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Immunomodulatory properties of subcellular fractions of a G+ bacterium, Bacillus firmus

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Abstract

Mucosal immunization with non-living antigens usually requires the use of an adjuvant. The adjuvant activity of Bacillus firmus in the mucosal immunization of mice was described by our laboratory previously. In the present study, subcellular localization of B. firmus activities was followed. After mechanical disintegration, subcellular components of bacterium were fractionated by differential centrifugation and salting out. Bacterial cell walls, cytoplasmic membrane fraction, soluble cytoplasmic proteins, and ribosomal fractions were isolated. Their effect on the mouse immune system was studied. Lymphocyte proliferation and immunoglobulin formation in vitro were stimulated by bacterial cell wall (BCW), cytoplasmic membrane (CMF), and ribosomal fractions. BCW and CMF increased antibody formation after intratracheal immunization of mice with influenza A and B viruses, and increased protection against subsequent infection with influenza virus. The BCW fraction even induced intersubtypic cross-protection: Mice immunized with A/California/7/04 (H3N2) + BCW were resistant to the infection by the highly pathogenic A/PR/8/34 (H1N1) virus.

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Abbreviations

BAL:

Bronchoalveolar lavage

BCW:

Bacterial cell wall fraction

CMF:

Cytoplasmic membrane fraction

ConA:

Concavalin A

EID50 :

50% egg infective dose

FTS:

Fetal calf serum

HT:

Hemagglutination titer

iNOS:

Inducible NO synthase

NAL:

Nasal cavity lavage

PWM:

Pokeweed mitogen

RiPF:

Purified ribosomal fraction

RiCF:

Crude ribosomal fraction

SCPF:

Fraction of soluble cytoplasmic proteins

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Acknowledgments

This work was supported by Grant of Ministry of Education, Youth and Sports of the Czech Republic MSM0021620806 and by Grant of the Czech Science Foundation 310/07/0675 and by Research Programme of Charles University in Prague PRVOUK P25/LF1/2.

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Authors and Affiliations

  1. First Faculty of Medicine, Institute of Immunology and Microbiology, Charles University in Prague, Studnickova 7, Prague 2, 128 00, Czech Republic

    Dana Cechova, Michaela Novakova, Olga Novotna, Jaroslav Julak, Peter Zanvit & Ludmila Prokesova

  2. Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic

    Karel Mikulik

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  1. Dana Cechova
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  2. Michaela Novakova
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  3. Karel Mikulik
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  4. Olga Novotna
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  5. Jaroslav Julak
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  6. Peter Zanvit
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  7. Ludmila Prokesova
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Correspondence to Ludmila Prokesova.

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Cechova, D., Novakova, M., Mikulik, K. et al. Immunomodulatory properties of subcellular fractions of a G+ bacterium, Bacillus firmus . Folia Microbiol 58, 111–121 (2013). https://doi.org/10.1007/s12223-012-0181-y

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  • DOI: https://doi.org/10.1007/s12223-012-0181-y

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