Trypanosoma cruzi: advantages of isolating bloodstream trypomastigotes by the carboxy methyl cellulose method.
Bloodstream trypomastigotes were isolated from blood of A/Sn mice 7 d after infection with 10(5) Trypanosoma cruzi Y strain. Red blood cells were removed by centrifugation and hypotonic shock and platelets and leucocytes by passage through a carboxy methyl cellulose column. Binding of trypomastigotes to the resin was prevented by including 10% normal mouse serum in the eluting buffer. In such conditions, more than 90% of the parasites applied to the column were recovered, free of white blood cells and platelets. A comparative study of the pre- and post-separation trypomastigotes showed that both had the same infecting capacity, ability to evade destruction by the complement system, and antigenic profile.
da Silva
A M
AM
PAHO/WHO Immunology Research and Training Centre, Butantan Institute, São Paulo, Brazil.
Costa
H H
HH
Takehara
H A
HA
Mota
I
I
D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis; D002266:Carboxymethylcellulose Sodium; D014355:Chagas Disease / Q000097:blood* / Q000469:parasitology; D003165:Complement System Proteins; D008297:Male; D008722:Methods; D051379:Mice; D014349:Trypanosoma cruzi / Q000276:immunology / Q000302:isolation & purification* / Q000472:pathogenicity
D000953:Antigens, Protozoan; D003165:Complement System Proteins; D002266:Carboxymethylcellulose Sodium