Journal: Infection and Immunity

Article Title: Immunogenicity and plasmid delivery pathways of non-invasive Lactococcus lactis -vectored mucosal DNA vaccination

doi: 10.1128/iai.00460-25

Figure Lengend Snippet: Plasmid DNA delivery from LL to different cell types. ( A ) Luciferase activity in RAW264.7, THP-1, CMT-93, and ARPE-19 cells co-cultured with LL/pLEC-Nanoluc at the indicated cell-to-bacteria ratios for 20 h. Data are mean ± SEM ( n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns, not significant by one-way ANOVA with Dunnett’s test (vs ratio 0 for each cell line). ( B ) Fluorescent and phase-contrast images of cells co-cultured with CFSE-labeled LL for 4 h. ( C ) Schematic of time-course experiments. RAW264.7 cells were co-cultured with LL/pLEC-Nanoluc or CFSE-labeled LL; after 2 h, medium was replaced with gentamicin-containing medium, and luciferase activity was measured at the indicated times. ( D ) Time course of luciferase activity in RAW264.7 cells (mean ± SEM, n = 5/time point; the symbols hide some error bars). ( E ) Representative percentage of EGFP-expressing RAW264.7 cells after 20 h co-culture with LL/pLEC-EGFP (mean ± SEM, n = 4). *** P < 0.001 by Student’s t -test. ( F ) Luciferase activity in adhered splenocytes co-cultured with LL/pLEC-Nanoluc for 24 h (mean ± SEM, n = 5). ** P < 0.01 by Mann-Whitney U-test. Cell-to-bacteria ratio was 1:1,000 for panels B, D, and E and 1:10 for panel F.

Article Snippet: The murine colonic epithelial cell line CMT-93 (ATCC) was cultured in low-glucose Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS at 37°C in 5% CO 2 .

Techniques: Plasmid Preparation, Luciferase, Activity Assay, Cell Culture, Bacteria, Labeling, Expressing, Co-Culture Assay, MANN-WHITNEY

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: Equal inoculums of CM TS1 , M7, or M8 were used to infect CMT93, C57, and McCoy cells, and IFU were counted at 24 hpi. The ratio of IFUs that the strains formed in (A) CMT93 versus McCoy cells and in (B) C57 versus McCoy cells were compared. Significance was determined by ordinary one-way ANOVA with Dunnett’s multiple comparisons posttest. Error bars represent standard deviation. **, P < 0.01; ****, P < 0.0001.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Standard Deviation

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: Equal inocula of CM TS1 , M7, or M8 were used to infect (A) CMT93 and (B) C57 cells + / − IFNγ, and IFU were counted at 24 hpi. IFUs that the strains formed in the cell lines + / − IFNγ were compared. Significance was determined by ordinary one-way ANOVA with Dunnett’s multiple comparison posttest. Error bars represent standard deviation. ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Comparison, Standard Deviation

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: M8, various recombinants, CM TS1 , or CM tsp were used to infect McCoy, ( A ) CMT93, and ( B ) C57 cells at MOIs of 1.0, and inclusions were counted at 24 hpi. Ratios of IFUs that the strains formed in CMT93 or C57 cells divided by the IFUs formed in McCoy cells are shown on the y-axis. Significance was determined by ordinary one-way ANOVA with Tukey’s multiple comparison test. Shared letters indicate groups with p-values greater than 0.05. Groups that do not share a letter have p-values less than 0.05. Error bars represent standard deviation.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Comparison, Standard Deviation

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: McCoy cells were infected with M8R4-pTC0237-FLAG at an MOI of 1.0 in +/ − aTc and the infected cells were lysed 24 hours later. The lysates, M8, M8R4 237 mut , M8R24 237 wt , CM TS1 , and CM tsp were used to infect fresh McCoy, (A) CMT93, or (B) C57 cells, and IFUs were determined at 24 hpi. (C) McCoy, CMT93, or C57 cells were infected with M8R4-pTC0237-FLAG at MOIs of 0.1, fresh medium + / − aTc was added at 2 hpi, and IFUs were determined at 24 hpi. The number of IFUs formed in each condition is on the y-axis and each dot represents the result of a technical replicate. Significance was determined by ( A-B ) ordinary one-way ANOVA with Tukey’s multiple comparison test or ( C ) unpaired t-test. Shared letters indicate groups with p-values greater than 0.05. Groups that do not share a letter have p-values less than 0.05. Error bars represent standard deviation. ns = not significant.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Infection, Comparison, Standard Deviation

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: McCoy (black circles) and CMT93 (white squares) cells were infected with Cmu wt , M8, M8R4 237mut , M8R24 237wt , and M8R4-pTC237-FLAG + / − aTc. The number of IFUs produced by these infections was compared to the input inoculum of the primary infection to determine the relative burst size (y-axis) at various hpi (x-axis). Relative burst sizes of infections in McCoy and CMT93 cells were compared by ordinary two-way ANOVA with Bonferroni correction. Graph shows the results from two independent trials, in technical triplicates. Error bars represent standard deviation. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Infection, Produced, Standard Deviation

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: McCoy (black circles) and CMT93 (white squares) cells were infected with Cmu wt , M8, M8R4 237mut , M8R24 237wt , and M8R4-pTC237-FLAG + / − aTc without centrifugation. The number of IFUs produced was compared to the input IFUs of the primary infection to determine the relative burst size (y-axis) at various hpi (x-axis). Relative burst sizes of infections in McCoy and CMT93 cells were compared by ordinary two-way ANOVA with Bonferroni correction. The graphs show the results from two independent experiment performed in technical triplicates. Error bars indicate standard deviation. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Infection, Centrifugation, Produced, Standard Deviation