Endotoxin-stimulated alveolar macrophage recruitment of neutrophils and modulation with exogenous surfactant

OBJECTIVE: To determine whether endotoxin-stimulated alveolar macrophages would attract neutrophils and whether exogenous surfactant treatment would modulate this chemoattraction. DESIGN: Alveolar macrophages were harvested from bronchoalveolar lavage fluid and neutrophils from the blood of anesthetized guinea pigs. SUBJECTS: Hartley guinea pigs. INTERVENTIONS: Alveolar macrophages were suspended in RPMI 1640 and stimulated with 1 microg/mL of lipopolysaccharide (LPS), the supernatant removed and the alveolar macrophages were incubated in either RPMI or RPMI with surfactant at two different doses (292 microg/mL or 875 microg/mL) for 16 hrs. MEASUREMENTS AND MAIN RESULTS: The supernatant was extracted from the alveolar macrophages and placed in a chemotaxis plate and the migration of neutrophils was measured. Chemotaxis of all cell types to be tested was measured by a change of absorbance on a microplate reader set at 492 nm. Results were compared with alveolar macrophages not stimulated with LPS, RPMI alone, and N formyl-methionyl-leucyl-phenylalanine (FMLP). The supernatant of the stimulated alveolar macrophages increased neutrophil chemotaxis as compared with unstimulated alveolar macrophages, and RPMI (p < .05). Surfactant treatment with 292 microg/mL significantly decreased LPS-stimulated alveolar macrophages induced neutrophil chemotaxis. Treatment with 875 microg/mL of surfactant did not alter neutrophil chemotaxis. CONCLUSIONS: Alveolar macrophages stimulation with LPS increased the chemotaxis of neutrophils. Treatment with surfactant at a concentration of 875 microg/mL did not alter neutrophil migration; however, treatment with 292 microg/mL significantly decreased neutrophil chemotaxis suggesting that at low concentrations, surfactant inhibits chemokine release and may reduce pulmonary neutrophil sequestration in vivo.     

Selective inhibition of T cell proliferation but not expression of effector function by human alveolar macrophages

BACKGROUND: Alveolar macrophages are thought to play an important part in regulating lung immune responses. While it is clear that human alveolar macrophages suppress T cell proliferation in vitro, the mechanisms by which this is achieved are not clear, nor is it known whether alveolar macrophages also inhibit other aspects of T cell function. METHODS: Peripheral blood mononuclear cells were stimulated with phytohaemagglutinin or house dust mite allergen, and cultured with variable numbers of autologous alveolar macrophages obtained by bronchoalveolar lavage from 20 normal subjects. RESULTS: Alveolar macrophages induced a reversible inhibition of T cell proliferation in response to both mitogen and allergen stimulation, with the latter being considerably more susceptible to inhibition. This was achieved via heterogenous mechanisms, involving both soluble factors derived from alveolar macrophages and cell-cell contact. Despite inhibiting proliferation, alveolar macrophages had little or no effect on T cell calcium flux, the characteristic changes in CD3, CD2, CD28 and interleukin-2 (IL-2) receptor expression which accompany normal T cell activation, and IL-2 and interferon gamma secretion. In contrast, alveolar macrophages inhibited the tyrosine phosphorylation of proteins which may be involved in IL-2 receptor-associated signal transduction. CONCLUSIONS: The immunoregulatory properties of alveolar macrophages are relatively selective, allowing T cell activation and cytokine secretion while inhibiting T cell proliferation within the lung.     

Serum human chorionic gonadotropin measurement in the diagnosis of ectopic pregnancy when transvaginal sonography is inconclusive

Transition from placental to pulmonary oxygenation at birth depends on a rapid removal of fetal lung fluid from the developing alveoli. Alveolar fluid clearance was examined in ventilated, anesthetized developing guinea pigs of the ages newborn, 2-d-old, 5-d-old, 30-d-old, and 60-d-old (adult). An isosmolar 5% albumin solution was instilled into the lungs of the guinea pigs; the guinea pigs were then studied for 1 h. Alveolar fluid clearance was measured from the increase in alveolar protein concentration as water was reabsorbed. Newborn guinea pigs had a very high alveolar fluid clearance rate that declined rapidly within the first 5 postnatal days towards adult levels. The high alveolar fluid clearance at birth was apparently mediated by the beta-adrenergic system as demonstrated by the elevated plasma epinephrine levels and the increased sensitivity to inhibition by the beta-adrenergic antagonist propranolol immediately after birth. Surprisingly, exogenous addition of epinephrine was not able to stimulate alveolar fluid clearance in the newborn lung, but exogenous epinephrine stimulation increased over time to adult levels. The elevated alveolar fluid clearance at birth was associated with a significantly greater amiloride sensitivity in the newborn guinea pig lung. Northern blot analysis of distal lung tissue as well as isolated alveolar epithelial type II cells showed and confirmed higher levels of the alpha-subunit of the epithelial sodium channel mRNA in the newborn lung that rapidly tapered off toward adult levels. In conclusion, these data demonstrate the importance of the beta-adrenergic system and amiloride-sensitive sodium transporting pathways for clearance of fetal lung fluid at birth.     

Immune adherence reactivity of rat alveolar macrophages following inhalation of crocidolite asbestos

The immune adherence phenomenon was used to demonstrate the in vivo deposition of complement on membranes of alveolar macrophages from rats chronically exposed to crocidolite asbestos dust. Pre-treatment of macrophage cualtures with anti-C3 antiserum greatly diminished the level of immune adherence reactivity. Alveolar macrophages exposed to crocidolite asbestos in vitro did not exhibit significant levels of immune adherence reactivity. These results may reflect an in-vivo antigen-antibody-complement interaction on the surface of a alveolar macrophages from animals which have inhaled asbestos dust.     

Quantitative differences in phagocytosis and degradation of Pneumocystis carinii by alveolar macrophages in AIDS and non-HIV patients in vivo

Bronchoalveolar lavage (BAL) specimens (n = 213) from AIDS and non-HIV immunosuppressed patients were investigated for the presence of Pneumocystis carinii infection by fluorescence microscopy of Papanicolaou-stained slides. Alveolar casts, extracellular pneumocysts and phagocytosed cysts and their degradation products in pulmonary alveolar macrophages were identified. The number of phagocytosed pneumocysts within human pulmonary alveolar macrophages was recorded and correlated with the number of extracellular cysts and alveolar casts, in both groups of patients. Both phagocytic and degradation capacity were depressed in AIDS patients. This observation may explain the large number of extracellular organisms found in BAL specimens of AIDS patients compared with non-HIV-positive immunocompromised individuals.     

Effect of alveolar lining material on phagocytic and bactericidal activity of lung macrophages against Staphylococcus aureau

Unstimulated rabbits were sacrificed and their lungs washed with heparinized saline. After alveolar macrophages were harvested, the cell-free lavage fluid was centrifuged at 47,000 X g to recover a small, whitish, surface-active pellet (F fraction.) The supernatant was concentrated 15-fold by vacuum dialysis (P fraction). Alveolar macrophages in a serum-free system were challenged with radiolabeled (32P) Staphylococcus aureus preincubated in either balanced salt solution or F or P fraction. A small increase in alveolar macrophage bacterial uptake occurred with P fraction-treated staphylococci. P fraction from locally immunized animals further enhanced phagocytosis. In bactericidial experiments, alveolar macrophages were allowed to phagocytize staphylococci preincubated in either balanced salt solution or F fraction. Intracellular bactericidal activity of alveolar macrophages was quantitated by lysotaphin lysis of extracellular bacteria and quantitation of viable intracellular bacteria. Enhanced lung macrophage bactericidal activity against F fraction-incubated staphylococci was noted.     

Bronchial adenoma resection with relief of hypoxic pulmonary vasoconstriction

Experimental airway obstruction is known to cause reflex pulmonary artery constriction, but clinical documentation of reversible bronchial obstruction and vasoconstriction is rare. A soft bronchial adenoma obstructed the left main bronchus, and scans showed minimal ventilation and perfusion on the left. Gas aspirated from beyond the tumor was hypoxic. The adenoma was removed and the lung left intact by means of a skin graft in the bronchial wall. Four months later, pulmonary function was normal, and both ventilation and perfusion of the left lung were normal. Reflex pulmonary vasoconstriction resulting from alveolar hypoxia minimizes systemic hypoxemia and also minimizes alveolar tissue hypoxia in the lung itself. The reflex is seen most frequently in perfusion scans in patients with chronic airways disease. This case in important in that it documents reversal of vasoconstriction after ventilation was restored.     

Perflubron decreases inflammatory cytokine production by human alveolar macrophages

OBJECTIVE: To determine whether inflammatory cytokine production by stimulated human alveolar macrophages is affected by perflubron exposure. DESIGN: Controlled laboratory investigation of alveolar macrophage function in vitro. SETTING: Research laboratory. SUBJECTS: Cultured alveolar macrophages obtained by bronchoalveolar lavage from eleven normal volunteers. INTERVENTIONS: Endotoxin-stimulated alveolar macrophages were treated with perflubron. MEASUREMENTS AND MAIN RESULTS: Alveolar macrophages were stimulated for 1 hr with lipopolysaccharide and then treated with perflubron for 23 hrs. Cell-free supernatants were collected and cytokines were assayed by enzyme-linked immunosorbent assay. Tumor necrosis factor-alpha, interleukin-1, and interleukin-6 were stimulated by lipopolysaccharide (endotoxin) and all of these cytokines were significantly (p < .05) inhibited by perflubron. Cell viability was not affected by perflubron. Basal cytokine concentrations from unstimulated alveolar macrophages were not altered by perflubron. CONCLUSIONS: Exposure of stimulated human alveolar macrophages to perflubron in vitro decreases cytokine production. This observation suggests that perflubron may have anti-inflammatory activity.     

Quantification of ammonia in human breath by the selected ion flow tube analytical method using H30+ and 02+ precursor ions

We show how our selected ion flow tube mass spectrometric technique for trace gas analysis can be used to determine the concentrations of ammonia in alveolar breath from single exhalations using both H30+ and 02+ precursor ions for chemical ionization. Thus, data are presented of the alveolar ammonia concentrations in the breath of six healthy volunteers following the ingestion of a liquid protein meal, which show that consistent values are obtained using these two precursor ions. Alveolar breath ammonia concentrations (which range from 200 to 1750 ppb in these individuals) are compared with those obtained from bag samples of breath from the same individuals.     

Distinct tumor necrosis factor-alpha responses in alveolar and peritoneal macrophages are associated with local levels of endotoxin

Tumor necrosis factor alpha (TNF-alpha) responses of alveolar macrophages (AMs) and peritoneal macrophages (PMs) were studied in rats after intravenous injection of lipopolysaccharide (LPS). High levels of plasma TNF-alpha, increased pulmonary myeloperoxidase activity, and leukopenia occurred within 2 h after LPS injection. Alveolar spaces exhibited a strict compartment property, as manifested by only slightly increased LPS and TNF-alpha levels in alveolar lavage fluid and an unchanged capacity of AMs to produce TNF-alpha. By contrast, the peritoneal cavity had greatly increased local LPS and TNF-alpha levels and a diminished PMs TNF-alpha response to LPS. The amount of LPS in the alveolar spaces was less than 0.2% of the level in peritoneal fluid. These results indicate that activation of resident macrophages is dependent on the amounts of local LPS and, in addition, suggest that resident AMs neither participate in the plasma TNF-alpha response nor contribute to neutrophil sequestration in the lung during the early stages of endotoxemia.     

Alveolar hemorrhage in mixed cryoglobulinemia associated with hepatitis C virus infection

Alveolar hemorrhage in mixed cryoglobulinemia associated with hepatitis C virus infection. A 61 year-old woman with type II mixed cryoglobulinemia associated to hepatitis C virus infection has suffered alveolar hemorrhage with multiple pulmonary infiltrates, purpura, glomerulonephritis and polyneuropathy. The respiratory and kidney findings resolved with prednisone, but glomerulonephritis reappeared when interferon-alpha treatment was started and prednisone was reduced. This is the third case of alveolar hemorrhage and glomerulonephritis associated with mixed cryoglobulinemia reported in the literature. The lung involvement in mixed cryoglobulinemia is reviewed. The clinic manifestations (asthma, pleural effusion, hemoptysis or pulmonary fibrosis) are uncommon, but the lung involvement is very frequent if roentgenographic signs and necropsy findings are assessed.     

Lung tissue plasticity: morphometric analysis of anisotropic strain in liquid filled lungs

Lungs of rats were fixed at different inflation pressures (Ptp) during liquid filling with the pulmonary vessels tied to prevent vascular volume change after fixation had begun. Morphometric analysis showed that alveolar surface varied as a alveolar volume (Va) to the power 0.82, while the arithmetic mean tissue thickness varied as Va-0.2. This is interpreted as evidence for anisotropic expansion. Capillary volume (Vc) was found to increase from zero Ptp to a maximum at Ptp = 2 cm H2O then decrease as Va increased. Morphometric diffusion capacity of the membrane component increased as Va0.59 while that for whole lung (DL)paralleled the change in Vc. Alveolar capillary tissue unfolding is described as the main factor accounting for anistropic expansion of alveolar surface and for capillary configuration. The absolute values of Vc and DL were lower by 60% and 50%, respectively, compared with values obtained by standard instillation fixation methods and this is suggested could account for previous discrepancres between morphometric and physiologic estimates of DL.     

Alveolar liquid clearance is increased by endogenous catecholamines in hemorrhagic shock in rats

The primary objective of this study was to test the hypothesis that hemorrhagic shock would stimulate alveolar liquid clearance by a catecholamine-dependent mechanism. Anesthetized rats were hemorrhaged to a mean arterial pressure of 30 mmHg for 90 min, but they were not resuscitated. Alveolar liquid clearance was measured by the concentration of labeled and unlabeled protein over 2 h in an isosmolar physiological solution of 5% albumin that had been instilled into one lung. Hemorrhaged rats developed a severe metabolic acidosis that was associated with a 5- to 10-fold rise in plasma epinephrine levels. There was a 60% increase in alveolar liquid clearance in the hemorrhaged rats compared with control rats (55 +/- 6 vs. 34 +/- 7%; P < 0.05). Amiloride (10(-4) M) or propranolol (10(-4)M) inhibited the increase in alveolar liquid clearance. Thus the endogenous release of catecholamines associated with hemorrhagic shock markedly stimulates alveolar fluid clearance by a beta-adrenergic-mediated stimulation of active sodium transport. These data suggest a new, previously unrecognized mechanism that may protect against alveolar flooding in the acute phase of hemorrhagic shock.     

Intraoperative modulation of alveolar macrophage function during isoflurane and propofol anesthesia

BACKGROUND: Alveolar macrophages are a critical part of the defense against pulmonary infection. Thus the authors determined time-dependent changes in alveolar macrophage functions in patients having surgery who were anesthetized with isoflurane or propofol. METHODS: Patients anesthetized with propofol (n = 30) or isoflurane (n = 30) during orthopedic surgery were studied. Alveolar macrophages were harvested by bronchoalveolar lavage immediately, and 2, 4, and 6 h after induction anesthesia and at the end of surgery. The fraction of aggregated and nonviable macrophages was determined. Then phagocytosis was measured by ingestion of opsonized and unopsonized particles. Finally, microbicidal activity was determined as the ability of the macrophages to kill Listeria monocytogenes directly. RESULTS: Demographic and morphometric characteristics of the patients given propofol and isoflurane were similar, as were their levels of pulmonary function and hemodynamic responses. The fraction of alveolar macrophages ingesting opsonized and unopsonized particles, and the number of particles ingested, decreased significantly over time, with the decrease slightly but significantly greater during isoflurane anesthesia. Microbicidal function decreased progressively during anesthesia and surgery, with the decrease almost twice as great during isoflurane compared with propofol anesthesia. The fraction of aggregated macrophages and recovered neutrophils increased over time in the patients given each anesthetic. CONCLUSIONS: Pulmonary immunologic function changed progressively during anesthesia and surgery. The data from this study suggest that pulmonary defenses are modulated by the type of anesthesia and by the duration of anesthesia and surgery.     

Noninvasive assessment of the direct action of oral nifedipine and nicardipine on left ventricular contractile state in patients with systemic hypertension: importance of reflex sympathetic responses

BACKGROUND: Alveolar macrophages from patients with sarcoidosis were analyzed for their ability to secrete tumor necrosis factor-alpha (TNF-alpha), interleukin-1-beta (IL-1-beta), and interleukin-6 (IL-6). RESULTS: Constitutive release of all three monokines in these patients was concomitantly increased in the active state of disease in comparison with inactive sarcoidosis or healthy control subjects. Alveolar macrophages from patients with inactive sarcoidosis compared with cells from healthy subjects showed increased spontaneous secretion of TNF-alpha and IL-6 only, whereas the constitutive release of IL-1-beta was similar as in healthy volunteers. In vitro stimulation of alveolar macrophages from healthy control subjects with lipopolysaccharide or pokeweed mitogen led to a time- and dose-dependent enhanced secretion of TNF-alpha, IL-1-beta, and IL-6. In a similar manner, with corresponding cells from patients with sarcoidosis the secretion of all three cytokines could be further increased by stimulation with lipopolysaccharide or pokeweed mitogen. CONCLUSIONS: The data presented indicate that an increased release of TNF-alpha, IL-1-beta, and IL-6 correlates to disease activity and may play a critical part in the pathogenesis of sarcoidosis.     

Beta 2-agonist treatment reduces beta 2-sensitivity in alveolar macrophages despite corticosteroid treatment

Alveolar macrophage beta 2-adrenoceptor sensitivity and bronchodilator responses to inhaled terbutaline were investigated before and after 2 wk of oral treatment with terbutaline 7.5 mg twice a day in healthy volunteers. The influence of corticosteroid treatment was examined by giving 10 subjects budesonide 400 micrograms twice a day by inhalation throughout the treatment period, and by giving 10 subjects 40 mg prednisolone and 10 subjects placebo orally 12 h before the second examination. Terbutaline treatment elicited marked attenuation (approximately 75% reductions) of isoprenaline-induced cyclic AMP accumulation in the alveolar macrophages. Responses to prostaglandin E1 were not influenced by treatment, suggesting homologous beta-adrenoceptor desensitization. Corticosteroid administration failed to either prevent (budesonide) or reverse (prednisolone) this desensitization. Bronchodilator responses to terbutaline were not altered by treatment in either group. We conclude that the beta 2-adrenoceptor sensitivity of human alveolar macrophages is markedly and selectively depressed by beta 2-agonist treatment and that corticosteroid treatment, contrary to previous hypotheses, fails to influence this phenomenon. Studies on the mechanisms involved are needed. The importance of alveolar macrophages in asthma is unclear, but the present data in humans are of interest in relation to possible effects of continuous beta 2-agonist treatment on inflammatory mechanisms in the airways.     

Submergence of roots for alveolar ridge preservation. A failure (4-year follow-up study)

A 4-year clinical and radiographic follow-up study of 20 cases of crown-resected root-filled roots, covered by mucosal flaps, distributed among 15 patients, has been carried out. Only cases of uneventful submergence of the root with intact healthy mucosa were regarded as successful. This study shows that: (1) The number of failures increases with the years; from 3 cases at the 1-year to 11 cases at the 4-year follow-up, corresponding to 53% (8/15) of the patients. In the failures, an exposure of the root-surface was seen, but without inflammation in the surrounding tissue. These roots could be extracted without bone loss. (2) Alveolar ridge atrophy is not prevented by retained roots, and is probably the primary reason for failure of coverage. The 11 failures showed alveolar ridge atrophy, which was observed before failure in 6 cases. None of the successful cases showed alveolar ridge atrophy. The present method cannot be recommended as a routine procedure.     

Tissue specificity of spontaneous point mutations in lambda supF transgenic mice

Abuse of nitrite inhalants, widespread among male homosexuals, has been identified by epidemiological studies as an independent risk factor for AIDS and for Kaposi's sarcoma. Subchronic exposure of mice to inhaled isobutyl nitrite was previously found to impair the tumoricidal activity of peritoneal macrophages. Because inhalants would be expected to have the greatest effects on cells in the lung, alveolar macrophages from exposed mice were examined in this study. Mice were exposed to 900 ppm isobutyl nitrite in an inhalation chamber for 45 min/day for 14 days. Following this treatment, the lungs of exposed mice had large increases in cellularity, both in the alveolar septa and within the alveoli. Bronchoalveolar lavages also contained increased numbers of cells. Alveolar macrophages collected from treated mice had increased tumoricidal activity compared with controls and produced higher levels of inducible nitric oxide and tumor necrosis factor-alpha (TNF-alpha). The frequency of alveolar cells secreting TNF-alpha was increased ninefold in mice exposed to the inhalant. Cell influx into the lung, as indicated by the presence of red blood cells in lung lavages, was evident after only a single 45-min exposure to inhaled isobutyl nitrite at doses as low as 300 ppm.     

Reversal of narcotic depression in the neonate by nalozone

Naloxone 40 mug was administered intravenously one minute after birth to 20 out of 44 neonates whose mother had been given pethidine in labour. These neonates were compared with 20 others whose mothers had had only lumbar epidural block. Alveolar PCO2, alveolar ventilation, and ventilatory rate were measured 10 and 30 minutes after birth. The untreated neonates of mothers who had had pethidine showed significant ventilatory depression compared with infants in the epidural and naloxone-treated groups. The naloxone-treated neonates were comparable with the epidural group, although the effects of naloxone were diminishing at 30 minutes. Naloxone is an effective narcotic antagonist which should be considered to be the drug of choice for treating narcotic depression in the neonate.     

Effect of zinc supplementation on cell-mediated immunity and lymphocyte subsets in preschool children

The pattern and distribution of periodontitis were investigated in 162 randomly selected dogs available for necropsy in veterinary practice. There were 82 males and 80 females of 50 different breeds (150 dogs were pure-bred and 12 were mongrels, aged between 7 months and 14 yr. Presence of periodontitis was determined by assessment of alveolar bone loss on radiographs of the skulls and jaws. Periodontitis occurred frequently with increasing age, although the prevalence varied markedly among and within different breeds. Of the breeds most represented in the sample, periodontitis was most frequently seen in poodles and dachshunds but was rarely recognized in German shepherd dogs. Regardless of age, the vast majority of the dogs displayed either one or both of two different radiographic patterns of alveolar bone loss. One pattern was characterized by slight, horizontal alveolar bone loss involving interradicular and interdental areas. The other pattern was one of predominantly crater-like, or narrow, vertical bone defects which, when advanced, often extended around a single root or tooth to surround the root apices. The two types of patterns did not seem to be breed-dependent. The posterior maxillary and mandibular premolars and molars were the most frequently affected teeth. Alveolar bone loss was most severe in the maxilla, while corresponding bone loss in the mandible was more often related to increasing age.