Proteomic Analysis of Human Cervical-Vaginal Fluids
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- 作者：Li-Jun Tang ; Francesco De Seta ; Federico Odreman ; Per Venge ; Caterina Piva ; Secondo Guaschino ; Rodolfo C. Garcia
- 刊名：Journal of Proteome Research
- 出版年：2007
- 出版时间：July 2007
- 年：2007
- 卷：6
- 期：7
- 页码：2874 - 2883
- 全文大小：474K
- 年卷期：v.6,no.7(July 2007)
- ISSN：1535-3907
文摘
The pathophysiology of vaginal conditions is stillill-defined at a molecular level. Because the proteome ofthe human cervical-vaginal fluid (CVF) has not beenreported to date, we undertook the identification ofproteins present in the cell-free fraction of these fluids.Proteins were separated bidimensionally (2-D) by isoelectrofocusing (pH 3-11) followed by SDS-polyacrylamideelectrophoresis. The proteins of 147 spots were identifiedby matrix-assisted laser desorption/ ionization-time-of-flight-mass spectrometry (MALDI-TOF/TOF). This approach was supplemented by immunoassays for markersof neutrophils (myeloperoxidase, MPO; neutrophil gelatinase-associated lipocalin, NGAL/HNL) and eosinophils(eosinophil cationic protein: ECP) and by immunoblotting(lactoferrin, calgranulins A and B and annexins A1 andA3. Nearly half of the proteins (69/147) and proteinfragments detected were found to be plasma components, on the basis of which the human CVF can bebroadly considered a plasma transudate. Although thepattern of protein spots was very similar for all fluidsanalyzed, a relative overabundance of major plasmaproteins such as albumin, transferrin, immunoglobulins,apolipoproteins, alpha-1-acid glycoprotein 1, and calgranulins was associated with the presence of a highnumber of polymorphonuclear leukocytes in the lavagesfrom which those cell-free fluids had been obtained.Instead, fluids from women experiencing vulvovaginalcandidiasis did not show differences in the protein mapscompared with asymptomatic individuals. Neutrophil andeosinophil granule secretion proteins were also detectedin variable amounts in the lavage fluids by both immunoassay and immunoblotting, indicating polymorphonuclear cell activation.