Leukocyte migration inhibition test in drug-induced pneumonitis]

The leukocyte migration inhibition test (LMIT) was performed in 8 cases of presumed drug-induced pneumonitis. The drugs involved were amiodarone in 3 cases, methotrexate in 2 cases, and fenofibrate, nadolol, and gold salt, each in one case. The agarose microdroplet technique for photoelectric readings of leukocyte migration inhibition was applied in the presence of a wide range of drug concentrations. LMIT was found to be positive in 6 of the 8 cases (75%). The presence of a positive LMIT indicates the elaboration of leukocyte migration inhibitory factor. These results suggest that cell-mediated immunity may play a role in the pathogenesis of drug-induced pneumonitis and that LMIT may be useful for the detection of causative drugs in patients with this condition.     

Leukocyte migration inhibition in methotrexate-induced pneumonitis. Evidence for an immunologic cell-mediated mechanism

Methotrexate-induced pneumonitis is a well-known clinical entity, but the mechanism for the induction of the pulmonary disease is ill defined. In three patients with this disorder, evidence was obtained for elaboration of a lymphokine, leukocyte inhibitory factor (LIF), by peripheral blood lymphocytes after incubation with methotrexate (MTX) in the direct leukocyte migration inhibition test. Control lymphocytes from normal subjects, as well as from patients receiving methotrexate but without pneumonitis, failed to elaborate LIF in the presence of the drug in this test. Along with these results, we obtained bronchoalveolar lavage (BAL) cell data displaying high grade lymphocyte alveolitis with a lymphocyte subset inverted ratio. This production of LIF suggests that pneumonitis associated with methotrexate therapy is also associated with a specific cellular immune response to the drug.     

Leukocyte migration inhibition in propranolol-induced pneumonitis. Evidence for an immunologic cell-mediated mechanism

About 20 cases of beta blocker-associated pneumonitis have been published in the mid-70s, and a case of interstitial pneumonitis has been attributed to propranolol. The pathogenesis of these cases of pneumonitis with or without pleural effusion is not clear. A 59-year-old man developed pneumonitis which showed all the characteristics of a drug-associated pneumonitis due to propranolol: BAL demonstrated a lymphocytosis, the variations of which closely correlated with a provocation test. The LIF appeared to be released by the patient's peripheral blood lymphocytes when cultured with optimal doses of propranolol. Production of LIF by the patients' lymphocytes suggests the existence of a drug-specific cellular immune response in propranolol-associated pneumonitis.     

Bronchoalveolar lavage cell data in amiodarone-associated pneumonitis. Evaluation in 22 patients.

To assess the value of bronchoalveolar lavage (BAL) for diagnosis, understanding, and treatment of amiodarone-associated pneumonitis, we examined the results of BAL total and differential cell counts and phenotyping of lymphocytes in 22 patients with this lung disorder and in 33 normal subjects. Overall, the total cell count was found to be almost the same as that seen in control subjects; the macrophage population was significantly reduced, and the lymphocyte, neutrophil, and eosinophil populations were increased in absolute number and percentage. When results were analyzed individually, BAL data appeared to be distributed according to two patterns. In the first pattern, there was no abnormal lymphocytosis. In the second pattern a lymphocyte alveolitis was found in percentage and in absolute number. This lymphocyte alveolitis was present either alone or associated with neutrophil alveolitis or with eosinophil alveolitis. In the first pattern, despite the normal level of the lymphocyte population, the percentage of CD4 T-lymphocytes and the CD4:CD8 T-lymphocyte ratio were significantly lowered. In the second pattern the CD8 T-lymphocyte count was increased in absolute number and percentage, with a low CD4:CD8 ratio. In six patients relavaged two to four months after amiodarone withdrawal, there was a significant fall in alveolar lymphocytosis, but the progressive increase in the neutrophil population over time seemed to be associated with the seriousness and progression of the disease. Finally, these findings closely resembled those obtained in patients with hypersensitivity pneumonitis due to inhalation of organic dust and suggest that an underlying immunologic cell-mediated mechanism may play a role in this iatrogenic pulmonary disease.     

Leukocyte migration and graft-versus-host disease

Graft-versus-host disease (GVHD) remains a significant complication of allogeneic bone marrow transplantation (allo-BMT). Acute GVHD is mediated by immunocompetent donor T cells, which migrate to lymphoid tissues soon after infusion, recognize host alloantigens, and become activated upon interaction with host antigen-presenting cells (APCs). Recent work from our group and others suggests that activated effector T cells exit lymphoid tissues and traffic to mucosal sites and parenchymal target organs such as the gastrointestinal (GI) tract, liver, lung, and skin where they cause tissue damage. The molecular interactions necessary for effector cell migration during GVHD have become the focus of a growing body of research, as these interactions represent potential therapeutic targets. In this review we discuss chemokine and chemokine receptor interactions and adhesion molecules that have been shown to play roles in effector cell migration in experimental GVHD models, and we discuss a potential model for the role of chemokines during the activation phase of GVHD.     

Acute necrotizing pneumonitis and hyperglycemia after amiodarone therapy: Case report and review of amiodarone-associated pulmonary disease

Amiodarone is a new and powerful antiarrhythmic agent currently under investigation in North America. In the past two years, there have been increasing reports of serious side effects associated with its use, including 14 cases of pneumonitis or pulmonary fibrosis. This report describes a case of acute necrotizing pneumonitis, a complication that has not been observed previously with amiodarone therapy. Amiodarone also appeared to alter carbohydrate metabolism in this patient. Metabolic changes induced by this drug may be mediated by Superoxide radicals. A high index of suspicion for pulmonary complications should be maintained in patients taking amiodarone, and nonspecific respiratory complaints should be investigated carefully.     

Pleural T-lymphocyte subsets in amiodarone-associated pleuropneumonitis

Two male patients presented with lung disorders with all the characteristics of amiodarone-related pneumonitis. Bilateral exudative pleural effusions were associated with pneumonitis. High lymphocytosis was present in the pleural fluid with a ratio of T-lymphocyte subsets close to that found in peripheral blood; in the blood T-lymphocyte subset ratio was nearly normal. By contrast, and as is usual in similar cases, lymphocytic alveolitis with T-lymphocyte subset imbalance was found in bronchoalveolar lavage fluid. These findings, never published so far to our knowledge, would favor a compartmentalization of the immune response inside the lung.     

Leukocyte migration inhibition induced by the combination of drug and a liver constituent in patients with drug-induced hepatitis.

The leukocyte migration inhibition test in agarose medium was performed in 23 cases of clinically diagnosed drug-induced hepatitis. When the test antigen was the combination of soluble phase of a liver homogenate fractionated by Sephadex G-100 which should have contained liver specific antigen and the offending drug the leukocyte migration was inhibited in 86% of cases. Whereas none of 12 cases of drug allergy without hepatic injury showed a positive result with the same combination of antigens. Other organ homogenate-muscle and kidney-never gave positive results when mixed with the offending drugs in cases of drug-induced hepatitis. It was concluded that in hypersensitivity type drug-induced hepatitis cell-mediated immunity might be established to the complex of liver specific antigen and the drug.     

Leukocyte migration inhibition in response to biliary antigens in primary biliary cirrhosis, sclerosing cholangitis, and other chronic liver diseases.

The leukocyte migration inhibition test has been used to investigate cellular immune responses to antigens in a protein fraction (BPF) of normal human gallbladder bile in patients with a variety of intra- and extrahepatic diseases. Inhibition of leukocyte migration in the presence of BPF was observed in 30 (81%) of 37 patients with PBC, in 8 (80%) of 10 patients with sclerosing cholangitis, and in 7 (26%) of 27 patients with chronic active hepatitis. Only 1 of 31 patients with other liver diseases or with uncomplicated ulcerative colitis showed a similar response to BPF. The BPF was found to contain three antigens which were distinct from plasma proteins. Immunofluorescence studies revealed that one of these antigens appears to be derived from that part of the hepatocellular membrane which forms the bile canaliculus and that a second appears to be associated with the epithelial cell membranes of interlobular and septal bile ducts. The site of origin of the third antigen could not be established. It is suggested that cellular immune responses to biliary antigens could be involved in the progressive bile duct destruction of chronic biliary disease.     

Leukocyte migration in agarose, a study on multiple sclerosis.

The effect of low concentrations of bovine encephalitogenic protein on the migration of human peripheral leukocytes in agarose was studied. A concentration of 0.3 mug/ml of the protein stimulated the migration of cells from many donors, including some healthy subjects. An indirect technique suggested that the migration enhancement is due to the production of soluble factor, possibly corresponding to the leukocyte migration enhancement factor described by others. The frequency of subjects whose cells could be stimulated and the recorded degree of stimulation tended to be higher in a group of patients with multiple sclerosis than in a group of healthy subjects. When the effect of some of the main peptide fragments of the protein was studied on cells that were stimulated by the intact protein, one or more of these peptides sometimes induced the opposite effect: a migration inhibition. There is, apparently, a complex balance between enhancing and inhibiting factors acting on leukocyte migration in vitro; and the character of the antigen seems to be one important factor.     

Leukocyte antigen-related inhibition attenuates palmitate-induced insulin resistance in muscle cells

Palmitate has been shown to induce insulin resistance in skeletal muscle cells. The aim of this study was to investigate the role of the leukocyte common antigen-related (LAR) gene in palmitate-induced insulin resistance in C2C12 cells. A stable C2C12 cell line was generated using LAR short hairpin RNA. The levels of LAR protein and phosphorylation of insulin receptor substrate-1 (IRS1) and Akt were detected by western blot analysis. 2-Deoxyglucose uptake was measured in LAR knockdown and control cells using d-[2-(3)H]glucose. LAR protein level was decreased by 65% in the stable cell line compared with the control cells. Palmitate (0.5?mM) significantly induced LAR mRNA (65%) and protein levels (40%) in myotubes compared with untreated cells. Palmitate significantly reduced insulin-stimulated glucose uptake in both the control and LAR knockdown cells by 33 and 51% respectively. However, LAR depletion improved insulin-stimulated glucose uptake in myotubes treated with palmitate. Furthermore, the inhibition of LAR prevented palmitate-induced decreases in phosphorylation of IRS1(Tyr632) and Akt(Ser473) in C2C12 cells. In conclusion, these results reveal that palmitate induces LAR expression in C2C12 cells. We also provided evidence that the inhibition of LAR attenuates palmitate-induced insulin resistance in myotubes.     

The serial changes of antibody production, macrophage migration inhibition test (MIT) and histopathologic examination in an experimental hypersensitivity pneumonitis in rabbits

We developed a hypersensitivity pneumonitis model by sensitizing rabbits intratracheally (IT) twice after two subcutaneous sensitizations (SS), and investigated the serial changes in the production of antibody against micropolyspora faeni (Mf), macrophage migration inhibition test (MIT), and histopathologic features. On the third day after the last IT, the proportions of neutrophils in bronchoalveolar lavage fluid (BALF) increased. The antibody levels of Mf specific-antigen in BALF decreased transiently, but those in serum began to increase. The number of AFC in spleen increased rapidly. Histopathologic examination revealed alveolitis and prominent acute pneumonia. On the seventh day after the last IT, the proportion of lymphocytes in BALF, the antibody levels of Mf specific-antigen in BALF, the number of AFC in BALF, and the percent inhibition of MIT were highest. Histopathologic features were characterized by the formation of prominent epithelioid granulomas. On the fourteenth day after the last IT, these features returned to the state of untreated group. These results indicate that the lesions in this HP model are formed by both type III and type IV immune reactions (Coombs-Gall). It was concluded that the seventh day after the last IT was most appropriate for the study of pathophysiology of human HP.     

The role of induced sputum in amiodarone-associated interstitial lung diseases

Amiodarone, a highly effective medication for suppressing cardiac rhythm disturbances, may cause pulmonary injury, such as chronic interstitial lung diseases, in 5-15% of the patients who take it. We applied induced sputum (IS), a non-invasive technique, for diagnosing amiodarone-induced pulmonary toxicity. Four patients with interstitial lung disease who were treated by amiodarone for ischemic heart diseases were evaluated by a conventional clinical workup. All four patients showed marked interstitial pattern on computerized tomography and decreased diffusion capacity (DLCO-SB 51-76%). IS showed lymphocytosis, a high CD4 or CD8 count, eosinophilia and amiodarone in 3 of 4 patients. IS may be a useful tool for assessing amiodarone toxicity in patients with ischemic heart diseases and concomitant pulmonary side effects.