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Entamoeba histolytica induces cyclooxy-genase-2 expression in macrophages isolated from liver abscess
Oral Sessions I Parasitology
inwrnational47
O-0162
(Suppl.) (1998) 133-281
o_o~~
EFFECT
OF IRON ON THE PATHOGENICITY
TRICHOMONAS &LJ_$,
‘Biotechnology Research Department, Hemofarm Concern, Vrsac, Yugoslavia and “Military Medical Academy, Institute for Microbiology, Belgrade, Yugoslavia
Axenic culture Monoxenic cunure Potyxenic culture
negative
I
3
1 1
3
I z
2 18 26
vary
during
menstrual
from lactoftin concenh-ation
cycle.The presentstudy
in vaginal known to
is
was
compared Normsl media was Diamond(l957)‘TY~vl wnc.:34.8/lM). Iron-supplemented C393~M) snd
undertaken
medium (iron iron-depleted
media(l.l~M)were made by adding ferrous sulfate or dipyridyl to TYM media respectively. Five Korean isolates and CDc85 shain of T. wginoris were used for the experiments. Trophozoites of normal TYM and iron-TYM media caused subcutaneous abscess formation in the mice. In contrast to trichomonads in dipyridyl-TYM didn’t make the abscess. Adhesions and cyotoxicity of hichomonads to HeLa cell were significantly decreased in trophozoites of dipyridyl-TYM in comparison with those of normal and iron-TYM media. Pmteinase activity using
1100.001100.00
Chemin J -School of Biological Sciences, University of Portsmouth King Henry Building, King Henry I Street, Portsmouth PO1 2DY, UK
DY’
to observe the effect of iron on the pathogenicity of T. wgimlis. Trophozoites cultivated in three sorts of TYM media were
1 11.64 ] 11.64 30.77 1 7.69 [ 1 69.23 1 69.23
THE IN-VITRO CULTURE OF HELMINTHS USING A THIXOTROPIC GEL
Aim MH’, Sbin MH”, Mii
T. wir&is obtain iron mainly mucus. However, vaginal la&of&
Persistent bacterial species have been identiied as Proteus mirabik, Escherichia co/i and Entemcoccus sp. Axenic cultiiatii of protozoan Bh can be achieved under controlled conditionsand with the use of appropriate antibiitics: _ respecting of the simplified methodologyof axenic cuitiiation, as it has been shown, does not guarantee a successful realization of the axenization, - certain Bh strains do not reproduce even during the first subcuitivation. while some other strains of this microorganism stop reproducing after initial success and four to five accomplished subcuitivatiins, -the surviving of Bh axenic culture is still an up-to-date problem, as axenic culture of Bh irreversibly degrade and die after six to eight subcuitivations. - monoxenic cultures of Bh represent the primary solution in preserving this protozoan in vitro.
o-@lG
OF
VAGINALIS
%epartment of Parasitology, College of Medicine, Hanyang University, Seoul 133-791 and “Deparbnent of Parasitology, College of Medicine, Ewha Womans’s University, Seoul 158-056, KOrea
Blastocystis hominis (Bh) is an eukaryotic organism of significant polymorphism and extraordinary diversity of size; Bh is an anaerobic microorganismand a gastrointestinal protozoan parasite of humans and primateswith certain pathogenic potential. The most important aim in our work has been the methodological working out of the simplifiedprocedure of Bh axenit cultivation. The axenic cultivation has been conducted on the basis of methodologies presented in references, along with specifically defined methodologicalmodificatiins. The axenic cultiiatii has been achieved wlh twenty-six strains of Bh. previcusiydiagnosed in human liiuid stool samples. The results of the axenic cultivation on twenty-six strains of Bh, after fourteen successive subcultivations, are shown in the table:
posit&e
1.53
Bz-Pro-Phe-Arg-Nan.
as substrate
of dipyridyl-TYM medium medium. Also trichomonads the gelatin
weakly
on immunoblot
on gelatin
as contrasted
These results suggest related with pathogenic&
were decreased
in trophozoites
in comparison with normal TYM of dipyridyl-TYM medium degraded SDS-PAGE
and showed
fewer band
with normal and iron-TYM that iron may of Trichmoms
affect on mgimlis.
media the
It
factors could be
proposed that aggravation of symtom and sign caused by 7: oo&faZs after menstration probably related with the iron.
O-0165
Entamoeba hisfolytica INDUCES CYCLOOXYGENASE-2 EXPRESSION IN MACROPHAGES ISOLATED FROM LIVER ABSCESS.
Sgnchez-Ramirez..*, Escalante, B.“, Resales-Encina, J.L.‘“. TalamBs-Rohana. P.*** Laboratory of lmmunoiogy Research, Faculty of Chemistry, Chihuahua State University. Apdo. Postal 1542-C, Chihuahua, Chih., MBxico. ** Departments of Pharmacology and Toxicology and ***Experimental Pathology. CINVESTAV-IPN, Mlxico. l
The majority of parasitic helminths can be kept alive in-vitro for varying lengths of time. There are examples where parasites have remained alive, viable and have show" varying degrees of development. However, in most cases a liquid culture medium has been used or a combined liquid semi-solid medium. Neither of these methods simulate closely the conditions found within the host, in particular those parasites that burrow into the tissues. An in-vitro culture medium using a thixotropic gel was prepared for maintaining both gut and tissue parasites. The results indicate that parasites are able to survive in or on this medium, the burrowing forms can be observed in medium. The survival rate of the parasites depends on the composition of the media added to the gel. The results indicate that this system may be suitable for in-vitro drug screening.
Enfamoeba histolytica infection curses with impairment of macrophage effector functions. Pretreatment of animals with the cyclooxygenase (COX) inhibitor indomethacin, reduced the hepatic abscess development in hamsters. Recently, the presence of a new inducible COX (COX-2) associated with inflammation has been described. To determinate if COX-2 was related to the inflammation process on amoebosis, we detected the enzyme by Western blot. COX-2 was expressed only by amoebic abscess derived macrophages but not by peritoneal macrophages from intraperitoneally infected (5 x IO’ trophozoites) animals. These results suggest that E. histolytica can stimulate COX-2 activity, this could down regulate macrophage functions. Alternatively, COX-2 induction might result from the effect of cytokines released during the inflammatory process. Studies are in progress to detenine whether liver COX-2 is being induced and to elucidate the modulatory role of PGE2 on the immune response during amoebosis.
inwrnational47
O-0162
(Suppl.) (1998) 133-281
o_o~~
EFFECT
OF IRON ON THE PATHOGENICITY
TRICHOMONAS &LJ_$,
‘Biotechnology Research Department, Hemofarm Concern, Vrsac, Yugoslavia and “Military Medical Academy, Institute for Microbiology, Belgrade, Yugoslavia
Axenic culture Monoxenic cunure Potyxenic culture
negative
I
3
1 1
3
I z
2 18 26
vary
during
menstrual
from lactoftin concenh-ation
cycle.The presentstudy
in vaginal known to
is
was
compared Normsl media was Diamond(l957)‘TY~vl wnc.:34.8/lM). Iron-supplemented C393~M) snd
undertaken
medium (iron iron-depleted
media(l.l~M)were made by adding ferrous sulfate or dipyridyl to TYM media respectively. Five Korean isolates and CDc85 shain of T. wginoris were used for the experiments. Trophozoites of normal TYM and iron-TYM media caused subcutaneous abscess formation in the mice. In contrast to trichomonads in dipyridyl-TYM didn’t make the abscess. Adhesions and cyotoxicity of hichomonads to HeLa cell were significantly decreased in trophozoites of dipyridyl-TYM in comparison with those of normal and iron-TYM media. Pmteinase activity using
1100.001100.00
Chemin J -School of Biological Sciences, University of Portsmouth King Henry Building, King Henry I Street, Portsmouth PO1 2DY, UK
DY’
to observe the effect of iron on the pathogenicity of T. wgimlis. Trophozoites cultivated in three sorts of TYM media were
1 11.64 ] 11.64 30.77 1 7.69 [ 1 69.23 1 69.23
THE IN-VITRO CULTURE OF HELMINTHS USING A THIXOTROPIC GEL
Aim MH’, Sbin MH”, Mii
T. wir&is obtain iron mainly mucus. However, vaginal la&of&
Persistent bacterial species have been identiied as Proteus mirabik, Escherichia co/i and Entemcoccus sp. Axenic cultiiatii of protozoan Bh can be achieved under controlled conditionsand with the use of appropriate antibiitics: _ respecting of the simplified methodologyof axenic cuitiiation, as it has been shown, does not guarantee a successful realization of the axenization, - certain Bh strains do not reproduce even during the first subcuitivation. while some other strains of this microorganism stop reproducing after initial success and four to five accomplished subcuitivatiins, -the surviving of Bh axenic culture is still an up-to-date problem, as axenic culture of Bh irreversibly degrade and die after six to eight subcuitivations. - monoxenic cultures of Bh represent the primary solution in preserving this protozoan in vitro.
o-@lG
OF
VAGINALIS
%epartment of Parasitology, College of Medicine, Hanyang University, Seoul 133-791 and “Deparbnent of Parasitology, College of Medicine, Ewha Womans’s University, Seoul 158-056, KOrea
Blastocystis hominis (Bh) is an eukaryotic organism of significant polymorphism and extraordinary diversity of size; Bh is an anaerobic microorganismand a gastrointestinal protozoan parasite of humans and primateswith certain pathogenic potential. The most important aim in our work has been the methodological working out of the simplifiedprocedure of Bh axenit cultivation. The axenic cultivation has been conducted on the basis of methodologies presented in references, along with specifically defined methodologicalmodificatiins. The axenic cultiiatii has been achieved wlh twenty-six strains of Bh. previcusiydiagnosed in human liiuid stool samples. The results of the axenic cultivation on twenty-six strains of Bh, after fourteen successive subcultivations, are shown in the table:
posit&e
1.53
Bz-Pro-Phe-Arg-Nan.
as substrate
of dipyridyl-TYM medium medium. Also trichomonads the gelatin
weakly
on immunoblot
on gelatin
as contrasted
These results suggest related with pathogenic&
were decreased
in trophozoites
in comparison with normal TYM of dipyridyl-TYM medium degraded SDS-PAGE
and showed
fewer band
with normal and iron-TYM that iron may of Trichmoms
affect on mgimlis.
media the
It
factors could be
proposed that aggravation of symtom and sign caused by 7: oo&faZs after menstration probably related with the iron.
O-0165
Entamoeba hisfolytica INDUCES CYCLOOXYGENASE-2 EXPRESSION IN MACROPHAGES ISOLATED FROM LIVER ABSCESS.
Sgnchez-Ramirez..*, Escalante, B.“, Resales-Encina, J.L.‘“. TalamBs-Rohana. P.*** Laboratory of lmmunoiogy Research, Faculty of Chemistry, Chihuahua State University. Apdo. Postal 1542-C, Chihuahua, Chih., MBxico. ** Departments of Pharmacology and Toxicology and ***Experimental Pathology. CINVESTAV-IPN, Mlxico. l
The majority of parasitic helminths can be kept alive in-vitro for varying lengths of time. There are examples where parasites have remained alive, viable and have show" varying degrees of development. However, in most cases a liquid culture medium has been used or a combined liquid semi-solid medium. Neither of these methods simulate closely the conditions found within the host, in particular those parasites that burrow into the tissues. An in-vitro culture medium using a thixotropic gel was prepared for maintaining both gut and tissue parasites. The results indicate that parasites are able to survive in or on this medium, the burrowing forms can be observed in medium. The survival rate of the parasites depends on the composition of the media added to the gel. The results indicate that this system may be suitable for in-vitro drug screening.
Enfamoeba histolytica infection curses with impairment of macrophage effector functions. Pretreatment of animals with the cyclooxygenase (COX) inhibitor indomethacin, reduced the hepatic abscess development in hamsters. Recently, the presence of a new inducible COX (COX-2) associated with inflammation has been described. To determinate if COX-2 was related to the inflammation process on amoebosis, we detected the enzyme by Western blot. COX-2 was expressed only by amoebic abscess derived macrophages but not by peritoneal macrophages from intraperitoneally infected (5 x IO’ trophozoites) animals. These results suggest that E. histolytica can stimulate COX-2 activity, this could down regulate macrophage functions. Alternatively, COX-2 induction might result from the effect of cytokines released during the inflammatory process. Studies are in progress to detenine whether liver COX-2 is being induced and to elucidate the modulatory role of PGE2 on the immune response during amoebosis.