Correlation between PGE2 production and suppressor activity of alveolar macrophages from patients with interstitial lung diseases

The suppressive activity of alveolar macrophages (AM) obtained from bronchoalveolar lavage (BAL), on PHA stimulation of autologous peripheral blood lymphocytes (APL) was evaluated. The effect on lymphocyte stimulation was evaluated by coculturing the AM and APL cells at a ratio of 1:1. PGE2 released by AM during the culture period was measured by a radioimmune assay. The patients included in the study were 11 cases with interstitial lung disease (ILD), 8 cases of lung cancer (CA), and 5 controls (CO). Addition of AM of patients from the CA group resulted in slight suppression of lymphocyte stimulation in 4 cases, slight enhancement in 3 cases and no effect in one case. AM from the CO group induced slight suppression in 4 out of 5 cases. AM from all 11 ILD cases exerted a significant high suppressive activity (65.6% +/- 18.2 - P less than 0.001 by comparison with the CO and CA groups). In ILD cases, a dichotomous pattern was found in regard to relation between high suppressive activity of AM and release of PGE2: in idiopathic pulmonary fibrosis (IPF) patients, high suppressive activity of AM (70.4% +/- 15.4) correlated well with elevated secretion of PGE2: 3.58 +/- 0.26 ng/ml/10(5) cells (P less than 0.02 compared to CO). AM from sarcoidosis patients suppressed PHA stimulation by 61.6% +/- 19.3 but secreted only 0.357 +/- 0.26 ng/ml/10(5) cells of PGE2 (P less than 0.02 compared with the idiopathic pulmonary fibrosis group). Therefore, it seems that other factors, in addition to PGE2, might be involved in the suppressive activity of AM from interstitial lung diseases.     

Detection of circulating soluble immune complexes in patients with various renal diseases.

Sera from patients with various types of glomerulonephritis (GN) as well as sera from rabbits with acute serum sickness were studied for the presence of circulating immune complexes (IC). The method used is based on the observation that IC inhibit the uptake of IgG aggregates by guinea-pig peritoneal macrophages. Inhibition significantly greater than with normal human sera was found with sera of patients with membranous GN, membranoproliferative GN, focal glomerular sclerosis, minimal change nephrotic syndrome, acute septicaemic glomerular diseases and systemic lupus erythematous. IC were also detected in rabbits with acute serum sickness during the period of immune elimination.     

Isolation and characterization of circulating immune complexes from patients with pneumococcal pneumonia.

Circulating immune complexes (CIC) were isolated from serum samples from patients with bacteremic and nonbacteremic pneumococcal pneumonia. Overall, 63% (26 of 41) of patients with pneumococcal pneumonia had elevated levels of immunoglobulin G (IgG)-containing CIC. IgM-containing CIC were identified in samples from only three patients. Serum samples from nonbacteremic patients contained significantly higher levels of IgG-containing CIC (96.6 +/- 111.7 micrograms/ml) than did samples from bacteremic patients (31.7 +/- 26.9 micrograms/ml) during week 1 in hospital (P less than 0.05). Immune complexes levels did not correlate with IgG concentrations in serum or anticapsular antibody levels. Immune complexes from nonbacteremic patients had sedimentation coefficients of greater than 19s by density gradient ultracentrifugation. In contrast, CIC from bacteremic patients had smaller coefficients, of between 9s and 14s. Pneumococcal capsular antigens were identified in concentrated dissociated CIC from both patient groups by counterimmunoelectrophoresis.     

Local immune complexes and inflammatory response in patients with chronic interstitial pulmonary disorders associated with collagen vascular diseases

Evidence is accumulating that the lung injury in collagen vascular diseases (CVD) is triggered by immune complexes (IC). These reactions are neutrophil- and complement-dependent. The direct, in vivo phagocytosis of IC by bronchoalveolar lavage polymorphonuclear leucocytes (BAL-PMN), was studied in 15 patients with CVD and chronic interstitial pulmonary disorders. A control group (NC) consisted of nine healthy, non-smoking volunteers. Concentrations of soluble IC were measured using a solid phase Clq ELISA assay, and an indirect, in vitro phagocytosis assay performed using healthy donor PMN. Local Ig and C3 concentrations were determined using laser nephelometry and Mancini techniques, respectively. In the patient group the total cell counts/ml recovered lavage fluid and the proportions of BAL-PMN were significantly increased (P less than 0.05 and P less than 0.001, respectively). The influxed PMN showed high scores of direct IC phagocytosis. Soluble IC concentrations were significantly increased compared with controls (all comparisons P less than 0.01), and in the BAL relatively higher than in the serum of the same patients. Concomitantly high local IgG concentrations were observed. Corticosteroid treatment gave rise to significantly decreased total cell counts (P less than 0.05), and proportions of BAL-PMN (P less than 0.001), a decrease in the in vivo IC phagocytosis (P less than 0.05), in the indirect, in vitro IC phagocytosis, in the Clq ELISA and in the local IgG concentrations (all comparisons P less than 0.001). We concluded that locally formed IC may induce an inflammatory response in the lungs of patients with CVD.     

Antigen expression of alveolar macrophages in smokers and patients with lung diseases

Alveolar macrophage function was studied immunocytochemically using three monoclonal antibodies—macrophage CD 68 KP 1 (M), protein CD 11C (P), and anti-elastin (EL)—and three polyclonal antibodies—lysozyme (LZ), alpha-1-antitrypsin (AAT), and alpha-1-antichymotrypsin (AACT). the material for study was smears obtained from bronchial washings from 15 healthy persons and 60 patients with respiratory infections or primary or secondary malignant lung infiltration. Eight of the healthy group and 40 of the patient group were smokers (SM). the percentage of cells obtained from the washings which were macrophages was also measured. The intensity of staining reactions for each of the six antigens was noted and in general more intense staining was noted in smokers than in non-smokers. More intense staining was observed in patients with pulmonary infections (group II PI) and metastatic pulmonary infiltrations (group IV MP Ca) than in controls (group IC), while patients with primary lung cancer (group III PP Ca) had highly reduced staining reactions. the number of macrophages was similarly increased in all groups in comparison with the IC group for non-smokers and in all groups except III PP Ca for smokers. It is concluded that smoking, pulmonary infections, and metastatic infiltration of the lung are associated with an increase in the number and activity of alveolar macrophages, while patients with primary lung cancer have an increase in the number of macrophages which are functionally incompetent.     

Determination of immune complexes in sera from dogs with various diseases by mastocytoma cell assay

Canine immunoglobulin G complexed with particulate or soluble antigen can bind to the Fc receptors on the mastocytoma cells. Attachment of immune complexes composed of immunoglobulin G and soluble antigen (ovalbumin) to mastocytoma cells was detected by an inhibition of rosette formation with indicator cells (sensitized sheep erythrocytes). Therefore, canine circulating immune complexes may also attach to mastocytoma cells and inhibit rosette formation (mastocytoma cell assay). Sera from 326 dogs with various diseases and from 50 clinically normal dogs were assayed for immune complexes. The incidence of immune complexes in sera from normal dogs was 6% as compared with 25% in dogs with various diseases. The immune complexes were demonstrated in 37% of sera from dogs with various neoplastic diseases, 40% of sera from dogs with diabetes, 24% of sera from dogs with hypothyroidism, 50% of sera from dogs with mycotic disease, 75% of sera from dogs with arthritis, 38% of sera from dogs with kidney disorders, 40% of sera from dogs with neurological diseases, 45% of sera from dogs with various parasitic diseases, and 27% of sera from dogs with liver disorders. Only 19% of sera from dogs admitted to the hospital for various surgeries gave positive results. The incidence of the positive sera from dogs with various diseases is discussed in regard to their counterparts of human diseases.     

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.     

Diagnosis of usual interstitial pneumonia and distinction from other fibrosing interstitial lung diseases

Usual interstitial pneumonia is an almost uniformly fatal form of fibrosing interstitial lung disease. It is the most common idiopathic interstitial pneumonia, and currently, there is no effective therapy. Lung biopsy is often needed for diagnosis, and pathologists must be able to recognize its features and distinguish it from other interstitial lung diseases that have a better prognosis and a more favorable response to therapy. This review is an attempt to clarify the diagnostic pathologic features of usual interstitial pneumonia and to provide guidelines for its distinction from other interstitial lung diseases that enter the differential diagnosis.     

The effects of some anti-arthritic drugs and cytokines on the shape and function of rodent macrophages

Non-steroidal antiinflammatory drugs (NSAID) enhanced the spreading of mouse and rat peritoneal macrophages attached to either plastic or glass. This was probably due to drug inhibition of prostaglandin E2 (PGE2) production since spreading was also inhibited by adding exogenous PGE2. Corticosteroids (dexamethasone, cortisol and prednisolone) and some immunosuppressants (6-mercaptopurine, methotrexate, but not cyclosporin-A) also enhanced in-vitro spreading of murine peritoneal macrophages. Some recombinant cytokines (human tumour necrosis factor alpha and beta, murine tumour necrosis factor alpha, and murine interferon gamma, but not human interferon gamma) also enhanced the spreading of mouse peritoneal macrophages in vitro. Scanning electron microscopy revealed significant differences in morphology of cells induced to spread by these drugs and cytokines. NSAID treatment also enhanced macrophage clumping in vitro, indicating that cell spreading may play an important role in the resolution of inflammatory processes and/or the formation of multinucleated giant cells.     

SETD4敲除对小鼠巨噬细胞功能及免疫细胞分化影响的初步研究#br#

目的　探讨SETD4（SET-domain containing protein 4）基因敲除对小鼠腹腔巨噬细胞（peritoneal macrophages,pMφ）分化成熟、功能的影响以及对小鼠外周血、脾脏免疫细胞分化,胸腺和脾脏发育的影响。 方法　(1)利用流式细胞术检测腹腔灌洗液中pMφ表面CD31分子的表达情况;(2)用液相芯片技术检测和比较SETD4-/-和SETD4+/+小鼠pMφ在受到脂多糖（LPS）刺激后对细胞因子TNF-α、IL-6释放的影响, 流式细胞术检测SETD4基因敲除对巨噬细胞吞噬细菌能力及Transwell检测对巨噬细胞迁移能力的影响;(3)流式细胞术分析和比较两组小鼠外周血及脾脏主要免疫细胞的数量及比例的差异;(4)HE染色对比两组小鼠胸腺和脾脏组织结构的差异。 结果　（1）与SETD4+/+小鼠相比,SETD4-/-小鼠pMφ受到LPS刺激后TNF-α、IL-6的释放明显减少,但是两组小鼠pMφ的比例及分化成熟程度无明显差异;（2）吞噬细菌能力和迁移能力两组无明显差异;（3）两组小鼠外周血及脾脏主要免疫细胞的数量及比例无明显差异;（4）SETD4基因敲除对小鼠胸腺和脾脏的结构无明显影响。 结论　（1）SETD4能促进炎性细胞因子TNF-α、IL-6的产生,但对pMφ的分化成熟及细菌吞噬、迁移能力无明显影响;（2）SETD4基因敲除对小鼠外周血及脾脏免疫细胞的分化没有明显影响,对胸腺和脾脏组织的发育无明显影响。     

A study of human interstitial lung diseases with special reference to immune complexes and hyaline membrane

To evaluate the pathogenetic roles of immune complexes and alveolar hyaline membrane in idiopathic interstitial pneumonia (IIP), immunohistological and ultrastructural studies of the kidney and lung were performed in 23 cases of IIP, 19 cases of autoimmune diseases, 17 cases of interstitial pneumonia other than IIP, and 11 cases of bronchopneumonia as a control group. None of the cases of IIP or interstitial pneumonia other than IIP showed immune complexes in the alveolar and glomerular capillary walls. On the other hand, one case of SLE was positive for IgG and components of complement along the alveolar and glomerular capillary walls. The alveolar hyaline membrane in the present cases revealed immunoglobulins as well as components of complement, which were poorly soluble in chaotropic solution or acidic buffer. These results indicate that circulating immune complexes play a minor role in the pathogenesis of IIP and other types of interstitial pneumonia, and that there is no relationship between immune complex deposition in alveoli and the alveolar hyaline membrane. It is necessary to further investigate factors other than immune complexes involved in alveolar tissue damage and to clarify the significance of the hyaline membrane in the processes occurring from acute changes to pulmonary fibrosis in IIP.     

Accessory function and costimulatory molecule expression of alveolar macrophages in patients with pulmonary tuberculosis

An effective immune response against M. tuberculosis requires a coordinated interaction of alveolar macrophages (AM) and lymphocytes. Secondary signals, such as accessory function (AF) of antigen presenting cells and interaction of costimulatory molecules are also important for T cell activation. In the present study we determined the AF and the expression of CD11a, CD54, CD58, CD80, CD86 and HLA-DR costimulatory molecules by AMs lavaged from patients with pulmonary tuberculosis and controls. We hypothesized that alterations in AF and costimulatory molecule expression may influence the presentation of tuberculosis. Therefore these parameters were also correlated with the radiographic extension of the disease. AMs of patients with tuberculosis exhibited an increased AF and a significantly increased expression of co-stimulatory molecules compared with controls. Furthermore, we observed that the expression of CD54 (ICAM-1) decreased with the course of the disease. We conclude that the infection by M. tuberculosis results in an increased AF of AMs and the activity of AMs remains uninfluenced by the extension of the disease. Clear-cut changes of patterns of costimulatory molecule expression by AMs could not be observed with the progression of tuberculosis.     

Interaction of alveolar macrophages from pulmonary patients and healthy subjects with liposomes

Formation of primary metabolites of oxidative burst in alveolar macrophages from healthy subjects and patients with chronic lung diseases is studied using the method of luminoldependent chemiluminescence. Incubation with liposomes composed from phosphatidylcholine and a phosphatidylcholine-fatty acid esters mixture induces similar activation of oxidative burst in alveolar macrophages from healthy subjects and patients with lung diseases. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 124, No. 11, pp. 520–523, November, 1997     

In vitro activation of rat neutrophils and alveolar macrophages with IgA and IgG immune complexes. Implications for immune complex-induced lung injury.

In the rat, both IgG and IgA immune complexes induce oxygen radical mediated lung injury that is partially complement-dependent. In vivo studies have suggested that the chief sources of oxygen radicals in IgG and IgA immune complex-induced lung injury are neutrophils and tissue macrophages, respectively. The current studies have been designed to provide additional insights into these two models of tissue injury. Preformed monoclonal IgG and IgA immune complexes stimulated dose-dependent O2-. and H2O2 production by alveolar macrophages. In contrast, neutrophils exhibited O2-. production and lysosomal enzyme secretion in response to IgG immune complexes, but not in response to IgA complexes. There is evidence that C5a significantly amplifies these responses. Purified human C5a enhanced the O2-. responses of neutrophils activated with IgG immune complexes and alveolar macrophages activated with either IgG or IgA immune complexes. Addition of C5a alone to neutrophils or alveolar macrophages had no direct stimulatory effect as measured by O2-. production. The observation that O2-. responses of immune complex-activated alveolar macrophages can be significantly enhanced by the presence of C5a and that C5a can also enhance O-2. responses of IgG immune complex-stimulated neutrophils suggests a potential amplification mechanism through which complement may participate in both IgG and IgA immune complex-induced lung injury. The present data corroborate in vivo studies which suggest that IgG immune complex lung injury is primarily neutrophil-mediated, whereas IgA complex lung injury is predominantly macrophage-mediated.