Comprehensive Proteomic Analysis of Human Cervical-vaginal Fluid Using Colposcopy Samples

Overview

Journal Proteome Sci

Publisher Biomed Central

Date 2009 Apr 21

PMID 19374746

Citations 46

Authors

Geert Zegels

Geert A A Van Raemdonck

Edmond P Coen

Wiebren A A Tjalma

Xaveer W M Van Ostade

Affiliations

Soon will be listed here.

Abstract

Background: Cervical-vaginal fluid (CVF) plays an important role in the prevention of gynecological infections, although little is known about the contribution of CVF proteins to the immunity of the lower female genital tract. In order to analyze the protein composition of human CVF, we used CVF samples that are routinely collected during colposcopy, but are usually discarded. Since these samples are available in large quantities we aimed to analyze their usefulness for proteomics experiments. The samples were analyzed using different prefractionation techniques (ultrafiltration and C4(RP)-LC protein separation) followed by C18(RP)-LC peptide separation and identification by MALDI-TOF-TOF mass spectrometry. To determine the reproducibility of this proteomics platform we analyzed three technical replicates. Using spectral counting, protein abundances were estimated in a semiquantitative way. We also compared the results obtained in this study with those from previous studies derived from patients with different physiological conditions in order to determine an overlapping protein set.

Results: In total, we were able to identify 339 proteins in human CVF of which 151 proteins were not identified in any other proteomics study on human CVF so far. Those included antimicrobial peptides, such as human beta-defensin 2 and cathelicidin, which were known to be present in CVF, and endometrial proteins such as glycodelin and ribonucleoprotein A. Comparison of our results with previously published data led to the identification of a common protein set of 136 proteins. This overlapping protein set shows increased fractions of immunological and extracellular proteins, confirming the extracellular immunological role of CVF.

Conclusion: We demonstrated here that CVF colposcopy samples can be used in proteomics experiments and hence are applicable for biomarker discovery experiments. The delineation of an overlapping set of proteins that is identified in most proteomics studies on CVF may help in the description of a reference proteome when performing proteomics studies on human CVF.

Citing Articles

Potential biomarkers to predict return to fertility after discontinuation of female contraceptives-looking to the future.

Cordova-Gomez A, Wong A, Sims L, Doncel G, Dorflinger L Front Reprod Health. 2023; 5:1210083.

PMID: 37674657 PMC: 10477712. DOI: 10.3389/frph.2023.1210083.

Point-of-care diagnosis of cervical cancer: potential protein biomarkers in cervicovaginal fluid.

Kose B, Takmaz O, Gungor M, Baykal A Turk J Biol. 2023; 46(3):195-206.

PMID: 37529256 PMC: 10387838. DOI: 10.55730/1300-0152.2608.

Rapid and visual detection of specific bacteria for saliva and vaginal fluid identification with the lateral flow dipstick strategy.

Yu D, Zhang J, Gao N, Huo Y, Li W, Wang T Int J Legal Med. 2023; 137(6):1853-1863.

PMID: 37358650 DOI: 10.1007/s00414-023-03051-9.

Metal piracy by Neisseria gonorrhoeae to overcome human nutritional immunity.

Liyayi I, Forehand A, Ray J, Criss A PLoS Pathog. 2023; 19(2):e1011091.

PMID: 36730177 PMC: 9894411. DOI: 10.1371/journal.ppat.1011091.

Roles of Antimicrobial Peptides in Gynecological Cancers.

Zhao C, Yan S, Song Y, Xia X Int J Mol Sci. 2022; 23(17).

PMID: 36077500 PMC: 9456504. DOI: 10.3390/ijms231710104.

References

Elias J, Gygi S . Target-decoy search strategy for increased confidence in large-scale protein identifications by mass spectrometry. Nat Methods. 2007; 4(3):207-14. DOI: 10.1038/nmeth1019. View

Urban C, Reichard U, Brinkmann V, Zychlinsky A . Neutrophil extracellular traps capture and kill Candida albicans yeast and hyphal forms. Cell Microbiol. 2006; 8(4):668-76. DOI: 10.1111/j.1462-5822.2005.00659.x. View

Ashburner M, Ball C, Blake J, Botstein D, Butler H, Cherry J . Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. Nat Genet. 2000; 25(1):25-9. PMC: 3037419. DOI: 10.1038/75556. View

Dennis Jr G, Sherman B, Hosack D, Yang J, Gao W, Lane H . DAVID: Database for Annotation, Visualization, and Integrated Discovery. Genome Biol. 2003; 4(5):P3. View

Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss D . Neutrophil extracellular traps kill bacteria. Science. 2004; 303(5663):1532-5. DOI: 10.1126/science.1092385. View

Bar-Ilan A, Pessah N, MAREN T . The effects of carbonic anhydrase inhibitors on aqueous humor chemistry and dynamics. Invest Ophthalmol Vis Sci. 1984; 25(10):1198-205. View

Gravett M, Thomas A, Schneider K, Reddy A, Dasari S, Jacob T . Proteomic analysis of cervical-vaginal fluid: identification of novel biomarkers for detection of intra-amniotic infection. J Proteome Res. 2007; 6(1):89-96. PMC: 2532920. DOI: 10.1021/pr060149v. View

Georgiou H, Rice G, Baker M . Proteomic analysis of human plasma: failure of centrifugal ultrafiltration to remove albumin and other high molecular weight proteins. Proteomics. 2001; 1(12):1503-6. DOI: 10.1002/1615-9861(200111)1:12<1503::aid-prot1503>3.0.co;2-m. View

Sjoberg I, Cajander S, Rylander E . Morphometric characteristics of the vaginal epithelium during the menstrual cycle. Gynecol Obstet Invest. 1988; 26(2):136-44. DOI: 10.1159/000293685. View

10.

Pollard A, Sparey C, Robson S, Krainer A, Europe-Finner G . Spatio-temporal expression of the trans-acting splicing factors SF2/ASF and heterogeneous ribonuclear proteins A1/A1B in the myometrium of the pregnant human uterus: a molecular mechanism for regulating regional protein isoform expression in vivo. J Clin Endocrinol Metab. 2000; 85(5):1928-36. DOI: 10.1210/jcem.85.5.6537. View

11.

van der Strate B, Beljaars L, Molema G, Harmsen M, Meijer D . Antiviral activities of lactoferrin. Antiviral Res. 2001; 52(3):225-39. DOI: 10.1016/s0166-3542(01)00195-4. View

12.

Buchanan J, Simpson A, Aziz R, Liu G, Kristian S, Kotb M . DNase expression allows the pathogen group A Streptococcus to escape killing in neutrophil extracellular traps. Curr Biol. 2006; 16(4):396-400. DOI: 10.1016/j.cub.2005.12.039. View

13.

Wolters D, Washburn M, Yates 3rd J . An automated multidimensional protein identification technology for shotgun proteomics. Anal Chem. 2002; 73(23):5683-90. DOI: 10.1021/ac010617e. View

14.

Clayton A, Turkes A, Navabi H, Mason M, Tabi Z . Induction of heat shock proteins in B-cell exosomes. J Cell Sci. 2005; 118(Pt 16):3631-8. DOI: 10.1242/jcs.02494. View

15.

Eschenbach D, Thwin S, Patton D, Hooton T, Stapleton A, Agnew K . Influence of the normal menstrual cycle on vaginal tissue, discharge, and microflora. Clin Infect Dis. 2000; 30(6):901-7. DOI: 10.1086/313818. View

16.

Quayle A . The innate and early immune response to pathogen challenge in the female genital tract and the pivotal role of epithelial cells. J Reprod Immunol. 2002; 57(1-2):61-79. DOI: 10.1016/s0165-0378(02)00019-0. View

17.

van Niel G, Raposo G, Candalh C, Boussac M, HERSHBERG R, Cerf-Bensussan N . Intestinal epithelial cells secrete exosome-like vesicles. Gastroenterology. 2001; 121(2):337-49. DOI: 10.1053/gast.2001.26263. View

18.

Klein L, Jonscher K, Heerwagen M, Gibbs R, McManaman J . Shotgun proteomic analysis of vaginal fluid from women in late pregnancy. Reprod Sci. 2008; 15(3):263-73. DOI: 10.1177/1933719107311189. View

19.

Lehrer R, Barton A, Daher K, Harwig S, Ganz T, Selsted M . Interaction of human defensins with Escherichia coli. Mechanism of bactericidal activity. J Clin Invest. 1989; 84(2):553-61. PMC: 548915. DOI: 10.1172/JCI114198. View

20.

Zybailov B, Florens L, Washburn M . Quantitative shotgun proteomics using a protease with broad specificity and normalized spectral abundance factors. Mol Biosyst. 2007; 3(5):354-60. DOI: 10.1039/b701483j. View