Oral dysbiosis enhanced Enterococcaceae expansion and translocation. (A) Relative abundance of oral microbiota (family level) in control and OLP mice at day 0 and day 21 after allogeneic (BALB/c recipients from B10.D2 donor grafts) HCT (n = 4-5 per group). (B) PCoA by AMOVA of family compositions of oral microbiota in the control and OLP group at day 0 and day 21 after allogeneic (BALB/c recipients from B10.D2 donor grafts) HCT (n = 4-5 per group). (C) α-Diversity (Shannon index) of oral microbiota in the control and OLP group at day 0 and day 21 after allogeneic (BALB/c recipients from B10.D2 donor grafts) HCT (n = 4-5 per group, ∗P < .05; and ∗∗P < .01 determined by the Mann-Whitney U test). Data represent means ± SDs. (D) Volcano plots of DESeq2 analysis showing the amplicon sequence variants identified to the family level features of oral and fecal microbiota that are differentially abundant between the control and OLP groups on day 21 after allo-HCT (BALB/c recipients from B10.D2 donor grafts, n = 4 to 5 per group). Blue- square (control) and orange-circle (OLP) dots represent family entities that are significantly abundant in each group with log₂-fold change of >1. The black dots represent the family features whose abundance is similar between the 2 groups for which the P value is not significant or the log₂-fold change is <1. (E) Quantitative polymerase chain reaction (qPCR) results for the absolute abundance of total microbiota attached to 1 ligature and fecal samples in the control and OLP groups on days 0 and 21 after allo-HCT (BALB/c recipients from B10.D2 donor grafts). Data show the mean ± SD (n = 4-5 per group, ∗P < .05; and ∗∗P < .01, estimated by the Mann-Whitney U test). Each dot indicates an individual mouse. (F) Representative Enterococcus immunofluorescence images in the cervical LNs of BALB/c recipients from B10.D2 donor grafts at day 0 and day 14 after HCT are shown. The top row shows nuclei stained with DAPI (4′,6-diamidino-2-phenylindole). The middle row shows Enterococcus stained with AF594 (arrowheads point to Enterococcus). The bottom row shows the merger of the top and middle rows (scale bar, 30 mm). (G) The number of Enterococcus in 1 cervical LN from control and ligature mice at day 0 and day 14 after allogeneic (BALB/c recipients from B10.D2 donor grafts) HCT, respectively, are shown (n = 4-5 per group, ∗P < .05 determined by the Mann-Whitney U test). Data represent means ± SDs. Panels F and G show data representative of 3 independent experiments.