CN1248631A - Fusion protein for immunoprophyaxis and immunotherapy of venereal disease and cancer - Google Patents
Fusion protein for immunoprophyaxis and immunotherapy of venereal disease and cancer Download PDFInfo
- Publication number
- CN1248631A CN1248631A CN 98112264 CN98112264A CN1248631A CN 1248631 A CN1248631 A CN 1248631A CN 98112264 CN98112264 CN 98112264 CN 98112264 A CN98112264 A CN 98112264A CN 1248631 A CN1248631 A CN 1248631A
- Authority
- CN
- China
- Prior art keywords
- hpv
- hsp
- fusion protein
- protein
- recombination fusion
- 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
- 102000037865 fusion proteins Human genes 0.000 title claims abstract description 31
- 108020001507 fusion proteins Proteins 0.000 title claims abstract description 31
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 19
- 201000011510 cancer Diseases 0.000 title claims abstract description 8
- 238000009169 immunotherapy Methods 0.000 title claims abstract description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims description 7
- 201000010099 disease Diseases 0.000 title claims description 6
- 241000701806 Human papillomavirus Species 0.000 claims abstract description 40
- 102000002812 Heat-Shock Proteins Human genes 0.000 claims abstract description 11
- 108010004889 Heat-Shock Proteins Proteins 0.000 claims abstract description 11
- 206010059313 Anogenital warts Diseases 0.000 claims abstract description 8
- 241000186366 Mycobacterium bovis Species 0.000 claims abstract description 7
- 230000006798 recombination Effects 0.000 claims description 26
- 238000005215 recombination Methods 0.000 claims description 26
- 150000001413 amino acids Chemical class 0.000 claims description 11
- 235000001014 amino acid Nutrition 0.000 claims description 10
- 235000014304 histidine Nutrition 0.000 claims description 10
- 230000004927 fusion Effects 0.000 claims description 9
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 5
- 150000002411 histidines Chemical class 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 241000894006 Bacteria Species 0.000 claims description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 4
- 230000000968 intestinal effect Effects 0.000 claims description 4
- 239000013600 plasmid vector Substances 0.000 claims description 4
- 108010067390 Viral Proteins Proteins 0.000 claims 2
- 101710104159 Chaperonin GroEL Proteins 0.000 claims 1
- 101710178376 Heat shock 70 kDa protein Proteins 0.000 claims 1
- 241000186359 Mycobacterium Species 0.000 claims 1
- 230000000694 effects Effects 0.000 claims 1
- 208000015181 infectious disease Diseases 0.000 abstract description 4
- 239000000427 antigen Substances 0.000 abstract description 3
- 102000036639 antigens Human genes 0.000 abstract description 3
- 108091007433 antigens Proteins 0.000 abstract description 3
- 241001467552 Mycobacterium bovis BCG Species 0.000 abstract description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 abstract description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 abstract description 2
- 230000001225 therapeutic effect Effects 0.000 abstract description 2
- 230000008105 immune reaction Effects 0.000 abstract 1
- 238000002347 injection Methods 0.000 abstract 1
- 239000007924 injection Substances 0.000 abstract 1
- 230000000069 prophylactic effect Effects 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 6
- 241000341655 Human papillomavirus type 16 Species 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 238000002649 immunization Methods 0.000 description 3
- 230000003053 immunization Effects 0.000 description 3
- 230000002265 prevention Effects 0.000 description 2
- 241000589158 Agrobacterium Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 101000767631 Human papillomavirus type 16 Protein E7 Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 101710132596 Protein E4 Proteins 0.000 description 1
- 101710132597 Protein E5 Proteins 0.000 description 1
- 101710132595 Protein E7 Proteins 0.000 description 1
- 101100084449 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) PRP4 gene Proteins 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 238000010009 beating Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000003763 carbonization Methods 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 230000008131 children development Effects 0.000 description 1
- 238000012407 engineering method Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 210000004392 genitalia Anatomy 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 229940114179 mycobacterium bovis Drugs 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 208000003154 papilloma Diseases 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000008521 reorganization Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 208000012991 uterine carcinoma Diseases 0.000 description 1
- 208000024719 uterine cervix neoplasm Diseases 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The present invention makes the heat shock protein of mycobacterium bovis var. BCG (M. Bovis BCG) fuse on tumor specific antigen, i. e. human papillomavirus (HPV) to form fusion protein. This fusion protein can be expressed in colibacillus, microzyme and plant, and (this fusion protein) is used to make immune injection, it can produce specific cell immune and humoral immune reaction of tumor specific antigen, not only possesses the immunoprophylaxis capacity for preventing human papillomavirus (HPV) from infection, but also can be used for immunoprophylaxis and immunotherapy of pointed condyloma, tumor and cancer caused by HPV, so that said invented recombinant fusion protein provides a simple and effective prophylactic immune preparation and therapeutic immune preparation.
Description
The invention belongs to biotechnology---the genetically engineered field.It is characterized in that: the albumen (as early protein E4 or E5 or the E6 or the E7 of the 16th type (HPV-16)) that merges one section or whole human papillomavirus's (HPV) early expression, heat shock protein(HSP) (Heat shock protein, i.e. Hsp at bacterium mycobacterium bovis var.BCG (M.BOVIS BCG).Heat shock protein(HSP) as 65kDa or 70kDa) on, constitutes fusion rotein.The fusion rotein of this type is expressed in intestinal bacteria, yeast or plant.This albumen not only has the ability that immunoprophylaxis HPV infects, and has the ability of pointed condyloma, tumour and cancer that immunoprophylaxis and immunotherapy cause by HPV.
It is the most extensive at present infectious venereal disease that human papillomavirus (HPV) infects.It is general that reproductive organ HPV infects, and take place in age 15-49 year the men and women of the U.S. about 10%, and most of such infection is subclinical (promptly not having clinical manifestation).In PCR detected, HPV-16 almost was present among all woman, and is irrelevant with the cytology result.Wherein 1% HPV-16 type male women will develop into diffustivity uterus carcinoma (in China, above-mentioned data are with much higher).No matter the man, or among the woman, infected at external genital by HPV and produce strumae, claim pointed condyloma.HPV infects also to bring out and produces tumor of cervix and cancer in the women.Serious HPV infects all will influence Fertility, also cause the rugged shape of infant development.
The pointed condyloma that infected by HPV to cause, existing treatment is an integrating laser carbonization in China, or the serious Interferon, rabbit of beating, yet efficient low, only controls table and does not control root, generally some months will recur.Because be virus infection, there is not specifics to control.
Cervical cancer is women's the most general second largest malignant diseases, is only second to mammary cancer.But,, all also only limit to excision of operation property and radiation treatment no matter be to infection disease or to tumor treatment between the more common cervical epithelial cell of having diagnosed out.
Therefore, in the therapeutic immunization mode cell that infects HPV is carried out the great potentiality of immunotherapy tool.In addition, to the HPV immunization can success induce secular immune response, thereby proof can prevent HPV to infect chronically, prevents the generation of pointed condyloma and tumour.
The objective of the invention is to, create a kind of fusion rotein, a kind of new simple and effective immunoprophylaxis and the novel method of immunotherapy are provided with gene engineering method.Not only can immunoprophylaxis HPV infect, and can treat pointed condyloma and the cancer that infects by HPV and bring out.
Embodiment
(1) structure of recombination fusion protein
Merging heat shock protein(HSP) (Hsp) goes up and the formation fusion rotein to the antigen (HPV) of tumour-specific.With recombination fusion protein Hsp-E7 is example, and 539 amino acid of its N-end come from the heat shock protein(HSP) of the 65kda (kilodalton) of mycobacteriumbovis var.BCG; Terminal 98 amino acid of C-come from the early protein E7 of human papilloma (HPV) Class1 6, and it is proteinic amino in proper order as Fig. 1, about 67737 dalton of the molecular weight of expectation.
The N-end of HspE7 can connect the leading order of a plurality of Histidines (Histidine), is convenient to purifying.
(2) expression of recombination fusion protein HspE7 in intestinal bacteria.
Recombination fusion protein is cloned on the pet28a carrier, and its promoter is T7, can be by this recombinant protein of IPTG abduction delivering.Recombination fusion protein also can be cloned on other colibacillary carrier, is expressed in the intestinal bacteria.
(3) fusion protein expression is in plant
Recombination fusion protein can be cloned on pBI121 or pBI221, changes in the plant by Agrobacterium and particle gun respectively, expresses in plant.Recombination fusion protein also can be cloned on other plasmid vector, is expressed in the plant.
(4) recombination fusion protein can be cloned on the zymic plasmid vector, expresses in yeast.
Embodiment:
(1) recombination fusion protein Hsp-E7 is cloned on the pet28a carrier, and its promoter is T7, at expression in escherichia coli.
(2) recombination fusion protein can be cloned on pBI121 or pBI221, expresses in plant.Its promoter is 2 * CaMV35S+AMV RNA4 promoter and enhanser.
(3) recombination fusion protein can be cloned on the zymic plasmid vector, expresses in yeast.
(4) with recombination fusion protein Hsp-E7 immunity mouse, not only inducing cell immunity but also induce humoral immunization.These immunity are specific to HPV-16-E7.Further, carry out epidemic prevention, be protected from the challenge of infecting of HPV and tumour with this recombination fusion protein in physiological saline; And curative immunity causes already present tumour to disappear.Although being (Histidine-tagged) recombination fusion protein (h) Hsp-E7 with the leading order of a plurality of Histidines of tool, most data obtains, but directly relatively recombination fusion protein (h) Hsp-E7 is with reorganization but there is not the fusion rotein Hsp-E7 of the leading order of Histidine (Histidine-tag), and the two all has the ability that the equal HPV of prevention infects, eliminates pointed condyloma and tumour.
Claims (11)
1. the present invention is a kind of fusion rotein for the treatment of the cancer that venereal disease and do as one likes disease cause.The method of claim recombination fusion protein, the heat shock protein(HSP) (Heatshock protein) that promptly merges mycobacterium bovis var.BCG is to human papillomavirus's (HPV) viral protein.
2. the method for claim 1, heat shock protein(HSP) a part of all be blended in human papillomavirus's viral protein a part of or all on, constitute fusion rotein.
3. as claim 1,2 described methods, the amino acid of recombination fusion protein N-end comes from the heat shock protein(HSP) of mycobacterium bovis var.BCG, and the C-end amino acid comes from the albumen (as E4 or E5 or E6 or the E7 of HPV-16) of the early expression of human papillomavirus (HPV) Class1 6 (HPV-16) or Class1 8 (HPV-18).
4. as claim 1,2 described methods, the amino acid of recombination fusion protein N-end comes from the albumen (as E4 or E5 or E6 or the E7 of HPV-16) of the early expression of human papillomavirus (HPV) Class1 6 (HPV-16) or Class1 8 (HPV-18), and the C-end amino acid comes from the heat shock protein(HSP) of mycobacterium bovisvar.BCG.
5. as claim 1,2,3 and 4 described fusion roteins, the N-end of recombination fusion protein can connect the leading order (Histidine-tagged) of a plurality of Histidines (Histidine), is convenient to purifying.
6. as claim 1,2,3,4,5 described recombination fusion proteins, can be expressed in intestinal bacteria, yeast and the plant by plasmid vector.
7. as claim 1,2,3 and 5 described recombination fusion proteins, the amino acid of its N-end comes from 65kDa or the 70kDa heat shock protein(HSP) of mycobacterium bovis var.BCG, and the C-end amino acid comes from the albumen (as E4 or E5 or E6 or the E7 of HPV-16) of the early expression of human papillomavirus (HPV) Class1 6 (HPV-16) or Class1 8 (HPV-18).
8. recombination fusion protein Hsp-E7 as claimed in claim 7, the amino acid that its N-is terminal 539 comes from the 65kDa heat shock protein(HSP) of mycobacterium bovis var.BCG, and terminal 98 amino acid of C-come from the albumen E7 of the early expression of human papillomavirus (HPV) Class1 6 (HPV-16).
9. recombination fusion protein Hsp-E7 as claimed in claim 8, its amino-acid sequence such as Figure of description 1.
10. as claim 8,9 described recombination fusion protein Hsp-E7, its N-end can connect the leading order of a plurality of Histidines (Histidine), is convenient to purifying.
11. can be with recombination fusion protein (h) Hsp-E7 of a plurality of Histidines (Histidine) as claim 8,9,10 described recombination fusion protein Hsp-E7 and N-end, not only have the ability that immunoprophylaxis human papillomavirus (HPV) infects, and have the effect of pointed condyloma, tumour and cancer that immunoprophylaxis and immunotherapy cause by human papillomavirus (HPV).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 98112264 CN1248631A (en) | 1998-09-24 | 1998-09-24 | Fusion protein for immunoprophyaxis and immunotherapy of venereal disease and cancer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 98112264 CN1248631A (en) | 1998-09-24 | 1998-09-24 | Fusion protein for immunoprophyaxis and immunotherapy of venereal disease and cancer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1248631A true CN1248631A (en) | 2000-03-29 |
Family
ID=5222134
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 98112264 Pending CN1248631A (en) | 1998-09-24 | 1998-09-24 | Fusion protein for immunoprophyaxis and immunotherapy of venereal disease and cancer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1248631A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001004344A3 (en) * | 1999-07-08 | 2001-11-15 | Stressgen Biotechnologies Corp | Induction of a th1-like response in vitro |
| US6797491B2 (en) * | 2000-06-26 | 2004-09-28 | Stressgen Biotechnologies Corporation | Human papilloma virus treatment |
| CN103146734A (en) * | 2013-03-12 | 2013-06-12 | 中国人民解放军军事医学科学院军事兽医研究所 | Anti-burn and scald infection multiple organ failure Pseudomonas aeruginosa toxin vaccine |
| CN101395174B (en) * | 2006-02-13 | 2014-03-19 | 艾比欧公司 | Hpv antigens, vaccine compositions, and related methods |
-
1998
- 1998-09-24 CN CN 98112264 patent/CN1248631A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001004344A3 (en) * | 1999-07-08 | 2001-11-15 | Stressgen Biotechnologies Corp | Induction of a th1-like response in vitro |
| US6797491B2 (en) * | 2000-06-26 | 2004-09-28 | Stressgen Biotechnologies Corporation | Human papilloma virus treatment |
| US7211411B2 (en) | 2000-06-26 | 2007-05-01 | Nventa Biopharmaceuticals Corporation | Human papilloma virus treatment |
| US7754449B2 (en) | 2000-06-26 | 2010-07-13 | Nventa Biopharmaceuticals Corporation | Human papilloma virus treatment |
| CN101395174B (en) * | 2006-02-13 | 2014-03-19 | 艾比欧公司 | Hpv antigens, vaccine compositions, and related methods |
| CN103146734A (en) * | 2013-03-12 | 2013-06-12 | 中国人民解放军军事医学科学院军事兽医研究所 | Anti-burn and scald infection multiple organ failure Pseudomonas aeruginosa toxin vaccine |
| CN103146734B (en) * | 2013-03-12 | 2014-10-01 | 中国人民解放军军事医学科学院军事兽医研究所 | Anti-burn and scald infection multiple organ failure Pseudomonas aeruginosa toxin vaccine |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6599508B1 (en) | Papilloma virus-like particles, fusion proteins as well as processes for their production | |
| Jagu et al. | Concatenated multitype L2 fusion proteins as candidate prophylactic pan-human papillomavirus vaccines | |
| RU2619187C2 (en) | Fusion proteins for application as immunogenic amplifying agents to induce antigen-specific t-cell response | |
| US7371391B2 (en) | Papilloma virus capsomere vaccine formulations and methods of use | |
| Lin et al. | Cottontail rabbit papillomavirus L1 protein-based vaccines: protection is achieved only with a full-length, nondenatured product | |
| Shi et al. | Papillomavirus pseudovirus: a novel vaccine to induce mucosal and systemic cytotoxic T-lymphocyte responses | |
| CN101518647A (en) | Human papilloma virus preventative vaccine, construction method and application | |
| CN102747047A (en) | Human papillomaviruse type hybrid virus-like particles and preparation method thereof | |
| EP0840747A1 (en) | Papillomavirus polyprotein constructs | |
| WO1993020844A1 (en) | Papillomavirus e7 protein | |
| AU6381794A (en) | Pharmaceuticals based on papillomaviruses | |
| CA2229955C (en) | Papilloma virus capsomere vaccine formulations and methods of use | |
| JPH06503559A (en) | Subunit papillomavirus vaccines and peptides used therein | |
| CN1248631A (en) | Fusion protein for immunoprophyaxis and immunotherapy of venereal disease and cancer | |
| KR20060053135A (en) | Fusion Proteins for Cervical Cancer Inhibition | |
| US7182947B2 (en) | Papillomavirus truncated L1 protein and fusion protein constructs | |
| CN106421774B (en) | Use of PreS1 for preparing hepatitis B vaccine and treating chronic hepatitis B | |
| CN114437236B (en) | Recombinant African swine fever virus multi-epitope fusion protein, preparation and application thereof | |
| CN102787119B (en) | Treat and/or prevent product and the method for virus infection | |
| CN108624611A (en) | The preparation and its application of infectious bursa of Fabricius virus virus-like particle | |
| CN103397036B (en) | Gene sequence for expressing recombinant human serum albumin by pichia yeast | |
| Davies et al. | Cancer of the cervix: prospects for immunological control | |
| CN114315962B (en) | A type of polypeptide that promotes the production of African swine fever virus antigen-specific immune response in pigs and its application | |
| CN1696152A (en) | LIE7 recombined protein of 16 type human papilomavirus expressed by bacillus coli | |
| CN113896771B (en) | A type of polypeptide that promotes the production of African swine fever virus antigen-specific immune response in pigs and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |