Fig. 3: Bacteriophages for therapy of M. chelonae.
From: Bacteriophage treatment of disseminated cutaneous Mycobacterium chelonae infection
A Ten-fold serial dilutions of each phage (left) were spotted onto top agar overlays of M. smegmatis mc2155 and M. chelonae GD153. Phages: 1, BPs∆33HTH_HRM10; 2, BPs∆33HTH_HRMGD03; 3, Muddy; 4, Muddy_HRMGD04; 5, Itos; 6, Adephagia∆41∆43; 7, ZoeJ∆45; 8, Fionnbharth∆45∆47; 9, Elmo_HRMmc2155; 10, Isca_cpm; 11, Fred313cpm_∆33; 12, CrimD∆41-43; 13- BPs∆33HTH_HRM10_REM1; 14, Faith1∆38-40; 15, Faith1∆38-40_HRMGD69; 16, MissWhite; 17, MissWhite-D29_Hybrid1, 18, Maco6; 19, TM4; 20, Wildcat; 21, D29; and 22, D29_HRMGD40. B As for panel A but using M. abscessus lytically induced prophages plated on M. chelonae GD153 and control strain M. abscessus GD40. Phages: 1, phiGD20-1; 2, phiGD22-1; 3, phiGD23-1; 4, phiGD21-1; 5, phiGD34-2; 6, phiGD89-1; 7, phiGD57-1; 8, phiGD17-1; 9, phiGD24-3. C Killing of M. chelonae GD153 by Muddy following infection in liquid culture (moi, 10); -Muddy, no phage control. D Efficient killing of M. chelonae GD153 over ranges of bacterial and phage concentrations. All rows contain 10-fold serial dilutions of bacterial culture. Top row (–), no phage control; rows 2-9, 10-fold serial diltions, with 1010 PFU in row 2. E ELISA responses for anti-Muddy IgG antibodies in sera from 3 days, 17 days or 16 weeks after phage treatment initiation. Negative controls (ctrl) for uncoated wells are also shown. n = 2 technical replicates are shown. Data points are the average of two independent sets of serum dilutions and measurements, with error bars ± one standard deviation. F Half-maximal IgG titers derived from ELISA curve fits in panel D. Duplicate measurements are shown as points; bar height is the mean titer with error bars±one standard deviation. Source data for panels E and F are provided as a Source Data file. G To test for antibody neutralization, Muddy was incubated with either plasma (p) or serum (s) from 2 days, 3 days, 17 days or 16 weeks after the start of phage administration for 2 h (top) or 24 h (bottom) and then 10-fold serial dilutions were spotted onto top agar overlays of M. smegmatis mc2155. Plates were incubated at 37 °C for 48 h.
