Fig. 6
Protection against HIV-1 challenge in CD34+ humanized mice. CD34+ hematopoietic stem cell (HSC)- reconstituted NSG mice were treated with NMDTG according to the scheme illustrated in a. HIV-1-infected mice without treatment served as positive controls. b Blood DTG concentrations were analyzed by UPLC-MS/MS. Dotted lines indicate the PA-IC90 (64 ng/mL) and four-times the PA-IC90 (256 ng/mL). c DTG concentrations were also analyzed in spleen, GALT, lung, and liver samples. d Plasma viral load was measured three-weeks after HIV-1 challenge. e DNA and f RNA semi-nested real-time PCR was performed on spleen, GALT, lung, bone marrow, and liver. g HIV-1 RNAscope was performed on spleen and lymph node sections and scored according to amount of positive staining [0 = no staining or <1 dot/10 cells, 1 = 1–3 dots/cell, 2 = 4–9 dots/cell with no, or very few, dot clusters, 3 = 10–15 dots/cell and <10% dots are in clusters, 4 = >15 dots/cell and >10% dots are in clusters]. Anything scoring less than or equal to one was considered as background. h Representative HIV-1 RNAscope staining (brown) of spleen (top) and lymph node (bottom) sections are shown. *P < 0.05, **P < 0. 01. Results are shown as the mean ± SEM of five positive control (5 female) and 7 NMDTG-treated (5 female, 2 male) animals. Results were analyzed by two-tailed Student’s t test (n = 5 HIV-1 control, 7 MDTG; degrees of freedom = 10)
