TABLE 5.

Agglutination and trypanolytic activity of the TcG1-TcG6-specific polyclonal sera

Antiseruma	% Agglutination activityb at serum dilution of:			% Trypanolytic activityc at serum dilution of:
	1:4	1:8	1:16	1:4	1:8	1:16
NMS	0	0	0	0	0	0
Y3	0	0	0	0	0	0
GP18	0	0	0	0	0	0
CMS	96	86	80	100	94	86
TcG1	90	82	68	92	68	42
TcG2	98	90	82	88	72	38
TcG3	92	85	76	42	24	0
TcG4	84	80	60	94	84	68
TcG5	86	74	68	0	0	0
TcG6	92	75	72	38	22	0

^aImmune sera were obtained from mice immunized with TcG1-TcG6-encoding plasmids. Parasites incubated with NMS, Y3 antibody to nonspecific MHC class 1 molecule, and polyclonal antibody to intracellular GPI8 protein of T. cruzi were used as controls. Parasites incubated with CMS were used as a positive control.

^bFor agglutination assay, epimastigotes (5 × 10⁴ in 25 μl of RPMI medium) were incubated for 4 h at 37°C with a 25-μl serum sample, and free, nonagglutinated epimastigotes were counted.

^cFor trypanolytic assay, trypomastigotes (5 × 10⁴ in 25 μl of RPMI medium) were incubated for 4 h at 37°C with a 25-μl serum sample plus 25 μl of human complement, and free, live trypomastigotes that did not take trypan blue stain were counted. No trypomastigote lysis was observed with heat-inactivated immune serum alone, complement alone, or heat-inactivated immune serum plus heat-inactivated complement.