Fig 1: Myeloid-specific Pkm2 deficiency protects mice against LPS-induced ALI. (A) Left, representative H&E staining of lung tissue sections in mice with or without exposure to intratracheal LPS. Right, the histology score of H&E staining. Lung injury was scored by examining the following parameters in a double-blind manner: pulmonary edema, inflammatory infiltration, hemorrhage, atelectasis, and hyaline membrane formation. n = 5–6. For each slide, 3–5 views were analyzed. (B) IHC staining of CD11b and MPO in mouse lungs with or without LPS treatment. (C) MPO level in mouse lungs with or without LPS treatment. (D) PMNs recovered in the BALF of mice after 24 h exposure to intratracheal LPS. (E) Mouse lung W/D ratio at 0 or 24 h post-LPS treatment. (F) The IL-1ß, IL-6, and TNF-a contents in BALF were measured using ELISA analysis. Data are represented as mean ± SEM. * p < 0.05, ** p < 0.01.
Fig 2: Myeloid-specific Pkm2-deficient mice showed less PMN infiltration. (A) Degranulation of secondary and tertiary granules represented by the release of lactoferrin and MMP9 in PMN isolated from WT and Pkm2-/-, respectively. (B) ROS production in neutrophils following fMLP and PMA induction. (C) Impaired degranulation of secondary/tertiary granules in PMN isolated from Pkm2-/- mice with PMA induction. Degranulation of secondary/tertiary granules was detected by measuring the surface CD11b level. (D) Impaired fMLP-induced trans-filter migration in PMN isolated from Pkm2-/- mice as measured by the MPO optical density (OD). (E) Impaired zymosan-induced PMN peritoneal infiltration in Pkm2-/- mice. Data are represented as mean ± SEM. NS, No significance. *, p < 0.05. **, p < 0.01. ***, p < 0.001.
Fig 3: Investigation of the anti-infective efficacy of TTCT in vivo.A Survival rates of acute KPN-infected and (B) TRKP-infected mice with pneumonia treated with TIG, TCT, TTCT, or Ts-TPGS/Cap nanorods for 5 days (n = 6 mice per group). Mice with pneumonia treated with saline were used as a negative control. C WBC and D neutrophil counts in blood samples from mice with pneumonia receiving various preparations (n = 5 mice for each group). E Total cell counts and F neutrophil percentages in BALF obtained from KPN- and TRKP-infected mice with pneumonia administered different preparations (n = 3 mice for each group). G Flow cytometry results of anti-Ly-6G/Ly-6C and anti-CD11b double-stained cells obtained from the BALF of mice with pneumonia. Double-positive cells represented neutrophils in BALF. H Levels of CRP in the BALF of mice with pneumonia after different treatments (n = 6 mice for each group). I Representative KPN and TRKP bacterial colonies formed on LB agar plates from the BALF of mice with pneumonia receiving various treatments. TIG free tigecycline, TIG(L) low dose of tigecycline (15 mg/kg), TIG (H) high dose of tigecycline (45 mg/kg), TCT tigecycline loaded TPGS/Cap nanorods, TTCT tigecycline-loaded Ts-TPGS/Cap nanorods, KPN Klebsiella pneumonia, TRKP tigecycline-resistant Klebsiella pneumonia. Data are presented as the mean ± SD. Log-rank (Mantel-Cox) test was performed in A and B. One-way analysis of variance (ANOVA) with post hoc Tukey tests were performed in C, D, E, F, and H. Source data are provided as a Source Data file.
Fig 4: Proportion of a CD11b+, b CD44+, c SCA-1+, d CD29+ BMSCs, and e relative expression of Cbfa1, OC, PPAR?2, Adipsin
Fig 5: pT181-Qß vaccination attenuates neuroinflammation and reduces CD3+ circulating T cells. Microglia morphology using the pan microglia marker (iba1) was modulated following pT181-Qß vaccination in the CX a and CA3 b, the phagocytic marker (CD45) was also reduced following pT181-Qß in the CX c and CA3 d. Isolated microglia from immunized brains (one week following vaccination) revealed that the percentage of resident microglia e (CD45lo/CD11b+, green box) were not affected following immunization (quantified in f). However, the presence of T-cells (CD45hi/CD3+) g, from the population of CD45hi/CD11b– cells (red box, e) was significantly decreased following pT181-Qß immunization h. All graphs display mean ± SEM, significance values were determined with a student’s t-test (p = 0.5 *; Qß (n = 4) or pT181-Qß (n = 4) rTg4510; scale bar is 20 µm
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