PMID- 9003228
OWN - NLM
STAT- MEDLINE
DCOM- 19970206
LR  - 20170214
IS  - 0022-0345 (Print)
IS  - 0022-0345 (Linking)
VI  - 75
IP  - 11
DP  - 1996 Nov
TI  - Chemokine expression in human oral keratinocyte cell lines and keratinized 
      mucosa.
PG  - 1827-34
AB  - Chemoattractant cytokines regulate the immune response within the tissue by 
      recruiting neutrophils and macrophages. These so-called chemokines include a 
      large family of peptide molecules encoded by distinct genes. Their expression is 
      controlled by a variety of microbial and host factors. Among host factors, 
      interleukin-1 (IL-1) is thought to be a key regulator of tissue destruction and 
      mediator of the local immune response. To study its influence on chemokine 
      expression, we used a highly sensitive, semi-quantitative method to assess gene 
      expression at the level of mRNA. RNA was extracted from human oral keratinocyte 
      cell lines after treatment with recombinant human IL-1. To test the method 
      further and possibly establish a chemokine mRNA expression pattern, we also 
      extracted RNA from healthy oral keratinized mucosa. Purified RNA was 
      reverse-transcribed and subsequently amplified in a polymerase chain reaction 
      (RT-PCR) by means of specific primer pairs. Amplified sequences were analyzed by 
      agarose gel electrophoresis, visualized by ethidium bromide staining, transferred 
      to nylon membranes, and hybridized to biotinylated oligonucleotide probes. 
      Detection was achieved by streptavidin-conjugated alkaline phosphatase, a 
      chemiluminescent substrate, and autoradiography. Autoradiographs were analyzed by 
      densitometric measurements. IL-1 stimulation resulted in an increase of the 
      chemokine mRNAs encoding interleukin-8 (IL-8), monocyte chemoattractant protein-1 
      (MCP-1), and GRO gamma. Macrophage inflammatory protein-1 alpha (MIP-1 alpha) 
      mRNA was not detectable in keratinocytes. In healthy oral mucosa, we found 
      considerable variation between the subjects. Detection of chemokine mRNAs by 
      RT-PCR proved to be sensitive, specific, and fast. It allows for the study of not 
      only cell-line-derived RNA, but also of RNA isolated directly from biopsy 
      material. The latter feature makes this method well-suited for diagnostic 
      purposes.
FAU - Bickel, M
AU  - Bickel M
AD  - Department of Removable Prosthodontics, University of Bern, Switzerland.
FAU - Nothen, S M
AU  - Nothen SM
FAU - Freiburghaus, K
AU  - Freiburghaus K
FAU - Shire, D
AU  - Shire D
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Dent Res
JT  - Journal of dental research
JID - 0354343
RN  - 0 (CXCL1 protein, human)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Chemokine CCL4)
RN  - 0 (Chemokine CXCL1)
RN  - 0 (Chemokines)
RN  - 0 (Chemokines, CXC)
RN  - 0 (Chemotactic Factors)
RN  - 0 (Growth Substances)
RN  - 0 (Intercellular Signaling Peptides and Proteins)
RN  - 0 (Interleukin-1)
RN  - 0 (Interleukin-8)
RN  - 0 (Macrophage Inflammatory Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Cell Line
MH  - Chemokine CCL2/biosynthesis
MH  - Chemokine CCL4
MH  - Chemokine CXCL1
MH  - Chemokines/*biosynthesis
MH  - *Chemokines, CXC
MH  - Chemotactic Factors/biosynthesis
MH  - Electrophoresis, Agar Gel
MH  - Gene Expression
MH  - Growth Substances/biosynthesis
MH  - Humans
MH  - Immunoenzyme Techniques
MH  - *Intercellular Signaling Peptides and Proteins
MH  - Interleukin-1/pharmacology
MH  - Interleukin-8/biosynthesis
MH  - Keratinocytes/drug effects/*immunology/metabolism
MH  - Macrophage Inflammatory Proteins/biosynthesis
MH  - Mouth Mucosa/drug effects/*immunology/metabolism
MH  - RNA, Messenger/analysis
MH  - Recombinant Proteins/pharmacology
EDAT- 1996/11/01 00:00
MHDA- 1996/11/01 00:01
CRDT- 1996/11/01 00:00
PHST- 1996/11/01 00:00 [pubmed]
PHST- 1996/11/01 00:01 [medline]
PHST- 1996/11/01 00:00 [entrez]
AID - 10.1177/00220345960750110301 [doi]
PST - ppublish
SO  - J Dent Res. 1996 Nov;75(11):1827-34. doi: 10.1177/00220345960750110301.