WO2016129724A1 - Vaccin antibactérine de mycoplasma hyopneumoniae et son procédé de préparation - Google Patents

Vaccin antibactérine de mycoplasma hyopneumoniae et son procédé de préparation Download PDF

Info

Publication number
WO2016129724A1
WO2016129724A1 PCT/KR2015/001464 KR2015001464W WO2016129724A1 WO 2016129724 A1 WO2016129724 A1 WO 2016129724A1 KR 2015001464 W KR2015001464 W KR 2015001464W WO 2016129724 A1 WO2016129724 A1 WO 2016129724A1
Authority
WO
WIPO (PCT)
Prior art keywords
recombinant protein
vaccine
hyopneumoniae
mycoplasma hyopneumoniae
present
Prior art date
Application number
PCT/KR2015/001464
Other languages
English (en)
Korean (ko)
Inventor
한태욱
아비히지트바레이트
Original Assignee
강원대학교산학협력단
우진 비앤지 주식회사
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 강원대학교산학협력단, 우진 비앤지 주식회사 filed Critical 강원대학교산학협력단
Publication of WO2016129724A1 publication Critical patent/WO2016129724A1/fr

Links

Images

Classifications

    • 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
    • A61K39/02—Bacterial 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
    • A61K39/02—Bacterial antigens
    • A61K39/0241—Mollicutes, e.g. Mycoplasma, Erysipelothrix
    • 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
    • A61K38/164—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/30—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Mycoplasmatales, e.g. Pleuropneumonia-like organisms [PPLO]
    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • 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
    • A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
    • A61K2039/552—Veterinary vaccine

Definitions

  • the present invention relates to a vaccine composition for preventing the infection of swine mycoplasma hyopneumoniae comprising a mycoplasma hyopneumoniae- derived recombinant protein and mycoplasma hyopneumoniae bacterin and a preparation method thereof.
  • Mycoplasma hyopneumoniae the causative agent of porcine mycoplasma pneumonia, is known as Porcine respiratory disease (PRDC). complex) and respiratory pathogens associated with enzootic pneumonia (EP), which cause infections beginning with colonization of respiratory ciliated cells. This pathogen destroys the mucous cilia, which act as a defensive defense from the outside, and causes acute inflammation in the airways. In chronic infections, pulmonary lesions can be enlarged to expose them to secondary infections caused by other bacteria. This leads to a decrease in pig productivity and significant economic losses for the pig industry (Maes, et al., 2008).
  • PRDC Porcine respiratory disease
  • EP enzootic pneumonia
  • the commercial vaccine currently being supplied worldwide to prevent infection is an inactivated vaccine, which consists of a typical whole cell bacterin and an adjuvant.
  • the productivity of pigs increased with the inoculation of these commercial vaccines, and their spread to lung lesions was reduced (Maes, et al., 2008), but the protective effect against natural infections is still ineffective (Thacker, et al., 1998, Sibila, et al., 2007, Maes, et al., 2008).
  • conventional commercial vaccines are not able to prevent the spread of natural infectious organisms and respiratory ciliary colonization (Maes, et al., 2008). Therefore, the need for an effective vaccine development that can solve these problems is important.
  • Attachment P97 is a membrane surface protein of M. hyopneumoniae and is known as a highly immunogenic antigen (Klinkert, et al., 1985, Zhang, et al., 1995, Minion, et al., 2000).
  • R1 and R2 sites consisting of repeating sections, of which the R1 site binds to the attachment site of the cilia (AAKPV / E) (Minion, et al., 2000). Bound M.
  • hyopneumoniae obtains the amino acids necessary for growth from the host and arranges the host's molecules in various ways to invade the wounded tissue (Raymond, et al., 2014). Hosts, on the other hand, may produce antibodies to prevent the attachment of M. hyopneumoniae to swine cilia (Zhang, et al., 1995) or to inhibit the growth of attached pathogens (Okamba, et al., 2010). However, previous studies have reported that existing commercial vaccines cannot induce antibodies against important antigens of P97 (Conceicao, et al., 2006, Okamba, et al., 2010).
  • the present inventors have developed a vaccine to prevent M. hyopneumoniae caused by natural infection in pigs.
  • the vaccine mixed with M. hyopneumoniae bacterin and recombinant P97 protein shows superior immune response compared to conventional commercial vaccines. It confirmed and completed this invention.
  • the present invention provides a recombinant protein having an amino acid sequence of SEQ ID NO: 1.
  • Recombinant proteins of the invention include fragments of P97 protein derived from Mycoplasma hyopneumoniae .
  • the fragment comprises a region R1 and R2 consisting of a repeating section at the C terminus of P97 (named rP97 in the sense of recombinant P97).
  • the present invention also provides a polynucleotide encoding the recombinant protein.
  • the polynucleotide may be one having a nucleotide sequence of SEQ ID NO: 2, but is not limited thereto.
  • the present invention also provides a vaccine composition for preventing mycoplasma hyopneumoniae infection comprising a mycoplasma hyopneumoniae bacterin and a recombinant P97 protein (rP97) having the amino acid sequence of SEQ ID NO: 1.
  • the recombinant protein comprises a fragment of P97 protein derived from Mycoplasma hypneumoniae.
  • the mycoplasma hyopneumoniae bacterin should be inactivated.
  • the vaccine composition is a vaccine composition for preventing diseases caused by infection of mycoplasma hyopneumoniae.
  • the disease may be swine pandemic, but is not limited thereto.
  • the present invention also provides a method for preparing a vaccine composition for preventing mycoplasma hyopneumoniae infection.
  • inactivating and concentrating a mycoplasma hyopneumoniae strain and mixing the concentrated mycoplasma hyopneumoniae with a recombinant protein having the amino acid sequence of SEQ ID NO: 1 It includes a step.
  • the recombinant protein having the amino acid sequence of SEQ ID NO: 1 preparing a vector comprising a polynucleotide encoding the recombinant protein; Transforming the host with the vector; Culturing the transformed host to express the recombinant protein; And it may be prepared by a method comprising the step of separating the recombinant protein.
  • the vector may be a plasmid vector, the host may be E. coli, but is not limited thereto.
  • the recombinant protein may be provided in the form of a lysate (lysate) isolated from the culture medium of the host containing the recombinant protein.
  • concentration of the lysate may be 0.3 mg / ml.
  • the concentrated mycoplasma hyopneumoniae may be mixed at a concentration of 2.5 ⁇ 10 8 CCU (Colour-changing units) / ml.
  • a pharmaceutical or veterinary acceptable excipient and / or carrier may be mixed together in the mixing process of the concentrated mycoplasma hyopneumoniae and the recombinant protein.
  • the recombinant protein of the present invention was tested for mycoplasma hyopneumoniae bacterin. Increasing the immune response was confirmed to show a superior protective effect than the existing vaccine. Therefore, the vaccine composition using the recombinant protein of the present invention can effectively prevent diseases caused by infection of mycoplasma hyopneumoniae, in particular swine pandemic pneumonia.
  • Figure 1 shows the M. hyopneumoniae specific antibody titer using the obtained serum from day 0 to day 63 after vaccination IDEXX M. hyo . It is the result evaluated by ELISA. All experimental data were expressed as mean SD, and the significant difference between the vaccine group and the PBS group was marked with * ( P ⁇ 0.05) and ** ( P ⁇ 0.01).
  • Figure 2 shows the results of evaluating rP97 specific antibody titers by indirect ELISA using serum obtained from day 0 to day 63 after vaccination. All experimental data are expressed as mean SD and Mycoflex Significant differences between the vaccine and PBS groups were indicated by *** ( P ⁇ 0.001).
  • Vaccination is a very important means of preventing M. hyopneumoniae .
  • Humoral immunity induced by the M. hyopneumoniae vaccine has been reported to protect against the development of pneumonia and pneumonia lesions in pigs (Lam & Switzer, 1971, Thacker, et al., 2000). Therefore, the present inventors investigated the immunogenicity against bacterial vaccines in which rP97 recombinant protein (SEQ ID NO: 1) and a lysate were mixed. As a result (see Fig. 1), 42 days after vaccination of the present invention showed a positive serum response was found that the humoral immune response was generated.
  • the pigs immunized with the commercial vaccine showed negative sera response up to 21 days after inoculation and showed similar results as previously reported literature (Kim, et al., 2011).
  • the vaccination group showed higher antibody value.
  • Differences in the M. hyopneumoniae specific antibody titers of the two vaccinated groups across the inoculation groups can be influenced by the diversity of strains used as antigens (Thacker, et al ., 1998).
  • M. hyopneumoniae P97 is known to induce an early immune response in immunized pigs (Klinkert, et al ., 1985, Zhang, et al ., 1995, Raymond, et al ., 2014).
  • the use of this single antigen reduces lung lesions by 60-90% (Okamba, et al ., 2010, Lee, et al ., 2014) and can reduce inflammation in the lungs and M. hyopneumoniae infection in the respiratory tract. (Okamba, et al ., 2010).
  • pigs immunized with a commercial vaccine did not induce rP97-specific antibody titers (FIG. 2). These results show similar results to previous reports that existing commercial vaccines are inadequate to produce rP97-specific antibody titers.
  • the lysate containing the vaccine and recombinant protein of the present invention showed a significant level of rP97-specific immune response. This may be because P97, which is involved in early infection and adhesion, improved the efficacy of the vaccine.
  • the vaccine of the present invention induces a high level of M. hyopneumoniae specific immune response, especially rP97-specific humoral immune response can be seen to be significantly higher than the commercial vaccine.
  • the present invention can provide a M. hyopneumoniae bacterin vaccine comprising the rP97 protein of M. hyopneumoniae and a method for preparing the same.
  • the recombinant rP97 protein lysate ie, cell lysate expressing rP97 protein alone, M. hyopneumoniae bacterin alone, and cell lysate expressing rP97 protein and M. hyopneumoniae Vaccine efficacy was analyzed in the mixture containing bacterin, and the vaccine was induced by cell lysate expressing rP97 protein and M. hyopneumoniae bacterin alone. The vaccine efficacy was found to be excellent.
  • the present invention can provide a vaccine composition for preventing mycoplasma hyopneumoniae infection comprising a mycoplasma hyopneumoniae bacterin and a recombinant protein having the amino acid sequence of SEQ ID NO: 1.
  • the mycoplasma hyopneumoniae bacterin that can be used in the present invention may be prepared using a mycoplasma hyopneumoniae strain, preferably isolated from the present invention. And the identified Mycoplasma hyopneumoniae strains Mhp 23-9 and Mhp 7-25. This is because other mycoplasma hyopneumoniae strains are not very good antibody generating ability, Mhp 23-9 and Mhp 7-25 is much better antibody production ability is excellent vaccine production is possible.
  • bacterin refers to bacterial debris or cultures that can induce protective immunity when combined with certain adjuvants to prevent disease or infection when administered to an animal, wherein the “adjuvant” refers to immunogenicity. Means a substance that enhances and enhances the efficacy of bacterin.
  • M. hyopneumoniae bacterin may be included in the vaccine composition of the present invention in an amount capable of providing immunity against mycoplasma hyopneumoniae (“immunity”), wherein the “immunity” is the species of the animal being administered, Age, size, health status, previous vaccine administration experience, and the like.
  • the recombinant protein according to the present invention there are two repeating regions R1 and R2 present in the C-terminal region of the M. hyopneumoniae P97 gene, the region attached to the cilia of the host is R1. Therefore, in this study, fusion of Escherichia coli Labile toxin B subunit (LTB) to enhance the adjuvant effect on R1 and R2, the main motifs of P97 protein, was to protect the early infection of M. hyopneumoniae . Fusion protein was prepared by fusion, and the recombinant protein prepared in the present invention is a protein consisting of the amino acid of SEQ ID NO: 1.
  • the vaccine composition of the present invention may include adjuvant mixture and one or more pharmaceutically or veterinarily acceptable carriers, excipients or diluents capable of enhancing the immunogenicity of the bacterin to induce protective immunity even with minimal administration.
  • pharmaceutically or veterinarily acceptable carriers capable of enhancing the immunogenicity of the bacterin to induce protective immunity even with minimal administration.
  • 'pharmaceutical or veterinary acceptable' refers to a composition which is physiologically acceptable and does not normally cause an allergic reaction such as gastrointestinal disorders, dizziness, or the like when administered to an animal.
  • Examples of such carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
  • fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers and preservatives may be further included.
  • Suitable carriers for use include, but are not limited to, saline, phosphate buffered saline, minimal essential medium (MEM), or an aqueous medium comprising MEM of HEPES buffer.
  • Adjuvant mixtures for use in the vaccine compositions of the present invention include oils that enhance the immune response and are metabolizable.
  • the vaccine compositions of the invention may be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.
  • the formulations may be in the form of powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders and the like.
  • the vaccine composition of the present invention may be administered by intramuscular, subcutaneous, transdermal, intravenous, intranasal, intraperitoneal or oral route, and preferably by intramuscular or subcutaneous route.
  • the vaccine may contain mycoplasma hyopneumoniae, metabolizable oil, polyoxyethylene-polyoxypropylene block copolymers and acrylic acid polymers inactivated as an oil-in-water emulsion form.
  • the dosage of the vaccine can be appropriately selected depending on several factors such as the route of administration, the age, sex, weight and severity of the animal.
  • the vaccine compositions of the present invention may be prepared by standard methods in the art, with the exception of methods specific to the present invention, such as the preparation of rP97 recombinant proteins.
  • an organism can be grown in a culture medium such as complete medium, and the growth of the organism can be monitored by standard techniques such as measuring color change units (CCUs) and harvested when sufficiently high titers are achieved.
  • CCUs color change units
  • the stock may be further concentrated or lyophilized by conventional methods prior to inclusion in the vaccine for formulation.
  • the rP97 recombinant protein of the present invention can be prepared by a method of transforming a host with a vector comprising a polynucleotide encoding the same.
  • the polynucleotide may be one or more codon sequences modified with codons suitable for expression of the host cell.
  • the host cell may be E. coli, yeast, or a combination thereof, but is not limited thereto.
  • the polynucleotide may be one having a nucleotide sequence of SEQ ID NO: 2.
  • the polynucleotide may be operably linked with a regulatory sequence (eg, promoter, operator, polyadenylate, ribosomal binding sequence, etc.) suitable for expression of a host cell.
  • the vector may include a replication origin that allows replication in the host cell.
  • the vector may be a plasmid vector, a viral vector or a combination thereof, but is not limited thereto.
  • M. hyopneumoniae strains were isolated from domestic pig farms. Among them, M. hyopneumoniae strains Mhp 23-9 and Mhp 7-25 were incubated for 3 weeks under conditions of 5% CO 2 , 37 °C using Friis medium. It was. The cultured bacterial solution was inactivated by raising the pH to about 7.8. Inactivation of the bacteria was carried out using an inactivating reagent such as Binary ethyleneimine (BEI) (US Pat. No. 5,565,205) (Petersen & Dayalu, 1996) .BEI was added to broth 2-bromoethylaminehydrobromide (BEA) (SigmaAldrich, St. Louis, MO, USA).
  • BEI Binary ethyleneimine
  • BEA 2-bromoethylaminehydrobromide
  • BEI was neutralized with sodium thiosulfate (SigmaAldrich, St. Louis, Mo., USA), a neutralization reagent.
  • sodium thiosulfate SigmaAldrich, St. Louis, Mo., USA
  • Some of the inactivated culture was added to fresh medium and incubated at 37 ° C., and incubated for at least one week to confirm inactivation.
  • the inactivated culture was concentrated by filtration or centrifugation.
  • the bacterin prepared using Mhp 23-9 was named "L1"
  • the bacterin prepared using Mhp 7-25 was named "L2”.
  • rP97 protein was constructed using the pET expression system (Barate, et al., 2014).
  • expression plasmids were inserted into E. coli BL21 competent cells by electroporation and cultured in 1 L Luria-Bertani (LB) medium with 100 ⁇ g / mL ampicillin. Thereafter, 1 mM of isopropyl- ⁇ -D-1-thiogalactopyranoside (IPTG) was added to induce protein expression for 3 hours, followed by sonication and centrifugation to obtain a lysate containing the recombinant protein.
  • IPTG isopropyl- ⁇ -D-1-thiogalactopyranoside
  • Genomic DNA was extracted using Genomic DNA Extraction Mini kit (RBC Biosciences) against M. hyopneumoniae 23-9 and 7-25 strains of domestic isolates. Extracted. Genomic DNA of enterococci E. coli was extracted by boiling method. E. coli heat-labile enterotoxin subunit B gene (eltb), P97 gene C-terminal repeat sequences (R1 and R1R2), and P97 genes (ltbR1 and ltbR1R2) fused with LTB were amplified by polymerase chain reaction (PCR). It became. Primers used for PCR are as described in Table 1 below.
  • PCR products amplified using LTBF and R1F primers containing four nucleotide sequences were inserted into the pENTR / SD vector (Gateway system, Invitrogen). Then, ltb and R1 were fused using LTBR and R1F primers containing BamHI restriction enzyme.
  • Each gene (R1, R1R2, ltbR1 and ltbR1R2) according to the manufacturer's method was inserted into pENTR / SD and pDEST-42 vector (Gateway system, Invitrogen), by using electroporation in order to express the respective recombinant proteins
  • E After transformation into coli BL21-competent cells, the cells were cultured in 1L LuriaBertani (LB) medium containing 100 ⁇ g / mL of ampicillin. The culture was induced protein expression for 3 hours by the addition of isopropyl- ⁇ -D-1-thiogalactopyranoside (IPTG) to a final concentration of 1 mM.
  • IPTG isopropyl- ⁇ -D-1-thiogalactopyranoside
  • the expressed protein was purified according to the manufacturer's method using HisPur Ni-NTA affinity chromatography (Thermo Scientific), the amino acid sequence of the recombinant protein rP97 of the present invention is set forth in SEQ ID NO: 1, the amino acid of SEQ ID NO: The base sequence encoding is set forth in SEQ ID NO: 2.
  • the concentrated M. hyopneumoniae was aseptically mixed with excipients, diluted in a sterile container as shown in Table 2 and mixed for at least 30 minutes.
  • Montanide Gel and Carbopol were inoculated with 2 ml of vaccine L1 and L2, respectively, prepared by mixing the ratios as shown in Table 2 above.
  • the PEP commercialized vaccine group was inoculated with 1 ml of Mycoflex (Boehringer Ingelheim Animal Health, St. Joseph, MO, USA), and the rlysate group contained a lysate solution containing the rP97 recombinant protein prepared in Example ⁇ 1-2>. ml inoculated.
  • the control group was inoculated with 2 ml of PBS alone, and a total of five groups were tested. All groups were boosted 21 days after initial inoculation. Serum samples were isolated at 0, 21, 42, and 63 days after initial vaccination and stored at ⁇ 20 ° C. until used in the experiment. The details of each experimental group are shown in Table 3 below.
  • M. hyopneumoniae specific antibodies are IDEXX M. hyo. Measurements were made using ELISA (IDEXX Laboratories Inc., Westbrook, ME, USA). Antibodies specific for rP97 were expressed and purified P97Rec as antigen, coated with rP97 (0.5 ⁇ g / well) on 96-well Maxi-Sorp microtiter plates (NUNC, Thermo Scientific, Penfield, NY, USA) and 4 Store at 16 ° C. for 16 hours. Plates were washed with PBS (PBST) added 0.05% Tween 20 and blocked with PBS (PBSM) containing 5% non-fat milk.
  • PBST PBS
  • PBSM PBS
  • HRP horseradish peroxidase
  • M. hyopneumoniae specific serological antibody levels induced by various vaccine candidates are shown in FIG. 1.
  • the PBS group was negative in the pre-inoculation section.
  • the groups vaccinated with the L1 and L2 vaccines were positive from day 42 to the end of the experiment.
  • the vaccinated group showed higher antibody levels than the commercial vaccine group over the entire experimental period, and L2 vaccine group showed similar levels as the commercial vaccine group at 63 days after vaccination.
  • the L1 and L2 vaccine groups induced the highest levels of antibodies at 42 and 63 days after inoculation and showed statistically high values when compared to the PBS group.
  • the control group and the commercial vaccine group did not show an rP97-specific immune response.
  • these results showed that the vaccine prepared in the present invention had a high antibody titer for rP97, while almost no vaccine had commercially available levels of the antibody against P97, which is an adhesion protein. Therefore, the vaccine prepared in the present invention was found to be more effective than the conventional commercial vaccine.
  • the control group not vaccinated showed high positive rate of cough, depression, nasal exudate, etc., but the clinical group of the vaccinated test group showed little clinical symptoms.
  • Gross lung lesion index also showed a significant difference between the unvaccinated control group and the vaccinated test group. Therefore, the vaccine prepared in the present invention was observed to alleviate respiratory symptoms and reduce lung lesions when inoculated on farms containing mycoplasma bacteria.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Microbiology (AREA)
  • Epidemiology (AREA)
  • Immunology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Mycology (AREA)
  • Wood Science & Technology (AREA)
  • Biophysics (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Physics & Mathematics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oncology (AREA)
  • Plant Pathology (AREA)
  • Communicable Diseases (AREA)
  • Pulmonology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)

Abstract

La présente invention concerne une composition de vaccin pour la prévention d'une infection par Mycoplasma hyopneumoniae (M. hyopneumoniae) chez les porcs et un procédé de préparation correspondant, la composition comprenant une protéine recombinante dérivée de M. hyopneumoniae et une bactérine de M. hyopneumoniae. La protéine recombinante de la présente invention permet au vaccin de présenter un effet protecteur supérieur à celui du vaccin classique par l'augmentation de la réponse immune contre la bactérine de M. hyopneumoniae et, de ce fait, la composition de vaccin de la présente invention permet de prévenir efficacement des maladies provoquées par une infection par M. hyopneumoniae chez les porcs, en particulier la pneumonie enzootique du porc.
PCT/KR2015/001464 2015-02-12 2015-02-13 Vaccin antibactérine de mycoplasma hyopneumoniae et son procédé de préparation WO2016129724A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2015-0021378 2015-02-12
KR1020150021378A KR20160099223A (ko) 2015-02-12 2015-02-12 마이코플라스마 하이오뉴모니애 박테린 백신 및 이의 제조방법

Publications (1)

Publication Number Publication Date
WO2016129724A1 true WO2016129724A1 (fr) 2016-08-18

Family

ID=56614408

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2015/001464 WO2016129724A1 (fr) 2015-02-12 2015-02-13 Vaccin antibactérine de mycoplasma hyopneumoniae et son procédé de préparation

Country Status (2)

Country Link
KR (1) KR20160099223A (fr)
WO (1) WO2016129724A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3498847A4 (fr) * 2016-08-12 2020-03-04 Innovac Composition vaccinale destinée à prévenir l'infection par les mycoplasmes porcins comprenant une protéine recombinée

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102228308B1 (ko) * 2018-12-19 2021-03-16 주식회사 이노백 마이코플라즈마 폐렴 및 흉막폐렴 예방용 백신 조성물

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20040030785A (ko) * 2001-07-02 2004-04-09 화이자 프로덕츠 인크. 마이코플라즈마 하이오뉴모니애로의 1회 투여량 예방접종
WO2007103042A2 (fr) * 2006-03-03 2007-09-13 Merial Limited Vaccin contre le mycoplasma hyopneumoniae
WO2010132932A1 (fr) * 2009-05-19 2010-11-25 Bioproperties Pty Ltd Souche de vaccin de mycoplasma hyopneumoniae sensible à la températureet ses utilisations
KR101225304B1 (ko) * 2007-11-06 2013-01-22 와이어쓰 엘엘씨 마이코플라스마 하이오뉴모니아에 비독성 항원보강된 살아있는 백신

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101329114B1 (ko) 2011-04-14 2013-11-14 원광대학교산학협력단 마이코플라즈마 하이오뉴모니에를 검출하기 위한 프라이머, 상기 프라이머를 이용한 마이코플라즈마 하이오뉴모니에 검출키트 및 방법, 및 상기 프라이머를 이용한 돼지 유행성 폐렴 진단키트 및 방법

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20040030785A (ko) * 2001-07-02 2004-04-09 화이자 프로덕츠 인크. 마이코플라즈마 하이오뉴모니애로의 1회 투여량 예방접종
WO2007103042A2 (fr) * 2006-03-03 2007-09-13 Merial Limited Vaccin contre le mycoplasma hyopneumoniae
KR101225304B1 (ko) * 2007-11-06 2013-01-22 와이어쓰 엘엘씨 마이코플라스마 하이오뉴모니아에 비독성 항원보강된 살아있는 백신
WO2010132932A1 (fr) * 2009-05-19 2010-11-25 Bioproperties Pty Ltd Souche de vaccin de mycoplasma hyopneumoniae sensible à la températureet ses utilisations

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BARATE, ABBIJIT K. ET AL.: "Immunogenicity of IMS 1113 Plus Soluble Subunit and Chimeric Proteins Containing Mycoplasma Hyopneumoniae P97 C-terminal Repeat Regions", FEMS MICROBIOLOGY LETTERS., vol. 352, no. 2, 13 February 2014 (2014-02-13), pages 213 - 220 *
DATABASE GenBank 31 March 2008 (2008-03-31), Database accession no. ACB38276.1 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3498847A4 (fr) * 2016-08-12 2020-03-04 Innovac Composition vaccinale destinée à prévenir l'infection par les mycoplasmes porcins comprenant une protéine recombinée

Also Published As

Publication number Publication date
KR20160099223A (ko) 2016-08-22

Similar Documents

Publication Publication Date Title
Oaks et al. Serum immune response to Shigella protein antigens in rhesus monkeys and humans infected with Shigella spp
Liu et al. Haemophilus parasuis vaccines
Fagan et al. Oral immunization of swine with attenuated Salmonella typhimurium aro A SL3261 expressing a recombinant antigen of Mycoplasma hyopneumoniae (NrdF) primes the immune system for a NrdF specific secretory IgA response in the lungs
Vanrompay et al. Protection of turkeys against Chlamydia psittaci challenge by gene gun-based DNA immunizations
US10519199B2 (en) Vaccine composition comprising recombinant protein for preventing swine Mycoplasma infection
Jorge et al. The Mycoplasma hyopneumoniae recombinant heat shock protein P42 induces an immune response in pigs under field conditions
Maas et al. Development of a DIVA subunit vaccine against Actinobacillus pleuropneumoniae infection
Hoelzle et al. Vaccination with the Mycoplasma suis recombinant adhesion protein MSG1 elicits a strong immune response but fails to induce protection in pigs
Fisch et al. Commercial bacterins did not induce detectable levels of antibodies in mice against Mycoplasma hyopneumoniae antigens strongly recognized by swine immune system
US10261092B2 (en) Cross-reactive determinants and methods for their identification
KR102243374B1 (ko) 소 코로나바이러스, 소 로타바이러스 및 소 바이러스성설사병바이러스에 대한 백신 조성물
Bao et al. Chaperonin GroEL: a novel phylogenetically conserved protein with strong immunoreactivity of Avian Pathogenic Escherichia coli isolates from duck identified by immunoproteomics
WO2020130224A1 (fr) Composition vaccinale pour la prévention de la pneumonie à mycoplasmes et de la pleuropneumonie
AU2002251192B2 (en) Bacteriophage-mediated immunisation
WO2016129724A1 (fr) Vaccin antibactérine de mycoplasma hyopneumoniae et son procédé de préparation
Macedo et al. Immune response to oligopeptide permease A (OppA) protein in pigs naturally and experimentally infected with Haemophilus parasuis
WO2011093783A1 (fr) Protéines recombinantes utilisables dans un vaccin, anticorps dirigés contre lesdites protéines et méthodes diagnostiques et thérapeutiques faisant appel à elles
Wu et al. Cross-protection of recombinant Pasteurella multocida toxin proteins against atrophic rhinitis in mice
Cauchard et al. Assessment of the safety and immunogenicity of Rhodococcus equi-secreted proteins combined with either a liquid nanoparticle (IMS 3012) or a polymeric (PET GEL A) water-based adjuvant in adult horses and foals—identification of promising new candidate antigens
Andoh et al. Identification of a major immunogenic region of equine herpesvirus-1 glycoprotein E and its application to enzyme-linked immunosorbent assay
Wang et al. Three novel immunogenic proteins determined through 2-Dimensional electrophoresis and mass spectrometry with immune serum confer protection against challenge with porcine Pasteurella multocida in mouse models
Throne Steinlage et al. Isolation and characterization of a 6/85-like Mycoplasma gallisepticum from commercial laying hens
WO2016173504A1 (fr) Protéine recombinante de toxine d'actinobacillus pleuropneumoniae et application correspondante
Liu et al. Development of a novel PmpD-N ELISA for Chlamydia psittaci infection
MX2012006303A (es) Procedimiento de identificacion de animales vacunados frente a brucella.

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15882087

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15882087

Country of ref document: EP

Kind code of ref document: A1