cell-based assay (cba  (EUROIMMUN)

Journal: Infection and Immunity

Article Title: Aspartate aminotransferase is required for Salmonella expansion in the inflamed gut via TCA anaplerosis

doi: 10.1128/iai.00161-26

Figure Lengend Snippet: S . Tm does not require AspC for systemic survival following intraperitoneal injection. CBA/J mice were infected with a competitive 1:1 ratio of WT and aspC -deficient S . Tm IR715 (Δ aspC ), via intraperitoneal injection at a dose of 10 4 CFU. Mice were allowed to carry the pathogen for predetermined time points prior to humane euthanasia and sample collection. ( A ) Infection schematic. ( B ) Weight-loss percentage relative to inoculation weight in mice sacrificed 3, 4, and 5 days post-infection. ( C ) Competitive index of inoculum at each time point based on the proportion of S . Tm in liver and spleen homogenates, as well as intraperitoneal lavage fluid. ( D ) S . Tm burden of each genotype in the liver. ( E ) S . Tm burden of each genotype in CFU/g in spleen. ( F ) S . Tm burden of each genotype in the intraperitoneal fluid following lavage. N = 6 per time point. Geometric mean with geometric SD. ns, not significant; ***, P < 0.001 using multiple t -test.

Article Snippet: Seven-week-old CBA/J mice were purchased from Jackson Labs Inc., infected with 10 4 CFU of each strain, and carried for 96 h post-infection before sacrifice.

Techniques: Injection, Infection

Journal: Infection and Immunity

Article Title: Aspartate aminotransferase is required for Salmonella expansion in the inflamed gut via TCA anaplerosis

doi: 10.1128/iai.00161-26

Figure Lengend Snippet: Deletion of aspC results in a significant expansion defect only in the cecum and colon during a murine model of gastroenteritis. To interrogate the necessity of AspC via the canonical fecal-oral route of infection, we gavaged CBA/J mice with a 1:1 competitive ratio of our WT S . Tm and isogenic Δ aspC mutant. ( A ) Schematic for competitive in vivo infection. ( B ) Competition index of inoculum over time in feces. ( C and D ) Competition index of inoculum 10 d.p.i. from systemic and gut lumen samples of infected mice. To corroborate our observations, we repeated our infection with either of our strains in isolation. ( E ) Schematic for single infection. ( F ) S . Tm burden in feces over time in mice infected with either WT S . Tm or Δ aspC mutant. N = 6 per condition. Geometric mean and geometric SD. *, P < 0.05; **, P < 0.01 using Mann-Whitney ( B ), paired-end ( E ), or unpaired t -test ( F ).

Article Snippet: Seven-week-old CBA/J mice were purchased from Jackson Labs Inc., infected with 10 4 CFU of each strain, and carried for 96 h post-infection before sacrifice.

Techniques: Infection, Mutagenesis, In Vivo, Isolation, MANN-WHITNEY

Journal: Infection and Immunity

Article Title: Aspartate aminotransferase is required for Salmonella expansion in the inflamed gut via TCA anaplerosis

doi: 10.1128/iai.00161-26

Figure Lengend Snippet: Genetic inactivation of T3SSs results in a colonization defect of Δ aspC but does not alter the overall expansion defect. We constructed isogenic S . Tm Δ aspC mutants in an avirulent background (Δ invA Δ spiB ) and repeated the competitive infection experiment in CBA/J mice, comparing the avirulent strains to the S . Tm WT background. ( A ) Infection schematic of competitive infection. ( B ) Competitive index of inoculum in feces over the course of the infection. Day 3 samples were not collected. ( C and D ) Competitive index of inoculum ( C ) and CFU counts ( D ) of S . Tm strains in cecal content at 10 d.p.i. ( E and F ) Competitive index of inoculum ( E ) and CFU counts ( F ) of S . Tm strains in colon content at 10 d.p.i. N = 6. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 using unpaired ( B ) or paired t -test ( E, F ).

Article Snippet: Seven-week-old CBA/J mice were purchased from Jackson Labs Inc., infected with 10 4 CFU of each strain, and carried for 96 h post-infection before sacrifice.

Techniques: Construct, Infection