1.Proinflammatory Cytokine and Nitric Oxide Production by Human Macrophages Stimulated with Trichomonas vaginalis.
Ik Hwan HAN ; Sung Young GOO ; Soon Jung PARK ; Se Jin HWANG ; Yong Seok KIM ; Michael Sungwoo YANG ; Myoung Hee AHN ; Jae Sook RYU
The Korean Journal of Parasitology 2009;47(3):205-212
Trichomonas vaginalis commonly causes vaginitis and perhaps cervicitis in women and urethritis in men and women. Macrophages are important immune cells in response to T. vaginalis infection. In this study, we investigated whether human macrophages could be involved in inflammation induced by T. vaginalis. Human monocyte-derived macrophages (HMDM) were co-cultured with T. vaginalis. Live, opsonized-live trichomonads, and T. vaginalis lysates increased proinflammatory cytokines, such as TNF-alpha, IL-1beta, and IL-6 by HMDM. The involvement of nuclear factor (NF)-kappaB signaling pathway in cytokine production induced by T. vaginalis was confirmed by phosphorylation and nuclear translocation of p65 NF-kappaB. In addition, stimulation with live T. vaginalis induced marked augmentation of nitric oxide (NO) production and expression of inducible NO synthase (iNOS) levels in HMDM. However, trichomonad-induced NF-kappaB activation and TNF-alpha production in macrophages were significantly inhibited by inhibition of iNOS levels with L-NMMA (NO synthase inhibitor). Moreover, pretreatment with NF-kappaB inhibitors (PDTC or Bay11-7082) caused human macrophages to produce less TNF-alpha. These results suggest that T. vaginalis stimulates human macrophages to produce proinflammatory cytokines, such as IL-1, IL-6, and TNF-alpha, and NO. In particular, we showed that T. vaginalis induced TNF-alpha production in macrophages through NO-dependent activation of NF-kappaB, which might be closely involved in inflammation caused by T. vaginalis.
Animals
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Cells, Cultured
;
Cytokines/*immunology
;
Humans
;
Macrophages/*immunology/parasitology
;
Nitric Oxide/*immunology
;
Trichomonas Infections/*immunology/parasitology
;
Trichomonas vaginalis/*immunology
2.Proinflammatory Cytokine and Nitric Oxide Production by Human Macrophages Stimulated with Trichomonas vaginalis.
Ik Hwan HAN ; Sung Young GOO ; Soon Jung PARK ; Se Jin HWANG ; Yong Seok KIM ; Michael Sungwoo YANG ; Myoung Hee AHN ; Jae Sook RYU
The Korean Journal of Parasitology 2009;47(3):205-212
Trichomonas vaginalis commonly causes vaginitis and perhaps cervicitis in women and urethritis in men and women. Macrophages are important immune cells in response to T. vaginalis infection. In this study, we investigated whether human macrophages could be involved in inflammation induced by T. vaginalis. Human monocyte-derived macrophages (HMDM) were co-cultured with T. vaginalis. Live, opsonized-live trichomonads, and T. vaginalis lysates increased proinflammatory cytokines, such as TNF-alpha, IL-1beta, and IL-6 by HMDM. The involvement of nuclear factor (NF)-kappaB signaling pathway in cytokine production induced by T. vaginalis was confirmed by phosphorylation and nuclear translocation of p65 NF-kappaB. In addition, stimulation with live T. vaginalis induced marked augmentation of nitric oxide (NO) production and expression of inducible NO synthase (iNOS) levels in HMDM. However, trichomonad-induced NF-kappaB activation and TNF-alpha production in macrophages were significantly inhibited by inhibition of iNOS levels with L-NMMA (NO synthase inhibitor). Moreover, pretreatment with NF-kappaB inhibitors (PDTC or Bay11-7082) caused human macrophages to produce less TNF-alpha. These results suggest that T. vaginalis stimulates human macrophages to produce proinflammatory cytokines, such as IL-1, IL-6, and TNF-alpha, and NO. In particular, we showed that T. vaginalis induced TNF-alpha production in macrophages through NO-dependent activation of NF-kappaB, which might be closely involved in inflammation caused by T. vaginalis.
Animals
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Cells, Cultured
;
Cytokines/*immunology
;
Humans
;
Macrophages/*immunology/parasitology
;
Nitric Oxide/*immunology
;
Trichomonas Infections/*immunology/parasitology
;
Trichomonas vaginalis/*immunology
3.Macrophages, myofibroblasts and mast cells in a rat liver infected with Capillaria hepatica.
Won Il JEONG ; Sun Hee DO ; Il Hwa HONG ; Ae Ri JI ; Jin Kyu PARK ; Mi Ran KI ; Seung Chun PARK ; Kyu Shik JEONG
Journal of Veterinary Science 2008;9(2):211-213
We trapped a rat (Rattus norvegicus) infected with Capillaria hepatica. At necropsy, grossly yellowish-white nodules (2-3 mm in diameter) were noted to be scattered on the liver's surface. Microscopically, granulomatous and fibrotic nodules that contained the eggs and/or adult worms of Capillaria hepatica were detected in the liver. Septal fibrosis was diffusely formed throughout the liver. There were a number of ED1-positive macrophages located in the sinusoids of the pseudolobules. On the double staining, myofibroblasts and mast cells were generally observed within the fibrous septa with the mast cells in close proximity to the myofibroblasts. We suggest that the interactions between macrophages, myofibroblasts and mast cells play a role in the septal fibrosis observed in rats infected by Capillaria hepatica.
Animals
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*Capillaria
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Enoplida Infections/immunology/parasitology/*veterinary
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Fibroblasts/immunology
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Liver/parasitology/pathology
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Macrophages/immunology
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Mast Cells/immunology
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Rats
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Rodent Diseases/*immunology/*parasitology/pathology
4.Involvement of MAP Kinases in Apoptosis of Macrophage Treated with Trichomonas vaginalis.
Yong Suk RYANG ; Jae Ho CHANG ; Ju Youn PARK
Yonsei Medical Journal 2004;45(4):751-754
A primitive protozoan parasite Trichomonas vaginalis selectively activates the signal transduction pathways in macrophages (RAW264.7). This study evaluated the correlation of these signaling pathways and T. vaginalis-induced cell apoptosis. In macrophages infected with T. vaginalis, apoptosis was assessed on the basis of DNA fragmentation on agarose gel electrophoresis. Infection of macrophages with T. vaginalis induced tyrosine phosphorylation of several proteins. Infected cells with T. vaginalis were shown to associate with phosphorylation of the extracellular signal-regulated (ERK) 1/2 kinase, p38, c-Jun N-terminal kinase (JNK) mitogen-activated protein (MAP) kinases on Western blot analysis. The present finding also demonstrated a link between the ERK1/2, JNK and p38 apoptotic pathways that was modulated by T. vaginalis infection.
Animals
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Apoptosis/*immunology
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Humans
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MAP Kinase Signaling System/immunology
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Macrophages/*cytology/enzymology/*parasitology
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Mitogen-Activated Protein Kinases/*metabolism
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Phosphorylation
;
Trichomonas Infections/*immunology
;
Trichomonas vaginalis/*immunology
5.Effects of specific monoclonal antibodies to dense granular proteins on the invasion of Toxoplasma gondii in vitro and in vivo.
Dong Yeob CHA ; In Kwan SONG ; Gye Sung LEE ; Ok Sun HWANG ; Hyung Jun NOH ; Seung Dong YEO ; Dae Whan SHIN ; Young Ha LEE
The Korean Journal of Parasitology 2001;39(3):233-240
Although some reports have been published on the protective effect of antibodies to Toxoplasma gondii surface membrane proteins, few address the inhibitory activity of antibodies to dense granular proteins (GRA proteins). Therefore, we performed a series of experiments to evaluate the inhibitory effects of monoclonal antibodies (mAbs) to GRA proteins (GRA2, 28 kDa; GRA6, 32 kDa) and surface membrane protein (SAG1, 30 kDa) on the invasion of T. gondii tachyzoites. Passive immunization of mice with one of three mAbs following challenge with a lethal dose of tachyzoites significantly increased survival compared with results for mice treated with control ascites. The survival times of mice challenged with tachyzoites pretreated with anti-GRA6 or anti-SAG1 mAb were significantly increased. Mice that received tachyzoites pretreated with both mAb and complement had longer survival times than those that received tachyzoites pretreated with mAb alone. Invasion of tachyzoites into fibroblasts and macrophages was significantly inhibited in the anti-GRA2, anti-GRA6 or anti-SAG1 mAb pretreated group. Pretreatment with mAb and complement inhibited invasion of tachyzoites in both fibroblasts and macrophages. These results suggest that specific antibodies to dense-granule molecules may be useful for controlling infection with T. gondii.
Animals
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Antibodies, Monoclonal/*pharmacology/therapeutic use
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*Antigens, Protozoan
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Female
;
Fibroblasts/parasitology
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Host-Parasite Relations
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Immunization, Passive
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Macrophages/parasitology
;
Mice
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Mice, Inbred BALB C
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Protozoan Proteins/*immunology
;
Support, Non-U.S. Gov't
;
Toxoplasma/*pathogenicity
;
Toxoplasmosis/parasitology/*therapy
6.Chronic Opisthorchis viverrini Infection and Associated Hepatobiliary Disease Is Associated with Iron Loaded M2-like Macrophages.
Moses T BILITY ; Banchob SRIPA
The Korean Journal of Parasitology 2014;52(6):695-699
Chronic Opisthorchis viverrini-induced hepatobiliary disease is associated with significant leukocyte infiltration, including activated macrophages; however, the polarization of infiltrating macrophages remains to be fully characterized. In this study, we characterized macrophage polarization and phenotype in chronic O. viverrini-induced hepatobiliary disease in humans and hamsters using gene expression and histochemical analysis. Chronic O. viverrini infection and associated hepatobiliary diseases were associated with iron loaded M2-like macrophages in both humans and hamsters. This study provides suggestive evidence that iron loaded M2-like macrophages promote hepatobiliary disease in chronic O. viverrini infection.
Animals
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Cricetinae
;
Gene Expression Profiling
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Histocytochemistry
;
Humans
;
Immunohistochemistry
;
Iron/*metabolism
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Liver Cirrhosis/*parasitology/*pathology
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Macrophages/*immunology/metabolism
;
Mesocricetus
;
Opisthorchiasis/*complications/*pathology
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Opisthorchis/*isolation & purification
7.Involvement of MAPK activation in chemokine or COX-2 productions by Toxoplasma gondii.
Ji Young KIM ; Myoung Hee AHN ; Hyun Ouk SONG ; Jong Hak CHOI ; Jae Sook RYU ; Duk Young MIN ; Myung Hwan CHO
The Korean Journal of Parasitology 2006;44(3):197-207
This experiment focused on MAPK activation in host cell invasion and replication of T. gondii, as well as the expression of CC chemokines, MCP-1 and MIP-1 alpha , and enzyme, COX-2/prostaglandin E2 (PGE2) in infected cells via western blot, [3H]-uracil incorporation assay, ELISA and RT-PCR. The phosphorylation of ERK1/2 and p38 in infected HeLa cells was detected at 1 hr and/or 6 hr postinfection (PI). Tachyzoite proliferation was reduced by p38 or JNK MAPK inhibitors. MCP-1 secretion was enhanced in infected peritoneal macrophages at 6 hr PI. MIP-1 alpha mRNA was increased in macrophages at 18 hr PI. MCP-1 and MIP-1 alpha were reduced after treatment with inhibitors of ERK1/2 and JNK MAPKs. COX-2 mRNA gradually increased in infected RAW 264.7 cells and the secretion of COX-2 peaked at 6 hr PI. The inhibitor of JNK suppressed COX-2 expression. PGE2 from infected RAW 264.7 cells was increased and synthesis was suppressed by PD98059, SB203580, and SP600125. In this study, the activation of p38, JNK and/or ERK1/2 MAPKs occurred during the invasion and proliferation of T. gondii tachyzoites in HeLa cells. Also, increased secretion and expression of MCP-1, MIP-1 alpha , COX-2 and PGE2 were detected in infected macrophages, and appeared to occur via MAPK signaling pathways.
Toxoplasmosis/*enzymology/*immunology
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Toxoplasma/*immunology/*metabolism
;
Mitogen-Activated Protein Kinases/*metabolism
;
Mice, Inbred BALB C
;
Mice
;
Macrophages, Peritoneal/enzymology/immunology/parasitology
;
Humans
;
Hela Cells
;
Enzyme Activation
;
Cyclooxygenase 2/*biosynthesis
;
Chemokines/*biosynthesis
;
Animals
8.Toxoplasma gondii: ultrastructural localization of specific antigens and inhibition of intracellular multiplication by monoclonal antibodies.
Boo Young LEE ; Myoung Hee AHN ; Hyun Chul KIM ; Duk Young MIN
The Korean Journal of Parasitology 2001;39(1):67-75
This experiment was focused on the characterization of anti-Toxoplasma monoclonal antibodies (mAbs) and the effect of mAbs on the parasite invasion of mouse peritoneal macrophages. Twenty eight mAbs including M110, M556, R7A6 and M621 were characterized by Ab titer, immunoglobulin isotyping and western blot pattern. Antibody titer (optical density) of 4 mAbs, M110, M556, R7A6 and M621, were 0.53, 0.67, 0.45 and 0.39 (normal mouse serum; 0.19) with the same IgG1 isotypes shown by Enzyme-linked immunosorbent assay (ELISA). Western blot analysis showed that M110, M556, R7A6 and M621 reacted with the 33 kDa (p30), 31 kDa (p28), 43 kDa and 36 kDa protein. Immunogold labelling of mAbs M110, M556, R7A6 and M621 reacted with the surface membrane, dense granules and parasitophorous vacuolar membrane (PVM), rhoptries and cytoplasm of tachyzoite, respectively. For in vitro assay, preincubation of tachyzoites with four mAbs, M110, M556, R7A6 and M621 resulted in the decrease of the number of infected macrophages (P < 0.05) and the suppression of parasite multiplication at 18 h post-infection. Four monoclonal antibodies including M110 (SAG1) were found to have an important role in the inhibition of macrophage invasion and T. gondii multiplication in vitro, and these mAbs may be suitable for vaccine candidates, diagnostic kit and for chemotherapy.
Animals
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*Antibodies, Monoclonal/pharmacology
;
*Antibodies, Protozoan/pharmacology
;
Antigens, Protozoan/*analysis/immunology
;
Cells, Cultured
;
Depression, Chemical
;
Macrophages, Peritoneal/parasitology
;
Mice
;
Mice, Inbred ICR
;
Support, Non-U.S. Gov't
;
Toxoplasma/growth & development/*immunology
9.Anti-leishmanial Effects of Trinitroglycerin in BALB/C Mice Infected with Leishmania major via Nitric Oxide Pathway.
Hossein NAHREVANIAN ; Mana NAJAFZADEH ; Reza HAJIHOSSEINI ; Habib NAZEM ; Mahin FARAHMAND ; Zahra ZAMANI
The Korean Journal of Parasitology 2009;47(2):109-115
This study investigated whether trinitroglycerine (TNG) as nitric oxide (NO) releasing agent had anti-leishmanial effects and mediated pathology in BALB/c mice infected with Leishmania major. Cutaneous leishmaniasis (CL), a zoonotic infection caused by leishmania protozoa is still one of the health problems in the world and in Iran. NO is involved in host immune responses against intracellular L. major, and leishmania killing by macrophages is mediated by this substance. Moreover, application of CL treatment with NO-donors has been recently indicated. In our study, TNG was used for its ability to increase NO and to modify CL infection in mice, in order to evaluate NO effects on lesion size and formation, parasite proliferation inside macrophages, amastigote visceralization in target organs, and NO induction in plasma and organ suspensions. Data obtained in this study indicated that TNG increased plasma and liver-NO, reduced lesion sizes, removed amastigotes from lesions, livers, spleens, and lymph nodes, declined proliferation of amastigotes, hepatomegaly, and increased survival rate. However, TNG reduced spleen-NO and had no significant effects on spelenomegaly. The results show that TNG therapy reduced leishmaniasis and pathology in association with raised NO levels. TNG had some antiparasitic activity by reduction of positive smears from lesions, livers, spleens, and lymph nodes, which could emphasize the role of TNG to inhibit visceralization of L. major in target organs.
Animal Structures/parasitology
;
Animals
;
Antiprotozoal Agents/chemistry/*therapeutic use
;
Female
;
Leishmania major/*drug effects/immunology
;
Leishmaniasis, Cutaneous
;
Macrophages/parasitology
;
Mice
;
Mice, Inbred BALB C
;
Nitric Oxide/blood/metabolism/*pharmacology
;
Nitroglycerin/*analogs & derivatives/*therapeutic use
;
Severity of Illness Index
;
Skin/pathology
;
Survival Analysis
10.Pentoxifylline inhibits liver fibrosis via hedgehog signaling pathway.
Hui LI ; Juan HUA ; Chun-Xia GUO ; Wei-Xian WANG ; Bao-Ju WANG ; Dong-Liang YANG ; Ping WEI ; Yin-Ping LU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2016;36(3):372-376
Infection of schistosomiasis japonica may eventually lead to liver fibrosis, and no effective antifibrotic therapies are available but liver transplantation. Hedgehog (HH) signaling pathway has been involved in the process and is a promising target for treating liver fibrosis. This study aimed to explore the effects of pentoxifylline (PTX) on liver fibrosis induced by schistosoma japonicum infection by inhibiting the HH signaling pathway. Phorbol12-myristate13-acetate (PMA) was used to induce human acute mononuclear leukemia cells THP-1 to differentiate into macrophages. The THP-1-derived macrophages were stimulated by soluble egg antigen (SEA), and the culture supernatants were collected for detection of activation of macrophages. Cell Counting Kit-8 (CCK-8) was used to detect the cytotoxicity of the culture supernatant and PTX on the LX-2 cells. The LX-2 cells were administered with activated culture supernatant from macrophages and(or) PTX to detect the transforming growth factor-β gene expression. The mRNA expression of shh and gli-1, key parts in HH signaling pathway, was detected. The mRNA expression of shh and gli-1 was increased in LX-2 cells treated with activated macrophages-derived culture supernatant, suggesting HH signaling pathway may play a key role in the activation process of hepatic stellate cells (HSCs). The expression of these genes decreased in LX-2 cells co-cultured with both activated macrophages-derived culture supernatant and PTX, indicating PTX could suppress the activation process of HSCs. In conclusion, these data provide evidence that PTX prevents liver fibrogenesis in vitro by the suppression of HH signaling pathway.
Animals
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Antigens, Helminth
;
isolation & purification
;
pharmacology
;
Cell Culture Techniques
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Cell Differentiation
;
drug effects
;
Cell Line
;
Culture Media, Conditioned
;
chemistry
;
pharmacology
;
Gene Expression Regulation
;
Hedgehog Proteins
;
agonists
;
antagonists & inhibitors
;
genetics
;
immunology
;
Hepatic Stellate Cells
;
cytology
;
drug effects
;
metabolism
;
Humans
;
Liver Cirrhosis
;
metabolism
;
parasitology
;
prevention & control
;
Macrophage Activation
;
drug effects
;
Macrophages
;
cytology
;
drug effects
;
immunology
;
Models, Biological
;
Monocytes
;
cytology
;
drug effects
;
metabolism
;
Pentoxifylline
;
pharmacology
;
Phosphodiesterase Inhibitors
;
pharmacology
;
RNA, Messenger
;
genetics
;
immunology
;
Schistosoma japonicum
;
chemistry
;
Signal Transduction
;
Tetradecanoylphorbol Acetate
;
pharmacology
;
Zinc Finger Protein GLI1
;
genetics
;
immunology
;
Zygote
;
chemistry