Papers by Shokoufeh Karimi
bioRxiv (Cold Spring Harbor Laboratory), Apr 2, 2024
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Physiological Reports, 2018
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<p>The signal stability was examined during 10 days of subculture using luciferase assay. T... more <p>The signal stability was examined during 10 days of subculture using luciferase assay. The signal intensity of the two strains was analyzed both in the presence and absence of induction peptide (P). Differences in signal intensities were analyzed by ANOVA and Tukey’s post-hoc test. Columns labelled with different letters are significantly different (p≤0.05).</p
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<p>Localization of 6475-CBRluc-mCherry, R2LC-CBRluc and wildtype strains was evaluated <... more <p>Localization of 6475-CBRluc-mCherry, R2LC-CBRluc and wildtype strains was evaluated <i>in vivo</i> and <i>ex vivo</i> using IVIS at 0, 60, 120 and 180 min after administration of a single dose of the bacteria (1x10<sup>10</sup> CFU/mouse) in 3 separate experiments. 6475-CBRluc-mCherry (n = 10), R2LC-CBRluc (n = 3), 6475-wild type (n = 3) and R2LC-wild type (n = 1) for luminescence and 6475-CBRluc-mCherry (n = 5) and 6475-wild type (n = 2) for fluorescence imaging. 6475-wildtype (<b>a, b</b>); 6475-CBRluc-mCherry 0 min (<b>c, d</b>); 60 min (<b>e, f</b>); 120 min (<b>g, h</b>); and 180 min post gavage (<b>i, j</b>); R2LC-wildtype (<b>k, l</b>); R2LC-CBRluc 60 min (<b>m, n</b>) and 180 min post gavage (<b>o, p</b>).</p
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<p>(A) Bright-field image. (<b>B</b>) Fluorescence imaging of the DAPI stained ... more <p>(A) Bright-field image. (<b>B</b>) Fluorescence imaging of the DAPI stained IPEC-J2 cells. (<b>C</b>) Fluorescence imaging of R2LC-mCherry. (<b>D</b>) Merging of the B and C images.</p
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<p>Bacterial cultures in the presence of antibiotics were induced and a fraction of cells a... more <p>Bacterial cultures in the presence of antibiotics were induced and a fraction of cells analyzed by flow cytometry after a short induction (ShI) (~4 hours) or long induction (LI) (~20 hours) period. Columns labelled with different letters are significantly different (<i>p</i>≤0.05). The error bars indicate the standard deviation of median values obtained from five independent biological replicates.</p
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<p>Bacterial strains and plasmids used in this study.</p
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<p>In 3 separate experiments for luminescence and fluorescent imaging, BALB/c male mice wer... more <p>In 3 separate experiments for luminescence and fluorescent imaging, BALB/c male mice were gavaged with fluorescent and luminescent strains: 6475-CBRluc-mCherry (n = 10), R2LC-CBRluc (n = 3) for luminescence and 6475-CBRluc-mCherry (n = 5) for fluorescence imaging. The signal intensities were compared 0 and 60 min after an intra-gastric inoculation. (<b>A</b>) Bioluminescence imaging of: (<b>a, b, c</b>) 6475-CBRluc-mCherry, 1x10<sup>5</sup> CFU/mouse; (<b>d, e, f</b>) 6475-CBRluc-mCherry, 1x10<sup>6</sup> CFU/mouse; (<b>g, h, i</b>) 6475-CBRluc-mCherry, 1x10<sup>8</sup> CFU/mouse; (<b>j, k, l</b>) 6475-CBRluc-mCherry, 1x10<sup>10</sup> CFU/mouse; (<b>m, n, o</b>) R2LC-CBRluc, 1x10<sup>8</sup> CFU/mouse; (<b>p, q, r</b>) R2LC-CBRluc, 1x10<sup>10</sup> CFU/mouse. (<b>B</b>) <i>In vivo</i> fluorescence imaging of: (<b>s, t</b>) 6475-CBRluc-mCherry, 1x10<sup>10</sup> CFU/mouse before and immediately post gavage.</p
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Physiological Reports, 2018
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<p>A. pSIP-CBRluc-mCherry. B. pSIP-CBRluc. C. pSIP-mCherry. The codon-optimized CBRluc-mChe... more <p>A. pSIP-CBRluc-mCherry. B. pSIP-CBRluc. C. pSIP-mCherry. The codon-optimized CBRluc-mCherry, CBRluc and mCherry fragments were cloned into the pSIP411 vector under control of the inducible promoter PsppQ, and the three constructs were introduced into <i>L</i>. <i>reuteri</i> 6475 and R2LC by electro-transformation. <b>D.</b> Map of the codon-optimized CBRluc::mCherry cassette under control of a constitutive promoter P11, and CBRluc and mCherry fragments prior to insertion into pSIP411.</p
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<i>Lactobacillus reuteri</i> is an inhabitant of the gastrointestinal (GI) tract of m... more <i>Lactobacillus reuteri</i> is an inhabitant of the gastrointestinal (GI) tract of mammals and birds and several strains of this species are known to be effective probiotics. The mechanisms by which <i><i>L. reuteri</i></i> confers its health-promoting effects are far from being fully understood, but protection of the mucosal barrier is thought to be important. Leaky gut is a state of abnormal intestinal permeability with implications for the pathophysiology of various gastrointestinal disorders. Enterotoxigenic<i><i> Escherichia coli </i></i>(ETEC) can invade the intestinal mucosa and induce changes in barrier function by producing enterotoxin or by direct invasion of the intestinal epithelium.Our hypothesis was that<i> L. reuteri </i>can protect the mucosal barrier, and the goal of the study was to challenge this hypothesis by monitoring the protective effect of <i>L. reuteri </i>strains on epithelial dysfunction caused by ETEC. This study examined the potential of <i><i>Lactobacillus reuteri</i></i> to attenuate the effect of ETEC strain 853/67 on mucosal permeability. Using an infection model based on the porcine intestinal cell line IPEC-J2, it was demonstrated that pre-treatment of the cells with human-derived <i>L. reuteri </i>strains (ATCC PTA 6475, DSM 17938 and 1563F) and a rat strain (R2LC) reduced the detrimental effect of ETEC in a dose-dependent manner, as monitored by permeability of FITC-dextran and transepithelial electrical resistance (TEER). Moreover, the results revealed that ETEC up-regulated pro-inflammatory cytokines IL-6 and TNFα and decreased expression of the shorter isoform of ZO-1 (187 kDa) and E-cadherin. In contrast, pre-treatment with<i> L. reuteri</i> DSM 17938 and 1563F downregulated expression of IL-6 and TNFα, and led to an increase in production of the longer isoform of ZO-1 (195 kDa) and maintained E-cadherin expression. Interestingly, expression of ZO-1 (187 kDa) was preserved only when the infected cells were pre-treated with strain 1563F. These findings demonstrate that < [...]
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Lactobacillus reuteri, a ubiquitous inhabitant of the mammalian gastrointestinal (GI) tract has k... more Lactobacillus reuteri, a ubiquitous inhabitant of the mammalian gastrointestinal (GI) tract has known health-promoting effects and various strains are commercially available as probiotics. Several probiosis mechanisms have been suggested in L. reuteri’s mode of action, but the mediators and factors involved are not well understood. This thesis examined the function of probiotics, particularly L. reuteri, in the GI tract by equipping L. reuteri ATCC PTA 6475 and R2LC with reporter genes (luminescence and fluorescence) and a mutant of strain 6475 was generated by inactivation of chaperon dnaK. Different in vitro and in vivo applications of fluorescent and luminescent strains were evaluated, and it was demonstrated that flow cytometry can be a powerful method for determination of plasmid persistence. Biophotonic imaging (BPI) enabled low doses (~1x10^5) of luminescent bacteria to be monitored in the GI tract and revealed retention of large numbers of bacteria in the stomach up to 3 hou...
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Journal for ImmunoTherapy of Cancer, 2021
BackgroundGlioblastoma (GBM) is refractory to immune checkpoint inhibitor (ICI) therapy. We sough... more BackgroundGlioblastoma (GBM) is refractory to immune checkpoint inhibitor (ICI) therapy. We sought to determine to what extent this immune evasion is due to intrinsic properties of the tumor cells versus the specialized immune context of the brain, and if it can be reversed.MethodsWe used CyTOF mass cytometry to compare the tumor immune microenvironments (TIME) of human tumors that are generally ICI-refractory (GBM and sarcoma) or ICI-responsive (renal cell carcinoma), as well as mouse models of GBM that are ICI-responsive (GL261) or ICI-refractory (SB28). We further compared SB28 tumors grown intracerebrally versus subcutaneously to determine how tumor site affects TIME and responsiveness to dual CTLA-4/PD-1 blockade. Informed by these data, we explored rational immunotherapeutic combinations.ResultsICI-sensitivity in human and mouse tumors was associated with increased T cells and dendritic cells (DCs), and fewer myeloid cells, in particular PD-L1+ tumor-associated macrophages. Th...
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PLOS ONE, 2016
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Papers by Shokoufeh Karimi