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Purification and partial characterization of a product from Yersinia pseudotuberculosis with the ability to activate
T Miyoshi-Akiyama1, K Imanishi, T Uchiyama
1Department of Microbiology and Immunology, Tokyo Women's Medical College, Japan.
Abstract:
We tried to purify a substance exhibiting mitogenicity for human peripheral blood lymphocytes from Yersinia pseudotuberculosis isolated from patients with Y. pseudotuberculosis infection manifesting acute and systemic clinical symptoms. The supernatant of a suspension of the bacteria disrupted by sonication was serially chromatographed on DEAE-Sepharose fast-flow, Sephacryl S-100 HR, and TSK-gel G2000SW high-pressure liquid chromatography columns. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that the purified mitogenically active substance migrated as a single band corresponding to a molecular mass of 21 kDa. We designated the purified substance Y. pseudotuberculosis-derived mitogen (YPM). YPM stimulated human peripheral T cells to proliferate and produce interleukin-2 at 0.1 ng/ml or more. YPM-induced T-cell activation required the expression of HLA class II molecules on accessory cells.
Insights
Researchers purified a novel Yersinia pseudotuberculosis-derived mitogen (YPM) that stimulates human T cells. This YPM enhances T-cell proliferation and interleukin-2 production, requiring HLA class II expression on accessory cells for activation.
Area of Science:
- Immunology
- Microbiology
- Biochemistry
Background:
- Yersinia pseudotuberculosis infections can cause severe systemic symptoms.
- Understanding bacterial components that modulate host immune responses is crucial.
Purpose of the Study:
- To isolate and characterize a mitogenic substance from Yersinia pseudotuberculosis.
- To investigate the immunomodulatory effects of the purified substance on human lymphocytes.
Main Methods:
- Bacterial sonication and supernatant collection.
- Sequential chromatography using DEAE-Sepharose, Sephacryl S-100 HR, and TSK-gel G2000SW.
- SDS-PAGE for molecular mass determination.
- Assays for T-cell proliferation and IL-2 production.
Main Results:
- A single mitogenically active substance, YPM, was purified.
- YPM has a molecular mass of 21 kDa.
- YPM stimulates human T-cell proliferation and IL-2 production at low concentrations (≥0.1 ng/ml).
- YPM-induced T-cell activation is dependent on HLA class II expression on accessory cells.
Conclusions:
- Yersinia pseudotuberculosis produces a potent mitogen, YPM.
- YPM is a valuable tool for studying T-cell activation and immune responses.
- The findings highlight YPM's role in Yersinia pathogenesis and immune evasion strategies.