Alveolar macrophages and the diagnosis of drowning

In the present study, we examined the number of alveolar macrophages in lung tissue from 17 cases of fresh water drowning, 22 cases of acute death and 6 cases of lung emphysema. When counting only the number of alveolar macrophages per alveolus without consideration of the alveolar size we found no relevant differences between the groups investigated. To exclude any influence of the alveolar size on the results the surface density of the alveolar macrophages and interstitial tissue was estimated and compared in the different groups. In cases of drowning, the lungs showed significantly lower values in both categories. The ratio of ‘alveolar macrophages/interstitial tissue’ was also reduced in cases of drowning in comparison to the other groups, however, without significant differences. These morphometrical results characterizing the ‘emphysema aquosum’ with almost ‘empty’ and dilated alveoli could be explained by a wash-out effect of the drowning fluid leading to a partial removal of the macrophages from the alveoli. This hypothesis was confirmed by the detection of alveolar macrophages in the drowning froth by immunohistochemical analysis. Even though alveolar macrophages were unambiguously identified in advanced putrefied lungs in HE-stained sections as well as by immunohistochemical staining, an estimation of the number of these cells cannot provide further information for the diagnosis of drowning in putrefied corpses due to the autolytic destruction of the lung architecture providing no reliable values

Lipid-laden alveolar macrophages and pH monitoring in gastroesophageal reflux-related respiratory symptoms

Lipid-laden alveolar macrophages and pH monitoring have been used in the diagnosis of chronic aspiration in children with gastroesophageal reflux (GER). This study was conducted to prove a correlation between the detection of alimentary pulmonary fat phagocytosis and an increasing amount of proximal gastroesophageal reflux. It was assumed that proximal gastroesophageal reflux better correlates with aspiration than distal GER. Patients from 6 months to 16 years with unexplained recurrent wheezy bronchitis and bronchial hyperreactivity, or recurrent pneumonia with chronic cough underwent 24-hour double-channel pH monitoring and bronchoscopy with bronchoalveolar lavage (BAL). Aspiration of gastric content was determined by counting lipid laden alveolar macrophages from BAL specimens. There were no correlations between any pH-monitoring parameters and counts of lipid-laden macrophages in the whole study population, even when restricting analysis to those with abnormal reflux index expressing clinically significant GER. Quantifying lipid-laden alveolar macrophages from BAL in children with gastroesophageal-related respiratory disorders does not have an acceptable specificity to prove chronic aspiration as an underlying etiology. Therefore, research for other markers of pulmonary aspiration is needed

Macrophage TNF-α mediates parathion-induced airway hyperreactivity in guinea pigs.

Organophosphorus pesticides (OPs) are implicated in human asthma. We previously demonstrated that, at concentrations that do not inhibit acetylcholinesterase activity, the OP parathion causes airway hyperreactivity in guinea pigs as a result of functional loss of inhibitory M2 muscarinic receptors on parasympathetic nerves. Because macrophages are associated with asthma, we investigated whether macrophages mediate parathion-induced M2 receptor dysfunction and airway hyperreactivity. Airway physiology was measured in guinea pigs 24 h after a subcutaneous injection of parathion. Pretreatment with liposome-encapsulated clodronate induced alveolar macrophage apoptosis and prevented parathion-induced airway hyperreactivity in response to electrical stimulation of the vagus nerves. As determined by qPCR, TNF-α and IL-1β mRNA levels were increased in alveolar macrophages isolated from parathion-treated guinea pigs. Parathion treatment of alveolar macrophages ex vivo did not significantly increase IL-1β and TNF-α mRNA but did significantly increase TNF-α protein release. Consistent with these data, pretreatment with the TNF-α inhibitor etanercept but not the IL-1β receptor inhibitor anakinra prevented parathion-induced airway hyperreactivity and protected M2 receptor function. These data suggest a novel mechanism of OP-induced airway hyperreactivity in which low-level parathion activates macrophages to release TNF-α-causing M2 receptor dysfunction and airway hyperreactivity. These observations have important implications regarding therapeutic approaches for treating respiratory disease associated with OP exposures

Hydrogen peroxide release by OKI A1 (anti DR-Monoclonal antibody) resustabt alveolar mnacrophages in tuberculosis

Phorbol myristate acetate (PMA) triggered hydrogen peroxide (H2O2) release from alveolar macrophages and corresponding blood monocytes were studied as a whole, in active tuberculosis, inactive tuberculosis (treated), non-tuberculous lung disease patients and normal individuals. Irrespective of the study subjects, the alveolar macrophages produced less H2O2 than the corresponding blood monocytes. The alveolar macrophages that were resistant to OKIa1 (Anti-DR monoclonal antibody and complement treatment) produced an increased level of H2O2 than the control ascites and complement treated alveolar macrophages. Moreover, such increase in H2O2 release was not seen with peripheral blood monocytes; more than 90% monocytes were OKIa1 resistant population. These OKIa1 resistant alveolar macrophages are probably important in their metabolic, microbicidal and the immunological functions

Alveolar macrophages lack CCR2 expression and do not migrate to CCL2

Background: The recruitment of mononuclear cells has important implications for tissue inflammation. Previous studies demonstrated enhanced CCR1 and CCR5 expression and decreased CCR2 expression during in vitro monocyte to macrophage differentiation. To date, no study examined the in vivo differences in chemokine receptor expression between human peripheral blood monocytes and alveolar macrophages. Methods: We examined the expression of these receptors in human peripheral blood monocytes and alveolar macrophages using microarray analysis, reverse-transcriptase PCR, flow cytometry and migration analyses. Results: In contrast to peripheral blood monocytes, alveolar macrophages did not express the CCL2 receptor, CCR2, and did not migrate toward CCL2. In contrast, monocytes and freshly isolated resident alveolar macrophages both migrated towards CCL3. However, up to 6-fold more monocytes migrated toward equivalent concentrations of CCL3 than did alveolar macrophages from the same donor. While peripheral blood monocytes expressed the CCL3 receptor, CCR1, alveolar macrophages expressed the alternate CCL3 receptor, CCR5. The addition of anti-CCR5 blocking antibodies completely abrogated CCL3-induced migration in alveolar macrophages, but did not affect the migration of peripheral blood monocytes. Conclusion: These data support the specificity of CCL2 to selectively drive monocyte, but not alveolar macrophage recruitment to the lung and CCR5 as the primary macrophage receptor for CCL3

Pulmonary giant cells and their significance for the diagnosis of asphyxiation

This study was performed to prove whether the detection of polynuclear giant cells in lungs is useful for the diagnosis of asphyxiation due to throttling or strangulation. Therefore, lung specimens of 54 individuals with different natural and unnatural causes of death were investigated. In most lungs examined numerous alveolar macrophages with 1-2 nuclei were found. Polynuclear giant cells, which were arbitrarily defined as alveolar macrophages containing 3 or more nuclei, were observed in all groups investigated except in the cases of hypoxia due to covering the head with plastic bags. Apparent differences between the other groups in particular an increased number in cases of throttling or strangulation, could not be observed. Immunohistochemical investigations confirmed the hypothesis that the observed polynuclear giant cells were derived from alveolar macrophages. The immunohistochemical analysis of the proliferation marker antigen Ki 67 revealed no positive reaction in the nuclei of polynuclear giant cells indicating that these cells had not developed shortly before death by endomitosis as an adaptative change following reduction in oxygen supply. The results provide evidence that the detection of pulmonary polynuclear giant cells cannot be used as a practical indicator for death by asphyxiation due to throttling or strangulation

Disparate oxidant gene expression of airway epithelium compared to alveolar macrophages in smokers

<p>Abstract</p> <p>Background</p> <p>The small airway epithelium and alveolar macrophages are exposed to oxidants in cigarette smoke leading to epithelial dysfunction and macrophage activation. In this context, we asked: what is the transcriptome of oxidant-related genes in small airway epithelium and alveolar macrophages, and does their response differ substantially to inhaled cigarette smoke?</p> <p>Methods</p> <p>Using microarray analysis, with TaqMan RT-PCR confirmation, we assessed oxidant-related gene expression in small airway epithelium and alveolar macrophages from the same healthy nonsmoker and smoker individuals.</p> <p>Results</p> <p>Of 155 genes surveyed, 87 (56%) were expressed in both cell populations in nonsmokers, with higher expression in alveolar macrophages (43%) compared to airway epithelium (24%). In smokers, there were 15 genes (10%) up-regulated and 7 genes (5%) down-regulated in airway epithelium, but only 3 (2%) up-regulated and 2 (1%) down-regulated in alveolar macrophages. Pathway analysis of airway epithelium showed oxidant pathways dominated, but in alveolar macrophages immune pathways dominated.</p> <p>Conclusion</p> <p>Thus, the response of different cell-types with an identical genome exposed to the same stress of smoking is different; responses of alveolar macrophages are more subdued than those of airway epithelium. These findings are consistent with the observation that, while the small airway epithelium is vulnerable, alveolar macrophages are not "diseased" in response to smoking.</p> <p>Trial Registration</p> <p>ClinicalTrials.gov ID: NCT00224185 and NCT00224198</p

Murine alveolar macrophages limit replication of vaccinia virus

AbstractBecause of concerns about zoonotic transmission of monkeypox to humans and the bioterrorism threat posed by orthopoxviruses, there is renewed interest in probing cellular and molecular mechanisms of host defense to these pathogens. In particular, it is essential to understand viral–host interactions in the respiratory tract, which is the route of infection for smallpox and a likely route of transmission for monkeypox. In this study, we analyze functions of alveolar macrophages in poxvirus infection, using a recombinant vaccinia virus expressing firefly luciferase to quantify infection in mice and cell culture. Depletion of alveolar macrophages with liposomal clodronate worsens the overall severity of infection in mice, including greater replication and systemic dissemination of vaccinia as determined by bioluminescence imaging. Absence of alveolar macrophages increases total numbers of granulocytes and granulocytes/monocyte progenitor cells in the lungs during vaccinia infection, indicating that protective effects of alveolar macrophages may be mediated in part by reducing the host inflammation. Alveolar macrophages also limit vaccinia infection in respiratory epithelium, as shown by a co-culture model of cell lines derived from alveolar macrophages and lung epithelium. Collectively, these data demonstrate that alveolar macrophages are key determinants of host defense against local and systemic infection with poxviruses

Quantitative GPCR and ion channel transcriptomics in primary alveolar macrophages and macrophage surrogates

Background: Alveolar macrophages are one of the first lines of defence against invading pathogens and play a central role in modulating both the innate and acquired immune systems. By responding to endogenous stimuli within the lung, alveolar macrophages contribute towards the regulation of the local inflammatory microenvironment, the initiation of wound healing and the pathogenesis of viral and bacterial infections. Despite the availability of protocols for isolating primary alveolar macrophages from the lung these cells remain recalcitrant to expansion in-vitro and therefore surrogate cell types, such as monocyte derived macrophages and phorbol ester-differentiated cell lines (e.g. U937, THP-1, HL60) are frequently used to model macrophage function.Methods: The availability of high throughput gene expression technologies for accurate quantification of transcript levels enables the re-evaluation of these surrogate cell types for use as cellular models of the alveolar macrophage. Utilising high-throughput TaqMan arrays and focussing on dynamically regulated families of integral membrane proteins, we explore the similarities and differences in G-protein coupled receptor (GPCR) and ion channel expression in alveolar macrophages and their widely used surrogates.Results: The complete non-sensory GPCR and ion channel transcriptome is described for primary alveolar macrophages and macrophage surrogates. The expression of numerous GPCRs and ion channels whose expression were hitherto not described in human alveolar macrophages are compared across primary macrophages and commonly used macrophage cell models. Several membrane proteins known to have critical roles in regulating macrophage function, including CXCR6, CCR8 and TRPV4, were found to be highly expressed in macrophages but not expressed in PMA-differentiated surrogates.Conclusions: The data described in this report provides insight into the appropriate choice of cell models for investigating macrophage biology and highlights the importance of confirming experimental data in primary alveolar macrophages. © 2012 Groot-Kormelink et al.; licensee BioMed Central Ltd

Tissue specific Cish expression supports alveolar macrophage homeostatic function

Macrophages play critical roles in defense against microbes and clearance of dead cells, but also perform tissue specific functions in homeostasis. Distinct gene expression signatures in macrophages isolated from varying tissues are largely determined by environmental signals. Specifically, the lung is highly susceptible to environmental changes, such as O2 pressure and inhalation of particulate and microbes. Alveolar macrophages are shaped by the lung environment and have critical tissue-specific functions in initiating and resolving lung inflammation, and in maintaining lung structure via surfactant and lipid catabolism. While research speculates lung specific factors form alveolar macrophage phenotype and homeostatic function, the specific signals and regulators remain largely unknown. Therefore, we sought to explore lung specific cytokine signals and downstream signaling regulators that shape homeostatic functions of alveolar macrophages. We found Cytokine Inducible SH2 Containing Protein (Cish), a SOCS family member known to regulate the JAK-STAT5 pathway, is basally expressed in a tissue-specific manner in alveolar macrophages. Further, we found that the STAT5 activating cytokine GM-CSF regulates Cish expression in alveolar macrophages and observed reduced alveolar macrophage Cish expression with GM-CSF blockade in the lung. Cish knockout mice exhibit enlarged “foamy” alveolar macrophages, impaired surfactant metabolism, and dysregulated response to GM-CSF, all hallmarks of pulmonary alveolar proteinosis. Thus, we show alveolar macrophage Cish expression is directly linked to lung specific factors, namely GM-CSF, and influences surfactant homeostasis in the lung, a critical homeostatic function of alveolar macrophages. The lung is an especially critical site of protection as it is a barrier site that is constantly exposed to inhaled particulate and microbes and possesses a fragile structure. Alveolar macrophages act as sentinels in the lung, protecting this sensitive tissue from challenge while maintaining proper homeostasis and structure. From a public health perspective, continuing to elucidate the specific mechanisms by which alveolar macrophages mediate lung homeostasis is essential to providing cutting edge health care and to continuing to develop therapeutic treatments that can provide cures instead of simply mitigating symptoms of pulmonary disease. Here, we highlight one of many yet to be uncovered regulators of lung homeostasis