TABLE 4.

Additive leishmanicidal effect of disulfiram combined with divalent metal ion salts^a

Control or salt (1 μM)	HepG2 or RAW 264.7 cellsb		L. major				L. donovani
			EC50		IVTI		EC50		IVTI
	Mean CC50 (μM) ± SE	P	Mean concn (μM) ± SE	P	Mean ± SE	P	Mean concn (μM) ± SE	P	Mean ± SE	P
Ex vivo systems
Control	36.0 ± 2.6		0.058 ± 0.013		665 ± 70		0.062 ± 0.007		591 ± 35
CuSO4	7.9 ± 0.6	***	0.018 ± 0.005	NS	483 ± 67	NS	0.052 ± 0.006	NS	155 ± 9	NS
ZnCl2	21.0 ± 1.5	***	0.003 ± 0.0008	***	8,337 ± 1,169	***	0.033 ± 0.004	NS	644 ± 38	NS
ZnSO4	20.1 ± 1.5	***	0.060 ± 0.016	NS	379 ± 53	NS	0.019 ± 0.002	***	1,091 ± 65	***
MgSO4	28.6 ± 2.1	*	0.010 ± 0.003	NS	3,140 ± 440	***	0.045 ± 0.005	NS	647 ± 39	NS
MgCl2	37.0 ± 2.7	NS	0.045 ± 0.012	NS	937 ± 131	NS	0.058 ± 0.007	NS	653 ± 39	NS
AgNO3	0.74 ± 0.05	***	0.152 ± 0.042	NS	5 ± 1	NS	0.076 ± 0.009	NS	10 ± 1	NS
MnCl2	19.0 ± 1.4	***	0.167 ± 0.046	NS	128 ± 18	NS	0.166 ± 0.019	NS	116 ± 7	NS
NiSO4	20.6 ± 1.5	***	0.060 ± 0.016	NS	620 ± 87	NS	0.060 ± 0.007	NS	350 ± 21	NS
LiCl	29.9 ± 2.2	NS	0.054 ± 0.015	NS	388 ± 55	NS	0.073 ± 0.008	NS	417 ± 25	NS
In vitro system
Control	14.1 ± 0.65		0.068 ± 0.006		209 ± 9		0.066 ± 0.008		217 ± 13
ZnCl2	22.1 ± 1.02	**	0.015 ± 0.002	***	1,540 ± 97	***	0.023 ± 0.016	*	1,917 ± 536	***
ZnSO4	22.1 ± 1.02	**	0.052 ± 0.020	NS	522 ± 88	***	0.029 ± 0.010	*	920 ± 157	***

^aThe data represent mean CC₅₀ and EC₅₀ values from three different experiments, using the HepG2 or RAW 264.7 cells and ex vivo or in vitro mouse macrophage systems for each Leishmania species (see details in Materials and Methods). The CC₅₀ and EC₅₀ were determined by regression analysis using GraphPad software (Prism 5.0). P values reflect comparisons of disulfiram with divalent salts versus disulfiram alone (control). *, P < 0.05; **, P < 0.01; ***, P < 0.001; NS, not significant.

^bHepG2 cells were used for ex vivo systems, and RAW 264.7 cells were used for the in vitro mouse macrophage system.