两性霉素B。而且，TA1增强了两性霉素B的疗效，正如用两种制剂联合治疗小鼠所表现出的小鼠存活率的增加和真菌载量的减少。TA1还能降低肺的病理特征。感染真菌的小鼠的肺切片显示出大量的曲霉菌菌丝侵入了肺实质，还伴有支气管壁的损伤和坏疽以及少量的炎症细胞的补充。相比较，在TA1治疗的小鼠上已观察不到这些特征，小鼠肺部的炎症细胞渗透已治愈，并且没有真菌生长和支气管壁损坏的迹象。可见，TA1具有治疗IA的疗效，并且还可能在与已知的具有降低BMT活性的抗真菌制剂的联合用药上有效。
TA1促进患有IA的小鼠的骨髓和Th1细胞恢复
TA1治疗后，循环系统中的淋巴细胞和嗜中性粒细胞的绝对数量显著增加。更重要的是血液中嗜中性粒细胞的水平并不能来预测个体易患曲霉病。在流式细胞分析中，用TA1治疗BMT小鼠后，其肺中CD4+和CD8+细胞以及嗜中性粒细胞显著增加。如抗原特异性增殖和IFN-γ生成所示，这些肺CD4+T淋巴细胞具有功能上的活性。接受治疗的小鼠中Thl细胞(产生IFN-γ)越多，Th2细胞(产生IL-4)就越少。而且，关于效应子巨噬细胞的抗真菌活性，TA1治疗后的小鼠体内巨噬细胞和嗜中性粒细胞中的分生孢子的(conidiocidal)活性较高。因此，TA1不仅能促进DC成熟，而且还能刺激局部效应子细胞来促进吞噬作用并杀死真菌。
仅使嗜中性粒细胞恢复，如用一已知剂量的G-CSF治疗小鼠来加快小鼠嗜中性粒细胞的恢复并不能足以介导抗真菌的能力达到TA1所获得的程度。相类似，尽管嗜中性粒细胞得到明显恢复，如果没有T细胞或产生IFN-γ的Th1细胞，则TA1的疗效还不够高。当Th1细胞增多时，如在IL-4缺陷型的小鼠中，TA1的疗效仍能得到改善。因此，在缺乏适应性Th1依赖性免疫的情况下，尽管嗜中性粒细胞在介导抗真菌中起着关键的作用，但是若要达到完全不受真菌侵入的状态，如用TA1治疗所达到的状态，则由先天性效应子巨噬细胞和保护性Th1细胞的协同作用所决定。
Claims (12)
1.一种治疗或预防哺乳动物中曲霉菌感染的方法，其包括给所述哺乳动物施用一种包含抗真菌有效量的胸腺素α1(TA1)的药物组合物。
2.根据权利要求1所述的方法，其中所述TA1的施用剂量能足以刺激树突状细胞产生促Th1细胞的细胞因子。
3.根据权利要求1所述的方法，其中所述TA1施加的剂量为200-400μg/kg体重/日。
4.根据权利要求1所述的方法，其中所述哺乳动物为免疫缺乏症的宿主。
5.根据权利要求4所述的方法，其中所述哺乳动物为人。
6.根据权利要求5所述的方法，其中所述人为骨髓移植接受者。
7.根据权利要求5所述的方法，其中施用所述的TA1来刺激树突状细胞产生促进Th1细胞的细胞因子。
8.根据权利要求5所述的方法，其中所述TA1的施用剂量为200-400μg/kg体重/日。
9.根据权利要求1所述的方法，其中所述方法进一步包括给所述人施用至少一种其它的抗真菌制剂。
10.根据权利要求9所述的方法，其中所述其它的抗真菌制剂为两性霉素B。
11.根据权利要求10所述的方法，其中所述两性霉素B的施用剂量为4000μg/kg体重/日。
12.根据权利要求1所述的方法，其中所述曲霉菌感染为侵入性曲霉病。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US45791103P | 2003-03-28 | 2003-03-28 | |
| US60/457,911 | 2003-03-28 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1767847A true CN1767847A (zh) | 2006-05-03 |
| CN100342907C CN100342907C (zh) | 2007-10-17 |
Family
ID=33131730
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004800084904A Expired - Lifetime CN100342907C (zh) | 2003-03-28 | 2004-03-29 | 胸腺素α1在制备用于治疗或预防哺乳动物中曲霉菌感染的药物组合物中的应用 |
Country Status (5)
| Country | Link |
|---|---|
| CN (1) | CN100342907C (zh) |
| CY (1) | CY1110218T1 (zh) |
| DE (1) | DE602004027127D1 (zh) |
| ES (1) | ES2345552T3 (zh) |
| PT (1) | PT1613340E (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103458681A (zh) * | 2011-02-09 | 2013-12-18 | 赛生制药有限公司 | 用于预防感染、降低感染的严重性、及治疗感染的胸腺素α肽 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TW224053B (zh) * | 1991-09-13 | 1994-05-21 | Paul B Chretien |
-
2004
- 2004-03-29 ES ES04749495T patent/ES2345552T3/es not_active Expired - Lifetime
- 2004-03-29 CN CNB2004800084904A patent/CN100342907C/zh not_active Expired - Lifetime
- 2004-03-29 DE DE602004027127T patent/DE602004027127D1/de not_active Expired - Lifetime
- 2004-03-29 PT PT04749495T patent/PT1613340E/pt unknown
-
2010
- 2010-07-22 CY CY20101100691T patent/CY1110218T1/el unknown
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103458681A (zh) * | 2011-02-09 | 2013-12-18 | 赛生制药有限公司 | 用于预防感染、降低感染的严重性、及治疗感染的胸腺素α肽 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100342907C (zh) | 2007-10-17 |
| PT1613340E (pt) | 2010-08-19 |
| DE602004027127D1 (de) | 2010-06-24 |
| ES2345552T3 (es) | 2010-09-27 |
| CY1110218T1 (el) | 2015-01-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bricknell et al. | The use of immunostimulants in fish larval aquaculture | |
| US8389680B2 (en) | Treatment of Aspergillus infections with alpha thymosin peptides | |
| ES2397854T3 (es) | Inmunoterapia para pacientes con inmunosupresión | |
| Johnson et al. | STING agonist mitigates experimental autoimmune encephalomyelitis by stimulating type I IFN–dependent and–independent immune-regulatory pathways | |
| MX2008000596A (es) | Inmunomodulacion inducida por la chaperonina 10. | |
| CN1723037A (zh) | 促单核细胞成熟的方法和组合物 | |
| CN102851295B (zh) | Toll 样受体调节性寡核苷酸及其用途 | |
| WO2021197381A1 (zh) | 具有免疫调节功能的CpG ODN及其应用 | |
| CN115120713A (zh) | 氢氧化铝-CpG寡核苷酸-多肽复合佐剂、疫苗及制备方法和用途 | |
| CN100342907C (zh) | 胸腺素α1在制备用于治疗或预防哺乳动物中曲霉菌感染的药物组合物中的应用 | |
| CN108558998A (zh) | 猪白细胞介素4/6与融合猪抗菌肽共表达重组酵母菌制剂的制备及应用 | |
| CN116162138A (zh) | 树枝状多肽及其用途 | |
| CN106075432A (zh) | 皮卡钙联合佐剂及含有皮卡钙联合佐剂的疫苗 | |
| Farber et al. | Effect of Corynebacterium acnes on interferon production in mice | |
| CN106075431A (zh) | 皮氨钙联合佐剂及含有皮氨钙联合佐剂的疫苗 | |
| CN112870224B (zh) | 一种放疗增敏剂及其制备方法 | |
| CN113817677B (zh) | 泛酸或其衍生物与α-D-葡萄糖-1,6-二磷酸或其衍生物在促进DC迁移中的用途 | |
| WO2024108955A1 (zh) | 乙型肝炎疫苗 | |
| Jin et al. | Goat milk extracellular vesicles: immuno-modulation effects on porcine monocyte-derived macrophages in vitro | |
| CN1889976A (zh) | 抗疱疹药物组合物、生产基于其的剂型的方法及其使用方法 | |
| CN101074266A (zh) | 转导肽－人源粒细胞集落刺激因子融合蛋白及其药物组合物 | |
| Wieland | Einfluss der interzellulären Kommunikation von Hepatozyten und Makrophagen auf die Reaktion von Makrophagen gegenüber Cytomegalieviren | |
| CN118086212A (zh) | 新冠病毒表位多肽致敏的树突状细胞及其应用 | |
| JPH0761950B2 (ja) | 抗腫瘍剤 | |
| Vecchiarelli et al. | Lymphocytes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: BEIJING SAISHENG PHARMACEUTICAL CO.,LTD. Free format text: FORMER NAME: SCICLONE PHARMACEUTICALS INC. |
|
| CP01 | Change in the name or title of a patent holder |
Address after: California, USA Patentee after: SCICLONE PHARMACEUTICALS, Inc. Address before: California, USA Patentee before: SCICLONE PHARMACEUTICALS, Inc. |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Treatment of aspergillus infections with thymosin alpha 1 Effective date of registration: 20180628 Granted publication date: 20071017 Pledgee: China Minsheng Banking Corp Hongkong branch Pledgor: SciClone Pharmaceuticals, Inc. Registration number: 2018990000502 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| CP01 | Change in the name or title of a patent holder |
Address after: California, USA Patentee after: Saisheng Pharmaceutical Co.,Ltd. Address before: California, USA Patentee before: SciClone Pharmaceuticals, Inc. |
|
| CP01 | Change in the name or title of a patent holder | ||
| CP01 | Change in the name or title of a patent holder |
Address after: California, USA Patentee after: SCICLONE PHARMACEUTICALS, Inc. Address before: California, USA Patentee before: Saisheng Pharmaceutical Co.,Ltd. |
|
| CP01 | Change in the name or title of a patent holder | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20200401 Granted publication date: 20071017 Pledgee: China Minsheng Banking Corp Hongkong branch Pledgor: SCICLONE PHARMACEUTICALS, Inc. Registration number: 2018990000502 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Treatment of aspergillus infections with thymosin alpha 1 Effective date of registration: 20200402 Granted publication date: 20071017 Pledgee: China Minsheng Banking Corp Hongkong branch Pledgor: SCICLONE PHARMACEUTICALS, Inc. Registration number: Y2020990000264 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20200925 Granted publication date: 20071017 Pledgee: China Minsheng Banking Corp Hongkong branch Pledgor: SCICLONE PHARMACEUTICALS, Inc. Registration number: Y2020990000264 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20201123 Address after: Greater Cayman, Cayman Islands Patentee after: Saisheng Pharmaceutical Co.,Ltd. Address before: California, USA Patentee before: SCICLONE PHARMACEUTICALS, Inc. |
|
| TR01 | Transfer of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Application of thymosin a 1 in the preparation of pharmaceutical compositions for the treatment or prevention of Aspergillus infection in mammals Effective date of registration: 20210119 Granted publication date: 20071017 Pledgee: China Minsheng Banking Corp Hongkong branch Pledgor: Saisheng Pharmaceutical Co.,Ltd. Registration number: Y2021990000079 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Granted publication date: 20071017 Pledgee: China Minsheng Banking Corp Hongkong branch Pledgor: Saisheng Pharmaceutical Co.,Ltd. Registration number: Y2021990000079 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| CX01 | Expiry of patent term | ||
| CX01 | Expiry of patent term |
Granted publication date: 20071017 |