Mutant bordetella strains and methods for use

A technology of Bordetella and Bacillus, applied in the fields of immunology, microbiology, allergy and medicine, can solve problems such as difficult production, troublesome attenuation, and poor immunogenicity

Active Publication Date: 2018-12-21
INSTITUT PASTEUR DE LILLE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Of these, despite modern molecular biology techniques and major advances in our understanding of microbiology and immunology, it remains difficult to produce sufficiently attenuated without causing significant pathology, while being sufficiently immunogenic to induce Vaccine products for potent and long-lasting immune responses to target pathogens
Achieving an optimal level of attenuation is particularly troublesome in the case of attenuated whole-cell live bacterial vaccines, as over-attenuation by reducing the amount or activity of virulence factors can lead to poor immunogenicity and / or poor immunogenicity after administration. Vaccines that do not survive or replicate in the body long enough to induce an immune response

Method used

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  • Mutant bordetella strains and methods for use
  • Mutant bordetella strains and methods for use
  • Mutant bordetella strains and methods for use

Examples

Experimental program
Comparison scheme
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example 1

[0065] Example 1 - Construction and characterization of a pertactin-deficient strain of Bordetella pertussis.

[0066] Escherichia coli DH5α, SM10 and Bordetella pertussis BPZE1, BPSM (Menozzi et al., Infect Immun [Infection and Immunity] 1994; 62:769-778) and B1917 (Bart et al. Bulletin] 2014; 2(6)). Bordetella strains were grown at 37°C on Bordet-Gengou agar (BG) supplemented with 1% glycerol and 10% defibrinated sheep blood. After growth, bacteria were harvested by scraping the plates and resuspended in phosphate-buffered saline (PBS) at the desired density. For liquid culture, Bordetella strains were incubated at 37°C in a modified Stainer- Grow in Scholte's medium (Imaizumi et al. Infect Immun [Infection and Immunity] 1983; 41:1138-1143). E. coli strains used in the cloning procedure were grown in LB broth or LB agar plates. Streptomycin (Sm) was used at 100 μg / ml, gentamicin (Gm) at 10 μg / ml, and ampicillin (Amp) at 100 μg / ml as needed.

[0067] To delete the gene p...

example 2

[0070] Example 2 - BPZE1P colonizes mice like BPZE1.

[0071] As previously described (Mielcarek et al., supra), with 10 6 Groups of 18 6-week-old mice were inoculated intranasally with 20 μl of PBS of live bacteria. At the indicated time points (3 hours, 3 days, 7 days, 14 days, 21 days and 28 days), 3 mice per group were sacrificed and lungs were harvested and homogenized to measure the total number of colony forming units (CFU) . Statistical analysis was performed by two-way ANOVA test using post hoc Bonferroni test (95% confidence interval). see figure 2 , BPZE1 and BPZE1P colonize animals equally well. Both strains exhibited a proliferation peak 3 days after vaccination and colonization continued for 4 weeks. No statistically significant differences were observed between these strains in their ability to colonize mouse lungs.

example 3

[0072] Example 3 - BPZE1P is as immunogenic as BPZE1 and protective against virulent B. pertussis challenge.

[0073] BPZE1P-induced immunity compared to BPZE1 was measured by antibody titration of mouse immune sera following nasal vaccination. use 10 5 Groups of 8 mice were inoculated intranasally with live BPZE1 or BPZE1P. After 4 weeks, mice were bled and total IgG titers were measured against total BPSM lysates. The blood was centrifuged at 5,000 xg for 5 minutes to separate the serum from the cells. Antibody titers against B. pertussis were estimated using enzyme-linked immunosorbent assay (ELISA) as previously described (Mielcarek et al., supra), lysed using total B. pertussis BPSM at 1 μg total protein per well thing. Statistical analysis was performed using GraphPad Prism software. Such as image 3 As shown, BPZE1 and BPZE1P vaccinated mice exhibited much higher antibody titers than naive control mice. No significant difference in antibody titers between BPZE1 a...

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Abstract

A method of reducing or preventing the development of airway inflammation in a subject includes the step of infecting the respiratory tract of a subject an amount of a composition including a pharmaceutically acceptable carrier and live attenuated pertactin-deficient Bordetella bacteria sufficient to colonize the respiratory tract of the subject. The step of infecting the subject with the live attenuated pertactin-deficient Bordetella bacteria results in reduction or prevention of the development of airway inflammation in the subject.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Patent Application Serial No. 62 / 314,843, filed March 29, 2016. [0003] sequence listing [0004] This application contains a Sequence Listing that has been filed electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on March 27, 2017, is named 7056-0073_SL.txt and is 2,134 bytes in size. [0005] Statement Regarding Federally Funded Research [0006] Not applicable. technical field [0007] The present invention relates generally to the fields of microbiology, immunology, allergy and medicine. More specifically, the present invention relates to live attenuated strains of Bordetella pertussis deficient in pertactin and their use as prophylactic and therapeutic agents in various disease settings. Background technique [0008] Microbes and their components have long been known to affect the immune system of...

Claims

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Application Information

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IPC IPC(8): A61K35/00A61K9/00A61P11/00
CPCA61K35/74A61P11/00A61P29/00A61P11/06A61P31/04A61K39/099C07K14/235C12N1/36A61K2039/51A61K2039/543A61K2039/575A61K35/00A61K2039/10A61K2039/58A61K35/42A61K9/007A61K2039/522
Inventor 路易斯·索兰斯卡米尔·洛赫特安妮·蒂斯科普洛斯萨利哈·艾特雅希亚森迪德
Owner INSTITUT PASTEUR DE LILLE
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