BACKGROUND: Endotoxin or lipopolysaccharide(LPS) can prime phagocytic cells such as polymorphonuclear leukocytes, monocytes or animal peritoneal macrophages to generate increased amounts of secretory products such as oxygen free radicals and tumor necrosis factor, which play an important role in developing adult respiratory distress syndrome in gram negative sepsis. Human alveolar macrophages(HAM) are continuously exposed to various stimuli inhaled into the alveoli, and the response to LPS might be different in HAM. Therefore, we investigated the effect of LPS pre-exposure on HAM adhered to plastic surface and A549 cell(type II human alveolar epithelial cell line) monolayer. METHODS: HAM were isolated from bronchoalveolar lavage fluid from normal lung of the patients with localized lung cancer and esophageal cancer. LPS was exposed to HAM for 2hrs before or after adherence to plastic surface of 24-well Linbro plate and A549 cell monolayer. And then HAM was stimulated with PMA(phorbol myristate acetate) or fMLP(N-formyl-methionyl- leucyl-phenylalanine). The amount of hydrogen peroxide(H2O2) production in the supernatant was measured on the principle of peroxidase-dependent oxidation of phenol red by hydrogen peroxide. RESULTS: LPS pre-exposure could not enhance H2O2 production in neither HAM adhered to plastic surface nor one to A549 cell monolayer. But LPS even in the absence of PMA or fMLP stimulation directly increased H2O2 release in HAM if added after the adherence to A549 cell monolayer. CONCLUSION: Endotoxin does not prime HAM, but may directly activate HAM adhered to alveolar epithelial cells. Further investagation will be necessary.
Animals ; Bronchoalveolar Lavage Fluid ; Epithelial Cells ; Esophageal Neoplasms ; Free Radicals ; Humans* ; Hydrogen ; Hydrogen Peroxide ; Lung ; Lung Neoplasms ; Macrophages, Alveolar* ; Macrophages, Peritoneal ; Monocytes ; Myristic Acid ; Neutrophils ; Oxygen ; Phagocytes ; Phenolsulfonphthalein ; Plastics ; Respiratory Distress Syndrome, Adult ; Sepsis ; Tumor Necrosis Factor-alpha

Recently, we have experienced two cases of xanthogranuloma in adults presenting either a solitary papule or multiple papules. Laboratory findings including blood lipid substances such as cholesterol and triglyceride were within normal limits. In both cases, we could not find any other extracutaneous manifestations. Granulomatous infiltrates containing foam cells, foreign body giant cells and Touton giant cells as well as histiocytes and lymphocytes were revealed by histopathologic findings.
Adult* ; Cholesterol ; Foam Cells ; Giant Cells ; Giant Cells, Foreign-Body ; Histiocytes ; Humans ; Lymphocytes ; Triglycerides

It is becoming increasingly clear that the inflammatory reaction can be ascribed to a complex array of mediators generated and released from activated phagocytes. In this study, the effect of PAF on interleukin-1(IL-1) activity by rat alveolar macrophages(AM) was examined using thymocyte proliferation assay in the supernate of sample obtained after 24 hr culture. When AM were cultured with PAF alone, no change in IL-1 activity was observed. However, the combined addition of PAF and muramyl dipeptide(MDP) or lipopolysaccharide(LPS) to AM cultures markedly enhanced IL-1 activity by 2-3 fold compared with AM cultures with the stimulant alone in a concentration dependent fashion. The peak effect was found at 10-8 M PAF with MDP and 10-14 M PAF with LPS. the effect of PAF was also tested in silica, toxic respirable dust, -added AM cultures as well as in the cultures containing bacterial compounds. Although silica did not stimulate the IL-1 activity, PAF could enhance IL-1 activity by 2 fold above the value of the silica-treated AM cultures with the peak response at 10-2 M PAF. Optimal enhancement of IL-1 activity occurred when MDP and PAF were present together at the initiation of the 24 hr AM cultures. Additionally, the biologically inactive precursor/metabolite of PAF, lyso-PAF failed to induce enhancement of IL-1 activity. When the specific, but structurally different PAF receptor antagonists, BN 5202 (10-5 M) and CV 3988(10-5M) was treated 15 min before addition of PAF(10-8 M) and MDP(10 microgram/ml) to the AM cultures, it markedly inhibited the enhancement of IL-1 activity induced by PAF. The effects of these PAF antagonists were also observed in LPS(1.0 microgram/ml)-stimulated cells. Collectively, these data suggest that PAF enhances IL-1 activity by interaction with a specific receptor.
Animals ; Dust ; Interleukin-1* ; Macrophages, Alveolar* ; Phagocytes ; Rats ; Silicon Dioxide ; Thymocytes

BACKGROUND: The immediate host response to LPS is the production of proinflammatory cytokines that act as intercellular mediators in inflammatory reactions, including acute lung injury. These "early response" cytokines transmit signals from recognition cells to target or effector cells. This host response is futher amplified by the expression of leukocyte chemoattractants, growth factors, and adhesion molecules, resulting in an array of proinflammatory events. This experiment was performed to define the lung origin of proinflammatory cytokines, such as TNF-alpha IL 6 in early periods of endotoxin induced acute lung injury (ALl). METHOD: The healthy male Sprague-Dawley, weighted 150-250g, were divided into saline control (NC) and endotoxemia-induced ALl (ETX-), and leukopenic endotoxemia-induced ALl (CPA-ETX-Group) which was induced by cyclophosphamide, 70 mg/kg i.p. injection. Acute lung injury was evoked by LPS, 5 mg/kg, intravenously administered. Bronchoalveolar lavage was performed at 0, 3, 6 h after LPS-treated to estimate the influx of phagocytes and concentration of total protein, and cytokines as TNF-alpha and IL 6 by a bioassy using MTT method. We also examined the localization of TNF-alpha and IL 6 protein in endotoxemia-challenged lung tissue by immunohistochemical stain (IH). RESULTS: The total cell, macrophage and PMN count in BALF were elavated in ETX group compared to NC (p<0.05). In CPA-ETX group, total cell and macrophage count in BALF were not changed compared to NC, but PMN count was markedly reduced and it took part. in less than 0.1% of total BAL cells (p<0.01). The protein concentration in BALF were significantly increased in ETX and CPA-ETX group compared to NC (p<0.05), but there was signifcant difference between ETX- and CPA-ETX group only at 6 h (p<0.05). This observation suggested that even if PMNs are involved in the pathogenesis of acute lung injury, their role cannot be viewed as essential. The concentration of TNF-alpha and IL 6 in BALF was significantly increased in the ETX- and CPA-ETX group compared to NC. There was no difference between ETX- and CPA-ETX group. In IH, anti-TNF-alpha- and anti-IL 6 antibody was strongly localized at interstitial monocytes and alveolar macro-phages in endotoxemia-challenged lung tissue. From above point of view, activated alveolar macrophage/monocyte considered as a prominent source of proinflammatory cytokines in endotoxemia-challenged lung injury. CONCLUSION: The prominent source of proinflammatory cytokines in early periods of endotoxemia-induced lung injury will be the activated resident macrophages like an alveolar macrophage and interstitial monocytes. The pulmonary macrophage/monocyte will impact the initiation and continuance of lung injury without PMNs s certain inflammatory role, particularly in endotoxemia-induced acute lung injury.
Acute Lung Injury ; Bronchoalveolar Lavage ; Chemotactic Factors ; Cyclophosphamide ; Cytokines* ; Endotoxemia* ; Humans ; Intercellular Signaling Peptides and Proteins ; Interleukin-6* ; Interleukins* ; Leukocytes ; Lung Injury ; Lung* ; Macrophages ; Macrophages, Alveolar ; Male ; Monocytes ; Phagocytes ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha*

Neutrophils are the most abundant white blood cells in the peripheral blood and have long been recognized as the major phagocytes in acute infection by destroying extracellular pathogens. Although research on neutrophils hampered by intractability in the experiments, the newly discovered effector functions of neutrophils includes granular proteins, and cytokines, extracellular traps. With all effector mechanism neutrophils play a critical role in the pathogenesis of acute and chronic infection, autoimmunity and cancer.
Autoimmunity ; Cytokines ; Inflammation ; Leukocytes ; Neutrophils ; Phagocytes ; Proteins

PURPOSE: We evaluated the easily-assessable risk factors to predict bacteremia in children with febrile neutropenia, who received anticancer chemotherapy. METHODS: We retrospectively reviewed 46 children who had febrile neutropenia caused by anticancer chemotherapy between March, 1993 and February, 1997. The patients with localized infection on presentation were not eligible for this study. We evaluated the correlation between bacteremia and some variables, including absolute neutrophil count (ANC), absolute monocyte count (AMoC) and absolute phagocyte count (APC). RESULTS: There was total of 147 consecutive episodes of fever in 46 children, with 90 episodes of fever were noted in neutropenic patients without localized infection. There were 20 episodes of bacteremia (22.2%) in 90 episodes of febrile neutropenia. The mean ANC of 365.5 +/- 448.3/microliter, mean AMoC 132.3 +/- 310.4/microliter and mean APC 502.0 +/- 603.3/microliter did not show significant correlation with bacteremia. There was no statistically significant correlation between bacteremia and ANC or AMoC. There was higher risk of bacteremia in patients with AMoC less than 100/microliter as compared with patients with AMoC above than 100/microliter (odds ratio : 1.39, 95%CI : 0.41-4.69). There were 17 episodes of bacteremia (28.8% of 59 febrile episodes) in patients with APC less than 500/microliter and 3 episodes of bacteremia (9.7% of 31 febrile episodes) in patients with APC above than 500/microliter (P=0.03, odds ratio : 3.78, 95%CI : 1.01-14.10). CONCLUSION: There was a statistically-significant correlation between bacteremia and APC with higher risk of bacteremia in patients with APC less than 500/microliter. Trials should be conducted to test whether APC may be used to assign some children to less intensive or outpatient antibiotic therapy at the time of presentation of febrile neutropenia.
Bacteremia* ; Child* ; Drug Therapy* ; Febrile Neutropenia* ; Fever ; Humans ; Monocytes ; Neutropenia ; Neutrophils ; Odds Ratio ; Outpatients ; Phagocytes ; Retrospective Studies ; Risk Factors*

BACKGROUND: Severe acute lung injury(ALI), also known as the adult respiratory distress syndrome(ARDS), is a heterogenous nature of dynamic and explosive clinical synrome that exacts a mortality of approximately 50%. Endotoxin(ETX) is an abundant component of the outer membrane of gram-negative bacteria capable of inducing severe lung injury in gram-negative sepsis and gram-negative bacterial pneumonia, which are among the most common predisposing causes of ARDS. The influx of PMNs into airway tissue is a pathological hallmark of LPS-induced lung injury. And th3re is a substantial evidence suggesting that cytokines are important mediators of lung injury in gram-negative sepsis. However, the kinetics of phagocytes and cytokines by an exact time sequence and their respective pathogenic importance remain to be elucidated. This study was performed to investigate the role of phagocytes and proinflammatory cytokines in ETX-induced ALl through a time course of changes in the concentration of protein, TNFa and IL-6, and counts of total and its differential cells in BALF. The consecutive histologic findings were also evaluated. METHOD: The experimental animals, healthy male Sprague-Dawley, weighted 200+/-50g, were divided into controland ALI-group. ALI was induced by an intravenous administration of ETX, 5mg/kg. Above mentioned all parameters were examined at 0(control), 3, 6, 24, 72 h after administration of ETX. TNFa and IL-6 conc. in BALE were measured by a bioassay. RESULTS: The protein concentration and total leukocyte count(TC) in BALF was significantly increased at 3h compared to controls(p<0.05). The protein conc. was significantly elavated during observation period, but TC was significantly decreased at 72h(p<0.05 vs. 24h). There was a close relationship between TC and protein cone. in BALF(r = 0.65, p <0.001). The PMN and monocyte count was well correlated with TC in BALF, and the correlation of PMN(r=0.97, p<0.001) appeared to be more meaningful than that of monoeyte(r = 0.61, p<0.001). There was also a significant correlation between protein cone. and PMN or monocyte count in BALF(PMN vs. monocyte r = 0.55, p<0.005 vs. r = 0.64, p<0.001). The count of monocyte was significantly elavated during observation period though a meaningful reduction of PMN count in BALF at 72h, this observation suggested that monocyte may, at least, partipate in the process of lung injury steadly. In this sudy, there was no relationship between IL-6 and TNFt conc., and TNFa but not IL-6 was correlated with TC(r 0.61, p <0.05) and monocyte(r = 0.67, p<0.05) in BALF only at 3, 6h after ETX introduced. In particular, the IL-6 cone. increased earlier and rapidly peaked than TNFz cone. in BALF. In histologic findings, the cell counts of lung slices were increased from 3 to 72h(p<0.001 vs. NC). Alveolar wallthickness was increased from 6 to 24h(p<0.001 vs. NC). There was a significant correlation between the cell counts of lung slices and alveolar wall-thickness(r= 0.61, p<0.001). This result suggested that the cellular infiltrations might be followed by the alterations of interstitium, and the edematous change of alveolar wall might be most rapidly recovered to its normal condition in the process of repair. CONCLUSION: We concluded that although the role of PMIN is partly certain in ETX-induced ALI, it is somewhat inadequate to its known major impact on ALL Alveolar macrophage and/or non-immune cells such as pulmonary endothelial or epithelial cells, may be more importantly contributed to the initiation and perpetual progression of ETX-induced ALI. The IL-6 in ETX-induced ALI was independent to TNFa, measured by a bioassay in BALF. The early rise in IL-6 in BALF implies multiple origins of the IL-6.
Acute Lung Injury* ; Administration, Intravenous ; Adult ; Animals ; Biological Assay ; Cell Count ; Cytokines ; Epithelial Cells ; Gram-Negative Bacteria ; Humans ; Interleukin-6 ; Kinetics ; Leukocytes ; Lung ; Lung Injury ; Macrophages, Alveolar ; Male ; Membranes ; Monocytes ; Mortality ; Phagocytes* ; Pneumonia, Bacterial ; Rats, Sprague-Dawley ; Sepsis

Nitric Oxide (NO) is an important mediator in various pathological conditions. The list of agents known to activate the NO pathway continues to expand and now includes bacterial products, cytokines, cAMP-elevating agents, trauma, and ozone. The activation of the L-arginine-dependent NO pathway via NO synthase is an important mechanism to stimulate both antimicrobial capability and cytotoxicity of phagocytes. NO has both beneficial and detrimental effects on host responses including lung injury. The effects of NO on the host were intensively investigated in lung injury, bovine pneumonic pasteurellosis (51). However, there was no description about the effect on the primary agent of the disease, Pasteurella haemolytica Al. Therefore, we investigated the effect of NO produced from bovine alveolar macrophages on the growth of Pasteurella haemolytica Al which is the primary agent of bovine pneumonic pasteurellosis. With the exogenous source of NO, sodium nitroprusside (SNP), the growth of the bacterium was dose-dependently inhibited by NO produced from SNP when measured by XTT colorimetric assay and standard plate count method. Also, same effect was observed in AM-derived NO. The effect was bacteriostatic rather than bactericidal.
Animals ; Cytokines ; Lung Injury ; Macrophages, Alveolar* ; Mannheimia haemolytica* ; Nitric Oxide Synthase ; Nitric Oxide* ; Nitroprusside ; Ozone ; Pasteurella* ; Pasteurellosis, Pneumonic ; Phagocytes ; Thiram

BACKGROUND: IFN-gamma is known to activate mononuclear phagocytes and to mediate host defense mechanism against some intracellular microorganisms, but litfle is known about anti-mycobacterial activity and mechanism of IFN-gamma in human. In this study, we investigated the role of IFN-gamma in the pathogenesis of tuberculosis by observing the effect of IFN-gamma on the phagocytosis of M.tuberculosis(MTB) and on the production of TNF-alpha by human pulmonary alveolar macrophage. METHOD: Pulmonary alveolar macrophage(PAM) were prepared with adhesion purification method from bronchoalveolar lavage fluid obtained from 8 persons without active lung lesion and cultured(1 x 106cells/ml) with MTB(3 x 107 bacteria/ml) with or without IFN-gamma(300U/ml), LPS(0.5ug/ml) and autologous serum(10%). After 2 hours, the percentage of PAM-phagocytosed MTh was counted after AFB staining(modified Kynion method). TNF-alpha production by PAM stimulated by IFN-gamma(300U/ml), MTB(1 x l06bacteria/ml) and LPS(0.5ug/ml) for 24hours was measured in culture supernatant using ELISA method. The degree of phagocytosis of MTh by PAM stimulated with IFN-gamma(300U/ml) and LPS(0.5ug/ml) for 24hours was also investigated. RESULTS: IFN-gamma did not influence the phagocytosis of MTB by PAM(percentage of PAM-phagocytosed MTB: control: 22.1+/- 4.9, IFN-gamma: 20.3+/- 5.3) and did not increase TNF-alpha production by PAM(controfl 21+/-38pg/ml, IFN-gamma : 87+/-106pg/ml), and the degree of phagocytosis of MTh by PAM pre-stimulated with IFN-gamma for 24 hours, was not increased (controL 24.5+/-9.5, IFN-gamma : 23.4+/-10.1). CONCLUSION: IFN-gamma does not influence on the phagocytosis of MTB and TNF-alpha production by PAM.
Bronchoalveolar Lavage Fluid ; Enzyme-Linked Immunosorbent Assay ; Humans* ; Lung ; Macrophages, Alveolar* ; Mycobacterium tuberculosis ; Phagocytes ; Phagocytosis* ; Tuberculosis* ; Tumor Necrosis Factor-alpha

Autophagy is a highly conserved intracellular degradation and energy-recycling mechanism that contributes to the maintenance of cellular homeostasis. Extensive researches over the past decades have defined the role of autophagy innate immune cells. In this review, we describe the current state of knowledge regarding the role of autophagy in neutrophil biology and a picture of molecular mechanism underlying autophagy in neutrophils. Neutrophils are professional phagocytes that comprise the first line of defense against pathogen. Autophagy machineries are highly conserved in neutrophils. Autophagy is not only involved in generalized function of neutrophils such as differentiation in bone marrow but also plays crucial role effector functions of neutrophils such as granule formation, degranulation, neutrophil extracellular traps release, cytokine production, bactericidal activity and controlling inflammation. This review outlines the current understanding of autophagy in neutrophils and provides insight towards identification of novel therapeutics targeting autophagy in neutrophils.
Autophagy ; Biology ; Bone Marrow ; Extracellular Traps ; Homeostasis ; Inflammation ; Neutrophils ; Phagocytes