Genetic and Virulence Characteristics of Linezolid and Pretomanid Dual Drug-Resistant Strains Induced from Mycobacterium tuberculosis in vitro

Figures & data

Figure 1 Experimental overview for generation of induced LZD and PA-824 dual drug-resistant MTB strains (LPDR mutants, marked as LP or PL strains) in vitro from M. tuberculosis H37Rv.

Table 1 Linezolid (LZD) and Pretomanid (PA-824) Mono- or Dual-Resistant Mutant Strains of M. tuberculosis

Strain	MIC(µg/mL)		rplC Mutation	rrl Mutation	ddn Mutation	fgd1 Mutation
	LZD	PA-824
H37Rv	0.25	0.07	WT	WT	WT	WT
L1	18.28	0.12	C154R	WT	WT	WT
L2	13.59	0.15	C154R	WT	WT	WT
L3	13.94	0.15	C154R	WT	WT	WT
L4	15.20	0.15	C154R	WT	WT	WT
L5	17.22	0.14	C154R	WT	WT	WT
L6	5.99	0.22	C154R	WT	WT	WT
L7	9.95	0.25	C154R	WT	WT	WT
P1	0.43	>20	WT	WT	V61G,Y89H	WT
P2	0.22	>20	WT	WT	Y89H	WT
P3	0.34	>20	WT	WT	Y89H	WT
P4	0.30	>20	WT	WT	Y89H	WT
P5	0.29	>20	WT	WT	Y89H	WT
LP1	12.37	>20	C154R	WT	88W→STOP	WT
LP2	5.37	>20	C154R	WT	WT	WT
LP3	9.58	>20	C154R	WT	WT	WT
LP4	4.91	>20	C154R	WT	G81S	WT
LP5	8.06	>20	C154R	WT	WT	57W→STOP
LP6	8.23	>20	C154R	WT	WT	WT
LP7	10.93	18.60	C154R	WT	Y89H	WT
LP8	8.69	>20	C154R	WT	WT	WT
LP9	8.51	>20	C154R	WT	WT	WT
LP10	4.92	>20	C154R	WT	WT	57W→STOP
LP11	9.88	>20	C154R	WT	WT	WT
LP12	17.04	>20	C154R	WT	WT	WT
LP13	14.55	>20	C154R	WT	88W→STOP	WT
LP14	4.88	>20	C154R	WT	WT	WT
LP15	6.93	>20	C154R	WT	WT	WT
LP16	6.01	>20	C154R	WT	WT	WT
LP17	12.98	>20	C154R	WT	88W→STOP	WT
LP18	9.09	>20	C154R	WT	I102T	WT
LP19	8.67	>20	C154R	WT	T56A	WT
PL1	6.41	>20	C154R	WT	V61G,Y89H	WT
PL2	12.78	>20	WT	WT	V61G,Y89H	WT
PL3	6.67	>20	C154R	WT	V61G,Y89H	WT
PL4	6.65	>20	C154R	WT	V61G,Y89H	WT
PL5	6.71	>20	C154R	WT	V61G,Y89H	WT
PL6	14.36	>20	C154R	WT	V61G,Y89H	R212Q
PL7	10.31	>20	C154R	WT	V61G,Y89H	WT

Abbreviation: MIC, the concentration required to inhibit the growth of 90% of the strains.

Figure 2 Growth of mutant strains through colony forming units (CFU) counts in an M. tuberculosis H37Rv background.

Figure 3 BALB/c mice survival after intravenous infection via the tail vein. Kaplan–Meier survival curves of groups of 16 mice infected with six representative mutants, or H37Rv strain. The median survival times of BALB/c mice were 27.5, undefined, 83, 63.5, 67, 68.5, and 57 days in the H37Rv, L1, L3, P1, P3, LP11 and PL7 groups, respectively.

Figure 4 Four weeks post-infection with M. tuberculosis in BALB/c mice. The body mass (A), organ mass ratio (B, C), and the total organ CFU counts (D, E) in each group. Eight mice each group were sacrificed at 4 weeks post-infection, of which body weight and weights of each lung and spleen were measured. Among them, lungs and spleens of 5 mice each group were removed for organ CFU counts. Statistically significant differences compared with infected groups are shown: *P< 0.05; **P< 0.01.