Molecular Studies of Cytokine Induction
Annual rept. no. 1, Feb-Nov 1989
UNIFORMED SERVICES UNIV OF THE HEALTH SCIENCES BETHESDA MD
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The immune system has evolved to respond to certain microbial products with a sequence of physiological events, termed the acute phase response, that appears to be adaptive. The molecular triggers for the acute phase response appear to be cell wall products such as endotoxin, in the case of bacteria and double stranded RNA, in the case of viruses. Both endotoxin and double stranded RNA induce a class of hormone like substances, cytokines, that drive the acute phase response. It is not known if bacterial and viral products induce overlapping sets of cytokines or if the virus response is completely dependent on interferon. Until now there has been no convenient way of assaying either serum for multiple cytokine activity, or detection of cytokine RNAs. To circumvent this problem I have developed a polymerase chain reaction method RNAPCR for the detection and semi-quantitation of specific mRNA species. This method will detect mRNA in a range of 10-4 to 10-5 copies per cell allowing for a systematic analysis for multiple cytokines simultaneously. Data on the double stranded RNA induced production of cytokine mRNAs by the mouse macrophage line RAW is shown. Because the RNAPCR is strand specific, the methodology can be used to monitor the production of double stranded RNA in infected tissues. The power of this technique is just beginning to be exploited.
- Medicine and Medical Research