دورية أكاديمية
In Vitro Semi-Quantitative Determination of Human Gamma-Interferon-mRNA Level by RT-PCR
| العنوان: | In Vitro Semi-Quantitative Determination of Human Gamma-Interferon-mRNA Level by RT-PCR |
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| المؤلفون: | Jalil Tavakol Afshari, Alireza Zamani, Javad Behravan, Behrouz Shisheeyan, Bita Saifi |
| المصدر: | پزشکی بالینی ابن سینا, Vol 10, Iss 2, Pp 5-11 (2003) |
| بيانات النشر: | Hamadan University of Medical Sciences, 2003. |
| سنة النشر: | 2003 |
| المجموعة: | LCC:Medicine |
| مصطلحات موضوعية: | ifn- ?, pha activated lymphocytes, semi-quantitative-rt-pcr, reverse transcriptase polymerase chain reaction, Medicine |
| الوصف: | Cells communicate with one another not only by cell-cell contact, but also via the elaboration of soluble mediators or cytokines. The functionally distinct repertoire of secreted cytokines of Th1/Th2 cells has been shown to play an important role in the pathogenesis of inflammatory diseases. Effector Th1 cells secretes predominantly IFN- γ and IL-2 and regulates cell-mediated immunity against intracellular pathogens. Regulatory cytokines are likely to be secreted in small quantities in response to specific stimuli and taken by responder cells. As a result, many scientists determine cytokine’s m-RNA expression by using a more sensitive technique, RT-PCR. The expression of IFN- γ -mRNA was studied using semiquantitative RT- PCR. Lymphocytes were stimulated by phytohaemagglutinin (PHA) (1μg/106) in culture medium (incubation time: 0,4,8,12,24,48 and72 hour) and total RNA extracted and cDNA synthesized. IFN- γ was detected by the RT-PCR (reverse transcription-polymerase chain reaction) and semi-quantitative-RT-PCR methods, in which sequences (273bp) between two oligonucleotide primers (chosen from two exons of the IFN- γgene sequences) were amplified using a heat-stable DNA polymerase. In semi-quantitative RT-PCR we used a serial dilution (1/2,1/4…) of cDNA in order to determine the titer of cDNA for that will give visible band in 2% agarose gel electrophoresis. Results showed that after 4h incubation express highest level of IFN- γ- mRNA and it is stable until 24 hour after that. Then it falls to baseline level. In order to study the IFN- γ gene expression kinetic compare results with other studies show that RT-PCR detection of IFN- γ level is more sensitive than other methods like Elisa and for maximum expression of IFN- γ gene 4h incubation of lymphocytes with PHA is sufficient. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | Persian |
| تدمد: | 2588-722X 2588-7238 |
| العلاقة: | http://sjh.umsha.ac.ir/article-1-664-en.htmlTest; https://doaj.org/toc/2588-722XTest; https://doaj.org/toc/2588-7238Test |
| الوصول الحر: | https://doaj.org/article/cf8583644bc540828531cb1a7b1a4ae9Test |
| رقم الانضمام: | edsdoj.f8583644bc540828531cb1a7b1a4ae9 |
| قاعدة البيانات: | Directory of Open Access Journals |
| تدمد: | 2588722X 25887238 |
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