Journal ArticleDOI
Continuous cultivation of Trypanosoma brucei blood stream forms in a medium containing a low concentration of serum protein without feeder cell layers.
Hiroyuki Hirumi,K. Hirumi +1 more
TLDR
Blood stream forms (BSF) of Trypanosoma brucei Brucei GUT were propagated in vitro in the absence of feeder layer cells at 37 C, using a modified Iscove's medium (HMI-18), and long slender BSFs increased in number from day 4 onward.Abstract:
Blood stream forms (BSF) of Trypanosoma brucei brucei GUT at 3.1 were propagated in vitro in the absence of feeder layer cells at 37 C, using a modified Iscove's medium (HMI-18). The medium was supplemented with 0.05 mM bathocuproine sulfonate, 1.5 mM L-cysteine, 1 mM hypoxanthine, 0.2 mM 2-mercaptoethanol, 1 mM sodium pyruvate. 0.16 mM thymidine, and 20% (v/v) Serum Plus (SP) (Hazleton Biologics, Lenexa, Kansas). The latter contained a low level of serum proteins (13 micrograms/ml). Each primary culture was initiated by placing 3.5-4 x 10(6) BSFs isolated from infected mice in a flask containing 5 ml of the medium (HMI-9) supplemented with 10% fetal bovine serum (FBS) and 10% SP. The cultures were maintained by replacing the medium every 24 hr for 5-7 days. During this period, many BSFs died. However, from day 4 onward, long slender BSFs increased in number. On days 5-7, trypanosome suspensions were pooled and cell debris was removed by means of diethylaminoethyl cellulose (DE52) column chromatography. Blood stream forms then were collected by centrifugation, resuspended in fresh medium at 7-9 x 10(5)/ml, and transferred to new flasks. Subcultures were maintained by readjusting the BSF density to 7-9 x 10(5)/ml every 24 hr. Concentrations of FBS were reduced gradually at 5-7-day intervals by alternating the amounts of FBS and SP in HMI-9 with 5% FBS and 15% SP, with 2% FBS and 18% SP, and finally with 20% SP (HMI-18). By this method, 2-3 x 10(6) VSFs/ml were obtained consistently every 24 hr. for more than 80 days.(ABSTRACT TRUNCATED AT 250 WORDS)read more
Citations
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Journal ArticleDOI
Anti-infective potential of natural products: how to develop a stronger in vitro 'proof-of-concept'.
TL;DR: This review provides a number of recommendations that will help to define a more sound 'proof-of-concept' for antibacterial, antifungal, antiviral and antiparasitic potential in natural products.
Journal ArticleDOI
A mechanism for cross-resistance to nifurtimox and benznidazole in trypanosomes.
TL;DR: It is reported that in trypanosomes, both drugs are activated by a NADH-dependent, mitochondrially localized, bacterial-like, type I nitroreductase (NTR), and that down-regulation of this explains how resistance may emerge.
Journal ArticleDOI
Hydrodynamic Flow-Mediated Protein Sorting on the Cell Surface of Trypanosomes
Markus Engstler,Markus Engstler,Thomas Pfohl,Stephan Herminghaus,Michael Boshart,Geert F. Wiegertjes,Niko Heddergott,Peter Overath +7 more
TL;DR: It is suggested that the hydrodynamic flow acting on swimming trypanosomes causes directional movement of Ig-VSG immune complexes in the plane of the plasma membrane, that is, immunoglobulins attached to VSG function as molecular sails.
Journal ArticleDOI
A VSG Expression Site-Associated Gene Confers Resistance to Human Serum in Trypanosoma rhodesiense
Hoang Van Xong,Luc Vanhamme,Mustapha Chamekh,Chibeka Evelyn Chimfwembe,Jan Van Den Abbeele,Annette Pays,Nestor Van Meirvenne,Raymond Hamers,Patrick De Baetselier,Etienne Pays +9 more
TL;DR: It is shown that the ETat 1.10 expression site is the one selectively transcribed in R variants, and contains SRA as an expression site-associated gene (ESAG) and is characterized by the deletion of several ESAGs.
Journal ArticleDOI
Four histone variants mark the boundaries of polycistronic transcription units in Trypanosoma brucei
T. Nicolai Siegel,Doeke R. Hekstra,Louise E. Kemp,Luisa M. Figueiredo,Joanna E. Lowell,David Fenyö,Xuning Wang,Scott Dewell,George A. M. Cross +8 more
TL;DR: It is suggested that histone modifications and histone variants play crucial roles in transcription initiation and termination in trypanosomes and that destabilization of nucleosomes by hist one variants is an evolutionarily ancient and general mechanism of transcription initiation, demonstrated in an organism in which general pol II transcription factors have been elusive.
References
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Journal ArticleDOI
Isolation of salivarian trypanosomes from man and other mammals using DEAE-cellulose.
Sheila M. Lanham,D.G. Godfrey +1 more
TL;DR: Salivarian trypanosomes were separated from infected blood by adsorbing the particulate blood components on to DEAE-cellulose columns and eluting the trypanOSomes, confirming previous observations, cyclically transmitted isolations behaving similarly to those which were mechanically transmitted.
Journal ArticleDOI
Cultivation in a semi-defined medium of animal infective forms of Trypanosoma brucei, T. equiperdum, T. evansi, T. rhodesiense and T. gambiense.
TL;DR: The cultured trypanosomes had all the characteristics of the in vivo bloodstream forms including: morphology, infectivity, antigenic variation and glucose metabolism.
Journal ArticleDOI
Hybrid formation between African trypanosomes during cyclical transmission.
L Jenni,S Marti,Jurg Schweizer,B Betschart,R. W. F. Le Page,Jerry M. Wells,Andrew Tait,Pascale Paindavoine,Etienne Pays,Maurice Steinert +9 more
TL;DR: Direct evidence of gene exchange between two different clones of trypanosomes after mixed infection and full cyclical development in the tsetse fly vector is reported.
Journal ArticleDOI
The epidemiology of Trypanosoma rhodesiense sleeping sickness in Alego Location, Central Nyanza, Kenya. I. Evidence that cattle may act as reservoir hosts of trypanosomes infective to man.
TL;DR: It is concluded that in an outbreak of sleeping sickness domestic cattle can act as natural reservoir hosts and therefore their mass treatment is strongly recommended during a control programme.
Journal ArticleDOI
Cysteine eliminates the feeder cell requirement for cultivation of Trypanosoma brucei bloodstream forms in vitro.
TL;DR: In all previous studies, bloodstream forms of Trypanosoma brucei could be grown in vitro only when supported by a feeder layer of mammalian fibroblasts and cysteine supported axenic growth if low concentrations of 2-mercaptoethanol were added at regular intervals.
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