RV: Lectin agglutination of promastigotes

[Fabio Aguiar Alves - Imunol.]

On Thu, 10 Jul 1997, Javier Moreno wrote:

DAER DR. JEAN, I'VE BEEN WORKING WITH METACYCLIC FORMS OF L.AMAZONENSIS,
AND WE'VE ALREADY DONE SOMETHING LIKE THAT USING COMPLEMENT LYSIS TEST.
THE FORMS ARE STILL AVAILABLE AFTER TREATMENT AND WE CAN USE THEM TO ANY
TEST. IF YOU'RE INTERESTED, I CAN SEND YOU THE STEPS OF DOING THAT. > 
> We are trying to separate procyclic from metacyclic promastigotes of L. infantum by agglutination of the procyclics with PNA (SIGMA). The promastigotes are from a young culture (3rd subculture) growing in RPMI. Till now lectin concentrations of up to 500 microgr./ml (in PBS) have had no visible effect (the promastigotes (10 millions/ml) were previously washed 4 x in PBS and the incubation lasted 1h at 23ºC.) What's wrong?
> If anybody has experience in handling lectins and would be so nice to help me I would be VERY grateful. If somebody has a better idea how to separate the pro- from the meta-cyclic promastigotes then don't hesitate to send me a message. Thanks a lot,
> 		
GOOD LUCK.

			
FABIO AGUIAR ALVES
FIOCRUZ-R.J- BRASIL.