Inhibition of phagocytosis and interleukin-1 production in pulmonary macrophages from rats with sialodacryoadenitis virus infection

To test whether or not sialodacryoadenitis virus (SDAV) infection in rats affects pulmonary macrophage function, we intranasally inoculated pathogen-free F344 rats with SDAV and collected alveolar and interstitial macrophages 5 d later. We assessed Fc receptor-mediated attachment and phagocytosis by phase-contrast microscopic examination of monolayers of alveolar and interstitial macrophages incubated with zymosan, nonopsonized sheep erythrocytes, or erythrocytes opsonized with rabbit antisheep-erythrocyte IgG. Alveolar macrophages from virus-infected rats had significantly (P less than or equal to .05) lower indices of attachment and phagocytosis of opsonized erythrocytes than control macrophages, but there was no difference in attachment of zymosan particles. Interstitial macrophages were not affected. Alveolar macrophages from SDAV-infected rats produced significantly less interleukin-1 than those from control rats, as assessed by testing supernatants from lipopolysaccharide-stimulated macrophage cultures for induction of mouse thymocytes to take up tritiated thymidine. Effects of SDAV infection on lung macrophages could increase host susceptibility to other pathogens or complicate studies of respiratory tract immunity.     

Superoxide generation by neonatal and adult rabbit alveolar macrophages

We compared the abilities of alveolar macrophages (AM) from neonatal and adult rabbits to generate and release superoxide (O2-) after exposure to soluble and particulate stimuli. Basal rates of O2- release, 0.1-0.2 nmol/10(6)AM/10 min, were modestly increased by exposing AM to phorbol myristate acetate, (dihydro)cytochalasin B, or cytochalasin C. Opsonized zymosan was a more effective stimulus, and maximal rates of O2- release were observed when AM were stimulated by a combination of opsonized zymosan and one of the aforementioned cytochalasins. Cytochalasins D and E were each potent activators of O2- production by AM in the absence of additional stimuli. Adult AM released 24.8 +/- 6.0, 20.4 +/- 4.4, and 33.5 +/- 6.4 nmol O2-/10(6)AM/10 min after stimulation by cytochalasin D, cytochalasin E, and dihydrocytochalasin B plus opsonized zymosan, respectively. Following exposure to the same stimuli, AM from 1, 3- and 7-day-old rabbits released O2- at rates that ranged from 45 to 75% of corresponding adult values. The discrepancy between O2- production by neonatal and adult AM was accentuated when comparisons were restricted to AM recovered from bacteriologically sterile respiratory tracts. Our data show the O2- generating capacity of neonatal AM to be substantially less than that of the adult AM. Immaturity of this response could predispose neonates to pulmonary infection.     

Oxidative metabolism of neonatal and adult rabbit lung macrophages stimulated with opsonized group B streptococci.

We compared the oxidative metabolism of alveolar macrophages (AM) from adult and neonatal (1- and 7-day-old) rabbits before and after their in vitro exposure to type Ia group B streptococci (GBS) opsonized with immune rabbit serum. Nonstimulated AM from 1-day-old, 7-day-old, or adult rabbits consumed O2 at a rate of 17 to 20 nmol/10(6) AM per 10 min under basal conditions and released minimal amounts of superoxide (O2-) into the medium. Approximately 80% of this basal respiration was of mitochondrial origin, based on its inhibition by NaCN. Exposure to GBS opsonized with immune rabbit serum stimulated O2 consumption approximately half as effectively in the neonatal AM as in the adult AM. Little O2- was released into the medium unless the cells were pretreated with dihydrocytochalasin B. Under such conditions, 1-day-old, 7-day-old, and adult AM released 3.6, 5.3, and 13.9 nmol of O2-/10(6) AM per 10 min, respectively. The uptake of opsonized GBS by 1-day-old AM was not affected by 1 mM NaCN, whereas phagocytosis by adult AM was substantially reduced under these conditions. Overall, our findings suggest that neonatal AM have less-well-developed postphagocytic oxidative metabolic responses and release less superoxide after exposure to opsonized GBS than do adult AM. They also demonstrate that the energy for phagocytosis is derived principally from mitochondrial metabolism in adult AM but not in neonatal AM. We conclude that metabolic differences between neonatal and adult AM may contribute to neonatal pulmonary susceptibility to GBS infections and account, in part, for the ability of GBS to succeed as neonatal pulmonary pathogens.     

In vitro stimulation of alveolar macrophage metabolic activity by polystyrene in the absence of phagocytosis

The technique of lucigenin-dependent phagocytic chemiluminescence was used to investigate the stimulation of metabolic activity in human alveolar macrophages on contact with polystyrene. The results were similar to those obtained using a spectrophotometric assay of superoxide release based on ferricytochrome C reduction. There was a marked stimulation of metabolic activity in the alveolar macrophages on incubation at 37 degrees in polystyrene vials which was shown to be due to contact with and adherence to the polystyrene. This could be reduced by the addition of gelatin or foetal calf serum without preventing the ability of the cells to respond to opsonized particles. By using several metabolic inhibitors it was shown that lucigenin-dependent chemiluminescence was associated with the release of superoxide at the time of adherence. The implications of these findings are discussed and it is suggested that the stimulation of alveolar macrophage metabolic activity by contact with polystyrene can contribute to the observed difference between alveolar macrophage and polymorphonuclear leucocyte oxygen consumption in the absence of phagocytosis.     

In vitro stimulation of alveolar macrophage metabolic activity by polystyrene in the absence of phagocytosis.

The technique of lucigenin-dependent phagocytic chemiluminescence was used to investigate the stimulation of metabolic activity in human alveolar macrophages on contact with polystyrene. The results were similar to those obtained using a spectrophotometric assay of superoxide release based on ferricytochrome C reduction. There was a marked stimulation of metabolic activity in the alveolar macrophages on incubation at 37 degrees in polystyrene vials which was shown to be due to contact with and adherence to the polystyrene. This could be reduced by the addition of gelatin or foetal calf serum without preventing the ability of the cells to respond to opsonized particles. By using several metabolic inhibitors it was shown that lucigenin-dependent chemiluminescence was associated with the release of superoxide at the time of adherence. The implications of these findings are discussed and it is suggested that the stimulation of alveolar macrophage metabolic activity by contact with polystyrene can contribute to the observed difference between alveolar macrophage and polymorphonuclear leucocyte oxygen consumption in the absence of phagocytosis.     

Role of the alveolar macrophage in host defense and immunity to Legionella micdadei pneumonia in the guinea pig

Guinea pigs develop a lethal pneumonia after intratracheal infection with Legionella micdadei, and the lung displays pathological changes similar to those observed in humans. To investigate the role of the resident alveolar macrophage in the pathogenesis of L. micdadei pneumonia, guinea pig alveolar macrophages obtained by bronchoalveolar lavage were cultured in vitro and infected with L. micdadei. In the absence of opsonins L. micdadei was phagocytized by, and multiplied within, alveolar macrophages with greater than a 100-fold increase in cell-associated colony forming units over 20 h. L. micdadei opsonized with complement or antibody multiplied within alveolar macrophages at the same rate as unopsonized bacteria. Guinea pigs which were treated with antimicrobials after infection with L. micdadei and recovered from the pneumonia were immune to challenge with an otherwise lethal inoculum of L. micdadei. However, the growth curve of both unopsonized and opsonized L. micdadei in the alveolar macrophages from immune animals was essentially identical to that in macrophages from susceptible animals. Thus, the resident alveolar macrophage is not capable of limiting the growth of Legionella. Rather, the alveolar macrophages appear to be the primary site of Legionella multiplication within the lung. Although alveolar macrophages may participate in other aspects of pulmonary immunity to the legionellae, these data indicate that the alveolar macrophage alone does not act as an effector cell in cell-mediated immunity to Legionella.     

Human bronchoalveolar lavage cells and luminol-dependent chemiluminescence.

Human bronchoalveolar lavage cells from several different disease states were examined by the technique of zymosan-stimulated, luminol-dependent chemiluminescence. Light production correlated well with polymorphonuclear leucocyte contamination of the alveolar macrophage suspension but not with lymphocyte contamination. Regression analysis indicated that human alveolar macrophages produce little if any luminol-dependent chemiluminescence. Further investigation of metabolic activity, using measurements of superoxide release, oxygen consumption, and lucigenin-dependent chemiluminescence, showed that "respiratory burst" activity in alveolar macrophages was stimulated by opsonised zymosan.     

Pulmonary surfactant protein A enhances the host-defense mechanism of rat alveolar macrophages

The effects of surfactant, surfactant lipids, and surfactant protein A (SP-A) on the surface phagocytosis of [3H]thymidine-labeled Staphylococcus aureus (SAE) by rat alveolar macrophages were studied. Alveolar macrophages only ingest SAE when the bacteria are opsonized with rat serum prior to incubation with alveolar macrophages. Preincubation or "opsonization" of the bacteria with surfactant did not result in phagocytosis by the macrophages. However, preincubation of the macrophages with surfactant increased the phagocytosis of rat serum-opsonized bacteria by approximately 70% when compared to the control macrophages. The factor present in surfactant causing the stimulation of the phagocytosis is probably SP-A. Preincubation of macrophages with human SP-A enhanced the phagocytosis to the same extent as whole surfactant, whereas preincubation with surfactant lipids had no effect on the phagocytosis. The SP-A-induced enhancement of the phagocytosis is time, temperature, and concentration dependent. Phagocytosis of opsonized SAE by alveolar macrophages was maximal after 15 min of incubation and at an SP-A concentration of 1 micrograms/ml. No phagocytosis occurred at 0 degrees C. In addition, whole surfactant and SP-A induce a lucigenin-dependent chemiluminescence response in alveolar macrophages. The chemiluminescence response is initiated after 15 min of incubation and reaches a maximum after 30 min. The concentration of SP-A needed for an optimal response is in the same order of magnitude as the concentration needed for maximal enhancement of the phagocytosis of SAE by alveolar macrophages.(ABSTRACT TRUNCATED AT 250 WORDS)     

Chemiluminescence of bronchoalveolar macrophages: effect of adherence to plastic cuvette.

The chemiluminescence of alveolar macrophages was determined in presence of luminol or lucigenin, either before or after the macrophage isolation by adherence to plastic. In presence of luminol, the purified and non-purified macrophages behave almost in the same way, and their chemiluminescence activity is stimulated both by zymosan and phorbol ester. On the other hand, the lucigenin chemiluminescence of non-purified macrophages is considerably higher than that obtained in presence of luminol, but is almost completely insensitive to the zymosan or phorbol ester stimulation. Moreover, the adherence purification considerably inhibits the lucigenin chemiluminescence. These results can be indicative of functionally different macrophage states.     

Hydrogen Peroxide Release by Rat Alveolar Macrophages: Comparison with Blood Neutrophils

Hydrogen peroxide release was examined using biochemical and cytochemical techniques in rat alveolar macrophages, at rest and during phagocytosis, and compared with rat blood neutrophils. Using biochemical techniques, alveolar macrophages released small amounts of hydrogen peroxide at rest, and no increase was observed after challenge with opsonized and nonopsonized zymosan particles at several particle-cell ratios (1:1 to 1:1,000). Neutrophils released similar quantities of hydrogen peroxide at rest but showed a 12-fold increase in hydrogen peroxide release following exposure to opsonized zymosan particles. Using cytochemical techniques to localize sites of hydrogen peroxide release, resting neutrophils showed little deposition of reaction product at the cell surface and occasional deposits in endocytotic vesicles. After exposure to latex particles, a dense reaction product was observed between the particle and the cell membrane, indicating significant increases in hydrogen peroxide release at the sites of particle contact with the neutrophil. The resting macrophage displayed a light, uniform precipitation of cerium over the cell surface and lining intracellular channels and endocytotic vesicles and vacuoles. Following particle exposure, there was no significant difference in the density or distribution of reaction product. These findings, together with previous studies of oxidative metabolism, suggest that alveolar macrophages do not release increased quantities of hydrogen peroxide during phagocytosis. In contrast to neutrophils, oxidative-dependent metabolic pathways may not be of primary importance for microbial killing by alveolar macrophages.     

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 RU 41740 aerosol treatment on mouse bronchoalveolar cells, and protection afforded against influenza virus infection

RU 41740, an immunomodulating compound extracted from Klebsiella pneumoniae, was previously shown to enhance mice resistance to bacterial and viral lung infections. To explore lung defense mechanisms, we studied the influence of RU 41740 aerosol treatment on the bronchoalveolar cell populations. Five successive daily RU 41740 aerosol treatments induced a large accumulation of leukocytes in the lungs 4h after the last treatment. Polymorphonuclear leukocytes predominated. The numbers of lymphocytes and monocytes rose significantly. A single RU 41740 aerosol treatment significantly raised the number of polymorphonuclears only. A luminol-dependent chemiluminescence assay was used to test the effect of RU 41740 on the opsonized zymosan induced response of alveolar macrophages. In vitro, addition of RU 41740 enhanced this chemiluminescence. After a single RU 41740 aerosol treatment of mice, the chemiluminescence of purified alveolar macrophages from these mice increased significantly. The protective effect of five daily RU 41740 aerosol treatments against influenza virus infection was believed to be due to the great intensity of the cellular response and the polymorphonuclear influx. The alveolar macrophage activation observed might also explain the enhanced resistance of mice to influenza virus infection.     

Candida albicans stimulates arachidonic acid liberation from alveolar macrophages through alpha-mannan and beta-glucan cell wall components.

Candida albicans is an increasingly important fungal pathogen. Alveolar macrophages respond to fungal components such as zymosan by releasing arachidonic acid (AA) and AA metabolites. However, few studies hypothesized that macrophages respond to C. albicans by releasing AA and generating AA metabolites as a consequence of interaction of mannose and beta-glucan receptors with fungal cell wall components. [14C]AA-labeled rabbit alveolar macrophages released AA following stimulation with either live or heat-killed C. albicans. High-pressure liquid chromatography analysis revealed that 55% of the AA released was metabolized via cyclooxygenase and lipoxygenase pathways. The metabolites consisted of prostaglandin E2, prostaglandin F2 alpha, 6-ketoprostaglandin F1 alpha, thromboxane B2, and leukotrienes B4 and D4. We further examined the roles of alpha-mannan and beta-glucan components of C. albicans in mediating these alterations of eicosanoid metabolism. Prior work in our laboratory has shown that soluble alpha-mannan and beta-glucan inhibit macrophage mannose and beta-glucan receptors, respectively. Incubation of alveolar macrophages with soluble alpha-mannan derived from C. albicans (1 mg/ml) resulted in 49.8% +/- 2.6% inhibition of macrophage AA release during stimulation with intact C. albicans (P = 0.0001 versus control). Macrophage AA release in response to C. albicans was also inhibited to a significant but lesser degree by soluble beta-glucan (36.2% +/- 1.3%; P = 0.008 versus control). These results indicate that C. albicans stimulates macrophage AA metabolism and that these effects are partly mediated by alpha-mannan and beta-glucan constituents of the fungus.     

Mycobacterium bovis BCG vaccination augments interleukin-8 mRNA expression and protein production in guinea pig alveolar macrophages infected with Mycobacterium tuberculosis

Alveolar macrophages are likely the first cell type to encounter Mycobacterium tuberculosis in a pulmonary infection, resulting in the production of chemokines. In order to evaluate this response, alveolar macrophages harvested from nonvaccinated and Mycobacterium bovis BCG-vaccinated guinea pigs were infected in vitro with live M. tuberculosis H37Ra or H37Rv (multiplicity of infection, 1:1) or cultured with lipopolysaccharide (10 micro g/ml) for 3, 12, and 24 h. Interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1) mRNA expression was determined by real-time PCR. Culture supernatants were assayed for guinea pig IL-8 protein by using a human IL-8 enzyme-linked immunosorbent assay kit. Alveolar macrophages harvested from BCG-vaccinated guinea pigs produced significantly more mRNA and protein for IL-8 than alveolar macrophages harvested from nonvaccinated guinea pigs at 12 and 24 h poststimulation or postinfection. Infection with attenuated M. tuberculosis (H37Ra) stimulated alveolar macrophages isolated from BCG-vaccinated guinea pigs to produce significantly more IL-8 mRNA than did alveolar macrophages infected with a virulent strain (H37Rv) at 12 and 24 h postinfection. Significant MCP-1 mRNA production was also detected in stimulated or infected alveolar macrophages; however, prior vaccination did not significantly affect levels of MCP-1 mRNA. Alveolar macrophages isolated from BCG-vaccinated guinea pigs produced significantly more IL-8 mRNA and protein when stimulated for 24 h with heat-killed H37Ra, heat-killed H37Rv, and H37Rv cell wall, but not mannose-capped lipoarabinomannan (ManLAM), than did cells stimulated with media alone. These observations indicate that prior vaccination may alter very early events in the M. tuberculosis-infected lung.     

Acquisition of peroxidase activity by rat alveolar macrophages during pulmonary inflammation.

The authors investigated the ability of rat alveolar macrophages to acquire peroxidase activity in the course of pulmonary inflammation. Granulomatous pulmonary inflammation was induced in bacille Calmette-Guérin (BCG)-immunized rats by intravenous injection of BCG in mineral oil. In contrast to normal alveolar macrophages, which are peroxidase-negative, alveolar macrophages lavaged from the BCG-treated rats showed significant peroxidase activity in large cytoplasmic inclusions compatible with internalized exogenous material. Alveolar macrophage uptake of intact peroxidase-positive neutrophils was also observed. Maximal numbers of peroxidase-positive alveolar macrophages were observed after the initial influx of neutrophils into the lungs, and peroxidase activity could be demonstrated in cell-free lavage fluid during the acute phase of lung injury. Normal alveolar macrophages acquired peroxidase activity after incubation with peritoneal exudate neutrophils, with purified soluble human myeloperoxidase, and with opsonized erythrocytes. It is concluded that alveolar macrophages acquire peroxidase activity from multiple sources during pulmonary inflammation. Internalization of peroxidase by the alveolar macrophage may serve to clear a potentially toxic enzyme(s) from the alveolar space and contribute to the resolution of pulmonary inflammation.     

The Alveolar Microenvironment of Patients Infected with Human Immunodeficiency Virus Does Not Modify Alveolar Macrophage Interactions with Streptococcus pneumoniae

We tested the hypothesis that HIV infection results in activation of alveolar macrophages and that this might be associated with impaired defense against pneumococcus. We compared alveolar macrophages and lymphocytes in 131 bronchoalveolar lavage samples from HIV-infected and healthy controls using inflammatory gene microarrays, flow cytometry, real-time PCR, and enzyme-linked immunosorbent assay (ELISA) to determine the pattern of macrophage activation associated with HIV infection and the effect of this activation on defense against pneumococcus. We used gamma interferon (IFN-γ) priming to mimic the cellular milieu in HIV-infected lungs. InnateDB and BioLayout 3D were used to analyze the interactions of the upregulated genes. Alveolar macrophages from HIV-infected adults showed increased gene expression and cytokine production in a classical pattern. Bronchoalveolar lavage from HIV-infected subjects showed excess CD8⁺ lymphocytes with activated phenotype. Toll-like receptor 4 (TLR4) expression was increased in macrophages from HIV-infected subjects, but function was similar between the groups; lung lavage fluid did not inhibit TLR function in transfected HeLa cells. Alveolar macrophages from HIV-infected subjects showed normal binding and internalization of opsonized pneumococci, with or without IFN-γ priming. Alveolar macrophages from HIV-infected subjects showed classical activation compared to that of healthy controls, but this does not alter macrophage interactions with pneumococci.     

Alveolar macrophage subpopulations'' responsiveness to chemotactic stimuli.

Recent data suggest that alveolar macrophages are a heterogeneous group of cells with several subpopulations. This study was undertaken to determine if there is heterogeneity among rat alveolar macrophage subpopulations ability to respond to chemotactic stimuli. Alveolar macrophages were harvested by bronchoalveolar lavage and fractionated into density-defined fractions by centrifugation through a continuous isoosmotic gradient of colloidal silica. Unfractionated and density-defined alveolar macrophages were then characterized as to their ability to migrate towards F-Met-Leu-Phen and zymosan-activated serum. Alveolar macrophages of density 1.083-1.097 gm/ml were found to have the greatest migrational movement toward F-Met-Leu-Phen, which was higher than the unfractionated population. In contrast, 2 peaks in alveolar macrophage subpopulations migrational movement towards zymosan-activated serum were noted that were lower than the unfractionated population. These results demonstrated that alveolar macrophages are heterogeneous in their migrational ability towards the chemotactic stimuli F-Met-Leu-Phen and zymosan-activated serum and that there may be a cooperative interaction between the subpopulations that affects macrophage migrational ability.     

Replication of parainfluenza (Sendai) virus in isolated rat pulmonary type II alveolar epithelial cells.

The major objectives of this study were to determine whether alveolar type II epithelial cells isolated from rat lung and maintained in tissue culture would support productive replication of parainfluenza type 1 (Sendai) virus and to determine whether isolated type II cells from neonatal (5-day-old) rats that are more susceptible to viral-induced alveolar dysplasia supported viral replication to a greater extent than those from weanling (25-day-old) rats. Isolated and cultured type II cells from neonatal and weanling rats that were inoculated with Sendai virus supported productive replication as indicated by ultrastructural identification of budding virions and viral nucleocapsids in type II cells and by demonstration of rising titers of infectious virus from inoculated type II cell cultures. Alveolar macrophages from neonatal and weanling rats also supported viral replication, although infectious viral titers in macrophage cultures were lower than those from type II cell cultures. Only minor differences were detected between viral titers from neonatal and weanling type II epithelial cell cultures. Higher densities of viral nucleocapsids were observed in neonatal type II cells than in those from weanling rats. The results indicate that isolated type II alveolar epithelial cells support productive replication of parainfluenza virus and that type II cells are probably more efficient in supporting productive viral replication than are alveolar macrophages.     

Effect of ketotifen on human alveolar macrophages

In order to examine the possibility of human alveolar macrophages being an effector site of an antiasthmatic drug, we studied the effect of ketotifen on chemiluminescence and Ca-activated K conductance at single channel level because they are associated with the activation of macrophages. Peak chemiluminescence response, induced by a single concentration of phorbol myristate acetate (0.5 micrograms/ml), was inhibited by 28 +/- 3, 51 +/- 8, and 76 +/- 7% by ketotifen concentrations of 0.02, 0.12, and 0.24 mmol/L, respectively. The inhibition of chemiluminescence by ketotifen concentrations of 0.12 and 0.24 mmol/L was also observed in macrophages stimulated with zymosan activated serum (54 +/- 7 and 80 +/- 6%, respectively). Moreover, ketotifen, 0.24 mmol/L, inhibited Ca-activated K channel currents by 44 +/- 7% (large currents) and 48 +/- 13% (small currents). These results may explain, at least partly, antiasthmatic effects of ketotifen.     

Oxygen free radical production by mouse peritoneal macrophages as a function of age

The ability of thioglycollate-elicited peritoneal macrophages (PM) from young and senescent C57BL/6J mice to produce oxygen free radicals was assessed by luminol-dependent chemiluminescence (CL) after introduction of phagocytic stimuli. A significant age-dependent variation in the CL response was detected. A 2-fold increase in the oxygen reactive species was produced by senescent PM in response to latex and zymosan stimulation; but, the capacity to ingest latex and zymosan A particles did not vary significantly between PM from young and senescent mice. Peritoneal macrophages from both age groups responded much more vigorously to opsonized zymosan. The response of the PM from young mice was, however, 2.8-fold higher than that of old ones. There was no age-related difference in oxygen free radical production after stimulation with phorbol myristate acetate (PMA). Also, no age-dependent differences were found in the relative contribution of the various oxygen reactive species O2.-, OH., 1O2 and H2O2) to the overall oxidative burst, with latex zymosan A or PMA.