Downregulation of Blood Monocyte HLA-DR in ICU Patients Is Also Present in Bone Marrow Cells

<div><p>Background</p><p>The downregulation of blood monocyte HLA-DR expression also occurs in tissue infiltrative cells in a context of acute clinical inflammation, especially sepsis. This context favors the development of secondary infections and results from various mechanisms. Little is known about HLA-DR expression on bone marrow (BM) cells of the monocyte lineage, the source of circulating monocytes. This study analyzed the BM HLA-DR expression in ICU patients compared to BM monocytes from non-ICU patients and to blood monocytes of control healthy donors. A potential dysfunction of myeloid differentiation was investigated in a sub-population of these ICU patients to characterize the phenotype of the immature forms of monocytes and granulocytes in BM.</p><p>Methods and Findings</p><p>BM and blood were drawn from 33 ICU and 9 non-ICU patients having a BM analysis to precise the etiology of abnormal low count in blood cells. The data were compared with blood cells of 28 control donors. Flow cytometry was used for both HLA-DR expression and phenotyping of immature forms of monocytes and granulocytes. HLA-DR expression was downregulated in both blood and BM monocyte in ICU patients compared to BM of non-ICU patients and blood of control donors. Amplitude of HLA-DR downregulation was comparable in septic and non-septic ICU patients. The phenotype of immature forms of monocytes and granulocytes in BM (n = 11) did not show abnormal myeloid (monocyte + granulocyte) differentiation.</p><p>Conclusion</p><p>The downregulation of HLA-DR in BM monocyte lineage is present in ICU patients without major changes in myeloid cells. It may result from a regulation mediated by soluble and/or neuro-endocrine factors present in BM cell microenvironment.</p></div

Monocyte differentiation stages in the BM.

<p>Monocyte differentiation stages were characterized by CD45, CD34, CD33 and CD14 expression. HLA-DR and CCR2 expression were measured in each stage and expressed as the number of sites per cell. a/ example of FACs analysis for monocyte differentiation (red: myelo/monoblasts, blue: promonocytes, green: monocytes), and the HLA-DR and CCR2 expression levels in each stage (patient 31, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0164489#pone.0164489.s003" target="_blank">S1 Table</a>); b/ individual values for HLA-DR expression (n = 10); c/ individual values for CCR2 expression (n = 10 for myelo/monoblasts, n = 11 for other cell populations). Wilcoxon comparisons.</p

Number of patients and study design.

<p>Number of patients and study design.</p

Analysis of CD16 positive and CD16 negative cells in BM monocyte lineage.

<p>a/ Percentage of CD16+ cells in blasts, promonocytes and monocytes. b/ HLA-DR expression in CD16neg and CD16pos cells in blasts, promonocytes, and monocytes (number of sites /cell). c/ CCR2 expression in CD16neg and CD16pos cells in blasts, promonocytes, and monocytes (number of sites /cell). Red filled circles corresponded to data obtained in rare events populations. Individual values and median. # p < 0.05 vs blasts p < 0.05 vs blasts, $ p < 0.05 vs promonocytes, * p < 0.05 vs CD16neg, Wilcoxon test.</p

Blood and BM mHLA-DR expression in patients and controls.

<p>a/ ICU patients (black), non-ICU patients (grey) and healthy controls (white). Individual values (circles) and the median (line) in AB/C. b/ septic shock ICU patients (dark blue), septic ICU patients (middle blue), non-septic ICU patients (light blue), non-ICU patients (grey) and healthy controls (white). Significant Kruskal-Wallis test led to perform Mann Whitney comparisons. No difference in each group between blood and BM (Wilcoxon test) were observed.</p

Granulocyte differentiation stages in the BM.

<p>Monocytes differentiation stages were characterized by CD45, CD66b, CD13, and CD16 expression. CD11b and CD62L expression were measured in each stage and expressed as the number of sites per cell. a/ example of FACs analysis for granulocyte differentiation (red: promyelo-myelocytes, blue: metamyelocytes, green: PMNs), and the expression of CD11b and CD62L in each stage (patient 30, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0164489#pone.0164489.s003" target="_blank">S1 Table</a>); b/ individual values for CD11b expression (n = 10); c/ individual values for CD62L expression (n = 10). Wilcoxon comparisons.</p

Clinical characteristics of the patients as well as the main diagnosis by the hematologist using BM analysis.

<p>Clinical characteristics of the patients as well as the main diagnosis by the hematologist using BM analysis.</p

Effect of Lipophilicity of Substrates and Surfactants on the Efficiency of Micellar Hydrolysis of p-N-nitrofenylalkanoates with Alkylpyridinium Bromides

Rigorosum Thesis 2010, Vendula Běhávková Abstract The subject of this work was to verify if a homologous series of N-alkylpyridinium substances and p-nitrophenylcarbonates possess the reciprocal influence of the length of alkyl chains on the rate of hydrolytic reaction. In a series of N-alkylpyridinium substances was tested pyridinium bromide with alkyl chain lengths of C10, 12, 14, 16 and C18. These surfactants were tested with p-nitrophenyl acetate, -butyrate, -caprylate, -caprate, -laurate, -myristate, -palmitate and-stearate. The highest values throughout the group results were achieved through a combination of p-nitrophenyl-caprate and octadecylpyridinium bromide. 5x10-3 M concentration of surfactant was able to accelerate the decomposition of p-nitrophenyl-caprate 120 times and almost reached the rate constant 0.0878 sec-1 , which corresponds to the half-life is approximately 8 seconds. octadecylpyridiniumbromide and hexadecylpyridinium bromide were proved as successful micellar catalysts. The best hydrolysable substrates were medium-length alkylated

MOESM1 of Impact of fluid challenge increase in cardiac output on the relationship between systemic and cerebral hemodynamics in severe sepsis compared to brain injury and controls

Additional file 1: Table S1. Systemic and cerebral hemodynamics in patients responders-responders (RR) vs responders-non responders (RnR) in the control group. Table S2. Systemic and cerebral hemodynamics in patients responders-responders (RR) vs responders-non responders (RnR) in the brain injury group

Allogenic T cell proliferation induced by DC derived from septic patients.

<p>MLR induced by non-sorted, CD1a+, and CD1a−negative DC purified at day 3 of monocyte differentiation (immature DC, iDC) and cultured an additional 48 h in the presence of LPS at 10 ng/ml (mature DC, mDC). The percentage of proliferation of CFSE-labeled T cells was determined as described in Methods. Patients, filled bars; controls, opened bars. Numbers (n) in each group are indicated. Histograms represent mean ± SD. § p<0.05 Wilcoxon non parametric test between iDC and mDC, 2 p<0.05 Wilcoxon non parametric test between non-sorted and either CD1a+ or CD1a−negative mDC.</p