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Philip E Silkoff, MD(*); Henry Milgrom, MD; Z V Tran, PhD; F Romero, BS; R Townley, MD; Niroo Gupta, MD; Patricia Rohane, MD and Robert B Fick, MD. National Jewish [...]

The sera from 32 patients with cystic fibrosis who were chronically colonized with mucoid strains of Pseudomonas aeruginosa(MPA) were tested for anti-MPA antibodies. Using an enzyme-linked immunosorbent assay we measured IgA, IgG, and IgM antibody titers to three MPA strains, extracts of those strains, and seaweed-derived sodium alginate, which is similar chemically to the exopolysaccharide of MPA. These titers were compared with identical tests performed on the sera of eight cystic fibrosis patients who never were colonized with MPA and 10 normal adults. The IgG titers were significantly higher in tests of sera from the colonized patients compared with the other two groups but the IgA and IgM titers were not significantly higher. In colonized patients antibody titers to the different antigens correlated with each other suggesting that the major antibody response was to common antigenic determinants. Using these titers as a data base, eight patients whose clinical status was unknown to the testers, had IgG-enzyme-linked immunosorbent assay tests of their sera and the four colonized patients with cystic fibrosis were correctly identified. Three of them had substantial titers of antibody in the bronchoalveolar lavage fluid.

ABSTRACT: The sera from 32 patients with cystic fibrosis who were chronically colonized with mucoid strains of Pseudomonas aeruginosa (MPA) were tested for anti-MPA antibodies. Using an enzyme-linked immunosorbent assay we measured IgA, IgG, and IgM antibody titers to three MPA strains, extracts of those strains, and seaweed-derived sodium alginate, which is similar chemically to the exopolysaccharide of MPA. These titers were compared with identical tests performed on the sera of eight cystic fibrosis patients who never were colonized with MPA and 10 normal adults. The IgG titers were significantly higher in tests of sera from the colonized patients compared with the other two groups but the IgA and IgM titers were not significantly higher. In colonized patients antibody titers to the different antigens correlated with each other suggesting that the major antibody response was to common antigenic determinants. Using these titers as a data base, eight patients whose clinical status was unknown to the testers, had IgG-enzyme-linked immunosorbent assay tests of their sera and the four colonized patients with cystic fibrosis were correctly identified. Three of them had substantial titers of antibody in the bronchoalveolar lavage fluid.

1. β-Adrenergic responsiveness may be decreased in cystic fibrosis. In order to determine whether this reflects an alteration in the human lymphocyte β-receptor complex, we studied 12 subjects with cystic fibrosis (six were stable and ambulatory and six were decompensated, hospitalized) as compared with 12 normal controls. 2. Lymphocyte β-receptor mediated adenylate cyclase activity (EC 4.6.1.1) was not decreased in the ambulatory cystic fibrosis patients as compared with controls. In contrast, decompensated hospitalized cystic fibrosis patients demonstrated a significant reduction in β-receptor mediated lymphocyte adenylate cyclase activity expressed as the relative increase over basal levels stimulated by the β-agonist isoprenaline compared with both normal controls and stable ambulatory cystic fibrosis patients (control 58 ± 4%; ambulatory cystic fibrosis patients 51 ± 7%; decompensated hospitalized cystic fibrosis patients 28 ± 5%; P < 0.05). 3. Our data suggest that defects in lymphocyte β-receptor properties in cystic fibrosis patients may be better correlated with clinical status than with presence or absence of the disease state.

ABSTRACT. Experiments performed in vitro have demonstrated that leukocyte neutral proteases produce an important mediator of inflammation, C5a, by proteolysis of the C5 component of the complement system. Cystic fibrosis (CF) lung fluids were characterized by high levels of neutrophils (39% of total cells versus 2% in normals) and contained significantly elevated amounts of elastolytic activity (mean 17.7 ng/µg total protein) compared to the lung fluids obtained from normal volunteers (0.2 ng elastolytic activity/µg protein, p = 0.001). The objective of these studies was to determine if complement activation and complement-derived chemotactic activity are present in CF lung fluids. C3c peptide representing activation of C3 could not be identified in the bronchial-alveolar lung lavage fluids of normal subjects but was readily identified by means of crossed immunoelectrophoresis in CF lung fluids (n = 9, mean 49% of C3); the mean level of C3 was decreased in CF lung specimens. Chemotactic activity was significantly elevated in lung fluids of the CF patients when compared to normal lung fluids. Using gel-filtration chromatography and a sensitive radioimmunoassay the chemotaxin present in CF specimens was identified as the anaphylatoxin C5a. C5a levels in the bronchial-alveolar lavage fluids of CF patients was inversely related to volume in liters expired in 1 s of a forced expiratory maneuver expressed as a percent of vital capacity determined from a forced expiratory maneuver (r = -0.72). Because there was a direct relationship between the total elastolytic activity present in CF airways and the concentration of C5a (r = 0.97, p = 0.03), it was postulated that airway proteases with elastolytic activity also cleave C5, nonimmunologically producing C5a. Detailed inhibition assays revealed that much of the total elastolytic activity had the inhibition profile of a serine proteinase. The levels of the serine proteinases were closely correlated with the numbers of neutrophilic leukocytes present per ml of lavage fluid (r = 0.7, p = 0.05). However, inhibitors of leukocyte serine proteases did not prevent the generation of additional chemotactic activity and the proteolysis of radiolabeled C5 substrate was not prevented by inhibitors of neutrophil elastase. Although the purified metalloelastase of Pseudomonas aeruginosa was active on cell-bound and free C5 yielding C5a, inhibition of this bacterial protease in CF lung fluids only partially blocked cleavage of the α- and β-chains of C5. Simultaneous inhibition of the serine and metalloprotease activities of CF lung fluids effectively blocked the in vitro generation of C5a leukocyte chemotactic activity. We conclude that CF lung fluids contain markedly increased levels of elastolytic activity which are closely linked to the extent of CF lung disease and which are capable of proteolytically generating C5a. Both a serine proteinase and a metalloenzyme in these lung fluids contribute to the nonimmunologic generation of C5a.

Experiments performed in vitrohave demonstrated that leukocyte neutral proteases produce an important mediator of inflammation, C5a, by proteolysis of the C5 component of the complement system. Cystic fibrosis (CF) lung fluids were characterized by high levels of neutrophils (39 of total cells versus2 in normals) and contained significantly elevated amounts of elastolytic activity (mean 17.7 ngµg total protein) compared to the lung fluids obtained from normal volunteers (0.2 ng elastolytic activity/µg protein, p = 0.001). The objective of these studies was to determine if complement activation and complement-derived chemotactic activity are present in CF lung fluids. C3c peptide representing activation of C3 could not be identified in the bronchial-alveolar lung lavage fluids of normal subjects but was readily identified by means of crossed immunoelectrophoresis in CF lung fluids (n = 9, mean 49 of C3); the mean level of C3 was decreased in CF lung specimens. Chemotactic activity was significantly elevated in lung fluids of the CF patients when compared to normal lung fluids. Using gel-filtration chromatography and a sensitive radioimmunoassay the chemotaxin present in CF specimens was identified as the anaphylatoxin C5a. C5a levels in the bronchial-alveolar lavage fluids of CF patients was inversely related to volume in liters expired in 1 s of a forced expiratory maneuver expressed as a percent of vital capacity determined from a forced expiratory maneuver (r = -0.72). Because there was a direct relationship between the total elastolytic activity present in CF airways and the concentration of C5a (r = 0.97, p = 0.03), it was postulated that airway proteases with elastolytic activity also cleave C5, nonimmunologically producing C5a. Detailed inhibition assays revealed that much of the total elastolytic activity had the inhibition profile of a serine proteinase. The levels of the serine proteinases were closely correlated with the numbers of neutrophilic leukocytes present per ml of lavage fluid (r = 0.7, p = 0.05). However, inhibitors of leukocyte serine proteases did not prevent the generation of additional chemotactic activity and the proteolysis of radiolabeled C5 substrate was not prevented by inhibitors of neutrophil elastase. Although the purified metalloelastase of Pseudomonas aeruginosawas active on cell-bound and free C5 yielding C5a, inhibition of this bacterial protease in CF lung fluids only partially blocked cleavage of the - and ß-chains of C5. Simultaneous inhibition of the serine and metalloprotease activities of CF lung fluids effectively blocked the in vitrogeneration of C5a leukocyte chemotactic activity. We conclude that CF lung fluids contain markedly increased levels of elastolytic activity which are closely linked to the extent of CF lung disease and which are capable of proteolytically generating C5a. Both a serine proteinase and a metalloenzyme in these lung fluids contribute to the nonimmunologic generation of C5a. (Pediatr Res201258–1268,1986)

Two patients were treated with bone marrow transplantation and subsequently developed nonbacterial thrombotic endocarditis (NBTE). Both patients died of embolic sequellae and in neither was NBTE suspected antefinem. It is clear that NBTE occurs with increased frequency in bone marrow transplant (BMT) recipients and through arterial embolization contributes significantly to the morbidity and mortality of this procedure. In those at greatest risk, including BMT recipients, detection of disseminated intravascular coagulation; soft, changing systolic cardiac murmurs; hematuria; and signs of central embolic events suggest the diagnosis of NBTE. Awareness of the diagnosis of NBTE is essential for those who must interpret neurologic, myocardial and renal abnormalities in BMT recipients.

A nonsurgical, less aggressive, less toxic chemotherapeutic protocol for the management of nontuberculous mycobacterial (NTB) pulmonary infections has been uniformly applied to patients in our institution between 1972 and 1985. Forty-three nonimmunocompromised patients with active lung disease caused by Mycobacterium avium-intracellulare(MAI) (n = 26), M kansasii(n = 16), and M xenopi(n = 1) were identified retrospectively. Eighteen MAI patients were treated with three or four antituberculosis agents resulting in sputum conversion and clinical improvement in 12 (67 percent). Additionally, 11 out of 16 (69 percent) patients completing therapy or still undergoing therapy for persistent MAI disease, achieved sputum conversion and clinical improvement after prolonged therapy (3.6 ± 0.5 years [SEM]). When M kansasiiwas identified as the etiologic agent, all patients were treated with four or fewer antituberculosis agents and 14 out of 16 patients (88 percent) achieved sputum conversion and clinical improvement throughout the follow-up period. We conclude that the use of three or four chemotherapeutic agents in the treatment of NTM lung disease provides an excellent probability of successful outcome even in MAI infections.