Abstract
BackgroundApproximately 114 million people are infected with Mansonella perstans in large proportions of Africa. In contrast to other filariae that infect humans, M. perstans-infected individuals show no distinct pathology or specific clinical picture, indicating a well-tuned adaptation to the host. In addition, since M. perstans adult worms reside in serous cavities which are difficult to access, research has been hindered and there is a paucity of knowledge about the biology of M. perstans, especially the development of the different life stages as well as M. perstans-driven immune responses. Thus in this study, an in vitro culture system was developed which allows an in-depth analysis of M. perstans.ResultsCulicoides species were caught in Ediki (Kumba), Southwest Region within Cameroon following a blood meal on a microfilaremic donor that had 1500 microfilariae/ml of peripheral blood and kept in captivity for 12 days at 23 °C. In a pilot experiment, 15 infective larvae were obtained from the midges and co-cultured with a confluent monolayer of monkey kidney epithelial cells (LLC-MK2) in DMEM medium supplemented with 10% FBS for up to 77 days. The resulting survival rates of 33% revealed that the cell-conditioned medium was suitable for long-term maintenance of M. perstans worms. To confirm these preliminary observations, 249 infective larvae were cultured for 50 days and their development was monitored daily and microscopically graded for motility. In total, 170 (68.3%) filariae survived and 124 (49.8%) larvae moulted between days 21–30 to become L5 stage larvae which were motile and showed continuous vigorous movement.ConclusionWe have established an in vitro culture system for the generation and long-term maintenance of viable M. perstans worms. This technique will be an important tool to study parasite biology and development, the role in host immunity, and might be helpful to discover novel treatment strategies against this filariae.
Highlights
114 million people are infected with Mansonella perstans in large proportions of Africa
We developed an in vitro culture to generate viable M. perstans worms from infective larvae (L3)
Long-term maintenance of infective M. perstans larvae was observed under these culture condition with 40% (n = 6) worm survival after 31 days and 33% (n = 5) surviving for up to 77 days in the in vitro culture (Fig. 1)
Summary
114 million people are infected with Mansonella perstans in large proportions of Africa. Since M. perstans adult worms reside in serous cavities which are difficult to access, research has been hindered and there is a paucity of knowledge about the biology of M. perstans, especially the development of the different life stages as well as M. perstans-driven immune responses. It is estimated that 110 million people in over 33 countries, especially in the tropical parts of Latin America as well as large portions of Africa are infected [1, 2]. The infective larvae (L3) develop into adult worms in an unknown manner and reside in serous body cavities [1,2,3]. Since M. perstans worms have only been recovered on rare occasions, research about the development of the different life stages remains limited. Fecund female worms can release numerous microfilariae (Mf), which circulate in the peripheral blood and can be taken up by another Culicoides midge during a blood meal [1,2,3]
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