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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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