Semi-quantitative analysis of cytokine gene expression in blood and cerebrospinal fluid cells by reverse transcriptase polymerase chain reaction

Research in Experimental Medicine. Zeitschrift Für Die Gesamte Experimentelle Medizin Einschliesslich Experimenteller Chirurgie
P RieckmannU Michel

Abstract

An easy, reproducible and semi-quantitative, non-radioactive method for the analysis of mRNA expression for various cytokines, (i.e., Interleukin (IL)-1 beta, IL-4, IL-6, tumor necrosis factor (TNF)-alpha, lymphotoxin (LT), transforming growth factor (TGF)-beta, interferon (IFN)-gamma and endothelin-1 (ET-1)) in cells from cerebrospinal fluid (CSF) and peripheral blood mononuclear cells (PBMC) has been established. By means of polymerase chain reaction primers that cover a splice junction, amplification of contaminating DNA was omitted. Densitometric scanning of ethidium bromide-stained agarose gels proved to be very sensitive for semiquantitative analysis of PCR products. Serial tenfold dilutions of cDNA revealed a log-linear regression from 10(6) to 10(2) cells under optimal cycle conditions. The intra- and inter-assay variability of the method was below 10%. With this assay, the cytokine expression pattern of as few as 10(4) mononuclear cells from blood or CSF was determined. This method made it possible to detect differences in the cytokine gene expression pattern of mononuclear cells from patients with different neurological diseases. CSF cells from 43 patients with various neurological diseases were analyzed. TNF-alpha, L...Continue Reading

References

Jan 1, 1992·Research in Experimental Medicine. Zeitschrift Für Die Gesamte Experimentelle Medizin Einschliesslich Experimenteller Chirurgie·H EhrenreichF D Goebel
Dec 1, 1992·The Journal of Investigative Dermatology·K Paludan, K Thestrup-Pedersen
Feb 1, 1992·Transplantation·O M MartinezN L Ascher
Jul 1, 1991·The Journal of Experimental Medicine·A E GoldfeldC Doyle
May 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·A N GlazerR A Mathies
Dec 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·A M WangD F Mark
Sep 1, 1989·Analytical Biochemistry·E A RibeiroD P Miller
Jan 1, 1986·Cold Spring Harbor Symposia on Quantitative Biology·A Cerami, B Beutler

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Citations

Mar 10, 2001·The American Journal of Pathology·A BagnatoP G Natali
Mar 31, 1998·Journal of Immunological Methods·N KruseP Rieckmann
Nov 5, 1998·Veterinary Immunology and Immunopathology·A GröneW Baumgärtner
Jul 27, 2005·Proceedings of the National Academy of Sciences of the United States of America·Horea RusPeter A Calabresi
Aug 1, 1995·Clinical Molecular Pathology·R D MalcomsonD J Harrison
Dec 30, 2014·Clinical and Experimental Medicine·Domenico RibattiMaria Trojano
Jul 23, 2014·Acta Neuropathologica Communications·Francesco GirolamoMaria Trojano
Apr 10, 2016·Journal of Neuroimmune Pharmacology : the Official Journal of the Society on NeuroImmune Pharmacology·Stephanie J BisselClayton A Wiley
Oct 8, 1998·Multiple Sclerosis : Clinical and Laboratory Research·P Monteyne, C J Sindic
Aug 4, 1998·International Journal of Cancer. Journal International Du Cancer·H HannigW Bodemer
Jan 2, 2007·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Ashok ChauhanAvindra Nath
Feb 2, 2008·International Review of Psychiatry·Bruce James Brew, Scott Lee Letendre

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