Heinz Vogelsang

Procalcitonin expression in human peripheral blood mononuclear cells and its modulation by lipopolysaccharides and sepsis-related cytokines in vitro.

Procalcitonin (PCT), the precursor of calcitonin, was recently put forward as a diagnostic marker of systemic bacterial infection and sepsis. The major PCT production site in sepsis still remains unclear. Because of a certain association between increased levels of PCT and leukocyte-derived cytokines during sepsis, we assessed the possible expression of PCT in human peripheral blood mononuclear cells (PBMCs) and the modulation of PCT by lipopolysaccharides (LPS) and various sepsis-related cytokines by reverse transcriptase-polymerase chain reaction (RT-PCR) by using a novel primer set and flow cytometric analysis with intracellular staining with antibodies to the PCT components calcitonin and katacalcin. RT-PCR and flow cytometric analysis demonstrated that PBMCs express PCT both on mRNA and on protein levels. LPS and various proinflammatory cytokines (interleukin-1beta (IL-1beta), IL-6, tumor necrosis factor-alpha (TNF-alpha), IL-2) had pronounced stimulatory effects on the expression of PCT mRNA. Under identical experimental conditions the anti-inflammatory cytokine IL-10 had no effect on the expression of mRNA for PCT. Flow cytometric analysis demonstrated increased intracellular amounts of PCT components after LPS stimulation. Thus we demonstrate for the first time that PCT is expressed in PBMCs. This expression is modulated by bacterial LPS and sepsis-related cytokines. Therefore PBMCs may be among the sources of elevated PCT levels in patients with sepsis.

Outcome prediction by traditional and new markers of inflammation in patients with sepsis.

Abstract Patients (n = 242) admitted to intensive care unit for longer than 48 hours were categorised for sepsis according to American College of Chest Physicians/Society of Critical Care Medicine (ACCP/SCCM) Consensus Conference criteria. Body temperature, leukocyte count, C-reactive protein (CRP) and procalcitonin (PCT) as well as tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-8, IL-10 and HLA-DR expression on monocytes were determined. Data of one randomly choosen day per patient entered analysis. Immunologic mediators contributing significantly to outcome were determined by logistic regression analysis. Area under the curves (AUC) of receiver operating characteristic curves of clinical markers of inflammation predicting prognosis were compared with AUC of relevant immunologic mediators. TNF-α, IL-6 and HLA-DR expression on monocytes were significantly associated with outcome; the AUCs were 0.835, 0.844 and 0.761 respectively. AUCs for clinical markers were 0.878, 0.811, 0.620 and 0.614 for PCT, CRP, leukocyte count and body temperature respectively. PCT had the highest AUC compared to other clinical markers. These data indicate that PCT might be a better marker than the classic criteria of inflammation, CRP, leukocyte count, and body temperature to identify patients endangered by severe infection or sepsis.

Lactobacillus acidophilus 74-2 and Bifidobacterium animalis subsp lactis DGCC 420 modulate unspecific cellular immune response in healthy adults.

Objective:It was determined whether a combination of Lactobacillus acidophilus (L. acidophilus) 74-2 and Bifidobacterium animalis subsp lactis DGCC 420 (B. lactis 420) affect the faecal microbiota as well as immunological parameters and blood lipids in healthy adults.Design:A placebo-controlled, double-blinded, randomized crossover trial was conducted.Subjects:Twenty-six healthy volunteers (mean age 25 years) were recruited by advertising in academical buildings. All of them completed the study.Methods:After 3-week run-in period, half of the volunteers consumed 300 g/day of yoghurt supplement containing probiotic strains L. acidophilus 74-2 and B. lactis 420, and the other half received the placebo product for a period of 5 weeks. The two groups were crossed during the following 5-week period. Blood and faecal samples were collected at the end of each period. The faecal content of probiotic bacteria, faecal short-chain fatty acids (SCFA), serum lipids and plasma immune system biomarkers were evaluated.Results:Faecal proportions of L. acidophilus and of B. lactis increased significantly from 0.02 to 0.19 and 0.4 to 1.4% (P<0.05), respectively. Percentages of granulocytes and monocytes showing phagocytic activity were significantly elevated from 92 to 95% during probiotic intervention, whereas their oxidative burst activity and specific immune parameters remained unaffected. Fecal SCFA and serum cholesterol levels were not influenced by the probiotics. However, serum concentrations of triacylglyceroles decreased significantly by 11.6% (P<0.05) in the probiotic supplementation period.Conclusions:L. acidophilus and B. lactis were recovered in faeces in significantly elevated numbers after supplementation. They are able to modulate unspecific cellular immune response indicated by the increased phagocytic activity.

The immune system in healthy adults and patients with atopic dermatitis seems to be affected differently by a probiotic intervention

Background Probiotic bacteria are proposed to alleviate atopic dermatitis (AD) in infants. There are few indications about the effect of probiotics on AD in adults.

Katacalcin and calcitonin immunoreactivity in different types of leukocytes indicate intracellular procalcitonin content

PURPOSE Procalcitonin is a new marker of severity of nonviral, in particular, bacterial infections. In respect of sepsis its site of production remains unknown. This study was carried out to determine whether subsets of human leukocytes contain procalcitonin. MATERIALS AND METHODS Blood samples of 17 patients who had demonstrated various degrees of serum procalcitonin levels on the day before evaluation were analyzed for serum procalcitonin by immunoluminometry and for intracellular reaction of monocytes, granulocytes, B-, and T-lymphocytes against katacalcin- or calcitonin-sensitive antibodies. Katacalcin and calcitonin are part of the procalcitonin molecule. Associations of these reactions with serum procalcitonin levels as well as differences between groups with a normal or elevated serum level were analyzed. RESULTS Intracellular antibody reaction against katacalcin was demonstrated in all cell types. We also found a lower rate of intracellular antibody reaction against calcitonin. Associations of serum procalcitonin with the two antibody reactions were demonstrated. Differences in intracellular reactions in the group with elevated serum procalcitonin were seen with both antibodies compared with a normal control. CONCLUSION Intracellular antibody reaction against katacalcin supports the notion that various types of leukocytes contain procalcitonin.

Influence of prebiotics and antioxidants in bread on the immune system, antioxidative status and antioxidative capacity in male smokers and non-smokers

Interest in functional foods is increasing. The aim of the present study was to investigate breads supplemented with functional components. One was bread supplemented with inulin, linseed and soya fibre (prebiotic bread). The other was a prebiotic antioxidant bread (pre-aox-bread), which additionally contained green tea powder, herbs and tomato paste. The effects of these two breads on immunological and antioxidative parameters were compared with control bread (placebo). Twenty smokers and eighteen non-smokers were enrolled in the randomised parallel study, which consisted of a control period and an intervention period, each lasting for 5 weeks. Daily intake of bread and nutrients did not differ between the intervention and the control period. Most of the twenty-three investigated immunological parameters measured in peripheral blood were unaffected. However, the percentage of CD19 increased after intervention with prebiotic bread, whereas intercellular adhesion molecule-1 (ICAM-1) and CD3+NK+ (P < 0.05) decreased in both intervention arms. The ferric reducing ability of plasma (FRAP) was increased after consumption of the pre-aox-bread for non-smokers (1256 v. 1147 micromol/l; P = 0.019) and remained unchanged for smokers consuming the pre-aox-bread. All analysed carotenoids (P <or= 0.001) in plasma were increased after the consumption of pre-aox-bread. The concentrations of uric acid and alpha-tocopherol rose after intervention with both breads. ICAM-1 as a marker of stress decreased after consuming the prebiotic bread. In conclusion, increased plasma concentrations of carotenoids and the responses observed with the FRAP assay after intervention with the pre-aox-bread indicate a unique response in terms of antioxidative potentials for this type of functional food.

Dietary supplementation with trans-11- and trans-12-18 : 1 increases cis-9, trans-11-conjugated linoleic acid in human immune cells, but without effects on biomarkers of immune function and inflammation.

Trans-fatty acid intake is associated with an increased risk of CHD and diabetes. The effects of single trans-fatty acid isomers are largely unexplored. The present study examined the effects of a 6-week supplementation with two trans-18 : 1 isomers (trans-11 and trans-12) in human subjects on immune cells, several inflammatory and immunological biomarkers (for example, IL, TNFalpha, C-reactive protein, adiponectin, intercellular adhesion molecule-1, prostacyclin, phagocytic process). Following a 2-week adaptation period without supplements, the test group (n 12) received vaccenic acid (trans-11-18:1) and trans-12-18 : 1 in equal amounts (6.0 g/d) for 6 weeks. The control group (n 12) consumed an oil without trans-fatty acids and conjugated linoleic acids (CLA). Samples were collected at the end of both periods. Trans-11- and trans-12-18 : 1 were significantly increased in cellular lipids. The endogenous synthesis of cis-9, trans-11-CLA from trans-11-18 : 1 was demonstrated via increased CLA in cellular lipids of the test group. Generally, trans-isomer supplementation did not affect either inflammatory biomarkers (for example, IL-6, IL-8, TNFalpha) or immune function (for example, phagocytosis) during the present study. The dietary supplementation of trans-11- and trans-12-18 : 1 (6 g/d) and their accumulation in leucocytes had no effects on biomarkers of inflammation and immune function. However, because of the limited data on the safety of trans-fatty acid intake and effects of individual trans isomers on human health (for example, trans-9-18 : 1, trans-10-18 : 1) at present, it is prudent to reduce trans-fat intake in general.

Flow Cytometry as a Diagnostic Tool for Hereditary Spherocytosis

Flow cytometric analysis of eosin-5′-maleimide-labeled red blood cells has been proposed as a new method of identifying hereditary spherocytosis (HS). The aim of the present study was to analyze sensitivity and specificity of this method. Red blood cells from patients with HS (n = 58) revealed significantly lower mean channel fluorescence values than red blood cells from normal subjects (n = 110), unaffected HS family members (n = 8), and patients with other anemias (n = 44). Taking a mean channel fluorescence of 400.0 units as the threshold value identified by logistic regression, sensitivity and specificity of the test for HS were 96.6 and 99.1%, respectively. Flow cytometric analysis is a valuable screening test for the diagnosis of HS.

Dietetic effects of oral intervention with mare's milk on the Severity Scoring of Atopic Dermatitis, on faecal microbiota and on immunological parameters in patients with atopic dermatitis

In a double-blind, placebo-controlled crossover trial, 23 patients consumed 250 ml mares milk or placebo for 16 weeks. The aim was to examine the effects of mares milk on the characteristics of atopic dermatitis (AD), on faecal microbiota and on clinical and immunological parameters. The intensity of AD was examined using the Severity Scoring of Atopic Dermatitis (SCORAD) index. During the mares milk period, the mean SCORAD value of patients (n=23; 17 females, 6 males) decreased from 30.1 to 25.3 after 12 weeks (P<0.05) and to 26.7 after 16 weeks (P<0.1). In a subgroup (n=7) the SCORAD index and especially the pruritus decreased by 30% through the mares milk period (P<0.01). In this subgroup, the faecal bifidobacteria increased during the mares milk period from 4.6% to 11.9% of eubacteria (P<0.05). The immunological parameters, except C-reactive protein, were unchanged.

ACCP/SCCM Consensus Conference definitions correlate better with procalcitonin than tumor-necrosis-factor-α and interleukin-6

ConclusionIn contrary to TNF-α and IL-6, PCT differs in all stages of systemic inflammation proportional to severity in accordance with the consensus conference definitions. PCT values are helpful in newly and severely infected patients, primarily of bacterial cause. After endotoxin injection PCT increases shortly after TNF-α and before IL-6 (1). Hence PCT might serve as an early and dependable indicator in patients suffering from severe infection/systemic inflammation. The results have yet to be confirmed by greater sample sizes.