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PMID: 15817763 Published · ppublish English Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S.

Real-time reverse transcription-PCR quantitation of substance P receptor (NK-1R) mRNA.

Clinical and diagnostic laboratory immunology ·Vol. 12 ·No. 4 ·2005-04-00 ·Pages 537-41

Lai JP, Douglas SD, Wang YJ, Ho WZ

Abstract

The substance P (SP)-preferring receptor, neurokinin-1 receptor (NK-1R), has an important role in inflammation, immune regulation, and viral infection. We applied a newly developed real-time reverse transcription (RT)-PCR assay to quantify NK-1R mRNA in human neuronal cell line (NT-2N), a human B-cell line (IM9), monocyte-derived macrophages (MDM), peripheral blood lymphocytes (PBL), and human astroglioma cells (U87 MG). The NK-1R real-time RT-PCR assay has a sensitivity of 100 mRNA copies, with a dynamic range of detection between 10(2) and 10(7) copies of NK-1R gene transcripts per reaction. This assay is highly reproducible, with an intraassay coefficient variation of threshold cycle (Ct) of less than 1.9%. The NK-1R real-time RT-PCR is highly sensitive for quantitative determination of NK-1R mRNA in human immune cells (MDM and PBL) that express low levels of NK-1R mRNA. In addition, the assay has the ability to accurately quantitate the dynamic changes in NK-1R mRNA expression in interleukin-1beta-stimulated U87 MG. These data indicate that the NK-1R real-time RT-PCR has potential for a wide application in investigation of NK-1R expression at the mRNA level under physiological and pathological conditions in both the central nervous system and the immune system.

MeSH Terms
Cells, Cultured Gene Expression Regulation Humans Linear Models RNA, Messenger/analysis Receptors, Neurokinin-1/analysis,genetics Reverse Transcriptase Polymerase Chain Reaction Sensitivity and Specificity
Chemicals
RNA, Messenger Receptors, Neurokinin-1
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lai Jian-Ping
Division of Allergy and Immunology, Joseph Stokes Jr. Research Institute at the Children's Hospital of Philadelphia, Department of Pediatrics, University of Pennsylvania Medical School, Philadelphia, Pennsylvania 19104, USA.
Douglas Steven D
Wang Yan-Jian
Ho Wen-Zhe
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Article Info
Journal
Clinical and diagnostic laboratory immunology
Abbr.
Clin Diagn Lab Immunol
ISSN
1071-412X
DOI
10.1128/CDLI.12.4.537-541.2005
Published
2005-04-00
Pages
537-41
Language
English
Region
United States
NLM ID
9421292
PMCID
PMC1074379
Subset
IM
Grants
NIMH NIH HHS · R01 MH049981 · United States
NIDA NIH HHS · NIH-DA112815 · United States
NIMH NIH HHS · NIH-MH 49981 · United States
External Links
PubMed source DOI: 10.1128/CDLI.12.4.537-541.2005 PMC Full Text
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