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Proteoglycan and proteome profiling of central human pulmonary fibrotic tissue utilizing miniaturized sample preparation : a feasibility study

Malmström, Johan LU ; Larsen, Kristoffer LU ; Hansson, Lennart LU ; Löfdahl, Claes-Göran LU ; Nörregard-Jensen, Ole ; Marko-Varga, György LU and Westergren-Thorsson, Gunilla LU (2002) In Proteomics 2(4). p.394-404
Abstract

The objective of this study was to isolate fibrotic cells from human lung biopsies taken from different central pulmonary locations. A comparison was made of cell morphology, proteoglycan- and protein-expression in mesenchymal cell cultures obtained from human bronchial biopsies from patients with asthmatic-like disorders. We isolated viable cells from 10 out of the 12 biopsies. The fibroblast-like cells were positive for the biomarker a-smooth muscle actin, indicating that the cells were in an activated state. Two different types of fibroblast-like cells were observed from human pulmonary connective tissue; one of contractile type with lamellipodia that facilitate migration and a second cell type with an increased cell size, which most... (More)

The objective of this study was to isolate fibrotic cells from human lung biopsies taken from different central pulmonary locations. A comparison was made of cell morphology, proteoglycan- and protein-expression in mesenchymal cell cultures obtained from human bronchial biopsies from patients with asthmatic-like disorders. We isolated viable cells from 10 out of the 12 biopsies. The fibroblast-like cells were positive for the biomarker a-smooth muscle actin, indicating that the cells were in an activated state. Two different types of fibroblast-like cells were observed from human pulmonary connective tissue; one of contractile type with lamellipodia that facilitate migration and a second cell type with an increased cell size, which most likely is of a synthetic phenotype. This is the first evidence of alterations in the proteoglycan expression pattern of versican, perlecan, biglycan and decorin which can be linked to the pathophysiological state of asthmatics proven by a combination of solid-phase extraction by reversed phase and by peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Protein expression analysis using two-dimensional electrophoresis was interfaced to miniaturized sample preparation techniques using microcapillary extraction. Four protein groups were identified; cytoskeletal, adhesion, scavenger and metabolic proteins. These patient's proteomes showed a high degree of heterogeneity between patients but larger homogeneity within biopsies derived from different locations of the same patient.

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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Actins, Cells, Cultured, Electrophoresis, Gel, Two-Dimensional, Fibroblasts, Humans, Lung, Miniaturization, Protein Array Analysis, Proteoglycans, Proteome, Pulmonary Fibrosis, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Journal Article, Research Support, Non-U.S. Gov't
in
Proteomics
volume
2
issue
4
pages
11 pages
publisher
John Wiley and Sons Inc.
external identifiers
  • wos:000175237800007
  • pmid:12164698
  • scopus:0036224578
  • pmid:12164698
ISSN
1615-9861
DOI
10.1002/1615-9861(200204)2:4<394::AID-PROT394>3.0.CO;2-6
language
English
LU publication?
yes
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Analytical Chemistry (S/LTH) (011001004), Respiratory Medicine and Allergology (013230111), Department of Experimental Medical Science (013210000)
id
a577142c-dd58-4c5a-b6f7-e9f4118a0731 (old id 109796)
date added to LUP
2016-04-01 12:16:07
date last changed
2020-01-22 02:48:18
@article{a577142c-dd58-4c5a-b6f7-e9f4118a0731,
  abstract     = {<p>The objective of this study was to isolate fibrotic cells from human lung biopsies taken from different central pulmonary locations. A comparison was made of cell morphology, proteoglycan- and protein-expression in mesenchymal cell cultures obtained from human bronchial biopsies from patients with asthmatic-like disorders. We isolated viable cells from 10 out of the 12 biopsies. The fibroblast-like cells were positive for the biomarker a-smooth muscle actin, indicating that the cells were in an activated state. Two different types of fibroblast-like cells were observed from human pulmonary connective tissue; one of contractile type with lamellipodia that facilitate migration and a second cell type with an increased cell size, which most likely is of a synthetic phenotype. This is the first evidence of alterations in the proteoglycan expression pattern of versican, perlecan, biglycan and decorin which can be linked to the pathophysiological state of asthmatics proven by a combination of solid-phase extraction by reversed phase and by peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Protein expression analysis using two-dimensional electrophoresis was interfaced to miniaturized sample preparation techniques using microcapillary extraction. Four protein groups were identified; cytoskeletal, adhesion, scavenger and metabolic proteins. These patient's proteomes showed a high degree of heterogeneity between patients but larger homogeneity within biopsies derived from different locations of the same patient.</p>},
  author       = {Malmström, Johan and Larsen, Kristoffer and Hansson, Lennart and Löfdahl, Claes-Göran and Nörregard-Jensen, Ole and Marko-Varga, György and Westergren-Thorsson, Gunilla},
  issn         = {1615-9861},
  language     = {eng},
  number       = {4},
  pages        = {394--404},
  publisher    = {John Wiley and Sons Inc.},
  series       = {Proteomics},
  title        = {Proteoglycan and proteome profiling of central human pulmonary fibrotic tissue utilizing miniaturized sample preparation : a feasibility study},
  url          = {http://dx.doi.org/10.1002/1615-9861(200204)2:4<394::AID-PROT394>3.0.CO;2-6},
  doi          = {10.1002/1615-9861(200204)2:4<394::AID-PROT394>3.0.CO;2-6},
  volume       = {2},
  year         = {2002},
}