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Sensors 2014, 14(5), 8869-8876; doi:10.3390/s140508869
Article

Quantitative Analysis of Staphylococcus Enterotoxin A by Differential Expression of IFN-γ in Splenocyte and CD4+ T-Cells

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Received: 28 March 2014; in revised form: 15 May 2014 / Accepted: 15 May 2014 / Published: 20 May 2014
(This article belongs to the Special Issue Live Cell-Based Sensors with Receptors)
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Abstract: Staphylococcus aureus is an important bacterial pathogen that produces a range of Staphylococcal Enterotoxins (SEs) which cause gastroenteritis and superantigen activation of T cells, the mechanism of which is not well understood. The ability to rapidly detect and quantify SEs is very important in order to learn the causes of staphylococcal outbreaks and to stop similar outbreaks in the future. Enzyme-linked immunosorbent assays (ELISAs) have been developed for detection of several SEs. However, these immunological methods cannot distinguish between active and inactive toxin. It is known that interferon-gamma (IFN-γ) expressed in response to stimulation by SEs contributes significantly to the pathogenesis of S. aureus infection. Nonetheless, the cellular source of IFN-γ is still unclear and the contributions of the specific splenocyte types. In our effort to understand the immunologic response to Staphylococcal Enterotoxin A (SEA) exposure, we studied IFN-γ production in mouse splenocytes. We demonstrated that short term ex vivo exposure of splenocytes or primary naïve CD4+ T-cells to biologically active SEA induces differential expression of IFN-γ mRNA in a time and dose dependent manner and the expression levels reflect the levels of IFN-γ secreted protein. Positive isolated CD4+ T-cells accounted for only 10% of IFN-γ production. We also demonstrate that expression of IFN-γ can be used for rapid quantitative analysis of active SEA with a detection limit of 1 ng/mL.
Keywords: staphylococcal enterotoxins; interferon-gamma; active toxin detection; CD4+ T-cell; splenocytes staphylococcal enterotoxins; interferon-gamma; active toxin detection; CD4+ T-cell; splenocytes
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Rasooly, R.; Hernlem, B.J. Quantitative Analysis of Staphylococcus Enterotoxin A by Differential Expression of IFN-γ in Splenocyte and CD4+ T-Cells. Sensors 2014, 14, 8869-8876.

AMA Style

Rasooly R, Hernlem BJ. Quantitative Analysis of Staphylococcus Enterotoxin A by Differential Expression of IFN-γ in Splenocyte and CD4+ T-Cells. Sensors. 2014; 14(5):8869-8876.

Chicago/Turabian Style

Rasooly, Reuven; Hernlem, Bradley J. 2014. "Quantitative Analysis of Staphylococcus Enterotoxin A by Differential Expression of IFN-γ in Splenocyte and CD4+ T-Cells." Sensors 14, no. 5: 8869-8876.



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