Successful long-term maintenance of Mansonella perstans in an in vitro culture system
Abstract Background Approximately 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...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2017-11-01
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| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13071-017-2515-8 |
| _version_ | 1811319838999576576 |
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| author | Abdel Jelil Njouendou Manuel Ritter Winston Patrick Chounna Ndongmo Chi Anizette Kien Gandjui Tchamatchoua Victor Narcisse Fanny Fri Fombad Dizzle Bita Tayong Kenneth Pfarr Laura E. Layland Achim Hoerauf Samuel Wanji |
| author_facet | Abdel Jelil Njouendou Manuel Ritter Winston Patrick Chounna Ndongmo Chi Anizette Kien Gandjui Tchamatchoua Victor Narcisse Fanny Fri Fombad Dizzle Bita Tayong Kenneth Pfarr Laura E. Layland Achim Hoerauf Samuel Wanji |
| author_sort | Abdel Jelil Njouendou |
| collection | DOAJ |
| description | Abstract Background Approximately 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. Results Culicoides 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. Conclusion We 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. |
| first_indexed | 2024-04-13T12:49:28Z |
| format | Article |
| id | doaj.art-8dc1c7d15fd645e5b8c3fe22bb6de46a |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-04-13T12:49:28Z |
| publishDate | 2017-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-8dc1c7d15fd645e5b8c3fe22bb6de46a2022-12-22T02:46:17ZengBMCParasites & Vectors1756-33052017-11-011011610.1186/s13071-017-2515-8Successful long-term maintenance of Mansonella perstans in an in vitro culture systemAbdel Jelil Njouendou0Manuel Ritter1Winston Patrick Chounna Ndongmo2Chi Anizette Kien3Gandjui Tchamatchoua Victor Narcisse4Fanny Fri Fombad5Dizzle Bita Tayong6Kenneth Pfarr7Laura E. Layland8Achim Hoerauf9Samuel Wanji10Parasite and Vector Research Unit (PAVRU), Department of Microbiology and Parasitology, University of BueaInstitute of Medical Microbiology, Immunology and Parasitology, University Hospital BonnParasite and Vector Research Unit (PAVRU), Department of Microbiology and Parasitology, University of BueaParasite and Vector Research Unit (PAVRU), Department of Microbiology and Parasitology, University of BueaParasite and Vector Research Unit (PAVRU), Department of Microbiology and Parasitology, University of BueaParasite and Vector Research Unit (PAVRU), Department of Microbiology and Parasitology, University of BueaParasite and Vector Research Unit (PAVRU), Department of Microbiology and Parasitology, University of BueaInstitute of Medical Microbiology, Immunology and Parasitology, University Hospital BonnInstitute of Medical Microbiology, Immunology and Parasitology, University Hospital BonnInstitute of Medical Microbiology, Immunology and Parasitology, University Hospital BonnParasite and Vector Research Unit (PAVRU), Department of Microbiology and Parasitology, University of BueaAbstract Background Approximately 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. Results Culicoides 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. Conclusion We 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.http://link.springer.com/article/10.1186/s13071-017-2515-8Mansonella perstansIn vitro cultureWormsL3 infective larvaeLong-term maintenance |
| spellingShingle | Abdel Jelil Njouendou Manuel Ritter Winston Patrick Chounna Ndongmo Chi Anizette Kien Gandjui Tchamatchoua Victor Narcisse Fanny Fri Fombad Dizzle Bita Tayong Kenneth Pfarr Laura E. Layland Achim Hoerauf Samuel Wanji Successful long-term maintenance of Mansonella perstans in an in vitro culture system Parasites & Vectors Mansonella perstans In vitro culture Worms L3 infective larvae Long-term maintenance |
| title | Successful long-term maintenance of Mansonella perstans in an in vitro culture system |
| title_full | Successful long-term maintenance of Mansonella perstans in an in vitro culture system |
| title_fullStr | Successful long-term maintenance of Mansonella perstans in an in vitro culture system |
| title_full_unstemmed | Successful long-term maintenance of Mansonella perstans in an in vitro culture system |
| title_short | Successful long-term maintenance of Mansonella perstans in an in vitro culture system |
| title_sort | successful long term maintenance of mansonella perstans in an in vitro culture system |
| topic | Mansonella perstans In vitro culture Worms L3 infective larvae Long-term maintenance |
| url | http://link.springer.com/article/10.1186/s13071-017-2515-8 |
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