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US20110189217A1 - Methods and materials for producing immune responses against polypeptides involved in antibiotic resistance - Google Patents

Methods and materials for producing immune responses against polypeptides involved in antibiotic resistance Download PDF

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US20110189217A1
US20110189217A1 US13/001,228 US200913001228A US2011189217A1 US 20110189217 A1 US20110189217 A1 US 20110189217A1 US 200913001228 A US200913001228 A US 200913001228A US 2011189217 A1 US2011189217 A1 US 2011189217A1
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polypeptide
antibiotic resistance
resistance
polypeptides involved
animal
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Michael A. Barry
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Mayo Clinic in Florida
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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
    • A61K39/02—Bacterial antigens
    • A61K39/04—Mycobacterium, e.g. Mycobacterium tuberculosis
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/085—Staphylococcus
    • 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
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • 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
    • A61K2039/5256—Virus 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
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    • A61K2039/53—DNA (RNA) vaccination
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    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
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    • C12N2710/10011—Adenoviridae
    • C12N2710/10041—Use of virus, viral particle or viral elements as a vector
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    • C12N2800/00—Nucleic acids vectors
    • C12N2800/22—Vectors comprising a coding region that has been codon optimised for expression in a respective host

Definitions

  • This document relates to methods and materials for producing immune responses against polypeptides involved in antibiotic resistance.
  • this document provides vaccines against polypeptides involved in antibiotic resistance as well as methods for vaccinating mammals against polypeptides involved in antibiotic resistance.
  • Mtb Mycobacterium tuberculosis
  • MDR multi-drug resistant
  • XDR extensively drug resistant
  • This document relates to methods and materials for producing immune responses against polypeptides involved in antibiotic resistance.
  • this document provides vaccines against polypeptides involved in antibiotic resistance as well as methods for vaccinating mammals against polypeptides involved in antibiotic resistance.
  • one aspect of this document features a method for inducing an immune response against a polypeptide involved in antibiotic resistance.
  • the method comprises, or consists essentially of, administering to an animal (e.g., a mammal) an amount of the polypeptide or a nucleic acid encoding the polypeptide effective for producing the immune response.
  • the polypeptide can be a blaZ polypeptide, a mecA polypeptide, a whiB7 polypeptide, a tap polypeptide, a RV1473 polypeptide, a katG polypeptide, an inhA polypeptide, a rpoB polypeptide, a gidB polypeptide, a pncA polypeptide, an embB polypeptide, or a gyrA polypeptide.
  • the antibiotic resistance can be penicillin-resistance.
  • the antibiotic resistance can be methicillin-resistance.
  • the antibiotic resistance can be vancomycin-resistance.
  • the animal can be a human.
  • the method can comprise administering the polypeptide to the animal.
  • the method can comprise administering the nucleic acid to the animal.
  • the nucleic acid can be a viral vector encoding the polypeptide.
  • the viral vector can be an adenoviral, vaccinia viral, measles, or adeno-associated virus vector.
  • the immune response can reduce the severity of an infection within said animal.
  • the infection can be a Mycobacterium tuberculosis infection.
  • the infection can be a Staphylococcus aureus infection.
  • FIG. 1 is a photograph of a western blot of cell lysates obtained from mammalian cells that were transfected with plasmids expressing either a BlaZ polypeptide or a MecA polypeptide. Proteins were detected with a anti-His6 antibody recognizing a His6 tag on the c-terminus of each of the proteins. The left band are normal cells. The right band of each set are cells expressing Rad23 that inhibits proteasome degradation of proteins. The negative control ( ⁇ ve control) is a lysate of cells transfected with GFP gene.
  • FIG. 2 is photograph of Western blot of BlaZ polypeptides purified from a cell lysate and from the media of bacterial cells.
  • CBD designates the cellulose binding domain fusion tag of the pET38 vector.
  • BlaZ-CBD is the fusion of BlaZ to the CBD tag.
  • This document provides methods and materials for producing immune responses against polypeptides involved in antibiotic resistance.
  • this document provides vaccines against polypeptides involved in antibiotic resistance as well as methods for vaccinating mammals against polypeptides involved in antibiotic resistance.
  • the vaccines provided herein can be in the form of recombinant polypeptides involved in antibiotic resistance or nucleic acid vectors (e.g., viral vectors) designed to express such recombinant polypeptides.
  • the vaccines provided herein can be used to immunize or treat any type of animal including, without limitation, humans, cows, pigs, poultry, dogs, and cats.
  • the vaccines provided herein can be used to induce an immune response against any type of pathogen including, without limitation, intracellular pathogens such as mycobacterium, chlamydia, rickettsiae, lysteria , and brucella as well as extracellular pathogens such as staphylococci, streptococci, enterococci, pneumococci, gram positive bacteria, and anthrax.
  • pathogens such as mycobacterium, chlamydia, rickettsiae, lysteria , and brucella
  • extracellular pathogens such as staphylococci, streptococci, enterococci, pneumococci, gram positive bacteria, and anthrax.
  • the vaccines provided herein can be designed to target ⁇ -lactamase and penicillin binding proteins to treat or prevent community and hospital-acquired bacterial infections such as methicillin-resistant Staphylococcus aureus.
  • polypeptides involved in antibiotic resistance include, without limitation, a blaZ polypeptide (see Entrez Gene ID: 2859819) from S. aureus ; a mecA polypeptide (Entrez Gene ID: 2861157) from S. aureus ; a whiB7 polypeptide from Mtb (Entrez Gene ID: 3205083); a tap polypeptide from Mtb (Entrez Gene ID: 924773); a RV1473 polypeptide from Mtb (Entrez Gene ID: 886549); a katG polypeptide from Mtb (Entrez Gene ID: 885638); an inhA polypeptide from Mtb (Entrez Gene ID: 886523); a rpoB polypeptide from Mtb (Entrez Gene ID: 925979); a gidB polypeptide from Mtb (Entrez Gene ID: 886243); a pncA polypeptide from Mtb (Entrez Gene ID: 888260); an
  • polypeptides can include the wild-type sequences of these proteins or variants acquired with mutations that confer drug-resistance.
  • a vaccine provided herein can be designed to induce an immune response against a polypeptide involved in antibiotic resistance and one or more polypeptides not involved in antibiotic resistance (e.g., Ag85a of Mtb).
  • the vaccines provided herein can be used as stand alone vaccines or as adjuvants to other vaccines that target one or more pathogens (e.g., a vaccine that targets multiple Mtb antigens).
  • a vaccine provided herein e.g., a vaccine that targets one or more polypeptides involved in antibiotic resistance
  • another vaccine e.g., a BCG vaccine
  • nucleic acid encoding one or more polypeptides involved in antibiotic resistance can be inactivated and introduced into another vaccine (e.g., a BCG vaccine). Genetic engineering can be used to inactivate polypeptides involved in antibiotic resistance by mutating active site amino acids or by expressing the polypeptide as a cocktail of fragments of the full length polypeptide.
  • nucleic acid encoding a polypeptide involved in antibiotic resistance can be delivered using plasmids or viral vectors such as adenoviral vectors, vaccinia viral vectors, measles, or adeno-associated virus vectors.
  • the polypeptides can be produced in bacteria or yeast as purified polypeptide vaccines.
  • a vaccine provided herein can be delivered as a prophylactic vaccine to assist in preventing the production of antibiotic resistance should an infection occur.
  • a vaccine provided herein can be applied therapeutically as an adjuvant to antibiotic treatment. For example, upon initiation of multidrug therapy against MDR or XDR, a patient can be vaccinated against Mtb and against one or more polypeptides involved in antibiotic resistance.
  • the vaccines provided herein can be designed to induce immune responses against one or more (e.g., two, three, four, five, six, seven, or more) polypeptides involved in antibiotic resistance.
  • a vaccine can be designed to be patient-specific.
  • a human known to have a particular antibiotic resistant pathogen can be given a vaccine that induces an immune response against the particular polypeptide or polypeptides involved in antibiotic resistance for that pathogen.
  • Any appropriate method can be used to determine the identity of the particular polypeptides involved in antibiotic resistance for a particular pathogen including, without limitation, PCR techniques.
  • polypeptides involved in antibiotic resistance can be used to generate antibodies (e.g., monoclonal antibodies) that can be used alone or in combination with antibiotics to deplete resistance proteins and enhance antibiotic efficacy.
  • antibodies e.g., monoclonal antibodies
  • a vaccine provided herein e.g., a vaccine that includes a recombinant polypeptide involve in antibiotic resistance
  • an adjuvant can be an immunological compound that can enhance an immune response against a particular antigen such as a polypeptide.
  • Suitable adjuvants include, without limitation, alum as well as other aluminum-based compounds (e.g., Al 2 O 3 ) that can be obtained from various commercial suppliers.
  • REHYDRAGEL® adjuvants can be obtained from Reheis Inc. (Berkeley Heights, N.J.).
  • REHYDRAGEL® adjuvants are based on crystalline aluminum oxyhydroxide, and are hydrated gels containing crystalline particles with a large surface area (about 525 m 2 /g). Their Al 2 O 3 content typically ranges from about 2 percent to about 10 percent. Rehydragel LG, for example, has an Al 2 O 3 content of about 6 percent, and flows readily upon slight agitation.
  • Rehydragel LG also has a protein binding capacity of 1.58 (i.e., 1.58 mg of bovine serum albumin bound per 1 mg of Al 2 O 3 ), a sodium content of 0.02 percent, a chloride content of 0.28 percent, undetectable sulphate, an arsenic level less than 3 ppm, a heavy metal content less than 15 ppm, a pH of 6.5, and a viscosity of 1090 cp.
  • Rehydragel LG can be combined with a polypeptide solution (e.g., a polypeptide in PBS) to yield Al(OH) 3 .
  • ALHYDROGELTM an aluminum hydroxy gel adjuvant, (Alhydrogel 1.3%, Alhydrogel 2.0%, or Alhydrogel “85”) obtained from Brenntag Stinnes Logistics can be used.
  • MN51 can be combined with a vaccine provided herein to form a composition that elicits an immune response when administered to a mammal.
  • MN51 (MONTANIDE® Incomplete SEPPIC Adjuvant (USA) 51) as well as MN720 are available from Seppic (Paris, France).
  • MN51 contains mannide oleate (MONTANIDE® 80, also known as anhydro mannitol octadecenoate) in mineral oil solution (Drakeol 6 VR).
  • MONTANIDE® 80 is a limpid liquid with a maximum acid value of 1, a saponification value of 164-172, a hydroxyl value of 89-100, an iodine value of 67-75, a maximum peroxide value of 2, a heavy metal value less than 20 ppm, a maximum water content of 0.35%, a maximum color value of 9, and a viscosity at 25° C. of about 300 mPas.
  • MONTANIDE® associated with oil e.g., mineral oil, vegetable oil, squalane, squalene, or esters
  • Drakeol 6 VR is a pharmaceutical grade mineral oil.
  • Drakeol 6 VR contains no unsaturated or aromatic hydrocarbons, and has an A.P.I. gravity of 36.2-36.8, a specific gravity at 25° C. of 0.834-0.838, a viscosity at 100° F. of 59-61 SSU or 10.0-10.6 centistokes, a refractive index at 25° C. of 1.458-1.463, a better than minimum acid test, is negative for fluorescence at 360 nm, is negative for visible suspended matter, has an ASTM pour test value of 0-15° F., has a minimum ASTM flash point of 295° F., and complies with all RN requirements for light mineral oil and ultraviolet absorption.
  • MN51 contains about 8 to 12 percent anhydro mannitol octadecenoate and about 88 to 92 percent mineral oil.
  • adjuvants include immuno-stimulating complexes (ISCOMs) that can contain such components as cholesterol and saponins ISCOM matrices can be prepared and conjugated to Cu 2+ using methods such as those described herein.
  • Adjuvants such as FCA, FIA, MN51, MN720, and Al(OH) 3 are commercially available from companies such as Seppic, Difco Laboratories (Detroit, Mich.), and Superfos Biosector A/S (Vedbeak, Demark).
  • immunostimulatory components include, without limitation, muramyldipeptide (e.g., N-acetylmuramyl-L-alanyl-D-isoglutamine; MDP), monophosphoryl-lipid A (MPL), formyl-methionine containing tripeptides such as N-formyl-Met-Leu-Phe, or a bacterial lipopolysaccarhide.
  • MDP muramyldipeptide
  • MPL monophosphoryl-lipid A
  • formyl-methionine containing tripeptides such as N-formyl-Met-Leu-Phe
  • bacterial lipopolysaccarhide bacterial lipopolysaccarhide.
  • Additional immunostimulatory components can include pneumovax (an approved human vaccine), CD40L, or IL-12.
  • an adjuvant can be Complete Freund's Adjuvant or Incomplete Freund's Adjuvant.
  • This document also provides methods for preparing a vaccine provided herein. Such methods can involve suspending an amount of a nucleic acid vector (e.g., viral vector) or a polypeptide in a suitable amount of a physiological buffer (e.g., PBS). The nucleic acid vector or polypeptide then can be combined with a suitable amount of an adjuvant/immunostimulatory compound.
  • the combining step can be achieved by any appropriate method, including, for example, stirring, shaking, vortexing, or passing back and forth through a needle attached to a syringe.
  • compositions can be prepared in batch, such that enough unit doses are obtained for multiple injections (e.g., injections into multiple mammals or multiple injections into the same mammal).
  • a “unit dose” of a composition provided herein refers to the amount of a composition administered to a mammal at one time.
  • a unit dose of the compositions provided herein can contain any amount of polypeptides involved in antibiotic resistance or nucleic acid encoding such polypeptides.
  • a unit dose of a composition can contain between about 0.1 ⁇ g and about 1 g (e.g., 1 ⁇ g, 10 ⁇ g, 15 ⁇ g, 25 ⁇ g, 30 ⁇ g, 50 ⁇ g, 100 ⁇ g, 250 ⁇ g, 280 ⁇ g, 300 ⁇ g, 500 ⁇ g, 750 ⁇ g, 1 mg, 10 mg, 15 mg, 25 mg, 30 mg, 50 mg, 100 mg, 250 mg, 280 mg, 300 mg, 500 mg, 750 mg, or more) of one or more polypeptides involved in antibiotic resistance.
  • 1 g e.g., 1 ⁇ g, 10 ⁇ g, 15 ⁇ g, 25 ⁇ g, 30 ⁇ g, 50 ⁇ g, 100 ⁇ g, 250 mg, 280 mg, 300 mg, 500 mg, 750 mg, or more
  • a unit dose of a composition can have a titer between about 10 3 to 10 10 (e.g., 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , or 10 10 ) viral particles or plaque forming units.
  • 10 3 to 10 10 e.g., 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , or 10 10
  • Methods for inducing a particular immune response in a mammal include, without limitation, administering to a mammal an amount of a vaccine provided herein that is effective for producing an antibody response against one or more polypeptides involved in antibiotic resistance.
  • the vaccines provided herein can be administered using any appropriate method.
  • Administration can be, for example, topical (e.g., transdermal, ophthalmic, or intranasal); pulmonary (e.g., by inhalation or insufflation of powders or aerosols); oral; or parenteral (e.g., by subcutaneous, intrathecal, intraventricular, intramuscular, or intraperitoneal injection, or by intravenous drip).
  • Administration can be rapid (e.g., by injection) or can occur over a period of time (e.g., by slow infusion or administration of slow release formulations).
  • Codon-optimized sequences for Mtb antigens are obtained from Genscript Corporation (Piscataway, N.J.) or generated using molecular cloning techniques. These sequences are codon-optimized for expression in mammalian cells for use as gene-based vaccines.
  • the following Mtb genes that are expressed by H37Rv, MDR Mtb, or XDR Mtb or those listed in Table 1 that are expressed by H37Rv, MDR Mtb, or XDR Mtb are synthesized and cloned into the pShuttle-CMV plasmid:
  • Ag85a Positive control protective antigen.
  • whiB7 Inducer of expression of a regulon of Mtb genes involved in antibiotic resistance (including tap, RV1473, and erm (Morris et al., Proc. Nat'l. Acad. Sci. USA, 102(34):12200-12205 (2005)).
  • Drug efflux pump conferring low level resistance to aminoglyosides and tetracycline.
  • RV1473 Putative macrolide transporter induced by whiB7 (Morris et al., Proc. Nat'l. Acad. Sci. USA, 102(34):12200-12205 (2005)).
  • erm Confers resistance to macrolide, lincosamide, and streptogramin.
  • Codon-optimized sequences for antigens from Staphylococcus aureus or other gram-positive bacteria are generated using molecular cloning techniques. These sequences are codon-optimized for expression in mammalian cells for use as gene-based vaccines.
  • Ad5 vectors are used to generate gene-based vaccines, which are used as an effective vaccine delivery vehicle in mice.
  • Any appropriate vaccine carrier including Bacillus Calmette-Guerin (BCG) or vaccinia is used as a vaccine delivery vehicle in humans.
  • BCG Bacillus Calmette-Guerin
  • vaccinia is used as a vaccine delivery vehicle in humans.
  • the recombinant polypeptides are delivered directly to the mammal (e.g., a human).
  • the pShuttle-CMV vectors are obtained, they are recombined into the Ad5 genome in bacteria and are used to generate CsCl-purified Ad5 vaccines.
  • the codon-optimized nucleic acid encoding a BlaZ polypeptide was cloned into a shuttle plasmid fusing the sequence to the alpha 1-anti-trypsin secretory leader. Expression in cell lines generated detectable amounts of polypeptide from mammalian cells ( FIG. 1 ).
  • Polypeptide vaccines can be produced from bacterial expression plasmids.
  • the sequence of SEQ ID NO:1 was cloned into a pET-38 plasmid fusing it to a cellulose binding domain (CBD) and a His6 tag for purification.
  • CBD cellulose binding domain
  • His6 tag for purification.
  • Expression of the polypeptide from bacteria followed by Western blotting revealed the presence of the polypeptide intracellularly in cell lysates and in a form secreted into the media ( FIG. 2 ).
  • amino acid sequence encoded by SEQ ID NO:3 is as follows: SKDK-EINNTIDAIEDKNFKQVYKDSSYISKSDNGEVEMTERPIKIYNSLGVKDINIQDRKIKK VSKNKKRVDAQYKIKTNYGNIDRNVQFNFVKEDGMWKLDWDHSVIIPGMQKDQSI HIENLKSERGKILDRNNVELANTGTAYEIGIVPKNVSKKDYKAIAKELSISEDYIKQQ MDQNWVQDDTFVPLKTVKKMDEYLSDFAKKFHLTTNETESRNYPLGKATSHLLGY VGPINSEELKQKEYKGYKDDAVIGKKGLEKLYDKKLQHEDGYRVTIVDDNSNTIAH TLIEKKKKDGKDIQLTIDAKVQKSIYNNMKNDYGSGTAIHPQTGELLALVSTPSYDV YPFMYGMSNEEYNKLTEDKKEPLLNKFQITTSPGSTQKILTAMI
  • the codon-optimized nucleic acid encoding a MecA polypeptide was cloned into a shuttle plasmid fusing the sequence to the alpha 1-anti-trypsin secretory leader. Expression in cell lines generated detectable amounts of polypeptide from mammalian cells ( FIG. 1 ).
  • mice are immunized intranasally with 10 10 virus particles of Ad vaccines expressing the following antigens: saline; GFP (negative control vaccine for Mtb); Ag85a (positive control for CD8 responses and protection); and whiB7, tap, RV1473 and erm (vaccines against polypeptides involved in antibiotic resistance). Additional groups are used for the genes listed in Table 1.
  • saline GFP (negative control vaccine for Mtb); Ag85a (positive control for CD8 responses and protection); and whiB7, tap, RV1473 and erm (vaccines against polypeptides involved in antibiotic resistance). Additional groups are used for the genes listed in Table 1.
  • ELISPOT intracellular cytokine staining.
  • the remaining mice of each group are challenged with 300 colony forming units of H37Rv/mouse by aerosolization to mimic the normal Mtb infection route.
  • H37Rv is not classified as MDR or XDR, but is resistant to a number of antibiotics (Morris et al., Proc. Nat'l. Acad. Sci. USA, 102(34):12200-12205 (2005)). In a separate study, legitimate MDR or XDR Mtb is used in place of H37Rv.
  • WPI 8 weeks post-infection
  • ten mice from each group are sacrificed, and Mtb is titered from the lung and the spleen. Histopathology and MST testing are also performed on the samples. The remaining mice are retained to estimate median survival time (MST). Control animals typically survive until 30 weeks with longer survival for successful vaccines. At 48 weeks, all animals are sacrificed for titering and histopathology.

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013112696A1 (fr) * 2012-01-27 2013-08-01 Los Angeles Biomedical Research Institute At Harbor-Ucla Medical Center Compositions et procédés pour l'immunisation contre des bactéries exprimant une carbapénèmase
US20170035873A1 (en) * 2008-06-26 2017-02-09 Mayo Foundation For Medical Education And Research Methods and materials for producing immune responses against polypeptides involved in antibiotic resistance

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013135274A1 (fr) * 2012-03-13 2013-09-19 Intercell Ag Composés d'aluminium et agents thérapeutiques destinés à être utilisés dans des vaccins
DK3785730T3 (da) * 2011-12-06 2024-05-06 Valneva Austria Gmbh Aluminiumforbindelser til anvendelse i terapeutiske produkter og vacciner
AU2012352455B2 (en) * 2011-12-12 2016-01-21 The Trustees Of The University Of Pennsylvania Proteins comprising MRSA PBP2a and fragments thereof, nucleic acids encoding the same, and compositions and their use to prevent and treat MRSA infections
US9895437B2 (en) 2012-04-18 2018-02-20 Valneva Austria Gmbh Aluminum compounds for use in therapeutics and vaccines
US11185555B2 (en) 2016-04-11 2021-11-30 Noah James Harrison Method to kill pathogenic microbes in a patient
CN110777154A (zh) * 2019-07-19 2020-02-11 华大生物科技（武汉）有限公司 用于结核分枝杆菌耐药检测的突变基因、及其检测方法和试剂盒
US12267062B2 (en) 2020-06-17 2025-04-01 Murata Manufacturing Co., Ltd. Transversely-excited film bulk acoustic resonators with three-layer electrodes

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5686590A (en) * 1993-05-14 1997-11-11 Agresearch, New Zealand Pastoral Agriculture Research Institute Ltd. Methods and compositions for detecting and treating mycobacterial infections using an INHA gene
US6353093B1 (en) * 1997-07-01 2002-03-05 Smithkline Beecham Corporation gidB
US20070037163A1 (en) * 1997-08-14 2007-02-15 Genome Therapeutics Corporation Nucleic acid and amino acid sequences relating to staphylococcus epidermidis for diagnostics and therapeutics
US20110014240A1 (en) * 2008-03-06 2011-01-20 Mayo Foundation For Medical Education And Research Single cycle replicating adenovirus vectors
US20130022997A1 (en) * 2011-07-21 2013-01-24 Raybiotech, Inc. Monoclonal antibody against pbp2a derived from mrsa with dual binding activities
WO2013090294A1 (fr) * 2011-12-12 2013-06-20 The Trustees Of The University Of Pennsylvania Protéines comprenant la pbp2a de sarm et des fragments de celle-ci, acides nucléiques codant pour celles-ci, et compositions et leur utilisation pour la prévention et le traitement d'infections par le sarm
US8546093B2 (en) * 2009-03-11 2013-10-01 Dinona Inc. Detection method for methicillin resistant staphylococcus aureus

Family Cites Families (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006516193A (ja) * 2002-12-06 2006-06-29 アイシス・ファーマシューティカルス・インコーポレーテッド ヒトおよび動物における病原体の迅速な同定方法
PL2457926T3 (pl) * 2005-04-29 2015-03-31 Glaxosmithkline Biologicals Sa Nowy sposób profilaktyki lub leczenia zakażenia m. tuberculosis
US9717788B2 (en) * 2007-03-02 2017-08-01 Glaxosmithkline Biologicals Sa Method of inducing an immune response against HIV employing HIV immunogens, adenoviral vectors encoding said immunogens, and adjuvant
US20110105531A1 (en) * 2007-06-22 2011-05-05 Ibis Biosciences, Inc. Compositions and methods for identification of subspecies characteristics of mycobacterium tuberculosis
US9598737B2 (en) * 2012-05-09 2017-03-21 Longhorn Vaccines And Diagnostics, Llc Next generation genomic sequencing methods
EP2231181B1 (fr) * 2007-12-17 2016-02-17 Marfl AB Nouveau vaccin pour le traitement de troubles liés à une mycobactérie
WO2009158284A2 (fr) * 2008-06-26 2009-12-30 Mayo Foundation For Medical Education And Research Méthodes et matériaux de production de réponses immunitaires contre des polypeptides impliqués dans la résistance aux antibiotiques
WO2010010179A1 (fr) * 2008-07-25 2010-01-28 Glaxosmithkline Biologicals S.A. Protéine rv2386c tuberculosis, compositions et utilisations correspondantes
US8367055B2 (en) * 2009-05-14 2013-02-05 Wisconsin Alumni Research Foundation Immunogenic compositions against tuberculosis
GB201002413D0 (en) * 2010-02-12 2010-03-31 Procarta Biosystems Ltd Nucleic acid complexes
GB201008719D0 (en) * 2010-05-25 2010-07-07 Nat Univ Ireland Diagnostic method
WO2015031385A1 (fr) * 2013-08-26 2015-03-05 The Translational Genomics Research Institute Analyse de lecture à chevauchement monomoléculaire pour la détection de mutation variante mineure dans des échantillons de pathogènes
US20160074480A1 (en) * 2015-11-24 2016-03-17 Totada R. Shantha Innovative methods of treatmenting tuberculosis

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5686590A (en) * 1993-05-14 1997-11-11 Agresearch, New Zealand Pastoral Agriculture Research Institute Ltd. Methods and compositions for detecting and treating mycobacterial infections using an INHA gene
US6353093B1 (en) * 1997-07-01 2002-03-05 Smithkline Beecham Corporation gidB
US20070037163A1 (en) * 1997-08-14 2007-02-15 Genome Therapeutics Corporation Nucleic acid and amino acid sequences relating to staphylococcus epidermidis for diagnostics and therapeutics
US20110014240A1 (en) * 2008-03-06 2011-01-20 Mayo Foundation For Medical Education And Research Single cycle replicating adenovirus vectors
US8546093B2 (en) * 2009-03-11 2013-10-01 Dinona Inc. Detection method for methicillin resistant staphylococcus aureus
US20130022997A1 (en) * 2011-07-21 2013-01-24 Raybiotech, Inc. Monoclonal antibody against pbp2a derived from mrsa with dual binding activities
WO2013090294A1 (fr) * 2011-12-12 2013-06-20 The Trustees Of The University Of Pennsylvania Protéines comprenant la pbp2a de sarm et des fragments de celle-ci, acides nucléiques codant pour celles-ci, et compositions et leur utilisation pour la prévention et le traitement d'infections par le sarm

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Berger-Bachi et al, Arch Microbiol (2002) 178 :165-171 *
Ito et al, ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, June 1999, p. 1449-1458 Vol. 43, No. 6 *
Ohwada et al, J. Antimicrobial Chemotherapy, 1999, 44:767-774 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170035873A1 (en) * 2008-06-26 2017-02-09 Mayo Foundation For Medical Education And Research Methods and materials for producing immune responses against polypeptides involved in antibiotic resistance
US10143736B2 (en) * 2008-06-26 2018-12-04 Mayo Foundation For Medical Education And Research Methods and materials for producing immune responses against polypeptides involved in antibiotic resistance
WO2013112696A1 (fr) * 2012-01-27 2013-08-01 Los Angeles Biomedical Research Institute At Harbor-Ucla Medical Center Compositions et procédés pour l'immunisation contre des bactéries exprimant une carbapénèmase
US9169477B2 (en) 2012-01-27 2015-10-27 Los Angeles Biomedical Research Institute At Harbor-Ucla Medical Center Compositions and methods for immunization against bacteria expressing a carbapenemase

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