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Bordetella parapertussis-containing whole-cell vaccine composition

A technology of vaccine composition and parapertussis, which is applied in the direction of antibacterial drugs, antibody medical components, bacterial antigen components, etc. It can solve the problems of difficulty in making an effective vaccine, unclear effect, and difficulty in the evaluation system of Bacillus parapertussis vaccine

Inactive Publication Date: 2010-08-04
DAIICHI SANKYO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, although the vaccine with the mutant strain of Bordetella as the immunogen (Patent Document 1), the vaccine belonging to the outer membrane protein expressed in B. parapertussis and related to pertactin, etc. are in progress ( Patent Document 2), but in fact their effect on pertussis caused by B. parapertussis is not clear, and vaccines that can effectively prevent pertussis caused by B. parapertussis infection are not yet known
In addition, it is very difficult to apply an effective vaccine evaluation system for B. parapertussis, and it has not been successful so far.
[0006] In addition, with regard to the new development of whole-bacteria vaccines, as the development of many pathogens such as Shigella, Neisseria gonorrhoeae, meningococcus, and malaria has ended in failure, in the development and manufacture of whole-bacteria pertussis vaccines , it is very difficult to make an effective vaccine (Non-Patent Document 6)

Method used

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Examples

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Embodiment

[0119] Hereinafter, the present invention will be more specifically described based on examples, but the present invention is not limited to these examples.

[0120] Composition of the culture medium used in the present invention

[0121]

[0122] Sodium chloride 5.5g

[0123] Agar 20g

[0124] Potato extract with 1% glycerin 1L

[0125] After sterilizing at 121° C. for 15 minutes, 150 mL of sheep decellulose blood was added. Add appropriate antimicrobials when using resistant strains. Streptomycin 200 μg / mL in the above case.

[0126]

[0127] Bovine Heart Extract 12g

[0128] Peptone 10g

[0129] Sodium chloride 5g

[0130] Soluble starch 10g

[0131] Yeast Extract 3.5g

[0132] Activated carbon powder 4g

[0133] Agar 18g

[0134] Ultrapure water 1L

[0135] After sterilizing at 121° C. for 15 minutes, 100 mL of sheep decellulose blood was added. Add appropriate antimicrobials when using resistant strains. Streptomycin 200 μg / mL in the above case.

[0136...

Embodiment 1

[0160] (Example 1) Bacillus parapertussis vaccine

[0161] Research on the cultivation method of the bacterial cells used as the Bacillus parapertussis vaccine

[0162] Suspend Bacillus parapertussis in the Stainer-Scholte liquid medium supplemented with casamino acids according to the method of reaching 1 billion cells / mL, take 10 mL of it and put it into a 75 cm 2 Culture flasks were cultured statically at 37°C. Under this condition, the depth of culture medium and static conditions are close to those of solid plate culture, and solid plate culture can quantify the difficult-to-determined bacterial cell proliferation. In addition, the Stainer-Scholte liquid medium to which casamino acids were added contained sufficient nutrients necessary for the growth of B. parapertussis similarly to the charcoal powder medium. On the other hand, in order to prepare a vaccine, the bacteria must be kept in a state of pathogenicity. In order to confirm whether pathogenicity is maintained,...

Embodiment 2

[0174] (Example 2) Detection of the infection defense activity (effectiveness) of the vaccine

[0175] The infection defense activity (efficacy) of each vaccine was judged using the mouse nasal infection method which has a strong correlation with the vaccine's effect on humans (Guiso, N. et al., Vaccine 17: 2366-2376, 1999.). That is, mice (3.5 weeks old) were subcutaneously inoculated with each vaccine at 0.125 mL (equivalent to 1 / 4 human single inoculation dose, SHD), and boosted (equal dose) 14 days later. 14 days after the booster immunization, the suspension of Bacillus parapertussis (about 10 8 CFU / mL, 50 μL / mouse) under pentobarbital anesthesia (0.5 mL intraperitoneally inoculated with 20-fold diluted Nembutal injection) for nasal infection. Mice were euthanized with pentobarbital after 0 (approximately 2 hours), 2, 5, and 8 days later, and lungs were aseptically removed and homogenized in 10 mL of sterile buffered saline. The homogenate was graded and diluted, spread...

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Abstract

It is intended to provide a whole-cell bacterial vaccine composition for preventing whooping cough derived from Bordetella parapertussis containing the whole cell of Bordetella parapertussis, a homogenate of the whole cell or a cell lysate as an immunogen, and a method for producing the same. In order to achieve the above object, the present inventors firstly cultured Bordetella parapertussis under a condition of 37 DEG C for 3 days, adjusted the cell density to 1010 cells / mL, added a formaldehyde solution thereto such that the final concentration of formalin was 0.2%, thereby inactivating the cells. The resulting cells were adjusted to 1010 cells / mL in terms of cell density by adding sterile phosphate-buffered saline, whereby a vaccine composition was obtained. The vaccine composition of the invention was inoculated into a mouse (3.5 weeks of age) and infected with a Bordetella parapertussis suspension. As a result, it was revealed that the mouse has a protective activity against infection with Bordetella parapertussis and further shows the protective activity against infection even at a dose one-thirtieth the common inoculum dose of whooping cough vaccine.

Description

technical field [0001] The present invention relates to a whole-bacteria vaccine composition for preventing pertussis caused by B. parapertussis, which contains whole bacteria, broken whole bacteria or lysate of bacteria of Bacillus parapertussis as immunogen and its manufacturing method . The present invention also relates to a method for preventing pertussis caused by parapertussis, which includes the step of administering whole cells of B. parapertussis, crushed whole cells, or cell lysates to a subject. Background technique [0002] Pertussis (whooping cough, pertussis) is a respiratory infectious disease with cough as the main symptom, which is caused by infection with Bordetella pertussis or Bordetella parapertussis. As the name of the disease suggests, the patient exhibits long-term (2 weeks to several months) coughing attacks. Since pertussis caused by Bacillus pertussis accounts for a relatively large proportion, vaccines targeting Bacillus pertussis have been wel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/10A61P31/04
CPCA61K39/099A61K2039/545A61K2039/521A61P31/04
Inventor 渡边峰雄小松荣司
Owner DAIICHI SANKYO CO LTD
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