Transfer of immunoglobulins through the mammary endothelium and epithelium and in the local lymph node of cows during the initial response after intramammary challenge with endotoxin-1

D in afferent (A) and efferent (B) lymph, respectively, depicting the transfer through the endothelium; and the percentage of the afferent lymph concentration that is simultaneously found in milk (C), depicting the transfer through the mammary endothelium. Each value represents the LS-mean. Samples were collected before endotoxin infusion (0 h) and at postinfusion hours (PIH) 2 and 4.<p><b>Copyright information:</b></p><p>Taken from "Transfer of immunoglobulins through the mammary endothelium and epithelium and in the local lymph node of cows during the initial response after intramammary challenge with endotoxin"</p><p>http://www.actavetscand.com/content/50/1/26</p><p>Acta Veterinaria Scandinavica 2008;50(1):26-26.</p><p>Published online 2 Jul 2008</p><p>PMCID:PMC2483282.</p><p></p

Transfer of immunoglobulins through the mammary endothelium and epithelium and in the local lymph node of cows during the initial response after intramammary challenge with endotoxin-0

<p><b>Copyright information:</b></p><p>Taken from "Transfer of immunoglobulins through the mammary endothelium and epithelium and in the local lymph node of cows during the initial response after intramammary challenge with endotoxin"</p><p>http://www.actavetscand.com/content/50/1/26</p><p>Acta Veterinaria Scandinavica 2008;50(1):26-26.</p><p>Published online 2 Jul 2008</p><p>PMCID:PMC2483282.</p><p></p

Immunogenic properties of the Rv2190c protein.

<p>N-terminal truncated Rv2190c was purified and analyzed for immunogenic properties by Dot-Blot ELISA against various sera. <b>A.</b> Immunogenic response in the human serum of an individual with latent TB (PPD-Pos.) (left panel) as opposed to a negligible response in a PPD negative human serum (right panel). <b>B.</b> Immunogenic response in serum from an <i>M. bovis</i> infected rabbit (left panel) and weak responses from sera raised against WhiB6 protein (middle panel) or pre-bleeds (right panel). i) 2ng of Rv2190c and ii) 2ng of BSA.</p

The <i>M. tuberculosis Rv2190c</i> mutant is attenuated <i>in vivo</i>.

<p><b>A. </b><i>M. tuberculosis</i> aerosol implantation in the lungs of BALB/c mice for the <i>Rv2190c</i> mutant, complement and WT (CDC1551) strains for the time-to-death experiment, determined day 1 post-infection. <b>B.</b> Survival curves for the time-to-death experiment. <b>C–D.</b> Lung (C) and spleen (D) CFU counts for BALB/c mice infected via aerosol with ∼3.4 log₁₀CFUs of WT, <i>Rv2190c</i> mutant and complement <i>M. tuberculosis</i> strains. <b>E.</b> Gross pathology of infected mouse lungs at 112 days post-infection.</p

The <i>M. tuberculosis Rv2190c</i> causes decreased lung histopathology in the mouse.

<p>Formalin-fixed lung sections from mice infected with WT, <i>Rv2190c</i> mutant and complement strains were stained with hematoxylin and eosin and visualized using light microscopy. Scale bar = 0.1 mm.</p

<i>Rv2190c</i> expression peaks at log phase and is involved in the response to cell wall disruption.

<p><b>A.</b> Fold change of <i>Rv2190c</i> expression (relative to level at the start of the growth curve) over increasing cell density in 7H9 broth in wild-type <i>M. tuberculosis</i> CDC1551, normalized to <i>sigA</i> expression. pc: post-clumping. <b>B.</b> Fold-change of <i>Rv2190c</i> expression after exposure to oxidative (cumene), disulfide (diamide) and detergent (SDS) stress conditions, relative to expression levels prior to addition of stressors and normalized to <i>sigA</i>. <b>C.</b> Survival of WT, the <i>Rv2190c</i> mutant and complement <i>M. tuberculosis</i> strains following 24 hours of lysozyme exposure. <b>D.</b> PDIMs for <i>M. tuberculosis</i> WT and Rv2190c mutant strains analyzed by thin layer chromatography using hexane∶diethyl ether∶acetic acid solvent. <b>E.</b> PDIMs analyzed using petroleum ether∶diethyl ether solvent.</p

Indole-2-carboxamide-based MmpL3 Inhibitors Show Exceptional Antitubercular Activity in an Animal Model of Tuberculosis Infection

Our team had previously identified certain indolecarboxamides that represented a new chemical scaffold that showed promising anti-TB activity at both an <i>in vitro</i> and <i>in vivo</i> level. Based on mutational analysis using bacteria found resistant to one of these indolecarboxamides, we identified the trehalose monomycolate transporter MmpL3 as the likely target of these compounds. In the present work, we now further elaborate on the SAR of these compounds, which has led in turn to the identification of a new analog, 4,6-difluoro-<i>N</i>-((1<i>R</i>,2<i>R</i>,3<i>R</i>,5<i>S</i>)-2,6,6-trimethylbicyclo­[3.1.1]­heptan-3-yl)-1<i>H</i>-indole-2-carboxamide (<b>26</b>), that shows excellent activity against drug-sensitive (MIC = 0.012 μM; SI ≥ 16000), multidrug-resistant (MDR), and extensively drug-resistant (XDR) <i>Mycobacterium tuberculosis</i> strains, has superior ADMET properties, and shows excellent activity in the TB aerosol lung infection model. Compound <b>26</b> is also shown to work in synergy with rifampin. Because of these properties, we believe that indolecarboxamide <b>26</b> is a possible candidate for advancement to human clinical trials

Intracellular cAMP levels increase within <i>M.tb.</i>-infected THP-1 human monocytic cells upon exposure to PDE-Is.

<p>The treated infected cells received 100 uM of PDE inhibitors (PDE-I 3, and 5 class) for 2 h followed by infection. UI: uninfected cells; the PDE5-I was 4-{[3′,4′-(Methylenedioxy)benzyl]amino}-6-methoxyquinazoline (MBM); and the PDE3-I was trequinsin. Results shown (mean and SD) represent two biological replicates, each with 2 technical replicates.</p

Administration of PDE-I monotherapy to <i>M.tb.</i>-infected mice shows therapeutic benefits by several parameters.

<p>(<b>A</b>) Time-to-death analysis; BALB/c mice infected with 3.6 log₁₀CFU <i>M.tb.</i> were treated daily by oral gavage (starting the day after infection) with 10 or 30 mg/kg of cilostazol (C) or sildenafil (S). Isoniazid at 1 or 25 mg/kg (INH1 or INH25 respectively) and sham (PBS) were used as positive and negative controls, respectively. (<b>B–D</b>) BALB/c mice infected with 3.1 log₁₀CFU <i>M.tb.</i> were treated daily by oral gavage (starting the day after infection) with 10 mg/kg cilostazol (C10), sildenafil (S10) or sham. At day 28 post-infection, lung CFU counts (<b>B</b>), spleen CFU counts (<b>C</b>) and body weight (<b>D</b>) were determined.</p

Interaction of PDE-Is with Rifampin.

<p>BALB/c mice were infected with 3.6 log₁₀CFU <i>M.tb.</i> and were treated daily by oral gavage (starting the day after infection) with 10 or 30 mg/kg cilostazol (C10 and C30, respectively), 10 mg/kg rifampin (R10), C10 plus R10, or C30 plus R10. Lung CFU counts were determined on days 14 and 28 post-infection.</p