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Published on: June 15, 2018
Detection, isolation and partial characterization of an immunostimulating glycoprotein from Rhodococcus fascians
Günter Butschak1, Uwe Karsten, Ute Schelhaas
1Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany. uwe.karsten@glycotope.com
Abstract:
In a search for novel immunostimulating substances we detected that culture supernatants of the gram-positive phytopathogenic bacterium, Rhodococcus fascians, were able to induce cytokine release (TNF(alpha)) from mouse peritoneal macrophages. Monoclonal antibodies were generated against the active principle, and were employed for its isolation and partial characterization as a high molecular (MW>100 kDa) glycoprotein. In addition, methods practicable for its biotechnological preparation and several ELISA variants for its determination were developed.
Insights
Rhodococcus fascians bacteria release a substance that stimulates immune cells. This high-molecular-weight glycoprotein was isolated and methods for its production and detection were developed.
Area of Science:
- Immunology
- Microbiology
- Biochemistry
Background:
- Phytopathogenic bacteria can produce bioactive compounds.
- Immune responses involve cytokine release, such as tumor necrosis factor-alpha (TNF-α).
Purpose of the Study:
- To identify novel immunostimulating substances from bacterial culture supernatants.
- To characterize the active principle responsible for immune cell activation.
Main Methods:
- Culturing Rhodococcus fascians and collecting culture supernatants.
- Using mouse peritoneal macrophages to assay for cytokine release (TNF-α).
- Generating monoclonal antibodies for isolation and partial characterization of the active substance.
Main Results:
- Rhodococcus fascians culture supernatants induced TNF-α release from macrophages.
- The active principle was identified as a high molecular weight (MW > 100 kDa) glycoprotein.
- Methods for biotechnological preparation and ELISA-based determination were developed.
Conclusions:
- Rhodococcus fascians produces an immunostimulating glycoprotein.
- The developed methods facilitate further research and potential applications of this substance.
