Isolation and propagation of Trypanosoma brucei gambiense from sleeping sickness patients in south Sudan

• Published in: Transactions of the Royal Society of Tropical Medicine and Hygiene Vol. 101; no. 6; pp. 540 - 546
• Main Authors: Maina, Naomi W.N., Oberle, Michael, Otieno, Charles, Kunz, Christina, Maeser, Pascal, Ndung’u, Joseph M., Brun, Reto
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01-06-2007; Royal Society of Tropical Medicine and Hygiene; Elsevier
• Subjects: Adolescent; Adult; Animals; Biological and medical sciences; Child; Child, Preschool; Cryopreservation; Female; Human protozoal diseases; Humans; Infant; Infectious diseases; Isolation; Male; Mastomys natalensis; Medical sciences; Mice; Mice, Inbred BALB C; Mice, SCID; Middle Aged; Murinae - parasitology; Parasitic diseases; Protozoal diseases; SCID mice; Sequence Analysis, DNA; Sleeping sickness; Sudan; Trypanosoma brucei gambiense; Trypanosoma brucei gambiense - genetics; Trypanosoma brucei gambiense - isolation & purification; Trypanosomiasis; Trypanosomiasis, African - blood; Trypanosomiasis, African - cerebrospinal fluid; Africa; Sudan; Sleeping sickness; Cryopreservation; SCID mice; Sudan; Isolation; Mastomys natalensis; Trypanosoma brucei gambiense; Animal model; Protozoal disease; Tropical medicine; Trypanosoma brucei; Trypanosomiasis; Kinetoplastida; Human; Protozoa; Rodentia; Parasitosis; African sickness; Infection; Vertebrata; Experimental disease; Mammalia; Mouse; Animal; Trypanosoma gambiense
• ISSN: 0035-9203; 1878-3503
• DOI: 10.1016/j.trstmh.2006.11.008

This study aimed at isolating
Trypanosoma brucei gambiense from human African trypanosomiasis (HAT) patients from south Sudan. Fifty HAT patients identified during active screening surveys were recruited, most of whom (49/50) were in second-stage disease. Blood and cerebrospinal fluid samples collected from the patients were cryopreserved using Triladyl
® as the cryomedium. The samples were stored at −150
°C in liquid nitrogen vapour in a dry shipper. Eighteen patient stabilates could be propagated in immunosuppressed
Mastomys natalensis and/or SCID mice. Parasitaemia was highest in SCID mice. Further subpassages in
M. natalensis increased the virulence of the trypanosomes and all 18 isolates recovered from
M. natalensis or SCID mice became infective to other immunosuppressed mouse breeds. A comparison of immunosuppressed
M. natalensis and Swiss White, C57/BL and BALB/c mice demonstrated that all rodent breeds were susceptible after the second subpassage and developed a parasitaemia >10
6/ml by Day 5 post infection. The highest parasitaemias were achieved in C57/BL and BALB/c mice. These results indicate that propagation of
T. b. gambiense isolates after initial isolation in immunosuppressed
M. natalensis or SCID mice can be done in a range of immunosuppressed rodents.