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Njouendou, Abdel Jelil, Kien, Chi Anizette, Esum, Mathias E., Ritter, Manuel, Chounna Ndongmo, Winston Patrick, Fombad, Fanny Fri, Gandjui, Narcisse Victor T., Njiokou, Flobert, Enyong, Peter, Pfarr, Kenneth, Turner, Joseph 
ORCID: https://orcid.org/0000-0002-2185-5476, Layland, Laura E., Hoerauf, Achim and Wanji, Samuel
(2019)
'In vitro maintenance of Mansonella perstans microfilariae and its relevance for drug screening'. Experimental Parasitology, Vol 206, p. 107769.
Abstract
Background:
Mansonellosis arises from infections with threadlike filarial nematodes in millions of individuals, especially in sub-Saharan Africa. Since infections present no overt clinical symptoms but attenuate immune responses that might lead to increased susceptibility and worsened disease course of concomitant infections, it is truly a neglected tropical disease. Nevertheless, only few studies focus on identifying suitable safe drugs for its control and little is known about the requirements for in vitro maintenance of the Mansonella perstans transmission stage. This study, therefore, evaluated the survival of M. perstans microfilariae (mf) using in vitro conditions that have been shown to promote survival of Loa loa, a closely related filarial nematode. Furthermore, the in vitro microfilaricidal effect of 15 agents was assessed on this helminth.
Methods:
The ability of two basic culture media; Dulbecco's Modified Eagle's Medium (DMEM) and Roswell Park Memorial Institute (RPMI-1640) supplemented with 10% fetal bovine serum (FBS) and a monkey kidney epithelial cell line (LLC-MK2) to support the survival of M. perstans microfilariae was investigated. Subsequently, 6 anti-helminthics, 5 anti-malarials, 1 anti-microbacterial, 2 trypanocidals and 1 anti-cancer agent were tested in vitro against mf. The suitability of the culture media as well as the effect of the anti-infective agents on mf survival was assessed by scoring their motility.
Results:
FBS supplement and additional LLC-MK2 cells significantly improved the survival of mf in DMEM and RPMI-1640 culture. In detail, RPMI-1640 supplemented with 10% FBS and LLC-MK2 cells sustained the maintenance of mf for at least 20 days (100.00 ± 0.00% survival). In co-cultures with LLC-MK2 cells without serum, M. perstans mf were maintained in DMEM and RPMI-1640 medium with a motility above 99% by day 5. Mefloquine displayed the highest microfilaricidal effect in vitro followed by artesunate.
Conclusion:
Both RPMI and DMEM in the presence of LLC-MK2 cells are suitable for the maintenance of M. perstans mf in vitro. In absence of the feeder cells, the addition of 10% FBS to RPMI-1640 medium improved the
parasite survival rate and motility. The microfilaricidal activity of mefloquine and artesunate on M. perstans mf
| Item Type: | Article |
|---|---|
| Subjects: | QX Parasitology > Insects. Other Parasites > QX 505 Diptera WC Communicable Diseases > Tropical and Parasitic Diseases > WC 880 Filariasis and related conditions (General) |
| Faculty: Department: | Biological Sciences > Department of Tropical Disease Biology |
| Digital Object Identifer (DOI): | https://doi.org/10.1016/j.exppara.2019.107769 |
| Depositing User: | Cathy Waldron |
| Date Deposited: | 07 Oct 2019 11:07 |
| Last Modified: | 20 Jan 2020 13:08 |
| URI: | https://archive.lstmed.ac.uk/id/eprint/12682 |
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