Document Detail


Technical problems with protein extraction of chemokines featuring RANTES.
MedLine Citation:
PMID:  10670033     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Chemokines are known to be one of the sources for eosinophilic tissue infiltration in eosinophilic inflammation. Detection of beta-chemokines such as RANTES was possible in nasal tissue with or without eosinophilic infiltration. The concentration of chemokines which has been measured in the same tissue differs often in the literature. Aim of this study was to compare the different techniques of protein extraction and help to understand and interpret the investigation on RANTES secretion. Tissue of nasal polyps, inferior and middle turbinate was cut into halves and every half on its own pulverized using liquid nitrogen. The protein extraction was performed either with citric acid solution (pH 2.5) or phosphate buffered saline (PBS). The samples were then lyophilized. The concentration of RANTES was measured by a specific double sandwich ELISA. Using the citric acid technique the average concentration of RANTES in middle turbinates was 1.3 ng/mg, in inferior turbinates 1.6 ng/mg and in polyps 2.6 ng/mg tissue, using the PBS technique respectively 0.6 ng/mg, 0.5 ng/mg and 0.8 ng/mg tissue. Our data revealed a mismatch of 3.3:1 for polyps (citric acid: PBS), 3.2:1 for inferior and 2.2:1 for middle turbinates, respectively. Consequent comparison between the results of different techniques was not possible. Of special interest was also the fact that different techniques had different efficiencies of protein extraction in different tissues. Present statements on RANTES concentrations as a prognostic factor in nasal tissues need a technically careful standardization as far as this study shows.
Authors:
S Maune; J E Meyer; B Spautz; M Sticherling; J M Schröder
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Rhinology     Volume:  37     ISSN:  0300-0729     ISO Abbreviation:  Rhinology     Publication Date:  1999 Dec 
Date Detail:
Created Date:  2000-02-24     Completed Date:  2000-02-24     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0347242     Medline TA:  Rhinology     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  179-81     Citation Subset:  IM    
Affiliation:
Dept. of Otorhinolaryngology, Head and Neck Surgery, University of Kiel, F.R.G.
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MeSH Terms
Descriptor/Qualifier:
Cells, Cultured
Chemokine CCL5 / secretion*
Enzyme-Linked Immunosorbent Assay
Humans
Nasal Mucosa / cytology,  metabolism
Rhinitis / metabolism*
Sinusitis / metabolism*
Turbinates / cytology,  metabolism
Chemical
Reg. No./Substance:
0/Chemokine CCL5

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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