A novel immortalized hepatocyte-like cell line (imHC) supports in vitro liver stage development of the human malarial parasite Plasmodium vivax
Abstract Background Eradication of malaria is difficult because of the ability of hypnozoite, the dormant liver-stage form of Plasmodium vivax, to cause relapse in patients. Research efforts to better understand the biology of P. vivax hypnozoite and design relapse prevention strategies have been ha...
Main Authors: | , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
BMC
2018-01-01
|
Series: | Malaria Journal |
Subjects: | |
Online Access: | http://link.springer.com/article/10.1186/s12936-018-2198-4 |
id |
doaj-0fbb9327683a43fc89f5a82044f62852 |
---|---|
record_format |
Article |
spelling |
doaj-0fbb9327683a43fc89f5a82044f628522020-11-25T01:37:08ZengBMCMalaria Journal1475-28752018-01-0117111410.1186/s12936-018-2198-4A novel immortalized hepatocyte-like cell line (imHC) supports in vitro liver stage development of the human malarial parasite Plasmodium vivaxYongyut Pewkliang0Siriwan Rungin1Kaewta Lerdpanyangam2Apisak Duangmanee3Phongthon Kanjanasirirat4Phichaya Suthivanich5Khanit Sa-ngiamsuntorn6Suparerk Borwornpinyo7Jetsumon Sattabongkot8Rapatbhorn Patrapuvich9Suradej Hongeng10Excellent Center for Drug Discovery (ECDD), Faculty of Science, Mahidol UniversityMahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol UniversityMahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol UniversityMahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol UniversityExcellent Center for Drug Discovery (ECDD), Faculty of Science, Mahidol UniversityExcellent Center for Drug Discovery (ECDD), Faculty of Science, Mahidol UniversityDepartment of Biochemistry, Faculty of Pharmacy, Mahidol UniversityExcellent Center for Drug Discovery (ECDD), Faculty of Science, Mahidol UniversityMahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol UniversityMahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol UniversityExcellent Center for Drug Discovery (ECDD), Faculty of Science, Mahidol UniversityAbstract Background Eradication of malaria is difficult because of the ability of hypnozoite, the dormant liver-stage form of Plasmodium vivax, to cause relapse in patients. Research efforts to better understand the biology of P. vivax hypnozoite and design relapse prevention strategies have been hampered by the lack of a robust and reliable model for in vitro culture of liver-stage parasites. Although the HC-04 hepatoma cell line is used for culturing liver-stage forms of Plasmodium, these cells proliferate unrestrictedly and detach from the culture dish after several days, which limits their usefulness in a long-term hypnozoite assay. Methods A novel immortalized hepatocyte-like cell line (imHC) was evaluated for the capability to support P. vivax sporozoite infection. First, expression of basic hepatocyte markers and all major malaria sporozoite-associated host receptors in imHC was investigated. Next, in vitro hepatocyte infectivity and intracellular development of sporozoites in imHC were determined using an indirect immunofluorescence assay. Cytochrome P450 isotype activity was also measured to determine the ability of imHC to metabolize drugs. Finally, the anti-liver-stage agent primaquine was used to test this model for a drug sensitivity assay. Results imHCs maintained major hepatic functions and expressed the essential factors CD81, SR-BI and EphA2, which are required for host entry and development of the parasite in the liver. imHCs could be maintained long-term in a monolayer without overgrowth and thus served as a good, supportive substrate for the invasion and growth of P. vivax liver stages, including hypnozoites. The observed high drug metabolism activity and potent responses in liver-stage parasites to primaquine highlight the potential use of this imHC model for antimalarial drug screening. Conclusions imHCs, which maintain a hepatocyte phenotype and drug-metabolizing enzyme expression, constitute an alternative host for in vitro Plasmodium liver-stage studies, particularly those addressing the biology of P. vivax hypnozoite. They potentially offer a novel, robust model for screening drugs against liver-stage parasites.http://link.springer.com/article/10.1186/s12936-018-2198-4MalariaPlasmodium vivaxSporozoiteLiver stageimHCs |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yongyut Pewkliang Siriwan Rungin Kaewta Lerdpanyangam Apisak Duangmanee Phongthon Kanjanasirirat Phichaya Suthivanich Khanit Sa-ngiamsuntorn Suparerk Borwornpinyo Jetsumon Sattabongkot Rapatbhorn Patrapuvich Suradej Hongeng |
spellingShingle |
Yongyut Pewkliang Siriwan Rungin Kaewta Lerdpanyangam Apisak Duangmanee Phongthon Kanjanasirirat Phichaya Suthivanich Khanit Sa-ngiamsuntorn Suparerk Borwornpinyo Jetsumon Sattabongkot Rapatbhorn Patrapuvich Suradej Hongeng A novel immortalized hepatocyte-like cell line (imHC) supports in vitro liver stage development of the human malarial parasite Plasmodium vivax Malaria Journal Malaria Plasmodium vivax Sporozoite Liver stage imHCs |
author_facet |
Yongyut Pewkliang Siriwan Rungin Kaewta Lerdpanyangam Apisak Duangmanee Phongthon Kanjanasirirat Phichaya Suthivanich Khanit Sa-ngiamsuntorn Suparerk Borwornpinyo Jetsumon Sattabongkot Rapatbhorn Patrapuvich Suradej Hongeng |
author_sort |
Yongyut Pewkliang |
title |
A novel immortalized hepatocyte-like cell line (imHC) supports in vitro liver stage development of the human malarial parasite Plasmodium vivax |
title_short |
A novel immortalized hepatocyte-like cell line (imHC) supports in vitro liver stage development of the human malarial parasite Plasmodium vivax |
title_full |
A novel immortalized hepatocyte-like cell line (imHC) supports in vitro liver stage development of the human malarial parasite Plasmodium vivax |
title_fullStr |
A novel immortalized hepatocyte-like cell line (imHC) supports in vitro liver stage development of the human malarial parasite Plasmodium vivax |
title_full_unstemmed |
A novel immortalized hepatocyte-like cell line (imHC) supports in vitro liver stage development of the human malarial parasite Plasmodium vivax |
title_sort |
novel immortalized hepatocyte-like cell line (imhc) supports in vitro liver stage development of the human malarial parasite plasmodium vivax |
publisher |
BMC |
series |
Malaria Journal |
issn |
1475-2875 |
publishDate |
2018-01-01 |
description |
Abstract Background Eradication of malaria is difficult because of the ability of hypnozoite, the dormant liver-stage form of Plasmodium vivax, to cause relapse in patients. Research efforts to better understand the biology of P. vivax hypnozoite and design relapse prevention strategies have been hampered by the lack of a robust and reliable model for in vitro culture of liver-stage parasites. Although the HC-04 hepatoma cell line is used for culturing liver-stage forms of Plasmodium, these cells proliferate unrestrictedly and detach from the culture dish after several days, which limits their usefulness in a long-term hypnozoite assay. Methods A novel immortalized hepatocyte-like cell line (imHC) was evaluated for the capability to support P. vivax sporozoite infection. First, expression of basic hepatocyte markers and all major malaria sporozoite-associated host receptors in imHC was investigated. Next, in vitro hepatocyte infectivity and intracellular development of sporozoites in imHC were determined using an indirect immunofluorescence assay. Cytochrome P450 isotype activity was also measured to determine the ability of imHC to metabolize drugs. Finally, the anti-liver-stage agent primaquine was used to test this model for a drug sensitivity assay. Results imHCs maintained major hepatic functions and expressed the essential factors CD81, SR-BI and EphA2, which are required for host entry and development of the parasite in the liver. imHCs could be maintained long-term in a monolayer without overgrowth and thus served as a good, supportive substrate for the invasion and growth of P. vivax liver stages, including hypnozoites. The observed high drug metabolism activity and potent responses in liver-stage parasites to primaquine highlight the potential use of this imHC model for antimalarial drug screening. Conclusions imHCs, which maintain a hepatocyte phenotype and drug-metabolizing enzyme expression, constitute an alternative host for in vitro Plasmodium liver-stage studies, particularly those addressing the biology of P. vivax hypnozoite. They potentially offer a novel, robust model for screening drugs against liver-stage parasites. |
topic |
Malaria Plasmodium vivax Sporozoite Liver stage imHCs |
url |
http://link.springer.com/article/10.1186/s12936-018-2198-4 |
work_keys_str_mv |
AT yongyutpewkliang anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT siriwanrungin anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT kaewtalerdpanyangam anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT apisakduangmanee anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT phongthonkanjanasirirat anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT phichayasuthivanich anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT khanitsangiamsuntorn anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT suparerkborwornpinyo anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT jetsumonsattabongkot anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT rapatbhornpatrapuvich anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT suradejhongeng anovelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT yongyutpewkliang novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT siriwanrungin novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT kaewtalerdpanyangam novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT apisakduangmanee novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT phongthonkanjanasirirat novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT phichayasuthivanich novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT khanitsangiamsuntorn novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT suparerkborwornpinyo novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT jetsumonsattabongkot novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT rapatbhornpatrapuvich novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax AT suradejhongeng novelimmortalizedhepatocytelikecelllineimhcsupportsinvitroliverstagedevelopmentofthehumanmalarialparasiteplasmodiumvivax |
_version_ |
1725059551466618880 |