1993 Fiscal Year Final Research Report Summary
Monitaring of rejection after transplantation by cytokine mRNA mapping
| Project/Area Number |
04454336
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Digestive surgery
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| Research Institution | Keio University |
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Principal Investigator |
TSUZUKI Toshiharu Keio Univ.Sch.Med, Professor, 医学部, 教授 (80051192)
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| Co-Investigator(Kenkyū-buntansha) |
SHITO Masaya Keio Univ.sch.Med., 医学部, 助手 (10215981)
SHIMIZU Soichi Keio Univ.Sch, Med., 医学部, 助手 (20206197)
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| Project Period (FY) |
1992 – 1993
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| Keywords | cytokine / mRNA / transplantation / monitaring / rejection / 末梢血 |
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| Research Abstract |
We developed a method of detecting cytokine mRNA from unfractionated whole blood using RT-PCR.IL-1beta and TNF gene expression could be detected from a 0.5ml blood sample. Total RNA was extracted by 4M guanidinium isothiocyanate mixture, 0.2M sodium acetate, phenol and chloroform and 10mug total RNA was obtained. The mRNA is reverse transcribed, and IL-1beta, TNF and IL-2R cDNA were selectively amplified by synthetic primers with PCR.Pheripheral blood was obtained from healthy volunteers and patients with severe infectons, operations and renal transplantation/Il-1beta, TNF and IL-2R mRNA were undetectable in blood of healthy volunteers. While in patients with severe infection and operation, only IL-1beta and TNF, but not IL-2R mRNA were detected. IL-2 mRNA was detectec in blood of the patients with renal transpantation. This method avoids artificial gene activation and may be applicable to monitoring cytokine gene espression in various pathophysiological state.
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