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Mutant Bordetella strains and methods of use thereof

A bacillus, subject technology, applied in the fields of immunology, microbiology, allergy and medicine, which can solve the problems of poor immunogenicity, troublesome attenuation, difficult to produce, etc.

Active Publication Date: 2022-03-18
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 of use thereof
  • Mutant Bordetella strains and methods of use thereof
  • Mutant Bordetella strains and methods of use thereof

Examples

Experimental program
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Effect test

example 1- 100

[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 6

[0078] Example 6 - Measurement of lung cell infiltration and cytokine profiles.

[0079] Evaluate Figure 6 Infiltration of airway cell populations in allergic mice from the experiments shown in and discussed immediately above. Following plethysmography measurements, bronchoalveolar lavage (BAL) fluid was collected to measure cellular infiltration in the airways. Cells from BAL fluid were harvested by centrifugation at 1,200 rpm for 5 minutes at 4°C, resuspended in PBS for cell counting using Shandon cytospin 4 (Thermo Fisher Scientific), and analyzed with May Grünwald Giemsa Staining is used for cell type counts. The total cell number was measured in the BAL fluid of mice ( Figure 7A ), and count eosinophils ( Figure 7B ), neutrophils ( Figure 7C ), lymphocytes ( Figure 7D ) and macrophages ( Figure 7E ) percentage. Statistical analysis was performed by applying a parametric one-way ANOVA test using a post-hoc Bonferroni comparison test (95% confidence interval)...

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Abstract

A method of reducing or preventing the development of airway inflammation in a subject comprising the step of infecting the subject's respiratory tract with an amount of a composition comprising a pharmaceutically acceptable carrier and sufficient to colonize the subject Respiratory attenuated B. pertussis adhesin-deficient live bacteria of the genus Bordetella. The step of infecting the subject with the live attenuated pertactin-deficient Bordetella bacterium results in reducing or preventing 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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/00A61K9/00A61P11/00
CPCA61K35/74A61P11/00A61P29/00A61P11/06A61P31/04A61K39/099C07K14/235C12N1/36A61K2039/51A61K2039/543A61K2039/575A61K35/00A61K2039/10A61K9/007A61K2039/522
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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