Mitogen-Induced Interferon Gamma Production in Human Whole Blood: The Effect of Heat and Cations
| Autor: | Jun-Young Park, Jeong-Ran Kim, Fukushi Abekura, Bomi Cha, Cheorl-Ho Kim, Ji-Hyun Nam |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Adult
Male 0301 basic medicine medicine.medical_specialty Hot Temperature T-Lymphocytes medicine.medical_treatment Pharmaceutical Science Enzyme-Linked Immunosorbent Assay chemical and pharmacologic phenomena Stimulation Lymphocyte Activation Interferon-gamma Young Adult 03 medical and health sciences 0302 clinical medicine Antigen Cations Internal medicine Concanavalin A medicine Humans Interferon gamma Phytohemagglutinins Tuberculosis Pulmonary Aged Whole blood biology Interferon-gamma production Chemistry Pokeweed mitogen Middle Aged 030104 developmental biology Cytokine Endocrinology Pokeweed Mitogens 030220 oncology & carcinogenesis biology.protein Female Biotechnology medicine.drug |
| Zdroj: | Current Pharmaceutical Biotechnology. 20:562-572 |
| ISSN: | 1389-2010 |
| Popis: | Background:Interferon-gamma release assays (IGRAs) are blood tests used to measure the amount of interferon-γ (IFN-γ) released by T lymphocytes after stimulation by antigens specific for the diagnosis of latent tuberculosis infection. A mitogen serves as a positive control to assess the immune function in IGRAs.Methods:This in vitro study was conducted to evaluate IFN-γ production by human whole blood stimulated with heat-treated and/or cation-supplemented phytohemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM), using QuantiFERON-TB Gold Kit ELISA tests.Results:The optimal concentrations of PWM, Con A and PHA for IGRAs were 2 µg/mL, 5 µg/mL and 10 µg/mL, respectively. The results showed that IFN-γ production in response to PWM was the highest and PHA was the lowest amount. The median values of three mitogens were in the following order: PWM≥Con A≥ positive control>PHA-P>negative control. PWM and PHA were heat stable, while Con A was heat sensitive. The mitogen response of lymphocytes to untreated or heat-treated PWM and heat-treated Con A was increased in 1 mM Ca2+-supplemented groups, whereas the response to heat-treated PHA was decreased. Exposure to 1 mM Mg2+ had no effect on untreated or heat-treated PWM, and a concentration of 1 mM Zn2+ inhibited the stimulation of un-treated PWM. We found that calcium supplementation improved the PWM-induced production of IFN-γ.Conclusion:Therefore, PWM is an appropriate mitogen for use as a positive control in IGRAs. It is a potential indicator of cytokine production in the diagnostic as well as research settings, and calcium supplementation improved stimulation. |
| Databáze: | OpenAIRE |
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