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Immunization against chlamydia infection

A chlamydia, immunogenic technology for use in immunology to address issues such as the increase in antibiotic-resistant microbes, where chemotherapy or antibiotic treatment is not a viable long-term strategy

Inactive Publication Date: 2007-01-31
圣诺菲·帕斯图尔公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chemotherapy or antibiotic treatment may not be a viable long-term strategy as increased antibiotic use has resulted in an increase in antibiotic-resistant microorganisms

Method used

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  • Immunization against chlamydia infection
  • Immunization against chlamydia infection
  • Immunization against chlamydia infection

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0094] Standard molecular biology techniques for preparing and purifying polynucleotides are used in the preparation of polynucleotide therapeutics of the invention. For use as a vaccine, the polynucleotides of the invention are formulated according to the various methods outlined below.

[0095] One method utilizes naked polynucleotides without any delivery vehicle. The polynucleotide is simply diluted in a physiologically acceptable solution, such as sterile saline or sterile buffered saline, with or without carrier. When present, the carrier is preferably isotonic, hypotonic or slightly hypertonic, and has a relatively low ionic strength, such as that provided by a sucrose solution, eg, a solution containing 20% ​​sucrose.

[0096] Alternative methods utilize polynucleotides in combination with agents that assist in cellular uptake. Examples of such agents are (i) chemical agents that modify cell permeability, such as bupivacaine (see, e.g., WO 94 / 16737), (ii) liposomes f...

Embodiment 1

[0148] This example illustrates the preparation of plasmid vectors for immunization.

[0149] Chlamydia trachomatis mouse pneumonia (MoPn) isolates were grown in Hela229 cells in Eagle MEM containing 10% fetal bovine serum and 2 mM L-glutamine. MoPn EBs were harvested and purified by a gradient density centrifugation step at 43,000 g for 60 min at 4°C. Purified EBs were washed twice with PBS, centrifuged at 30,000g for 30 minutes, resuspended in sucrose-phosphate-glutamate (SPG) buffer and frozen at -70°C until use.

[0150] The nucleic acid molecule encoding the Mgp002 gene is cloned in-frame into the eukaryotic expression plasmid pCAMycHis, and the Myc-His tag exists in the vector. This vector was constructed from pcDNA3.1(-)Myc-His C (Invitrogen, San Diego) and plasmid VR1012 (Vical). Details of the construction are disclosed in PCT Publication WO 00 / 55326, published September 21,2000. Briefly, plasmid pcDNA3.1(-)Myc-His C (Invitrogen) was digested with Spe I and Bam HI ...

Embodiment 2

[0155] This example shows the results of immunization studies using nucleic acid vectors.

[0156] To investigate whether the immune response elicited by nucleic acid immunization was functionally effective, in vivo protective efficacy was assessed according to previously described methods (ref. 20). Briefly, female Balb / c mice (4 to 5 weeks old) were purchased from Charles River Canada (St. Constant, Canada). Mice were immunized intramuscularly and intranasally with plasmid DNA prepared according to the method described in Example 1, see FIG. 3 at three times of 0, 2 and 4 weeks. For each immunization, 200 μl of a total of 200 μg DNA was injected into both quadriceps using a 27 gauge needle (100 μg DNA per injection site). At the same time, 50 μl of 50 μg DNA was delivered to the nostrils of the mice using a micropipette. The droplets were then inhaled by the mice.

[0157] Fourteen days after the last immunization, 2 × 10 3 IFU of Chlamydia trachomatis MoPn EB challenged...

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Abstract

The present invention provides nucleic acids, proteins and vectors for a method of nucleic acid, including DNA, immunization of a host, including humans, against disease caused by infection by a strain of Chlamydia, specifically C. trachomatis. The method employs a vector containing a nucleotide sequence encoding a Mgp002 polypeptide of a strain of Chlamydia operably linked to a promoter to effect expression of the gene product in the host. Truncated forms of the full-length Mgp002 gene are useful immunogens for protecting against disease caused by infection with Chlamydia. The invention further provides recombinant Mgp002 protein useful for protecting against disease caused by infection with Chlamydia.

Description

field of invention [0001] The present invention relates to immunology, and in particular to the use of nucleic acid molecules to immunize a host to provide protection against Chlamydia infection. Background of the invention [0002] Nucleic acid immunization is a method of generating protective immunity against infectious disease (Reference 1 - Throughout this application, various references are cited in parentheses to more fully describe the state of the art to which this invention pertains. (Cited The full bibliographic information of the present invention is listed at the end of the present invention book, before the claims. The contents disclosed by these references are hereby incorporated by reference in this disclosure). Unlike protein or peptide-based subunit vaccines, nucleic acid or DNA immunization is achieved through Host cells express exogenous proteins to provide protective immunity, thus allowing antigen presentation to the immune system ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31A61K39/118A61K31/7088A61P31/04C07H21/00C07K14/295C07K16/12C12P21/02A61K38/16A61K39/00
CPCA61K2039/55555A61K2039/55577A61K2039/53C07K14/295A61K39/00A61K39/118A61P31/04A61P31/10C12N15/11A61K31/7088
Inventor R·C·布伦哈姆A·劳多尼基尼S·加利钱A·穆尔丁
Owner 圣诺菲·帕斯图尔公司
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