30. 針刺麻酔に関する文献的ならびに基礎的研究(第519回千葉医学会例会 第8回佐藤外科例会)

<p>Cytokine response in lung homogenates, BALF and plasma of WT and <i>Trem-2</i>^<i>-/-</i> mice during experimental melioidosis.</p

Effect of TREM-1 deficiency on bacterial clearance, pulmonary neutrophil influx and organ damage during experimental melioidosis.

<p>WT (closed circles/black bars) and <i>Trem-1/3</i>^<i>-/-</i> mice (open circles/ white bars) were intranasally infected with 5 x 102 CFU of <i>B</i>. <i>pseudomallei</i> and sacrificed 24 and 72 h post-infection, followed by determination of bacterial loads in lung homogenate <b>(</b><i>A</i><b>),</b> BALF <b>(</b><i>B</i><b>),</b> blood <b>(</b><i>C</i><b>)</b> and liver <b>(</b><i>D</i><b>).</b> Neutrophil influx as determined by % Ly6G positive surface of lung slides was calculated for WT and <i>Trem-1/3</i>^<i>-/-</i> mice <b>(</b><i>E</i><b>).</b> Lung <b>(</b><i>F</i><b>)</b> and spleen <b>(</b><i>G</i><b>)</b> pathology was scored as described in the Methods section. Aspartate transaminase (AST; <i>H</i><b>),</b> alanine transaminase (ALT; <i>I</i>), lactate dehydrogenase (LDH; <i>J</i><b>)</b> and blood urea nitrogen (BUN; <i>K</i>) were measured as a marker for end organ damage. Data are expressed as mean ± SEM. n = 7–8 mice per group. *<i>P</i> < 0.05; **<i>P</i>< 0.01 (Mann-Whitney <i>U</i> test).</p

No effect of TREM-1 deficiency on the cellular responsiveness and phagocytosis or intracellular killing of <i>B</i>. <i>pseudomallei</i>.

<p>Whole blood (<i>A</i>), bone marrow derived macrophages (BMDM; <i>B</i>) and alveolar macrophages (AM; <i>C</i><b>)</b> of WT and <i>Trem-1/3</i>^<i>-/-</i> mice were stimulated with medium, <i>E</i>.<i>coli</i> LPS(100 ng/ml) or heat-inactivated wild type <i>B</i>. <i>pseudomallei</i> (107 CFU/ml at a MOI of 50). TNF-α levels were measured in the supernatant obtained after 20 h of stimulation. BMDM (<i>D</i>) and AM (<i>E</i>) of WT and <i>Trem-1/3</i>^<i>-/-</i> mice were incubated at 37°C with FITC labeled heat-inactivated <i>B</i>. <i>pseudomallei</i> after which time-dependent phagocytosis was determined. Data are expressed as mean ± SEM and are representative of two or three independent experiments. n = 4 or 8 (for the whole blood assay) per group. *<i>P</i>< 0.05 (Mann-Whitney <i>U</i> test).</p

Increased TREM-1 and TREM-2 expression in experimental melioidosis.

<p>TREM-1 and TREM-2 mRNA expression was determined in wild type (WT) mice prior to infection or at 24 or 72h post-infection with 5 x 102 CFU <i>B</i>.<i>pseudomallei</i> intranasally. TREM-1 mRNA expression in lung (<i>A</i>) and liver (<i>B</i>) was determined. Likewise, TREM-2 mRNA expression was measured in lung (<i>C</i>) and liver (<i>D</i>) tissue. Data are presented as fold induction compared to the mRNA expression in uninfected mice (all RNA data are normalized to GAPDH). Data are mean ± SEM, n = 4–5 mice/group. * <i>P</i>< 0.05, ** <i>P</i> < 0.01, compared to gene-expression at t = 0h (Mann-Whitney <i>U</i> test).</p

Reduced neutrophil influx in lungs of <i>Trem-2</i> ^<i>-/-</i> mice, without affecting lung pathology.

<p>Lung pathology was determined in wild-type (WT; black bars) and <i>Trem-2</i>^<i>-/-</i> mice (white bars) infected with 5 x 102 CFU <i>B</i>. <i>pseudomallei</i> at 72h post-infection as described in the Methods section (<i>A</i>). Representative lung slides of WT (<i>B</i>) and <i>Trem-2</i>^<i>-/-</i> mice (<i>C</i>) (original magnification 10x). Neutrophil influx was defined by Ly6G positivity (expressed as % of total lung surface; <i>D</i>). Representative photographs of Ly6G-immunostaining for granulocytes on lung slides of WT (<i>E</i>) and <i>Trem-2</i>^<i>-/-</i> mice (<i>F</i>) (original magnification 10x). Data are expressed as mean ± SEM, n = 5–6 mice per group per time point. * <i>P</i> < 0.05. (Mann-Whitney <i>U</i> test).</p

Survival of <i>Trem-2</i>^<i>-/-</i> mice, but not of <i>Trem-1/3</i>^<i>-/-</i> mice, is enhanced in experimental melioidosis.

<p>Survival was observed for every 4-6h, up to a maximum of 14 days after intranasal inoculation with 5 x 102 CFU <i>B</i>. <i>pseudomallei</i> in wild-type (WT; closed circles) and <i>Trem-1/3</i>^<i>-/-</i> mice (open circles; <i>A</i>). Similarly, survival of WT (closed circles) and <i>Trem-2</i>^<i>-/-</i> mice (open circles) was determined (<i>B</i>) (n = 20 per group). The <i>P</i> value indicates significance of the difference in survival between <i>Trem-2</i>^<i>-/-</i> and WT mice (Kaplan-Meier analysis, followed by a log-rank test). ns = not significant. In addition, WT (closed circles) and <i>Trem-2</i>^<i>-/-</i> mice (open circles) were infected with 5 x 102 colony forming units (CFU) of <i>B</i>. <i>pseudomallei</i> intranasally (n = 5–6 mice per group) and sacrificed 72 h post-infection, in order to determine bacterial loads in lung homogenates (<i>C</i>), broncho-alveolar lavage fluid (BALF) (<i>D</i>), whole blood (<i>E</i>), liver (<i>F</i>) and spleen (<i>G</i>). Data are expressed as mean ± SEM, n = 5-6/group. ** <i>P</i>< 0.01. BC+ denotes positive blood cultures (Mann-Whitney <i>U</i> test).</p