Simultaneous measurement of adherence and chemiluminescence by polymorphonuclear leukocytes.

Oxygen radical release from adhering polymorphonuclear leukocytes (PMN) has been implicated as an important feature of many vascular diseases. We developed a technique by which adherence and production of O2 radicals by PMN can be measured simultaneously. The technique combines the conventional nylon fiber assay for measuring adherence of PMN with concurrent scintillation counter measurement of chemiluminescence (CL) to assess O2 radical production by PMN. We found that adherence of PMN to nylon fiber is associated with increases in CL. Moreover, increases in CL appear to be dependent on generation of O2 radicals from PMN since they are not seen with PMN from a patient with chronic granulomatous disease (CGD) or in the presence of O2 radical scavengers, superoxide dismutase, or catalase. Furthermore, agents which increase the adherence of PMN to nylon fiber are associated with increases in CL. Use of this approach may facilitate simultaneous evaluation of adherence and O2 radical generation by PMN.

Surgical management of severe thoracic lordosis. A new technique to restore normal kyphosis.

A technique of combined anterior and posterior surgery for correction of severe thoracic lordosis with the restoration of normal kyphosis is described. The results in three patients are presented. Preliminary results suggest that this is a valid procedure for obtaining a normal thoracic kyphosis, and that correction of thoracic lordosis may result in improved pulmonary function.

The value of bronchoscopy in establishing the etiology of pneumonia in renal transplant recipients.

Fifty-two episodes of fever and new pulmonary infiltrates were evaluated prospectively in 51 renal allograft recipients. Thirty-nine flexible fiberoptic bronchoscopies were performed in the diagnostic evaluation of these infiltrates. Specific etiologic diagnoses were obtained in 30 (77%) of the patients. This information was clinically useful, as defined by preset criteria, in 21 (54%) of the patients and definitive but not clinically useful in an additional 9 (23%). In the remaining 9, it was neither definitive nor clinically useful. Microbiology brush specimens were useful in establishing etiologic diagnoses in 12 (44%) of the 27 patients in whom it was performed. Transbronchial lung biopsies yielded specific etiologic diagnoses in 9 (53%) of the 17 biopsies obtained. Complications related to the bronchoscopic procedure occurred in 2 patients (5% of total bronchoscopies). No prolonged morbidity was noted. We conclude that fiberoptic bronchoscopy is a safe, useful procedure, and should be considered early in the diagnostic evaluation of pulmonary infections in renal transplant recipients.

Platelets increase neutrophil adherence in vitro to nylon fiber.

The effect of platelets on the adherence of neutrophils to nylon fiber was assessed in whole blood samples and purified neutrophil suspensions in the presence or absence of plasma. In whole blood samples, increasing numbers of platelets were associated (r = 0.47, p less than 0.02) with increasing adherence of neutrophils. Addition of platelets in plasma to purified neutrophil suspensions increased (p less than 0.05) neutrophil adherence from 76.2% +/- 1.4 to 88.0% +/- 2.0. Similarly, addition of washed platelets without plasma also increased (p less than 0.05) neutrophil adherence from 67.9% +/- 5.8 (without added platelets) to 94.2% +/- 1.6 (with 300,000 platelets/mm3 added). In contrast, no augmentation of neutrophil adherence occurred if platelets had their aggregation response suppressed by pretreating platelet donors with aspirin. SEM supported these findings by showing platelets in close association with neutrophils adhering to nylon fiber. These findings emphasize the importance of platelet numbers and reactivity on the adherence of neutrophils.

Serum from patients with invasive fungal infections inhibits the adherence of polymorphonuclear leukocytes and alveolar macrophages.

Adherence of various combinations of polymorphonuclear leukocytes (PMN) or alveolar macrophages (AM) and serum from nonimmunosuppressed patients wit culture-proved, invasive fungal infections or control subjects was evaluated in vitro using the standard nylon fiber pipette technique. In autologous serum, adherence of PMN from patients with wide variety of untreated fungal infections, including blastomycosis, histoplasmosis, cryptococcosis, coccidiodomycosis, and spirotrichosis, was significantly (p less than 0.001) decreased when compared with PMN from uninfected control subjects or patients with untreated bacterial infections. Studies using various combinations of PMN and serums from patients with blastomycosis suggested that the adherence defect was due to a serum disorder rather than an intrinsic cellular abnormality. Preincubation in serum from patients with blastomycosis decreased the adherence of control PMN. Preincubation in control serum corrected the decreased adherence of PMN from patients with blastomycosis. Additional studies revealed that the inhibitor was heat-stable and reversible, being present before, but not after, treatment or spontaneous resolution of the patient's infections. Adherence of AM from patients with fungal infection was also normal except when AM were preincubated in serum from patients with untreated fungal infections. We concluded that the intrinsic adherence of PMN and AM from patients with untreated fungal infections is normal, but that these patients do have an extrinsic heat-labile serum factor that can decrease the adherence of PMN and AM.

Isolation of Chlamydia trachomatis from the lower respiratory tract of adults.

Chlamydia trachomatis was isolated from the lower respiratory tract of six patients who had pulmonary infection, the severity varying from acute bronchitis to severe diffuse interstitial pneumonia. The latter appeared in four immunosuppressed patients, from three of whom cytomegalovirus was also isolated. Therapy directed against C. trachomatis resulted in prompt clinical improvement in three patients. Two patients died. C. trachomatis causes disease in the adult respiratory tract, the prevalence of which has yet to be determined, and it may be an important cause of morbidity and mortality in the immunosuppressed patient.

Spontaneous regression of diffuse tracheobronchial amyloidosis.

Diffuse tracheobronchial amyloidosis which underwent an extensive spontaneous regression is described in a 39-year-old man. This regression was documented by serial roentgenograms of the chest, studies of pulmonary function, and bronchoscopic procedures with mucosal biopsies. The clinical implications of this spontaneous change are discussed.

Reversible impairment of the adherence of alveolar macrophages from cigarette smokers.

The nylon fiber adherence in vitro of alveolar macrophages (AM) from cigarette smokers was uniformly decreased. The mean adherence of AM from 16 cigarette smokers was 53 +/- 3.0 per cent, compared to a mean adherence of 77.2 +/- 1.7 per cent for AM from control nonsmokers. The defect was not present after the subjects quit smoking, was not a result of factors in lavage fluids from smokers, and was not apparent in polymorphonuclear leukocytes. The morphologic features of the surface of AM were examined with scanning electron microscopy to determine whether morphologic changes paralleled the decreased adherence of AM from smokers. Marked alterations in the surface of AM from cigarette smokers, which could affect the ability of AM to adhere optimally, were demonstrated before attachment to the fiber. In summary, there exists a reversible, intrinsic defect in the structure and adherence of AM from cigarette smokers that may influence their function and may accout, in part, for the increased yield of AM from the lavage fluid of cigarette smokers.

Comparison of the metabolism of alveolar macrophages from humans, rats, and rabbits: phorbol myristate acetate.

Metabolic activities of unstimulated or stimulated AMs from humans, rats, and rabbits were examined and compared in vitro. Rates of oxygen consumption, chemiluminescence, and glucose (1- or 6-14C) oxidation by unstimulated AMs from these three species increased following stimulation of the AMs by bacteria or PMA. Although the absolute metabolic responses of AMs from humans, rats, or wild rabbits were different, the metabolic activities from each species were nearly identical when compared on the basis of protein content of the cells. In contrast to the enhanced biochemical responses of AMs from humans, rats, or wild rabbits, stimulated AMs from certain commercially supplied rabbits failed to increase their metabolism. The failure of AMs from these rabbits to respond metabolically was probably due to an acquired abnormality resulting from their care and storage at the supplier. The defect was associated with the presence of large numbers of Bordetella bronchiseptica organisms in the lavage effluents from these commercially supplied rabbits. This abnormality in metabolism of AMs was reversed following prolonged residence of the rabbits in the laboratory, and the correction of the defect was accompanied by a disappearance of B. bronchiseptica from the lavage fluid. The results comprehensively compare and contrast the metabolism of AMs from humans and animals and emphasize the need to document the appropriateness of animal models before using them to predict biologic reactions of humans.

Defective neutrophil locomotion in human blastomycosis: evidence for a serum inhibitor.

Unstimulated or stimulated locomotion, bactericidal, and metabolic activities of polymorphonuclear leukocytes (PMN) from 12 nonimmunosuppressed patients with invasive fungal infections proved by culture, were evaluated before and after treatment of the patients with antimicrobial drugs. The major observation was that PMN from patients with blastomycosis had a defect in stimulated locomotion. The specificity of the defect for blastomycosis was substantiated by the normal stimulated locomotion of PMN from uninfected control subjects or untreated patients with histoplasmosis, cryptococcosis, coccidioidomycosis, or sporotrichosis. The defect was due to a heat-stable, cell-directed, reversible serum inhibitor. In unheated or heated serum from untreated patients with blastomycosis, control PMN had decreased stimulated locomotion. Multiple washing followed by addition of control serum corrected locomotion of PMN from untreated patients with blastomycosis. The abnormality was not present in PMN from patients who had been treated with amphotericin B or had spontaneous resolution of their infections. Inhibition was not due to absence of chemoattractant activity because zymosan-activated patient serum or mixtures of patient and control serum stimulated PMN locomotion normally. The defect did not correlate with age, sex, neutrophil count, nitroblue tetrazolium reduction, serologic reactivity, or duration or severity of infection. No defect was found in bactericidal or metabolic activities of various combinations of PMN and serum from untreated or treated patients with blastomycosis or the 4 other fungal infections tested, indicating that the inhibitor was specific for stimulated locomotion.

The effect of phorbol myristate acetate on the metabolism and ultrastructure of human alveolar macrophages.

In the present investigation we examined the influence of the surface-active agent phorbol myristate acetate (PMA) and opsonized heat-killed bacteria (HKB) on oxygen consumption, superoxide release, and glucose oxidation of human alveolar macrophages (AM). Both PMA and HKB produced a surge in oxygen consumption, superoxide release, and oxidation of 1-14C-glucose and 6-14C-glucose by human AM. Examination of AM by electron microscopy following stimulation by these two agents demonstrated membrane ruffling, loss of microvilli, and increased vacuolization in PMA-treated cells and phagocytic vacuoles containing bacteria in HKB-treated cells. The vacuolization produced by PMA-treated AM was much less striking than the vacuolization produced in PMA-treated leukocytes. The similarity in the metabolic and some of the physical responses of AM stimulated by PMA and HKB suggest that PMA may be a useful agent for evaluating cell-membrane-related events of phagocytosis in AM.

Chemiluminescence by human alveolar macrophages: stimulation with heat-killed bacteria or phorobol myristate acetate.

Chemiluminescence of human alveolar macrophages (AM) was evaluated in vitro. Unstimulated AM generated chemiluminescence that remained constant during incubation. Addition of heat-killed Staphylococcus aureus 502A (HKB) or a chemical agent, phorbol myristate acetate, produced high rates of chemiluminescence that were significantly (P less than 0.05) increased over unstimulated AM. Phorbol myristate acetate-and HKB-stimulated increases in AM chemiluminescence were completely blocked by the enzyme superoxide dismutase. In comparison with unstimulated polymorphonuclear leukocytes, unstimulated AM had significantly (P less than 0.005) greater levels of chemiluminescence. However, after stimulation by phorbol myristate acetate or HKB, AM showed less chemiluminescence than similarly treated polymorphonuclear leukocytes.