Cystatin A protease inhibitor and cysteine proteases in human cervicovaginal fluid in term pregnancy and labor

Published:December 17, 2010DOI:


      The purpose of this study was to investigate the temporal changes in immunoreactive cystatin A and the enzymatic activity of cathepsins B, H, L, and S in human cervicovaginal fluid (CVF) in late pregnancy and spontaneous labor.

      Study Design

      CVF was collected weekly (n = 95 women) from 36 weeks gestation until spontaneous term labor. Cystatin A was quantified using enzyme-linked immunosorbent assay. The enzyme activity of cathepsins B, H, L, and S was measured with fluorometric enzyme assay kits.


      Cystatin A significantly decreased towards (P = .016, 2-way analysis of variance) and during labor (P < .001, 2-way analysis of variance). Enzymatic activity of cathepsins B, H, and S did not change with labor onset (P = .452, P = .703, P = .411, respectively, 2-way analysis of variance).


      In late gestation, CVF-decreased expression of the cysteine protease inhibitor, cystatin A, is associated with labor. Although the role and contribution of cystatin A to increased extracellular matrix remodeling has yet to be elucidated, the data that were obtained are consistent with this hypothesis.

      Key words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to American Journal of Obstetrics & Gynecology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Rinne A.
        Epidermal SH-protease inhibitor (ACPI, cystatin A) in cancer: a short historical review.
        Pathol Res Pract. 2010; 206: 259-262
        • Davies M.E.
        • Barrett A.J.
        Immunolocalization of human cystatin in neutrophils and lymphocytes.
        Histochemistry. 1984; 80: 373-377
        • Rinne A.
        • Järvinen M.
        • Räsänen O.
        A protein reminiscent of the epidermal SH- protease inhibitor occurs in squamous epithelia of man and rat.
        Acta Histochem. 1978; 63: 183-192
        • Abrahamson M.
        • Barrett A.J.
        • Salvesen G.
        • Grubb A.
        Isolation of six cysteine proteinase inhibitors from human urine: their physicochemical and enzyme kinetic properties and concentrations in biological fluids.
        J Biol Chem. 1986; 261: 11282-11289
        • Di Quinzio M.K.
        • Oliva K.
        • Holdsworth S.J.
        • et al.
        Proteomic analysis and characterisation of human cervico-vaginal fluid proteins.
        Aust N Z J Obstet Gynaecol. 2007; 47: 9-15
        • Turk B.
        • Turk V.
        • Turk D.
        Structural and functional aspects of papain-like cysteine proteinases and their protein inhibitors.
        Biol Chem. 1997; 378: 141-150
        • Rudenskaya G.N.
        • Pupov D.V.
        Cysteine proteinases of microorganisms and viruses.
        Biochem (Mosc). 2008; 73: 1-13
        • Barrett A.J.
        • Kirschke H.
        Cathepsin B, cathepsin H, and cathepsin L.
        Methods Enzymol. 1981; 80: 535-561
        • Yasothornsrikul S.
        • Greenbaum D.
        • Medzihradszky K.F.
        • et al.
        Cathepsin L in secretory vesicles functions as a prohormone-processing enzyme for production of the enkephalin peptide neurotransmitter.
        Proc Natl Acad Sci U S A. 2003; 100: 9590-9595
        • Shi G.P.
        • Webb A.C.
        • Foster K.E.
        • et al.
        Human cathepsin S: chromosomal localization, gene structure, and tissue distribution.
        J Biol Chem. 1994; 269: 11530-11536
        • Guinec N.
        • Dalet-Fumeron V.
        • Pagano M.
        “In vitro” study of basement membrane degradation by the cysteine proteinases, cathepsins B, B-like and L: digestion of collagen IV, laminin, fibronectin, and release of gelatinase activities from basement membrane fibronectin.
        Biol Chem Hoppe Seyler. 1993; 374: 1135-1146
        • Felbor U.
        • Dreier L.
        • Bryant R.A.
        • Ploegh H.L.
        • Olsen B.R.
        • Mothes W.
        Secreted cathepsin L generates endostatin from collagen XVIII.
        EMBO J. 2000; 19: 1187-1194
        • Maciewicz R.A.
        • Etherington D.J.
        A comparison of four cathepsins (B, L, N and S) with collagenolytic activity from rabbit spleen.
        Biochem J. 1988; 256: 433-440
        • Novinec M.
        • Grass R.N.
        • Stark W.J.
        • Turk V.
        • Baici A.
        • Lenarcic B.
        Interaction between human cathepsins K, L, and S and elastins: mechanism of elastinolysis and inhibition by macromolecular inhibitors.
        J Biol Chem. 2007; 282: 7893-7902
        • Mason R.W.
        • Johnson D.A.
        • Barrett A.J.
        • Chapman H.A.
        Elastinolytic activity of human cathepsin L.
        Biochem J. 1986; 233: 925-927
        • Etherington D.J.
        • Evans P.J.
        The action of cathepsin B and collagenolytic cathepsin in the degradation of collagen.
        Acta Biol Med Ger. 1977; 36: 1555-1563
        • Riese R.J.
        • Wolf P.R.
        • Brömme D.
        • et al.
        Essential role for cathepsin S in MHC class II-associated invariant chain processing and peptide loading.
        Immunity. 1996; 4: 357-366
        • Reddy V.Y.
        • Zhang Q.Y.
        • Weiss S.J.
        Pericellular mobilization of the tissue-destructive cysteine proteinases, cathepsins B, L, and S, by human monocyte-derived macrophages.
        Proc Natl Acad Sci U S A. 1995; 92: 3849-3853
        • Di Quinzio M.K.
        • Georgiou H.M.
        • Holdsworth-Carson S.J.
        • et al.
        Proteomic analysis of human cervico-vaginal fluid displays differential protein expression in association with labor onset at term.
        J Proteome Res. 2008; 7: 1916-1921
        • Shaw J.L.
        • Smith C.R.
        • Diamandis E.P.
        Proteomic analysis of human cervico-vaginal fluid.
        J Proteome Res. 2007; 6: 2859-2865
        • Dasari S.
        • Pereira L.
        • Reddy A.P.
        • et al.
        Comprehensive proteomic analysis of human cervical-vaginal fluid.
        J Proteome Res. 2007; 6: 1258-1268
        • Pereira L.
        • Reddy A.P.
        • Jacob T.
        • et al.
        Identification of novel protein biomarkers of preterm birth in human cervical-vaginal fluid.
        J Proteome Res. 2007; 6: 1269-1276
        • Shah S.J.
        • Yu K.H.
        • Sangar V.
        • Parry S.I.
        • Blair I.A.
        Identification and quantification of preterm birth biomarkers in human cervicovaginal fluid by liquid chromatography/tandem mass spectrometry.
        J Proteome Res. 2009; 8: 2407-2417
        • Martin J.R.
        • Craven C.J.
        • Jerala R.
        • et al.
        The three-dimensional solution structure of human stefin A.
        J Mol Biol. 1995; 246: 331-343
        • Japelj B.
        • Waltho J.P.
        • Jerala R.
        Comparison of backbone dynamics of monomeric and domain-swapped stefin A.
        Proteins. 2004; 54: 500-512
        • Watari M.
        • Watari H.
        • DiSanto M.E.
        • Chacko S.
        • Shi G.P.
        • Strauss 3rd, J.F.
        Pro-inflammatory cytokines induce expression of matrix-metabolizing enzymes in human cervical smooth muscle cells.
        Am J Pathol. 1999; 154: 1755-1762
        • Al-Zaid N.S.
        • Gumaa K.A.
        • Bou-Resli M.N.
        Changes in amniotic fluid cathepsins with gestational age.
        J Dev Physiol. 1989; 12: 273-275
        • Grubb J.D.
        • Koszalk T.R.
        • Drabick J.J.
        • Metrione R.M.
        The activities of thiol proteases in the rat visceral yolk sac increase during late gestation.
        Placenta. 1991; 12: 143-151
        • Schick C.
        • Pemberton P.A.
        • Shi G.P.
        • et al.
        Cross-class inhibition of the cysteine proteinases cathepsins K, L, and S by the serpin squamous cell carcinoma antigen 1: a kinetic analysis.
        Biochemistry. 1998; 37: 5258-5266
        • Heng Y.J.
        • Di Quinzio M.K.
        • Permezel M.
        • Ayhan M.
        • Rice G.E.
        • Georgiou H.M.
        Temporal proteomic analysis of human cervicovaginal fluid with impending term labor.
        J Proteome Res. 2010; 9: 1344-1350
        • Venkataraman N.
        • Cole A.L.
        • Svoboda P.
        • Pohl J.
        • Cole A.M.
        Cationic polypeptides are required for anti-HIV-1 activity of human vaginal fluid.
        J Immunol. 2005; 175: 7560-7567
        • Klein L.L.
        • Jonscher K.R.
        • Heerwagen M.J.
        • Gibbs R.S.
        • McManaman J.L.
        Shotgun proteomic analysis of vaginal fluid from women in late pregnancy.
        Reprod Sci. 2008; 15: 263-273
        • Zegels G.
        • Van Raemdonck G.A.A.
        • Coen E.P.
        • Tjalma W.A.A.
        • Van Ostade X.W.M.
        Comprehensive proteomic analysis of human cervical-vaginal fluid using colposcopy samples.
        Proteome Sci. 2009; 7: 17