US20070154491A1 - Edible vaccine - Google Patents

Edible vaccine Download PDF

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Publication number
US20070154491A1
US20070154491A1 US11/684,062 US68406207A US2007154491A1 US 20070154491 A1 US20070154491 A1 US 20070154491A1 US 68406207 A US68406207 A US 68406207A US 2007154491 A1 US2007154491 A1 US 2007154491A1
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Prior art keywords
hpv16
protein
vaccine
yeast
hpv
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Abandoned
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US11/684,062
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English (en)
Inventor
Toshiyuki SASAGAWA
Hideki Tohda
Yuko Hama
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Kanazawa University NUC
AGC Inc
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Asahi Glass Co Ltd
Kanazawa University NUC
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Assigned to ASAHI GLASS COMPANY, LTD., NATIONAL UNIVERSITY CORPORATION KANAZAWA UNIVERSITY reassignment ASAHI GLASS COMPANY, LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: SASAGAWA, TOSHIYUKI, HAMA, YUKO, TOHDA, HIDEKI
Publication of US20070154491A1 publication Critical patent/US20070154491A1/en
Priority to US12/116,501 priority Critical patent/US8282936B2/en
Abandoned legal-status Critical Current

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    • 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
    • A61K39/12—Viral antigens
    • 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
    • A61K39/12—Viral antigens
    • A61K39/23—Parvoviridae, e.g. feline panleukopenia virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/52—Bacterial cells; Fungal cells; Protozoal cells
    • A61K2039/523—Bacterial cells; Fungal cells; Protozoal cells expressing foreign proteins
    • 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
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A61K2039/5258—Virus-like particles
    • 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
    • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
    • A61K2039/541—Mucosal route
    • A61K2039/542—Mucosal route oral/gastrointestinal
    • 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
    • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
    • A61K2039/541—Mucosal route
    • A61K2039/543—Mucosal route intranasal
    • 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
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55544—Bacterial toxins
    • 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
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/20011—Papillomaviridae
    • C12N2710/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • the present invention is an edible vaccine effective against human papilloma virus (sometimes hereinafter referred to simply as “HPV”).
  • HPV is a small del DNA virus having an icosahedral structure and no envelope.
  • the genome of the virus contains open reading frames (ORFs) called E1-E7 and L1 and L2: “E” means early, and “L” means late.
  • L1 and L2 encode capsid proteins of the virus.
  • the early (E) genes are associated with functions such as virus replication and cell transformation.
  • the L1 protein is the major capsid protein having a molecular weight of from 55 to 60 kDa when measured by polyacrylamide gel electrophoresis.
  • the L2 protein is the minor capsid protein which also has an estimated molecular weight of from 55 to 60 kDa and an apparent molecular weight of from 75 to 100 kDa.
  • HPV high-risk types of HPV which can induce cancer.
  • HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 67 and 68, possibly some other types as well, are considered to be high-risk types.
  • intranasal vaccination is also problematic like injection vaccination, because it requires preparation of relatively large amounts of purified HPV-VLPs.
  • Stimulation of the gut-associated lymphoid tissue (GALT) with edible human papilloma virus vaccines was attempted to induce strong mucosal immunity in the vagina.
  • Edible HPV vaccines Two groups have produced edible HPV vaccines from tobacco and potato plants that express the HPV11 (non-patent document 7) and HPV16 (non-patent document 8) L1 genes.
  • HPV-VLPs Purification of HPV virus-like particles (HPV-VLPs) was disclosed in patent document 1.
  • Patent documents 2-4 and patent document 5 disclose HPV vaccine preparations from expression systems in baculovirus and in insect cells, respectively.
  • a nucleic acid vaccine for immunotherapy of HPV was also disclosed (patent document 6).
  • the object of the present invention is to provide an edible HPV vaccine which is available in large amounts inexpensively.
  • the present inventors pursued their research for solutions to the above-mentioned problems by examining usefulness of a recombinant fission yeast expressing HPV proteins as a vaccine and have accomplished the present invention.
  • the present invention provides the following:
  • An edible human papilloma virus vaccine obtained by culturing a transformant of an avirulent fission yeast host, wherein the transformant carries a gene encoding an antigenic protein of human papilloma virus introduced therein and accumulates the expressed antigenic protein in it.
  • FIG. 1A Digestion of a freeze-dried yeast in the stomach (Test Example 1)
  • FIG. 1B Digestion of a freeze-dried yeast in the abdominal cavity (Test Example 1)
  • FIG. 1C Digestion of a freeze-dried yeast in the intestine (Test Example 1)
  • FIG. 2 Induction of antibodies by immunization with HPV16 vaccines (Test Example 2)
  • FIG. 3 Induction of antibodies by immunization with HPV16 vaccines followed by intranasal boosting with HPV16-VLPs Test Example 3)
  • FIG. 4 Change in antibody responses to immunization HPV16 vaccines followed by intranasal boosting with HPV16-VLPs (Test Example 5)
  • FIG. 5 Changes in antibody responses to immunization with HPV16 vaccines followed by intranasal boosting with HPV16-VLPs (Test Example 5)
  • the present invention provides a cultured transformant which carries a gene encoding an antigenic protein of HPV and expresses and accumulates the protein, as an edible HPV vaccine.
  • the host into which a gene encoding an antigenic protein of HPV is introduced to obtain the transformant is an avirulent fission yeast, specifically the fission yeast S. pombe.
  • the antigenic protein of HPV specifically means a capsid protein of HPV, preferably L1.
  • the HPV is preferably a high-risk type of HPV with the danger of cervical cancer, specifically HPV16.
  • any inducible expression vector for S. pombe into which a foreign gene is inserted such as multicloning vectors disclosed in JP-A-7-163373 and JP-A-11-192094, may be used to introduce a gene encoding an antigenic protein of HPV into the host S. pombe without any restrictions.
  • a vector having a HPV16-L1 gene insert under the control of a thiamine-repressible promoter may be used to synthesize virus-like fragments recombinantly (non-patent document 3).
  • the presence of the 55-kDa L1 protein in the virus-like fragments can be confirmed by Western blotting or the like.
  • the transformant which accumulates an antigenic protein of HPV in it is a transformant of an avirulent fission yeast host which accumulates the antigenic protein of HPV, and there are any particular restrictions.
  • the transformant (hereinafter referred to “recombinant S. pombe” ) is cultured at 23 to 37° C. for 6 to 192 hours in a known culture medium, preferably YPD medium under appropriate conditions to accumulate the expressed antigenic protein of HPV in it. After culturing, the culture medium containing the recombinant S. pombe is centrifuged under appropriate conditions, for example, at 0 to 50° C. for 1 to 60 minutes at an acceleration of 500- 3000 g, to collect the precipitate before oral administration.
  • the centrifugally collected recombinant S. pombe is called a yeast pellet and may be used as the “transformant which accumulates an antigenic protein of HPV in it” of the present invention.
  • the yeast pellet may be processed appropriately, if necessary, for example, by freeze-drying, and the “transformant which accumulates an antigenic protein of HPV in it” of the present invention covers such a processed pellet. Freeze-drying may be carried out under ordinary conditions, for example, at a maximum shelf temperature of ⁇ 20° C. overnight, though there are no particular restrictions as long as S. pombe is freeze-dried.
  • the edible HPV vaccine of the present invention covers a transformant having accumulated an antigenic protein of HPV thus obtained.
  • Oral immunization has some advantages over prophylactic immunization through other routes.
  • edible vaccines are readily administered and acceptable to vaccine recipients.
  • edible vaccines may contain active ingredients at lower concentrations than injection vaccines and therefore, can be produced at low production costs.
  • the edible HPV vaccine of the present invention may be formulated into preparations with at least one medicinally acceptable additional ingredient such as a carrier, a diluent, an adjuvant and/or a buffer. Further, the edible HPV vaccine of the present invention may be used as a mixture with food or the like.
  • Known adjuvants such as an E. coli -derived mucosal adjuvant LT (R192) may be used.
  • the edible HPV vaccine of the present invention is administered at a dose of yeast (on a wet basis) selected within the range of from 10 to 500 mg/kg, preferably from 20 to 200 mg/kg, or at a dose of the HPV16-L1 protein selected within the range of from 0.05 to 5 mg/kg, preferably from 0.1 to 2 mg/kg.
  • the edible HPV vaccine may be administered only once or repeatedly.
  • the edible HPV vaccine of the present invention is useful as a prophylactic vaccine against HPV16.
  • the edible HPV vaccine of the present invention may be co-administered with conventional injection HPV vaccines and/or intranasal HPV vaccines in ordinary vaccination regimens for them, for example, as “a concomitant vaccine” to be used in combination with known vaccines. It may also be used as a “booster vaccine” to keep induced antibody titers.
  • a recombinant S. pombe strain was obtained in accordance with non-patent document 3.
  • the HPV16-L1 gene (B27;
  • the recombinant S. pombe strain was cultured in 2 L of YPD medium to allow the expression of the HPV16-L1 protein. It was confirmed by Western blotting that the recombinant S. pombe (pTL2-HPV16-L 1 ) expressed high levels of the 55-kDa L 1 protein. It was confirmed by electron microscopy that the expressed protein assembled into virus-like particles.
  • the total amount of the proteins expression by the yeast was about 10% of the wet weight of the yeast and about 50% of the freeze-dried yeast, and the L 1 protein accounted for from 5 to 10% of the expressed proteins.
  • a recombinant S. pombe strain was cultured in 2 L of YPD medium and collected by centrifugation at 4° C. for 10 minutes at 2000 g.
  • the yeast pellet was washed with phosphate buffered saline (PBS) and resuspended in PBS at 150 mg/mL on a wet basis.
  • PBS phosphate buffered saline
  • the S. pombe suspension was freeze-dried overnight at a maximum shelf temperature of ⁇ 20° C.
  • the freeze-dried recombinant S. pombe was sealed in air-tight plastic tubes and stored at 4° C. until use.
  • the resulting edible vaccine is referred to as “HPV16-L 1 yeast” for the sake of simplicity.
  • freeze-dried HPV16-L1 yeast was resuspended in more than ten volumes of 70% ethanol in water and incubated at 4° C. for 30 minutes, in order to inactivate the yeast, then separated by filtration and dried, before use as an edible vaccine. Separate aliquots of the freeze-dried S. pombe strain was treated with ethanol similarly before oral administration. Further, “fresh-live yeast cells” were treated with ethanol similarly before oral administration.
  • a recombinant S. pombe strain expressing a red fluorescent protein (RFP) was obtained in the same manner as in Example 1.
  • the recombinant S. pombe strain was cultured as described above to allow it to express the two proteins. After culturing, the yeast strain was centrifuged at 40° C. for 10 minutes at 2000 g, and the yeast pellet was collected.
  • RFP red fluorescent protein
  • the Escherichia coli heat-labile toxin LT (R192G), provided in accordance with Curr Top Microbiol Immunol 1999; 236: 216-36, was used as a mucosal adjuvant.
  • the powdery adjuvant was suspended in PBS and stored at ⁇ 30° C. until use.
  • HPV16-VLP protein was purified from the recombinant S. pombe , pTL2-HPV16-L1, obtained in Example 1 was purified by cesium chloride gradient ultracentrifugation (Giga-Hama Y, et. Biotechnology (N Y), 1994; 12: 400-4).
  • the cells were centrifuged at 4° C. at 2000 g for 5 minutes, resuspended in 50 mM potassium phosphate buffer [50 mM KH 2 PO 4 (pH 6.5) containing 20 mM ethylenediaminetetraacetic acid (EDTA) and cooled on ice.
  • the suspension was centrifuged again, and the yeast pellet was resuspended in 10 ml KKC buffer [20 mM KPO 4 (pH 6.5), 800 mM KCl, 0.1 mM CaCl 2 , 1.5 mM MgCl 2 containing 5 mg/ml Novozyme] and incubated at 32° C.
  • the supernatant was collected and layered on 40% sucrose VLP buffer and centrifuged at 4° C. for 2 hours at 27000 g in a Beckman SW28 rotor (Beckman Coulter). The pellet was resuspended in VLP buffer and centrifuged at 4 ° C. for 20 hours at 27000 g in CsCl-equilibrated VLP buffer in the SW rotor. The appropriate fractions were diluted to a density of 1.29 g/ml and centrifuged at 4° C. for 2.5 hours at 27000 g.
  • the pellet was resuspended in VLP buffer and stored at ⁇ 30° C.
  • HPV16-VLP protein was confirmed by enzyme-linked immunosorbent assay (ELISA) using two anti-HPV16 monoclonal antibodies, Camvir-5 and Camvir-6 (provided from Margaret Stanley, Cambridge University), which recognize conformation-dependent epitopes.
  • ELISA enzyme-linked immunosorbent assay
  • HPV16-VLP protein was used for intranasal immunization.
  • the intranasal vaccine is called “HPV16-VLP” for the sake of simplicity.
  • mice Four 9-week-old BALB/c mice were fed 20 mg (on a wet basis) of the fresh-live or freeze-dried yeast expressing a fluorescent protein (RFP) obtained in Example 3, 6 times at hourly intervals after 12 hours of starvation. After the final feed, the mice were dissected and examined for yeast digestion. The digestive tracts of the mice were cut, and touch smears were obtained on glass slides. Fluorescent yeast cells were observed on each slide under Axiovert S-100 Microscope (Carl Zeiss, Germany) and photographed with Fuji 3CCD camera (Fuji Film).
  • RFP fluorescent protein
  • the fresh-live yeast cells were not digested because they were observed at most parts of the digestive tract and excreted in stools.
  • freeze-dried yeast cells were not disrupted in the stomach ( FIG. 1A ) or the jejunum ( FIG. 1B ), but cells decreased from the ileum to the large 5 intestine and were very few in the rectum ( FIG. 1C ). This suggests that freeze-dried yeast cells are digested in the intestine where the gut-associated lymphoid tissue is located.
  • mice Female 9-week-old BALB/c mice were fed the freeze-dried yeast suspended in PBS after 12 hours of starvation. The mice were divided into 6 groups as shown in Table 1 and vaccinated at 4-week intervals and examined for antibody production. Group 1 is a negative control, and Group 2 is a positive control. The doses are expressed on a wet basis. TABLE 1 Ad- Vaccination Mouse ID Vaccine Dose juvant route Group 1 Nos. 1-3 Wild-type 50 mg — Oral yeast Group 2 Nos. 4-6 Purified 5 ⁇ g 10 ⁇ g Intranasal HPV16-L1 Group 3 Nos. 7-9 HPV16-L1 50 mg — Oral yeast Group 4 Nos. 10-12 HPV16-L1 50 mg 10 ⁇ g Oral yeast Group 5 Nos. 13-18 HPV16-L1 150 mg — Oral yeast Group 6 Nos. 19-24 HPV16-L1 150 mg 10 ⁇ g Oral yeast
  • mice serum samples and vaginal samples were collected from the mice.
  • the serum samples were obtained from blood collected from the mouse tail, and the vaginal samples were obtained by washing the vaginae with 100 ⁇ l of PBS using a micropipette.
  • the vaginal samples were collected twice with a 5-day interval, and the two samples were mixed and used for analysis.
  • the samples were collected a few days before the first oral immunization and four weeks after each immunization. All the samples were divided into aliquots and stored at ⁇ 30° C. until use to avoid repeated thawing.
  • HPV16-specific antibodies IgG and IgA were evaluated by ELISA ( FIG. 2 ).
  • HPV16-VLP protein purified from insect cells in accordance with Rose J Virol. 1933; 67: 1936-44., J Gen Virol 1994; 75: 2445-9. was used as the HPV16-VLP antigen for ELISA.
  • HPV16-VLP antigen 100 ng and 300 ng of HPV16-VLP antigen was incubated in PBS on ELISA plates (NUNC Immunoplate Maxisorp; Nalgene Nunc International) at 4° C. overnight.
  • the coated plates were washed with PBST (PBS, 0.1% Tween-20) once and incubated with blocking buffer (3% albumin, 0.5% FCS in PEST) at room temperature (RT; 20-24° C.) for 1 hour. All the subsequent washes were carried out with PEST.
  • PBST PBS, 0.1% Tween-20
  • blocking buffer 3% albumin, 0.5% FCS in PEST
  • reaction buffer (1.5% bovine albumin, 0.25% FCS in PEST)
  • a biotinylated anti-mouse IgA or IgG antibody (diluted with the reaction buffer at a ratio of 1:1500 for IgA and 1:1000 for IgG) was added to the plates and incubated at room temperature for 1 hour.
  • strepavidin-horseradish conjugate (DAKO, Germany) diluted with PEST at a ratio of 1:5000 was added to the plates and incubated for 30 minutes.
  • mice showed high OD values for serum IgG, vaginal IgG and vaginal IgA, although no serum IgA responses were detected.
  • mice 11%) (No. 7 in Group 3 and No. 14 in Group 5) showed transient weak serum IgG responses after the second immunization ( FIG. 2A ). In contrast, no vaginal IgG or IgA responses were observed in any orally vaccinated mice ( FIGS. 2B and 21C ).
  • a suboptimal dose (1 ⁇ g) of the HPV16-VLP was intranasally administered as a booster to all the mice, including the positive controls, negative controls and those orally immunized with the HPV16-L1 yeast, 12 weeks after the final immunization.
  • the antibody-positive rates in the HPV16-L1 yeast-vaccinated mice did not differ at a low HPV16-L1 yeast dose (50 mg) and a high HPV16-L1 yeast dose (100 mg).
  • the mucosal adjuvant LT (R192G) was administered with the HPV16-L1 yeast to some mice to enhance the antibody response.
  • the adjuvant did not cause serious side effects in mice.
  • the adjuvant did not any difference in serum IgG responses among groups orally vaccinated with the HPV16-L1 yeast, while the positive rates were two-fold higher for vaginal IgG and 2.5-fold higher for vaginal IgA in the adjuvant groups than in the non-adjuvant groups, though there were no statistically significant differences.
  • HPV16-VLPs were boiled in bicarbonate buffer (100 g for IgG and 300 ng for IgA) for 10 minutes and used as the denatured HPV16-L1 protein antigen in the ELISA.
  • the reactivities to the VLP antigen or the denatured HPV16-L1 antigen expressed in OD were compared.
  • the sera and the vaginal washings were serially diluted, and the antibody titers were measured.
  • mice orally vaccinated with the HPV16-L1 yeast were transiently positive but eventually negative after boosting.
  • the antibodies in one (No. 7) of the two reacted more strongly with the denatured HPV16-L 1 antigen, while the antibodies in the other (No.14) reacted equally with both types of HPV16-VLP antigen throughout the time course of the experiment ( FIG. 4B ).
  • the edible HPV16-L1 yeast vaccine elicits HPV16-specific vaginal IgA and IgG antibodies, as well as serum IgG.
  • oral administration of the freeze-dried HPV16-L1 yeast alone did not induce anti-HPV16 antibodies, but when it was followed by intranasal boosting with a suboptimal amount of HPV16-VLPs, serum IgG, vaginal IgG and vaginal IgA were elicited in 50%, 33% and 39% of the mice, respectively.
  • no anti-HPV16 antibodies were elicited even after the same intranasal boosting.
  • HPV16-VLPs Intranasally administered HPV16-VLPs increased antibody production probably by enhancing the activity of primed memory B cells that recognize HPV16. All of the induced antibodies reacted more strongly with HPV16-VLP antigen than with the denatured HPV16-L1 antigen, which suggests that these antibodies recognize conformation-dependent HPV16-L1 epitopes and are neutralizing.
  • the edible HPV16-L1 yeast vaccine of the present invention induces neutralizing antibodies and is functional as a vaccine. Therefore, a vaccine is provided which is less painful for patients than intranasal vaccines and injectable vaccines to be used singly.
  • the edible vaccine needs no purification and therefore, is available in large amounts inexpensively.
  • the vaccine may be used in various ways, primarily to induce anti-HPV antibodies in unimmunized people, and also as a booster to keep induced antibody titers.

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EP (1) EP1790332B1 (fr)
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KR (1) KR101187625B1 (fr)
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CN1869215B (zh) * 2006-05-19 2012-07-04 长春百克生物科技股份公司 一种制备人乳头瘤病毒的病毒样颗粒的方法
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EP1790332A1 (fr) 2007-05-30
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CA2579882C (fr) 2012-07-17
CA2579882A1 (fr) 2006-03-16
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US8282936B2 (en) 2012-10-09
ATE474596T1 (de) 2010-08-15
EP1790332B1 (fr) 2010-07-21
CN101010099B (zh) 2011-11-09
KR20070050465A (ko) 2007-05-15
DE602005022458D1 (de) 2010-09-02
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