Tomasz Skirecki

Sepsis immunopathology: perspectives of monitoring and modulation of the immune disturbances

Septic syndromes are the main cause of death in the intensive care units and although the mortality rates is slowly decreasing, the occurrence of the disease has been increasing. The pathogenesis of sepsis includes countless disturbances of the host immune system starting with a harmful, infection-triggered exaggerated inflammatory cascade, followed by the development of an immunoparalysis state. The latter contributes to the failure in pathogen eradication and leads to secondary infections, which are often the cause of fatal complications. In this review, we consider different novel therapeutic strategies for restoration of immune function. The use of glucocorticoids, intravenous immunoglobulins, heparin, recombinant human activated protein C, granulocyte macrophage colony-stimulating factor, granulocyte colony-stimulating factor, interferon-γ, statins, macrolides and high-volume hemofiltration are discussed. Even though some clinical trials of these regimens are promising, the key to their successful application seems to be the precise monitoring of the status of immune system followed by implementation of the adequate therapy. Thus, in this paper we present disturbances in the immune system in the course of human sepsis, with special attention to the parameters that could be monitored and serve as markers for immunomodulatory therapies. We conclude by briefly presenting the current sepsis treatment strategy.

Bronchoalveolar lavage total cell count in interstitial lung diseases--does it matter?

Bronchoalveolar lavage (BAL) is a useful technique for differential diagnosis of various interstitial lung diseases (ILDs) and is usually realized by analysis of the differential cell count. This study was conducted to estimate the value of bronchoalveolar lavage fluid (BALF) total cell count (TCC) in the diagnosis of ILD. We analyzed 237 BAL samples from patients with ILD: sarcoidosis (SA), idiopathic pulmonary fibrosis (IPF), cryptogenic organizing pneumonia (COP), hypersensitivity pneumonitis (HP), chronic eosinophilic pneumonia (CEP), and smoking-related ILD (sr-ILD). The control group consisted of 30 healthy volunteers. The statistical analysis revealed significant differences in the BALF TCC between healthy controls and patients with SA, IPF, HP, COP, sr-ILD, and eosinophilic disorders (mean values 6.9 vs. 14.5, 22.5, 22.8, 20.7, 64.5, and 27.3 × 106, respectively). Logistic regression revealed a significant relation between the TCC and ILD diagnosis. We conclude that the TCC, as well as the value of total number of inflammatory cells, should be reported in the description of BAL.

T, B, and NKT Cells in Systemic Inflammation in Obstructive Sleep Apnoea

Background. Obstructive sleep apnoea syndrome (OSAS) brings risk of serious complications. The study objective was to assess elements of the cellular immune response in the course of OSAS. Methods. Peripheral blood (PB) lymphocytes: T, B, NK, NKT-like, Th, Tc, and HLA DR+ T cells were evaluated by flow cytometry of 48 OSA patients; the concentration of adiponectin, interleukin 1β, and TNFα was measured by ELISA method. The OSA complication score was developed and used for statistical analysis. Results. The proportion of B cells and Th/Tc ratio were significantly lower in the BP of OSA patients when compared with control subjects (median 7.9 versus 10.9%, 0.9 versus 1.5, p < 0.05). The proportion of Tc, NK, NKT-like, and HLADR positive T cells were elevated in OSA patients when compared with healthy subjects (36.4 versus 26.8, 15.5 versus 8.5, 5.7 versus 3.0, and 8.4 versus 4.5%, p < 0.05, resp.) and were more pronounced in patients with metabolic syndrome. The grade of OSA complication score correlated with systemic inflammation markers and the proportion of B cells. The value of adiponectin/BMI ratio correlated significantly with SpO2 (r = 0.31, p < 0.05), CRP (r = −0.35, p < 0.05), TNFα concentration (r = −0.36, p < 0.05), and proportion of B cells (r = 0.32, p < 0.05). Conclusion. Lymphocytes B, Tc, NK, NKT-like, and adiponectin are involved in systemic immune response in OSA patients possibly predisposing them to cardiovascular and metabolic complications.

Absolute counts of peripheral blood leukocyte subpopulations in intraabdominal sepsis and pneumonia-derived sepsis: a pilot study

The leading pathophysiological changes during sepsis include systemic abnormalities in the immune response. Due to the general character of these disturbances, sepsis is usually studied as a homogenous clinical condition. We aimed to compare the immune response in intraabdominal sepsis (IAS) and pneumonia-derived sepsis (PDS). The following cell populations were examined: white blood cell count (WBC), monocytes, lymphocytes: CD3+, CD4+ and CD8+ T cells, B cells, and NK cells. In both studied groups (i.e. IAS and PDS), the WBC was elevated. However, it was significantly higher in the IAS group than in the PDS group. The difference was due to a lower granulocyte count, as well as a lower monocyte count in PDS. We found no significant correlation between the total lymphocyte number and CD3+CD8+ T cells in either form of sepsis. Similarly, we observed no correlation between the total lymphocyte number and the NK cells subset in IAS. However, the numbers of CD3+CD8+ and NK cells correlated similarly in both types of sepsis. Both studied types of sepsis induced profound lymphocytopenia, with marked loss of CD8+ T cells and the NK cells. However, the similar relation between them, which was independent of the infection type, suggests that the NK and CD3+CD8+ cells have shared mechanisms of regulation. The primary site of infection has an impact on the global immune reaction. These alternations include especially myeloid cells: granulocytes and monocytes which disappear from peripheral blood during PDS, but increase in IAS.

Monitoring of procalcitonin but not interleukin-6 is useful for the early prediction of anastomotic leakage after colorectal surgery.

Abstract Background: Early recognition of patients who have developed anastomotic leakage (AL) after colorectal surgery is crucial for the successful treatment of this complication. The aim of this study was to assess the usefulness of the assessment of procalcitonin (PCT) and interleukin-6 (IL-6) levels in the prognosis of AL. Methods: This observational study included 157 patients who underwent major elective colorectal surgery. The most common indications for surgery were cancer and inflammatory bowel diseases. Serum samples were obtained directly before surgery (D0) and 1 day (D1) after surgery, and the relationships between the serum concentrations of PCT and IL-6 and development of AL were assessed. Results: In total, 10.2% of patients developed post-surgical infections due to AL. PCT levels on D1 were significantly higher in patients who developed AL [2.73 (1.40–4.62)] than in those who recovered without complications [0.2 (0.09–0.44)]. The area under the ROC curve for PCT on D1 was 0.94, 95% CI (0.89–0.98). The sensitivity and specificity of the prediction of an infection were 87% and 87%, respectively, for PCT on D1, which was higher than 1.09 ng/mL. The increase in PCT concentration between D0 and D1 was significantly higher in patients with AL (p<0.001). Patients who developed AL had higher concentrations of IL-6 on D1, but the difference was not significant (p=0.28). Conclusions: This study confirms that surgical trauma increases serum PCT concentrations and that the concentration of PCT on D1 can predict AL after colorectal surgery. However, IL-6 is not a good early marker for developing AL.

Fas-positive lymphocytes are associated with systemic inflammation in obstructive sleep apnea syndrome

PurposeObstructive sleep apnea syndrome (OSAS) is associated with alterations in immune system which may lead to serious complications. The aim of this study was to explore lymphocyte populations in OSAS with special attention to the Fas-positive cells.MethodsFifty-one patients with confirmed OSA and 20 healthy subjects were investigated. The OSA severity indices, data concerning comorbidities, and markers of inflammation and metabolic disorders were collected. Flow cytometry was used to analyze the lymphocyte profile and expression of Fas receptors (CD95). Concentration of adiponectin, IL-1β, TNF-α, and sFas were measured.ResultsProportions of Fas-positive cells in the pool of CD4+ and Fas-positive in the pool of CD8+ cells in the blood of patients were significantly increased when compared with healthy subjects (74.5% vs. 65.6% and 78.8% vs.70.9%, respectively, p < 0.05). No correlation with OSA severity was found. However, the proportion and number of Fas+ cells were elevated in obese patients, in non-smokers, and in patients suffering from COPD and hypertension. There were several significant relations of Fas+ cells with inflammatory markers of systemic inflammation.ConclusionLymphocytes with the expression of Fas receptor are associated with systemic inflammation in OSAS.

The Early Expression of HLA-DR and CD64 Myeloid Markers Is Specifically Compartmentalized in the Blood and Lungs of Patients with Septic Shock

Identification of reliable biomarkers is key to guide targeted therapies in septic patients. Expression monitoring of monocyte HLA-DR and neutrophil CD64 could fulfill the above need. However, it is unknown whether their expression on circulating cells reflects the status of tissue resident cells. We compared expressions of HLA-DR and CD64 markers in the circulation and airways of septic shock patients and evaluated their outcome prognostic value. The expression of CD64 on neutrophils and HLA-DR on monocytes was analyzed in the peripheral blood and mini-bronchoalveolar lavage fluid cells by flow cytometry. Twenty-seven patients with septic shock were enrolled into the study. The fluorescence intensity of HLA-DR on circulating monocytes was 3.5-fold lower than on the pulmonary monocytes (p = 0.01). The expression of CD64 on circulating and airway neutrophils was similar (p = 0.47). Only the expression of CD64 on circulating neutrophils was higher in nonsurvivors versus survivors (2.8-fold; p = 0.031). Pulmonary monocytes display a higher level of HLA-DR activation compared to peripheral blood monocytes but the expression of neutrophil CD64 is similar on lung and circulating cells. Death in septic patients was effectively predicted by neutrophil CD64 but not monocytic HLA-DR. Prognostic value of cellular activation markers in septic shock appears to strongly depend on their level of compartmentalization.

Stem Cells in Infection and Sepsis

Severe infections, sepsis, and septic shock remain a major cause of death in the critically ill patients. Activation of systemic response to infection and disruption of most regulatory axes in the body lead to tissue injury and the development of organ failure. Moreover, reprogramming of the immune system results in the unsuccessful fight with pathogens and is the cause of vulnerability to nosocomial infections. Stem cell therapies may be a tempting option for treatment of this septic-induced organ injury. Also, a growing body of evidence supports the notion that different stem cell subpopulations may directly sense the pathogens and interact with the components of the immune system. This chapter covers the known interactions between hematopoietic, endothelial, and mesenchymal stem cells and the immune response during sepsis. We discuss potential applications of different stem cells in the therapy of sepsis and review available experimental data are reviewed.

Characterization of a new small cell lung cancer (SCLC) cell line STP54 derived from a metastatic bioptate of a combined type of SCLC with Non-SCLC component.

Small cell lung cancer constitutes 15-20% cases of lung cancers, currently the leading cause of death from malignant diseases. It also causes the demise of >90% of affected individuals in 5 years. We have established a new SCLC cell line STP54 derived from fine needle aspirate of metastatic supraclavicular lymph node of 54 -year-old women for model experiments. The primary tumor was diagnosed by histopathological examination as combined type of small cell lung cancer with a non-small cell component. We cultured the cancer cells in the RPMI 1640 medium. In the long-term culture only the small cell component survived. The cell line was established after 30 passages and then characterized by performing cell morphology, cell growth analysis, tumorigenicity in vitro and flow cytometry analysis of selected markers (like NCAM, cytokeratines, HLA-ABC, Fas, Bcl-2, p53, CXCR4, CD210). The cells were growing in floating aggregates and show features suggesting its invasiveness. We suggest that this new cell line may serve as a valuable tool for further studies on lung tumor biology, molecular pathogenesis and metastatic mechanism.