Differential Neutrophil Research Articles

Continuous antibiotic treatment for experimental abdominal sepsis: effects on organ inflammatory cytokine expression and neutrophil sequestration.

Tumour necrosis factor (TNF) alpha and interleukin (IL) 1 beta are produced in the lung after peritonitis and may contribute to neutrophil-mediated organ injury. It was hypothesized that, during experimental peritonitis, continuous rather than intermittent antibiotic therapy would reduce lung expression of TNF-alpha and IL-1 beta messenger RNA (mRNA) and neutrophil sequestration. After caecal ligation and puncture, mice received either intermittent or continuous cefoxitin, or continuous metronidazole or aztreonam. Cytokine mRNAs were determined by reverse transcription differential polymerase chain reaction and lung neutrophil content by myeloperoxidase (MPO) assay. Continuous cefoxitin reduced median (interquartile range (i.q.r.)) lung IL-1 beta mRNA expression ((ratio to beta-actin): continuous 0.18 (0.14-0.34), intermittent 0.46 (0.44-0.49), saline 0.43 (0.38-0.53), P < 0.05) and median (i.q.r.) lung MPO content (continuous 22.5 (9.7-40), intermittent 65 (57.5-76), saline 47 (41-64), P < 0.05) compared with intermittent therapy and saline controls. Continuous infusion was also associated with reduced bacteraemia (P < 0.05) but not serum TNF-alpha or endotoxin levels. Both continuous metronidazole and aztreonam reduced lung MPO concentration (P < 0.05) and TNF-alpha and IL-1 beta mRNA expression (P < 0.05) compared with those in saline controls. These effects were dependent on a reduction in the number of susceptible bacteria rather than serum TNF-alpha or endotoxin levels. The stimulus for organ inflammatory cytokine production and neutrophil sequestration during peritonitis is the level of bacteraemia present, which is more effectively controlled with continuous antibiotic therapy.

Physical Feature, Physiological Character and Behavior Study of Gayal (Bos frontalis)

The physical feature, physiological character and behavior studies were conducted with fifteen newly collected gayals in Bandarban hill tract area of Bangladesh. Their morphology is different from domestic cattle. The range of pulse rate, body temperature and respiration rate were 47 to 75 per minute, 37.78 to 38.88°C and 20 to 40 per minute, respectively. These physiological values vary with different age group and seasonal variation. In hematological feature, the average findings were RBC 7.01±0.52 million/cu.mm, WBC 14.3±3.69 thousand/cu.mm, hemoglobin concentration 9.81±2.25 gm%, PCV 35.86±3.68%. In differential WBC count neutrophils 28.23±1.75 %, lymphocytes 62±2.05 %, monocytes 4.4±1.34%, eosinophils 5±2.49% and basophils 0.4±0.51%. In behavior study, the animal shows browsing nature on hill slopes. They are watchful in new environment, become excited and nervous with strangers. Heated female gayals response for mating with domestic bull. (Asian-Aust. J. Anim. Sci. 2003. Vol 16, No. 11 : 1599- 1603)

Differential neutrophil response to sepsis in large and small bowel anastomoses

Differential neutrophil response to sepsis in large and small bowel anastomoses

Differential neutrophil traffic in gut and lung after scald injury.

Neutrophil recruitment to the lung after thermal injury has been reported by various laboratories. Changes in neutrophil populations in the gut and lung have not been examined simultaneously after burn injury. Mice aged 8 to 10 weeks were anesthetized and subjected to 15% topical scald injury. Animals were sacrificed at 30 minutes and 1, 2, 4, 8, 16, and 32 hours after injury with harvesting of terminal ileum and lung for myeloperoxidase (MPO) assay. Lungs were evaluated after bronchoalveolar lavage and lavage of the vascular bed to remove neutrophils in these compartments. Myeloperoxidase activity was compared between groups of sham-injured and burned animals. Although pulmonary neutrophil recruitment was obvious after scald burn; in the ileum, burned animals showed diminished MPO activity. Histology and bronchoalveolar lavage revealed no evidence of gross organ injury apart from obvious changes in cellular content in the lung. Thermal injury is associated with differential neutrophil movement in the lung and the gut in this model. Pulmonary neutrophil recruitment is confirmed, whereas the gut seems to lose neutrophils as indicated by diminished MPO activity in the initial hours after dorsal scald injury.

Alpha9beta1 integrin is expressed on human neutrophils and contributes to neutrophil migration through human lung and synovial fibroblast barriers.

Accumulation of leukocytes in inflamed tissue involves their migration through vascular endothelium and then in the connective tissue. Recently we utilized a barrier of human synovial, dermal, and lung fibroblasts (HSF, HDF, and HLF) grown on polycarbonate filters as a model of human polymorphonuclear leukocyte (PMN) migration through connective tissue. The beta2 integrins (CD 11/ CD18) and alpha4, alpha5, and alpha6beta1 (VLA-4, -5, and -6) integrins each contributed to this PMN migration. Here we report that on human blood leukocytes, alpha9beta1 (VLA-9) is expressed only on PMNs and that it is up-regulated after PMN activation. Based on monoclonal antibody (mAb) blocking studies, alpha9beta1 integrin contributed to C5a-induced PMN migration through fibroblast (HLF and HSF) barriers. This role was apparent only when alternate mechanisms such as CD18, alpha4, alpha5, and alpha6beta1 integrins were blocked and then mAb to alpha9beta1 integrin inhibited the residual PMN migration (by 40-50%) through the HLF or HSF barrier, resulting in > or = 75% inhibition overall. In contrast, PMN migration across interleukin-1-activated endothelium (HUVEC) in response to a C5a gradient, which is partly (30-40%) via CD11/CD18-independent mechanisms, was not inhibited by adhesion blocking by mAbs to alpha4, alpha5, alpha6, and alpha9beta1 even in combination. These results indicate that alpha9beta1 integrin on PMN may have a special role, in conjunction with other beta1 integrins, in mediating PMN migration in the extravascular space, and may contribute to differential neutrophil function within tissues.

Influx of Neutrophils into the Airway Lumen at 4 h after Segmental Allergen Challenge in Asthma

Background: Segmental allergen challenge is a powerful tool to study inflammatory reactions in asthmatic airways. There is little information on the early events at 5 min and 4 h after allergen challenge with respect to the cell influx and the chemokine interleukin–8 (IL–8). Methods: Seven mild to moderate allergic asthmatics (AA group), 5 allergic nonasthmatics (ANA group) and 5 nonallergic controls underwent segmental allergen challenge, with allergen doses based upon skin reactivity. Bronchoalveolar lavage (BAL) samples were obtained before, 5 min and 4 h postchallenge, and were analyzed for cell numbers and differential counts, eosinophil and neutrophil chemotactic activity, and levels of IL–8. Results: At 5 min postchallenge, no changes were observed compared to baseline. At 4 h postchallenge, an increase was found in the number of neutrophils and the levels of IL–8, which was dependent on the dose of allergen in the AA and ANA group. At the same allergen dose, the increases in neutrophils and levels of IL–8 were calculated to be 91 and 67 times higher, respectively, in AA than in ANA. Levels of IL–8 correlated with the number of neutrophils and with the in vitro neutrophil chemotactic activities in BAL fluid. Conclusions: Neutrophil chemotactic activity is increased in BAL fluid at 4 h after segmental allergen challenge. We suggest that apart from IgE–mediated mast cell degranulation, additional local factors in the airways determine the degree of IL–8 increase and neutrophil influx.

EFFECT OF ALBENDAZOLE ON SOME HORMONAL, HAEMATOLOGICAL AND BIOCHEMICAL PROFILES IN SHEEP

In this study, the effects of albendazole (10 mg/kg b.wt.) on some hormonal, haematological and biochemical profiles of 20 mature sheep of both sex were studied after its oral administration as 2.5 % suspension. Albendazole exerted a significant increase in the levels of serum globulins, alanine aminotransferase (ALT),aspartate aminotransferase (AST), superoxide dismutase (SOD), total leukocytic count (WBCS) and their differential count, particularly lymphocytes and neutrophils. Moreover, it evoked a significant decrease in serum testosterone of rams. Meanwhile, it demonstrated no significant effect on the levels of estradiol and progesterone in ewes.

Mapping Quantitative Trait Loci for Immune Capacity in the Pig

Immune capacity traits show considerable genetic variation in outbred populations. To identify quantitative trait loci (QTLs) for immune capacity in the pig, various measures of immune function (total and differential leukocyte counts, neutrophil phagocytosis, mitogen-induced proliferation, IL-2 production, and virus induced IFN-alpha production in whole blood cultures, and Ab responses to two Escherichia coli antigens) were determined in 200 F2 animals from a wild pig-Swedish Yorkshire intercross. The pedigree has been typed for 236 genetic markers covering all autosomes, the X chromosome and the X/Y pseudoautosomal region. Through interval mapping using a least-squares method, four QTLs with significant effects were identified; one for total leukocyte counts, one for mitogen-induced proliferation, one for prevaccination levels of Abs to E. coli Ag K88, and one for Ab response to the O149 Ag. In addition, several putative QTLs were indicated. The results from the present study conclusively show that it is possible to identify QTLs for immune capacity traits in outbred pig populations by genome analysis.

Effect of inhaled endotoxin on induced sputum in normal, atopic, and atopic asthmatic subjects

BACKGROUNDInhalation of lipopolysaccharide (LPS) causes an inflammatory response in the lungs. To explore this response, inflammatory indices were measured in induced sputum from atopic asthmatic patients and compared with atopic...

Exhaled nitric oxide in human lung transplantation. A noninvasive marker of acute rejection.

Acute allograft rejection in animals and humans has been associated with increased nitric oxide production in the graft. Exhaled nitric oxide (ENO) measurement is a noninvasive method of assessing inflammation in airway diseases, e.g., asthma, which might be applicable to lung transplant recipients. Over 12 months, ENO of lower respiratory origin was measured in 108 lung transplant recipients with a mean time after transplant of 1,083 d. ENO (mean +/- SEM; ppb) in stable patients (19.5 +/- 1.1; p < 0.001) was not different from that of healthy controls (23.8 +/- 3.2). ENO was significantly higher in episodes of clinical acute rejection (51.1 +/- 6.3) compared with stable patients but not elevated in bronchiolitis obliterans syndrome (18.6 +/- 1.5) or pulmonary infection (25.9 +/- 4.0). A retrospective analysis of bronchoscopy findings and concurrent ENO (n = 99) showed that ENO did not vary according to histological findings (normal, acute rejection grade I, nonspecific inflammatory change) or with a positive BAL culture. ENO was not correlated with differential lymphocyte and neutrophil counts. ENO appears to be a valid marker of clinical acute rejection in human lung transplantation as distinct from infection or bronchiolitis obliterans. Furthermore, bronchoscopic findings in the absence of a clinical illness were not associated with a rise in ENO.

CHANGES IN WBC COUNT IN FEMORAL VEIN AND RADIAL ARTERY BLOOD IN PATIENTS UNDERGOING REPAIR OF ABDOMINAL AORTIC ANEURYSM

White blood cells (WBC) have been shown to be sequestered in ischemic tissue following reperfusion and to contribute to reperfusion injury in models of cardiac, intestinal, and skeletal muscle. Several studies have shown that the efferent venous WBC count was lower than the afferent arterial WBC count in ischemic tissue following reperfusion. As the patient undergoing abdominal aortic aneurysm repair is subject to temporary clamping of the abdominal aorta, and consequently to ischemia-reperfusion of the lower limb, we speculate that WBCs are sequestered in the lower limib such that the femoral WBC count becomes lower than the systemic WBC count. Eight patient with abdominal aortic aneurysms undergoing arterial reconstruction were subjected to a study in which WBC counts in the femoral vein and radial artrery were measured before and during the first 48 hr after declamping of the abdominal aorta. Both neutrophils and monocytes were significantly increased in both venous and arterial blood, but lymphocytes were significantly decreased after declamping of the abdominal aorta. There were no significant differences in neutrophils and lymphocytes between the femoral vein and radial artery blood. It is thought that the WBC sequestration in the lower limb was not very marked so and that the femoral venous WBC count differed from the radial arteial WBC count in patients undergoing repair of an abdominal aortic aneurysm.

Effects of inotropes on human leucocyte numbers, neutrophil function and lymphocyte subtypes

We have investigated the effects of inotropes with different adrenergic receptor specificity on differential white cell count, lymphocyte subtypes and neutrophil function in healthy volunteers. Six healthy, male volunteers were enrolled into this randomized, placebo-controlled pilot study. Each volunteer was studied on four separate occasions during a 2-h infusion of various agents, and for 2 h after stopping the infusion. The agents investigated were adrenaline 0.1 microgram kg-1 min-1, dobutamine 5 micrograms kg-1 min-1, dopexamine 2 micrograms kg-1 min-1 and 5% glucose 0.5 ml kg-1 h-1. Venous blood was sampled at 0, 30, 120 and 240 min. Haemodynamic monitoring was continued throughout the study. Full blood count, white cell differential count and enumeration of lymphocyte subtypes were performed. Neutrophil function tests included chemoluminescence, and assessment of neutrophil chemotaxis, phagocytosis and adhesion. The Wilcoxon signed rank test was used to compare differences between placebo and active drugs at each time compared with baseline. There was a significant increase in white cell count, lymphocyte count and neutrophil count with adrenaline, and a small but significant decrease in these variables with dobutamine and dopexamine. These changes were also apparent for absolute CD3+, CD4+ and CD8+ lymphocyte counts. Neutrophil respiratory burst in response to f-methionyl-leucyl-phenylalanine increased significantly only with adrenaline at 30 min (P = 0.046). There were no other significant changes in tests of neutrophil function. Infusion of inotropes was associated with changes in white cell numbers, lymphocyte subtypes and neutrophil respiratory burst. In healthy volunteers, adrenaline had effects different from those of dobutamine and dopexamine. The clinical relevance of such effects requires further investigation in critically ill patients.

Cellular and mediator responses twenty-four hours after local endobronchial allergen challenge of asthmatic airways

The effects of acute allergen exposure on bronchoalveolar lavage cells and mediators and mucosal inflammatory cells were evaluated in 10 subjects with atopic asthma who underwent lavage and biopsy 24 hours after segmental endobronchial allergen challenge. Increased numbers of bronchoalveolar lavage eosinophils were retrieved from the allergen-challenged sites compared with the saline-challenged sites (mean 21.4 vs 1.5 × 10 3 cells/ml; p < 0.02). Numbers of neutrophils and proportions of CD4 +, CD8 +, CD25 +, and HLA-DR + T cells were similar at the saline- and allergen-challenged sites. In contrast to the bronchoalveolar lavage findings, eosinophil numbers were not increased in the bronchial submucosa or epithelium. There was also no significant difference in neutrophils, mast cells, CD3 +, CD4 +, or CD8 + T cells in the submucosa after allergen challenge, but the number of activated (CD25 +) T lymphocytes in the mucosa did increase after allergen challenge. Allergen challenge did not induce any significant change in endothelial expression of P-selectin, E-selectin, intercellular adhesion molecule-1, or vascular cell adhesion molecule-1. CD11a + and very late antigen-4 + cell numbers were similar in the saline- and allergen-challenged sites. This study suggests that in patients with very mild asthma, local allergen challenge induces persistent bronchoalveolar lavage eosinophilia, but the recruitment process seems to have diminished or ceased by 24 hours. (J A LLERGY C LIN I MMUNOL 1996;98:133-43.)

Acute versus chronic administration of phosphodiesterase inhibitors on allergen-induced pulmonary cell influx in sensitized guinea-pigs.

1. The aims of this study were to determine which phosphodiesterase (PDE) isoenzymes are involved in the control of eosinophil accumulation in the airways of ovalbumin (OVA)-immunized guinea-pigs by the use of isoenzyme selective inhibitors and to compare the effects of acute versus chronic administration of PDE isozyme inhibitors on pulmonary cell influx in ovalbumin-immunized guinea-pigs. 2. Guinea-pigs were sensitized and subsequently challenged with aerosolized OVA. Twenty four hours later bronchoalveolar lavage (BAL) was performed to permit assessment of inflammatory cell accumulation. A significant increase in the number of eosinophils was observed in the lavage fluid from OVA-immunized (13.6 +/- 1.4 x 10(4) ml-1 in acute experiments and 10.1 +/- 1.4 x 10(4) ml-1 in chronic experiments) animals compared with sham-treated controls (5.6 +/- 0.6 x 10(4) ml-1 in acute experiments and 5.1 +/- 0.6 x 10(4) ml-1 in chronic experiments). There was no difference in neutrophil, mononuclear cell or total cell numbers between the two groups. 3. Acute administration of a high dose of selective and non-selective PDE inhibitors by the i.p. route had no significant effect on eosinophil accumulation in the airways. 4. Chronic administration of a low dose (3 mg kg-1, i.p., twice daily for 7 days) of the type IV PDE inhibitor, RO 20-1724, and the PDE III/IV inhibitor, zardaverine, produced a significant inhibition of eosinophil accumulation (46% and 59% respectively). 5. These results suggest that the type IV PDE isoenzyme plays a role in the control of allergen-induced eosinophil infiltration into the airways, but indicate that a period of low dose chronic treatment with a type IV or mixed type III/IV PDE inhibitor is necessary for eosinophil accumulation in the airways to be reduced.

Pulmonary immunopathology of sudden infant death syndrome

Sudden infant death syndrome (SIDS) is the most common cause of postneonatal mortality in the UK. Pathological investigations have shown evidence suggestive of respiratory obstruction with subsequent hypoxia leading to death. We examined 48 infants who died of SIDS and 30 who died of other, non-pulmonary, causes and identified pulmonary eosinophil and neutrophil leucocytes, mast cells, and T and B lymphocytes by immunocytochemistry. Positively stained cells were counted in the parenchyma and around the bronchi without knowledge of the tissue source. The results showed three times more eosinophils in the lungs of infants who died of SIDS (27 61 vs 7·91 [99% Cl 1 76-5 87] cells/mm2 for parenchyma) accompanied by increased T lymphocytes and B lymphocytes. There were more peribronchial mast cells in the SIDS group (22·1 vs 14·7 [1·03-2·10] cells/mm2) and insignificant differences in neutrophils and parenchymal mast cells. There were significant associations between eosinophil, B lymphocyte, and T lymphocyte numbers. These findings provide evidence for an abnormal T lymphocyte-mediated pulmonary inflammatory response in SIDS. Products of eosinophil degranulation can cause epithelial damage and pulmonary oedema, which could cause the respiratory obstruction and hypoxia associated with SIDS.

Evaluation of the Sysmex NE-8000:Automated Hematology Analyzer in a High-Volume Outpatient Laboratory

An evaluation of SYSMEX NE-8000 (Toa Medical Electronics Co., Ltd., Kobe, Japan) hematology analyzer, including its automated five-part white blood cell (WBC) differential count, was performed in a high-volume outpatient laboratory. Precision, mixing studies, stability, carry-over, and linearity were all within the limits stated by the manufacturer. Evaluation of the five-part WBC differential revealed excellent correlation with manual differential neutrophil and lymphocyte counts. Monocyte, eosinophil, and basophil data were acceptable given the known poor precision of 200 cell manual WBC differential counts when low percentages of individual cell types are present. Overall, flagging for morphologic abnormalities displayed a sensitivity of 87% and a specificity of 97%. The combination of the flagging system with the visual record provided by the NE-8000 WBC histogram represents an effective tool for separating normal from abnormal specimens, and allows considerable selectivity in the performance of follow-up manual WBC differential counts.

Autologous marrow transplantation for patients with acute myeloid leukemia in untreated first relapse or in second complete remission.

This study compares outcomes of autologous bone marrow transplantation (ABMT) in patients with acute myeloid leukemia (AML) in untreated first relapse (REL1) or in second complete remission (REM2). Forty-seven patients with AML in REL1 (n = 21) or in REM2 (n = 26) were treated with busulfan (BU) and cyclophosphamide (CY) with or without total-body irradiation (TBI) followed by ABMT. All REL1 patients and four REM2 patients had marrow stored during first remission (REM1). Twenty-seven had marrow stored with and 20 without treatment in vitro with 4-hydroperoxycyclophosphamide (4-HC). Eighteen patients received BU and CY and 29 received BU, CY, and TBI. REL1 patients relapsed within a median of 9 months (range, 2 to 26) after marrow harvest and were transplanted a median of 30 days (range, 9 to 87) from detection of relapse. With a median follow-up of 2.1 years (range, 0.4 to 5.3), 19 patients survive in remission (10 of 21 in REL1; nine of 26 in REM2). The actuarial probabilities of relapse-free survival at 2 years for patients transplanted in REL1 and REM2 were 45% +/- 22% and 32% +/- 18%, respectively (P = .33). The corresponding probabilities of relapse were 30% +/- 26% and 44% +/- 23%, respectively (P = .45). No conclusions could be drawn about the benefits of adding TBI to BU plus CY. There were no significant differences in neutrophil or platelet recovery or in posttransplant probabilities of relapse and nonrelapse mortality between patients who received marrow treated or not treated with 4-HC. These results suggest that ABMT may produce long-term leukemia-free survival in approximately one third of patients with AML in REL1 or in REM2. There is no apparent clinical advantage in attempting to obtain second remissions in relapsed patients before ABMT if marrow has been cryopreserved during REM1. Although a strategy of transplantation in REL1 has advantages for the patient, such an approach involves the storage of marrow, which may not be used, and is impractical without the coordinated support of the treating physician, the patient, and the marrow transplant center.

Notes on age-related changes in differential leucocyte counts of the Charles River outbred albino SD rat and CD1 mouse

Comprehensive baseline data from differential leucocyte counts are essential for the evaluation of leucocyte status in long-term carcinogenicity studies. Little information on lifetime variations of leucocyte types in the Charles River outbred albino rat and mouse has been published and this report describes the differential leucocyte count in rats and mice, held in our environment, sampled at 52, 78 and 104 weeks. Data revealed an age-related increase in differential neutrophil count and a corresponding decrease in differential lymphocyte count. Monocyte, eosinophil and basophil counts were unchanged during the lifetime of the animals.

Cytology of Bronchoalveolar Lavage Fluid in Bacterial Pneumonia

The findings of Drs. Castella and Artigas are very interesting. We agree that the differences in methods used are probably insufficient to account for the differences in neutrophils found in patients with pneumonia. Cytology of Bronchoalveolar Lavage Fluid in Bacterial PneumoniaCHESTVol. 97Issue 6PreviewTo the Editor: Full-Text PDF

Increased neutrophil receptors for and response to the proinflammatory bacterial peptide formyl-methionyl-leucyl-phenylalanine in Crohn's disease

Neutrophils have an important role in mediating tissue injury in inflammatory bowel disease. The proinflammatory peptide formyl-methionyl-leucyl-phenylalanine (FMLP), which is produced by intestinal bacteria, is a potent chemotactic factor for human neutrophils and has been used experimentally to induce acute colitis in animal models. Its action involves specific receptors on the surface of the neutrophil. Increased mucosal permeability, which has been described in Crohn's disease, might contribute to the pathogenesis of the intestinal lesions in this disease by means of increased absorption of FMLP and other gut-derived bacterial products. To evaluate the interaction between FMLP and neutrophils in patients with inflammatory bowel disease, we have studied the binding characteristics and responsiveness of circulating neutrophils to this peptide. Neutrophils from patients with Crohn's disease but not those with ulcerative colitis were found to have significantly increased numbers of receptors for FMLP with similar affinity constants. This was associated with a significantly increased functional response to FMLP in chemiluminescence assays. Differences in neutrophils between Crohn's disease and ulcerative colitis are probably not intrinsic to the neutrophil but reflect their conditioning by the different inflammatory milieu of these diseases.