IN-SITU IMMUNOASSAY FOR THE ASSESSMENT OF TRYPANOSOMA-CRUZI INTERIORIZATION AND GROWTH IN CULTURED-CELLS

Interiorization and multiplication of Trypanosoma cruzi within its hos t cells are usually assessed by counting parasites in fixed and staine d cover slip preparations, a subjective and time-consuming method. Her e we describe an immunoenzymatic assay (ELISA) for assessing the numbe r of internalized parasites in infected LLC-MK(2) seed on chamber slid es (NUNC). ELISA was performed employing a rabbit polyclonal serum aga inst trypomastigote components (MOP) and anti-rabbit IgG conjugated to peroxidase. The bottom of the chamber slide was then detached and pro cessed for quantification of internalized parasites by the conventiona l method. Data analysis showed a linear relationship between optical d ensities and number of internalized parasites (r(2) = 93.99, p < 0.001 ). The assay was also efficient to assess inhibition of parasite inter iorization induced by the monosaccharide NAc-D-glucosamine.