Macrophage development: IV. Effects of blood factors on macrophages from prenatal rat lung cultures

Effects of colony-stimulating factors M-CSF, GM-CSF, G-CSF, and IL-3 were assessed on cells of macrophage lineage present in organ cultured 14-day prenatal rat lungs. Treatment groups were compared between one another and against control lungs grown on standard medium containing 40% fetal bovine serum without added factors, where a monoculture of macrophages rapidly develops from precursors present at explantation, leading to appearance of a large mature population on the pleural surface outside the lungs. Studies were carried out in living cultures and by light and electron microscopy using peroxidase-coupled isolectin B₄ of Griffonia simplicifolia to identify macrophages and their precursors. In the first experiment, 14-day prenatal lung explants (14+0 days) containing macrophage precursors but not matured cells were exposed to individual CSFs for 7 days in an attempt to determine whether precursors are committed irrevocably to the macrophage line or can be altered by exposure to factors promoting significant granulocyte development. In succeeding experiments, 4- and 7-day-old cultures (14+4, 14+7 days) containing matured macrophages were targeted to see whether macrophage survival can be extended beyond expectations in controls and whether mitotic activity is stimulated. Recombinant CSFs were used at dosage levels known to promote colony formation in vitro (200–1,000 CFU/ml). Cultures exposed from prenatal day 14 to M-, GM-, G-CSF, or IL-3 yielded a monoculture of macrophages without exception. Populations developed in the presence of M- or GM-CSF were much larger than in controls or cultures grown with the other blood factors. GM-CSF-exposed cultures produced by far the largest macrophages, among them many multinucleate giant cells. Macrophages developed in the presence of G-CSF were also significantly larger than controls. Growth of the mature macrophage population was greatly stimulated by exposure to M-CSF or GM-CSF but not by IL-3 or G-CSF. Mitotic figures were noted in the coronas of emerged cells surrounding stimulated cultures, compared to none in the controls. Ultrastructurally, macrophages stimulated by M-CSF retained a mature appearance like macrophages in control, IL-3, and G-CSF treatment groups, whereas many in the GM-CSF group became less differentiated. As to long-term survival, a single 14-day explant was grown for 8 days on standard medium (the equivalent date for birth), then placed in a soft agar medium containing M-CSF. Supplemented irregularly by M-CSF and GM-CSF, the culture remained viable until fixed on the 137th “postnatal” day and retained a small population of macrophages. Conclusions: (1) the macrophage lineage from embryonic rat lungs can be manipulated in culture; (2) macrophage precursors in these lungs seem committed to the macrophage line; (3) replication of both immature and mature macrophages is stimulated by M-CSF and GM-CSF; (4) with M-CSF, however, retention of mature characteristics and longevity are favored, whereas with GM-CSF maturity is partly lost and formation of giant cells emphasized. © 1992 Wiley-Liss, Inc.     

Alveolar macrophages. II. Inhibition of lymphocyte proliferation by purified macrophages from rat lung.

Macrophages were prepared from the lung, peritoneal cavity and blood of normal, unstimulated rats from a number of strains. The macrophages were purified by adherence, and characterized via surface markers, enzyme activity and phagocytic capacity, and subsequently tested for activity in cultures of mitogen-stimulated syngeneic lymphocytes. Peritoneal macrophages and blood monocytes were mildly stimulatory, or ineffective in modulating mitogen-induced DNA synthesis; peritoneal macrophages reconstituted the blastogenic responses of macrophage-depleted lymph node cell cultures to normal limits. In contrast, alveolar macrophages were markedly inhibitory to lymphocyte proliferation; in some instances inhibitory activity was demonstrable when added alveolar macrophages comprised only 0.04% of the total cells in culture. Lymphocyte proliferation induced by T-cell mitogens was more susceptible to this inhibition than was proliferation induced by the B-cell mitogen LPS. Alveolar macrophages recovered from SPF rats, while less in number, exhibited comparable inhibitory activity. These results form part of an emerging picture picture of the normal alveolar macrophage as a potential 'suppressor' of T-cell activity in the lung.     

Macrophage subpopulations regulate intrathymic T-cell development. I: Ia-positive macrophages augment thymocyte proliferation

The contribution of H2-Ia-positive thymic macrophages (Ia 1 thymic M phi) to intrathymic lymphopoiesis was investigated. An isolation method yielding cell suspensions highly enriched for Ia+ thymic M phi was performed. Cocultivation of these Ia+ thymic M phi with thymocytes showed that, while not affecting spontaneously proliferating thymocytes, the Ia+ thymic M phi strongly augmented the mitogen-induced proliferation of thymocytes by about 200%. This effect was dependent on the number of Ia+ M phi added as well as on the degree of thymocyte maturity: stronger augmentation occurred at higher concentrations of M phi and immature thymocytes showed highest susceptibility to the Ia+ thymic M phi-mediated effect. Cytochalasin B was employed to prove that cellular interaction is an important prerequisite for the proliferation amplifying effect of Ia+ thymic M phi. Additionally, humoral factors produced by Ia+ thymic M phi after induction with LPS are also involved in the described phenomenon. Furthermore, the use of interleukin preparations in the thymocyte-Ia+ M phi cocultures demonstrated that humoral factors support or probably regulate the interaction of these cells. The implications of these findings in view of the proliferation and differentiation events of thymocytes within the thymus are discussed.     

Suppressive activity of alveolar macrophages and blood monocytes from interstitial lung diseases: role of released soluble factors

Two groups of patients suffering from interstitial lung diseases (ILD) namely sarcoidosis (SA) and idiopathic pulmonary fibrosis (IPF) were investigated for alveolar macrophages (AM), secretion of prostaglandin E2 (PGE2) and interleukin 1 (IL-1), together with superoxide anion (O2-) production. Peripheral blood monocytes (PBMO) of the same patients were examined concomitantly for suppressive activity. Consistent with previous results, AM obtained by bronchoalveolar lavage (BAL) from ILD patients markedly suppressed the effects of PHA stimulation of autologous peripheral blood lymphocytes (APL): 61.8 +/- 9.7% suppressive activity compared to 15.5 +/- 15.4% in the control group (CO) P less than 0.001. The AM suppressive activity was correlated with an increase in PGE2 secretion: 3.861 +/- 2.194 ng/10(5) cells/ml in the IPF group, but not in the sarcoid group: 0.217 +/- 0.116 ng/10(5) cells/ml (P less than 0.001 between them). On the other hand, IL-1 secretion by AM was greatly increased in sarcoid patients (308 +/- 196 U/ml) but was within the normal limits in IPF (27.3 +/- 28.8 U/ml, P less than 0.01 between them). Therefore, an inverse correlation was found between degree of PGE2 secretion and IL-1 release by AM in ILD. O2-production by AM was markedly increased in all ILD patients but this mechanism is apparently not involved in suppressive activity. PBMO originating from ILD patients were less suppressive than the corresponding AM.     

Macrophage colony development: properties of colony stimulating factors from murine embryo and pregnant uterus.

Extracts from embryonic and uterine tissue of mice, operationally defined as colony stimulating factor (CSF), promoted the growth of macrophage-granulocyte colonies in vitro. Uterine CSF focusses from pH 5.15 to 6.00 and embryonic CSF from pH 3.60 to 5.20, although both forms have similar biological activity. CSF is relatively resistant to denaturation but it is inactivated by periodate and dithiothreitol. Gel filtration indicates a molecular weight of 45,000 which is unchanged following treatment with insolubilized trypsin, a procedure which affords a useful purification (240-fold). Trypsin-sensitive material in CSF preparations modifies colonial form under certain conditions of culture, probably by increasing the motility of macrophages. Diaminoethane derivatives of CSF were prepared and retained biological activity at isoelectric points above pH 9.0. These derivatives may be covalently linked to Sepharose providing an insolubilized form of CSF to study interactions of CSF with the cell surface.     

Macrophage development: III. Transformation of pulmonary macrophages from precursors in fetal lungs and their later maturation in organ culture.

The fate of macrophage precursors residing in 14-day prenatal rat lungs was followed in organ cultures to obtain a detailed, ultrastructurally resolved picture of the sequence and timing of events accompanying their transformation into typical pulmonary macrophages. Cultures were examined at close intervals during the first day (1, 2, 3, 4, 6, 9, 12, 15, 18, and 24 hr) and at wider intervals thereafter (2, 4, 5, 7, 9, and 13 days) to yield a developmental series of cells identified as in the macrophage line based on binding of peroxidase-coupled isolectin B4 of Griffonia simplicifolia (GSA I-B4) to cell membranes and on negligible content of peroxidase-positive granules in the cytoplasm. Organ culturing stimulated virtually all precursors to develop into macrophages. GSA-positive cells in explants occurred outside vessels in pulmonary connective tissue, and at the outset none were typical macrophages: 71% were angular cells, resembling unlabeled mesenchymal cells around them, 16% were undifferentiated leukocytes, and the remainder were irregularly shaped cells with few vacuoles intermediate between the preceding and the macrophages. During the first 12 hr in culture the proportion of angular cells and leukocytes fell to zero, and that of intermediate cells first rose, then receded. In the same interval the proportion of macrophages rose to 87.5%, and by 24 hr all GSA-positive cells were typical macrophages generally engorged with phagocytosed material; about 8 hr appear necessary for converting half the population. Notable ultrastructural changes during this period of transformation involved the centrioles and cytoskeleton, reflecting enhanced cell mobility and phagocytosis. A period of maturation followed, marked by disappearance of cellular debris from phagosomes and an increased prevalence of cells with elaborate lamellipodia. This accords with earlier work showing that macrophage Fc receptor density increases sharply during the first 24 hr, but elevated levels of histochemically demonstrable acid phosphatase appear only later. Mitotic activity was conspicuous in GSA-positive cells throughout both periods. 3H-thymidine labeling indices for precursors and macrophages, determined at six intervals between 1 hr and 24 hr, remained steady at approximately 34%, whereas indices of other categories of lung cells (GSA-negative stromal cells, pleural cells, and airway epithelium) began at this level but rapidly declined, indicating that the GSA-positive cells constitute a single population distinct from others in the lungs. Macrophages found outside the lung cultures after 4-5 days qualify as a mature population, but having migrated away from direct contact with the lung stroma, they survive only a week or two and no longer divide.     

In vivo prenatal chlordane exposure induces development of endogenous inflammatory macrophages.

Macrophages (m phi s), important cells in host resistance, undergo a series of biochemical changes during their progression from the resident to the fully activated stage. Both resident and inflammatory m phi s are characterized by some unique properties. In the present study, female BALB/c mice were prenatally treated with 8 mg/kg body weight of chlordane, a cyclodiene poly-chlorinated hydrocarbon that appears to reduce immunocompetence by selectively impairing m phi function. Therefore, we examined functions in m phi s from chlordane-treated mice that had been stimulated with thioglycollate. The 5'-nucleotidase activity, present in high levels in resident m phi s but low levels in inflammatory m phi s was elevated in resident m phi s from vehicle-exposed animals. Conversely, inflammatory m phi s from these animals showed significantly diminished levels of this function. Moreover, chlordane-exposed m phi s, regardless of whether they were resident or inflammatory, exhibited decreased 5'-nucleotidase responses. When a second function, transferrin receptor binding, was analyzed, vehicle-treated inflammatory m phi s displayed high levels of activity whereas the resident m phi s showed very little transferrin binding. However, both resident and inflammatory m phi s from the chlordane-exposed group demonstrated transferrin binding activity similar in magnitude to that of the vehicle-treated inflammatory m phi s. Finally, two-dimensional polyacrylamide gel electrophoresis analysis of m phi s from chlordane-exposed mice have characteristics of normal m phi s that have advanced to the inflammatory stage.     

Anti-fungal susceptibility and virulence factors of Candida spp. isolated from blood cultures

Candidemia is one of the most common fungal nosocomial infections worldwide. It causes high mortality and morbidity rate with significant hospital costs due to increased length of hospital stay and costs for anti-fungal treatment. This study aims to investigate anti-fungal drug susceptibility, enzymatic activity and biofilm formation of the Candida spp. isolated from blood cultures. In 2016, a total of 84 clinical Candida isolates were analyzed for minimum inhibitory concentration (MIC) against fluconazole and amphotericin B by agar diffusion E-test (E-strips). Three enzymatic activity tests for phospholipase, proteinase and esterase were performed by using egg yolk agar, bovine serum albumin medium and Tween 80 opacity medium, respectively. Biofilm formation was determined by crystal violet staining. To describing the various Candida distributions cultured, C. albicans was the most frequent species (n = 37, 44.1%), followed by C. tropicalis (n = 30, 35.7%), C. parapsilosis (n = 8, 9.5%), C. glabrata (n = 6, 7.1%) and C. guilliermondii (n = 3, 3.6%). Regarding anti-fungal drug susceptibility, C. albicans was susceptible to fluconazole (100%). In addition, all clinical Candida isolates were fully susceptible to amphotericin B (100%). The predominant enzyme activity of C. albicans included medium to high levels of phospholipase, proteinase and esterase activities. C. tropicalis displayed esterase activity, while C. glabrata and C. guilliermondii had no phospholipase and proteinase activity. Non-albicans Candida (NAC) i.e. C. tropicalis formed a biofilm at a higher rate than C. albicans. This study revealed the production of virulent factors in Candida strains from candidemia patients.     

Macrophage development: III. Transformation of pulmonary macrophages from precursors in fetal lungs and their later maturation in organ culture

The fate of macrophage precursors residing in 14-day prenatal rat lungs was followed in organ cultures to obtain a detailed, ultrastructurally resolved picture of the sequence and timing of events accompanying their transformation into typical pulmonary macrophages. Cultures were examined at close intervals during the first day (1, 2, 3, 4, 6, 9, 12, 15, 18, and 24 hr) and at wider intervals thereafter (2, 4, 5, 7, 9, 12, and 13 days) to yield a developmental series of cells identified as in the macrophage line based on binding of peroxidase-coupled isolectin B₄ of Griffonia simplicifolia (GSA I-B₄) to cell membranes and on negligible content of peroxidase-positive granules in the cytoplasm. Organ culturing stimulated virtually all precursors to develop into macrophages. GSA-positive cells in explants occurred outside vessels in pulmonary connective tissue, and at the outset none were typical macrophages: 71% were angular cells, resembling unlabeled mesenchymal cells around them, 16% were undifferentiated leukocytes, and the remainder were irregularly shaped cells with few vacuoles intermediate between the preceding and the macrophages. During the first 12 hr in culture the proportion of angular cells and leukocytes fell to zero, and that of Intermediate cells first rose, then receded. In the same interval the proportion of macrophages rose to 87.5%, and by 24 hr all GSA-positive cells were typical macrophages generally engorged with phagocytosed material; about 8 hr appear necessary for converting half the population. Notable ultrastructural changes during this period of transformation involved the centrioles and cytoskeleton, reflecting enhanced cell mobility and phagocytosis. A period of maturation followed, marked by disappearance of cellular debris from phagosomes and an increased prevalence of cells with elaborate lamellipodia. This accords with earlier work showing that macrophage Fc receptor density increases sharply during the first 24 hr, but elevated levels of histochemically demonstrable acid phosphatase appear only later. Mitotic activity was conspicuous in GSA-positive cells throughout both periods. ³H-thymidine labeling indices for precursors and macrophages, determined at six intervals between 1 hr and 24 hr, remained steady at ～ 34%, whereas indices of other categories of lung cells (GSA-negative strimal cells, pleural cells, and airway epithelium) began at this level but rapidly declined, indicating that the GSA-positive cells constitute a single population distinct from others in the lungs. Macrophages found outside the lung cultures after 4–5 days qualify as a mature population, but having migrated away from direct contact with the lung stroma, they survive only a week or two and no longer divide.     

Effects of prenatal procarbazine administration on intrauterine development in rats.

The effects of prenatal procarbazine (PCZ) administration on the intrauterine development of rat fetuses were investigated. Gravid rats were treated on day 14 of gestation (GD14) with 25 mg or 50 mg/kg body weight PCZ via stomach tube. Controls received normal saline in the same dosis and manner. On GD20, all fetuses were collected by caesarian section. Live and dead fetuses as well as resorptions were counted. In the live fetuses, the following investigations were conducted: measurement of body weight, occipito-coccygeal-lenght (OCL), tail length (TL), placental weight and diameter, external macroscopic and binocular microscopic examination, and sectional analysis of the animals using the razorblade sectioning technique. Both PCZ doses caused a significant reduction in the number of live fetuses and a significant increase in resorptions. Mean body weight in PCZ groups was antidromic affected. OCL and TL were significantly depressed. Placental weight and diameter as well as number of dead fetuses were comparable to those of controls. External and sectional investigations revealed no PCZ-related deviations. In the light of our findings we conclude that PCZ in the doses used in this experimental study significantly affects the intrauterine development in rats in terms of fetal toxicity but displays no teratological properties.     

Effects of prenatal morphine exposure on rat heterotypical sexual behavior.

Prenatal exposure to morphine inhibits ovarian steroid-dependent lordosis behavior in female rats, and enhances certain components of male sexual behavior in male rats. In the present study, the effects of mid to late gestational morphine exposure on male sexual behavior in females and on female sexual behavior in males were examined in adult offspring. Gonadectomized male rats were injected at weekly intervals with 30 or 60 microg estradiol benzoate and 1.0 mg progesterone and tested for female sexual behavior with stimulus males on 2 consecutive weekly tests. Ovariohysterectomized (OVX) females were injected with 500 microg testosterone propionate (TP) daily for 15 days and tested for male sexual behavior with stimulus females on the last day of TP injection and 1 week later, after TP withdrawal. Prenatal morphine exposure increased the expression of male sexual behaviors in female rats, but it did not increase lordosis behavior in male rats. Thus, exposure to morphine during gestation alters male and female sexual behavior in young adult animals. Because prenatal morphine exposure both defeminized and masculinized adult sexual behavior in female rats, it is possible that female brain development is more vulnerable to prenatal insult such as opiate exposure.     

Effect of interferon-gamma on the 5-lipoxygenase pathway of rat lung macrophages.

In view of conflicting reports concerning the effect of macrophage activation on arachidonic acid metabolism, we examined the effect of the macrophage activator, interferon-gamma (IFN-gamma), on the 5-lipoxygenase pathway in rat lung macrophages. Rat lung macrophages were conditioned in the presence or absence of 10(2) U/ml IFN-gamma for 4 h before stimulation with 1 microM A23187 for 15 min or 100 micrograms/ml opsonized zymosan for 60 min at 37 degrees C as well as other stimuli. Lipoxygenase products in extracted cell supernatants were identified and analyzed by high-pressure liquid chromatography and ultraviolet spectroscopy. The predominant lipoxygenase products included leukotriene (LT) B4, LTC4, and 5-hydroxyeicosatetraenoic acid (5-HETE). These products were not qualitatively altered by conditioning with IFN-gamma. However, 5-lipoxygenase pathway activity, as measured by LTB4 release, was maximally increased 2-fold after conditioning with IFN-gamma and stimulating with either A23187 or opsonized zymosan. IFN-gamma-conditioned macrophages, stimulated with A23187, released greater quantities of lipoxygenase products in comparison with control cells (307.6 +/- 13.3 versus 167.6 +/- 3.9 pmol LTB4/10(6) cells) (mean +/- SEM) (P less than 0.05). Similar results were obtained with the less potent stimulus, opsonized zymosan. IFN-gamma had no direct stimulatory effect on the 5-lipoxygenase pathway. No effect was observed with a variety of other stimuli with or without IFN-gamma conditioning.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of maternal prenatal stress on offspring development: a commentary

Pregnancy is associated with major physiological changes and adaptation to these changes is crucial for normal fetal development. Heightened emotional stress during pregnancy may interfere with the necessary adaptation and lead to dysregulation of the two major stress response systems: the Hypothalamic-Pituitary-Adrenal (HPA) Axis and the Autonomic Nervous System (ANS). Negative effects on the fetus of such maladaptation have been documented in both animals and humans and range from poor birth outcomes to negative impacts on neurodevelopment, as well as long term emotional and behavioural disturbances. Conversely, it has been hypothesized that low levels of maternal prenatal stress may actually have an adaptive value for the offspring. Investigation of these associations employing physiological markers and repeated measures throughout pregnancy and postpartum of both the mother and the offspring, is required in order to understand the various effects of prenatal stress on the development of the offspring. It is also crucial to explore the possibility of variable periods of vulnerability throughout gestation. The aim of this commentary is to reexamine the current literature on the ill-effects of maternal stress during pregnancy on the offspring and to explore avenues for future treatment and prevention.     

Functional immaturity of rat alveolar macrophages during postnatal development.

Alveolar macrophages (AM) are important in the regulation of immune responses in the lung, through their role as scavenger cells and through the production of many bioactive factors. Because in early infancy pulmonary infections are a recurrent problem, we studied the postnatal functional maturation of AM in a rat model. AM were isolated from rat lungs by bronchoalveolar lavage at several time intervals after birth and tested for their ability to ingest Escherichia coli in the presence of surfactant protein A (SP-A). Furthermore, their capacity to produce nitric oxide (NO) and interleukin-1 beta (IL-1 beta) after in vitro lipopolysaccharide (LPS) stimulation was analysed, as well as their capacity to downregulate proliferation of T cells from both mature and neonatal rats. SP-A-mediated phagocytosis of E. coli by AM was reduced in 14-day-old neonatal rats, as compared with mature rats (P < or = 0.05). Also the IL-1 beta production by rat AM after LPS stimulation was impaired at 14 days of age, as compared with IL-1 beta production by AM from mature rats (P < or = 0.05). In contrast, the LPS-induced NO production by rat AM as well as the capacity to inhibit T-cell proliferation were well developed at all ages tested. In conclusion, during postnatal development the rat AM is functionally immature, with respect to phagocytosis and secretion of inflammatory mediators. These differences may underly the enhanced susceptibility to pulmonary infections as found in human neonates.     

Structural analysis of fetal rat lung development.

The primary aim of this morphological investigation was to elaborate a concept allowing us to coherently define reference spaces for morphometric analysis of fetal lung development. Beyond this quantitative goal, morphological analysis of cell types, definition of compartments, and reflection about the prospective fate of their constituents provided per se new insights into the developmental processes. Lungs of rat fetuses aged 17-23 days and newborns aged 20 hours were fixed with an osmium tetroxide and glutaraldehyde mixture and their volume determined. Left lungs were embedded in Epon and investigated by light and electron microscopy. The right lung of one animal per group was embedded in methacrylate and step sections obtained to precisely locate the airways within the mesenchyme. The various cell types, their topographical relationships, and their morphological alterations with ongoing development were analyzed with regard to their prospective potentials of differentiation. The developing lung could be partitioned into four zones further subdivided into defined compartments. Zone I forms a superficial mantle around the lobes and the future acini. Consisting of primitive mesenchymal cells, it represents a zone of growth which disappears with the onset of the saccular stage. Zone II is mainly a zone of differentiation. Its interstitium stains intensely due to a dense population of dark cells. Up to gestational day 19, zone II contains future conductive airways with their vessels. After day 21, it comprises the whole prospective gas exchange region. Zones III and IV contain the elements of the airway tree and vascular system, zone IV corresponding to the most proximal generations with an adventitial layer. For all differentiation processes, a centrifugal directionality is manifested.     

Cellular organization and development of slice cultures from rat visual cortex

Summary Slice cultures from the visual cortex of young rats were prepared using the roller culture technique (Gähwiler 1984). After 10 days in vitro the cortical cultures flattened to 1–3 cell layers, surviving for up to 12 weeks. The cultures were organotypically organized, the typical layered structure of the cortex was preserved. The neuronal composition of slice cultures was studied using intracellular staining, Golgi impregnation and GABA immunohistochemistry. Both pyramidal cells and several types of nonpyramidal cells were identified in the slice cultures. Electrophysiological recordings showed that the electrical properties of cells in culture were similar to those measured in acute slice preparations; for some cells, however, the spontaneous activity was higher. The maintained activity was strongly increased by application of the GABA antagonist bicuculline and decreased by GABA, suggesting that GABAergic inhibition is present in these preparations. We could observe the postnatal maturation of some characteristic morphological features in culture. For example, pyramidal cells in 6 day-old rats in situ have very short basal dendrites with growth-cones, and the dendrites are free of spines. After 2–3 weeks in culture growth-cones were no longer observed. Instead, the cells had developed a large basal dendritic field and the dendrites were covered with spines. Slice cultures therefore may provide a useful tool for physiological, anatomical, pharmacological and developmental studies of cortical neurons in an organotypical environment.     

Superoxide anion release from blood monocytes and alveolar macrophages in patients with diffuse lung fibrosis.

Superoxide anion release (O2-) after stimulation with phorbol myristate acetate was measured in alveolar macrophages (AM) obtained by bronchoalveolar lavage and in blood monocytes from 47 patients with diffuse interstitial lung disease: idiopathic pulmonary fibrosis (N = 15), hypersensitivity pneumonitis (N = 7), pneumoconiosis (N = 6) and sarcoidosis (N = 19). Differential cell counts demonstrated a lymphocyte predominance in patients with hypersensitivity pneumonitis (HP) and sarcoidosis while the other groups had neutrophil predominance. No correlation between O2- activity in alveolar macrophages (AM) or blood monocytes (BM) compared to lung function (VC and diffusing capacity) could be demonstrated. Smoking pneumoconiotics had significantly decreased BM O2- release (1.25 +/- 0.25 (SEM) nmol/min/10(6) cells) and significantly increased AM/BM O2- ratios (2.04 +/- 0.26) compared to smokers with idiopathic pulmonary fibrosis (IPF) who had the following mean values: BM O2- release = 2.58 +/- 0.25 and AM/BM O2- ratio = 0.86 +/- 0.23. When matched for sex and smoking, a significantly increased AM/BM O2- ratio was seen among patients with HP (2.19 +/- 0.98) in comparison with patients who had sarcoidosis (0.40 +/- 0.18). Patients with either HP or pneumoconiosis had generally elevated AM O2- release and reduced BM O2- release. These results suggest that environmentally related interstitial lung disorders (HP and pneumoconiosis) may be associated with elevated AM O2- release relative to BM O2- release in comparison to non-environmentally related disorders (IPF or sarcoidosis).     

Pg-LPS对兔单核/巨噬细胞炎症因子表达的影响

目的比较Pg-LPS对新西兰兔不同部位单核/巨噬细胞炎症因子(IL-1β、IL-6、TNF-α)表达的影响。方法分离新西兰兔不同部位(血液、肺、腹腔、肝脏)的单核/巨噬细胞(Mo、AM、PM、KC),将每一个部位的细胞分别用E.coli-LPS、Pg-LPS刺激。运用RT-PCR法检测各组Mo、AM、PM、KC中IL-1β、IL-6、TNF-αmRNA的表达情况。结果各实验组的4个部位(Mo、AM、PM、KC)相比,IL-1β、IL-6、TNF-αmRNA的表达均存在部位差异(P﹤0.05)。E.coli-LPS组和Pg-LPS组IL-1β、IL-6、TNF-αmRNA的表达较对照组总体上均明显升高(P﹤0.05),并且E.coli-LPS组的升高作用强于Pg-LPS组(P﹤0.05)。结论不同部位的单核/巨噬细胞对Pg-LPS的刺激存在敏感性差异。Pg-LPS可以增强单核/巨噬细胞炎症因子基因的表达水平。