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Korean J Parasitol > Volume 35(1):1997 > Article

Original Article
Korean J Parasitol. 1997 Mar;35(1):39-46. English.
Published online Mar 20, 1997.  http://dx.doi.org/10.3347/kjp.1997.35.1.39
Copyright © 1997 by The Korean Society for Parasitology
Degradation of human immunoglobulins and cytotoxicity on HeLa cells by live Trichomonas vaginalis
D Y Min,1J S Ryu,*1S Y Park,1M H Shin,2 and W Y Cho1
1Department of Parasitology, College of Medicine, Hanyang University, Seoul 133-791, Korea.
Received February 28, 1997; Accepted March 03, 1997.

Abstract

The present study was undertaken to determine whether live T. vaginalis degrades human secretory IgA, serum IgA and IgG molecules. Human immunoglobulins were exposed to live trophozoites, parasite lysate, and excretory-secretory product (ESP) of T. vaginalis. To determine the fragmentation of immunoglobulins, the reaction sample was subjected to SDS-PAGE and EITB, and peroxidase conjugated antihuman IgA and IgG were used as probes. Live trophozoites degraded secretory IgA. Serum IgA and IgG, and degradation were pressed forward by the prolongation of the incubation time and by increasing the number of trichomonads respectively. Also the lysates and ESP of trichomonads degraded IgA and IgG. The cysteine and serine proteinase inhibitors such as E-64, antipain, iodoacetic acid, iodoacetamide, TLCK reduced the ability of cleaving immunoglobulins. The proteinase activity and cytotoxicity of T. vaginalis to HeLa cells were decreased when live T. vaginalis was treated with metallo-proteinase inhibitor as well as cysteine and serine proteinase inhibitors. These results suggest that proteinase secreted from live T. vaginalis may play a part role in host pathogenesis by T. vaginalis, and the cleaving ability of host immunoglobulins by the proteinase may contribute as a one of immune evasion mechanism for parasite survival in the host.

Figures


Fig. 1
Degradation of secretory IgA by live T.vaginalis. Live T. vaginalis digest secretory IgA in a dose-dependent manner. Lane 1, secretory IgA alone; Lane 2-5, secretory IgA incubated with 2 × 105, 5 × 105, 2 × 105 and 5 × 106 of T. vaginalis. (▸; heavy chain)


Fig. 2
Degradation of serum IgA by live T. vaginalis. Live T. vaginalis digest serum IgA in a time-dependent and dose-dependent manner. Lane 1, serum IgA alone; Lane 2-4, serum IgA incubated with T. vaginalis (2 × 106) for 30 min, 1 hr and 2 hrs. Lane 5-8 serum IgA incubated for 2 hrs with 5 × 104, 2 × 105, 5 × 105 and 1.5 × 106 of T. vaginalis. (▸; heavy chain)


Fig. 3
Degradation of serum IgG by live T. vaginalis. Live T. vaginalis digest serum IgG in a time-dependent and dose-dependent manner. Lane 1, serum IgG alone; Lane 2-4, serum IgG incubated with T. vaginalis (2 × 106) for 30 min, 1 hr, 2 hrs and 22 hrs. Lane 6-9 serum IgG incubated for 2 hrs with 5 × 104, 2 × 105, 5 × 105 and 1.5 × 106 of T. vaginalis. (▸; heavy chain)


Fig. 4
Degradation of human immunoglobulins by T. vaginalis. lysate (50 µg) and excretory-secretory product (ESP) (50 µg). A: Degradation of secretory IgA by T. vaginalis lysate (lane 2) and ESP (lane 3). B: Degradation of serum IgA by T. vaginalis lysate (lane 2) and ESP (lane 3). C: Degradation of IgG by T. vaginalis lysate (lane 2) and ESP (lane 3).


Fig. 5
Degradation of secretory IgA by live T. vaginalis treated with following proteinase inhibitors. Lane 1, secretory IgA alone; Lane 2, DTT; Lane 3, bestatin; Lane 4, EDTA; Lane 5, TLCK; Lane 6, E-64, Lane 7, leupeptin; Lane 8, antipain; Lane 9, IAA; Lane 10, iodoacetamide. (▸; heavy chain)

Tables


Table 1
Effect of proteinase inhibitor on the cytotoxicity of Trichomonas vaginalis to HeLa cells


Table 2
Effect of proteinase inhibitor on the of proteinase activity of Trichomonas vaginalis

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