Leonard C. Altman

Severe recurrent bacterial infections associated with defective adherence and chemotaxis in two patients with neutrophils deficient in a cell-associated glycoprotein

We studied two patients with delayed umbilical cord detachment, recurrent bacterial infections, inability to form pus, rapidly progressive periodontitis, and persistent leukocytosis. The phagocytes of both patients were strikingly abnormal in their ability to adhere to surfaces. The adherence of polymorphonuclear leukocytes to endotoxin-coated glass coverslips, glass beads, or nylon wool was markedly reduced. Scanning electron microscopy of the few adherent polymorphonuclear leukocytes from both patients showed a failure to flatten and form fine pseudopods. In vivo polymorphonuclear leukocyte and monocyte chemotaxis assessed by skin window and skin chamber methods was dramatically impaired, and in vitro chemotaxis was severely depressed. Chemiluminescence of zymosan- but not phorbol-stimulated polymorphonuclear leukocytes was markedly reduced. Allogeneic polymorphonuclear leukocytes transfused into these patients functional normally, indicating that the defect is intrinsic to the cells and not a secondary phenomenon. A 180-kilodalton glycoprotein normally present in the particulate fraction of polymorphonuclear leukocytes was found to be completely absent in Patient 1 and present in low concentration in Patient 2. We postulate that the glycoprotein deficiency interferes with the migration of polymorphonuclear leukocytes from the bloodstream into the interstitial space and to the site of infection.

Cytokines and eosinophil-derived cationic proteins upregulate intercellular adhesion molecule-1 on human nasal epithelial cells

BACKGROUND Allergic and nonallergic rhinitis with eosinophilia syndrome are characterized by tissue eosinophilia and nasal mucosal injury. Recently, it has been shown that the adherence of eosinophils and other leukocytes to epithelial cells is mediated by intercellular adhesion molecule-1 (ICAM-1) and related adherence-promoting glycoproteins. METHODS In this study we examined the constitutive expression of ICAM-1 on human nasal epithelial cells (HNECs), and the effects of interferon-gamma, tumor necrosis factor-gamma eosinophil major basic protein, and eosinophil cationic protein on the regulation of ICAM-1 expression on these cells. Similar studies were performed with A549 pneumocytes as comparative epithelial cells. RESULTS Constitutive expression of ICAM-1 was significantly higher on cultured HNECs than on A549 cells, although nasal epithelial cells in tissue specimens did not demonstrate detectable levels of ICAM-1. This spontaneous expression of ICAM-1 on cultured HNECs may explain the unique susceptibility of the nasal mucosa to rhinovirus infection, because ICAM-1 is the epithelial cell receptor for most rhinoviruses. Physiologic concentrations of major basic protein and eosinophil cationic protein stimulated significant upregulation of ICAM-1 on HNECs, which was comparable to that produced by interferon-gamma and tumor necrosis factor-alpha. In contrast, these eosinophil constituents did not stimulate ICAM-1 upregulation on A549 alveolar epithelial cells, although A549 cells did respond to interferon-gamma and tumor necrosis factor-alpha. CONCLUSION The observation that eosinophil products upregulate ICAM-1 on HNECs suggests a positive feedback mechanism, in which the products released from migrating eosinophils might promote additional HNEC-leukocyte adherence by enhancing interactions between leukocyte beta 2 integrins (CD11/18) and nasal epithelial ICAM-1.

Synthesis of interleukin-1α, interleukin-6, and interleukin-8 by cultured human nasal epithelial cells

Nasal epithelium forms the initial barrier between the environment and the respiratory system and may be a potential source of proinflammatory interleukins, which contribute to the pathophysiology of allergic and nonallergic rhinitis. To explore this possibility, epithelium and cultured human nasal epithelial cells from nasal turbinates of patients undergoing surgery for treatment of upper airway obstruction were examined for the spontaneous expression of interleukin (IL)-1 alpha, IL-1 beta, IL-6, and IL-8. Human nasal epithelial cell lysates and culture supernatants were assayed by two-site ELISAs specific for IL-1 alpha, IL-1 beta, IL-6, or IL-8. Maximum concentrations of these cytokines in supernatants ranged from approximately 0.2 to 2 ng/ml for IL-1 alpha, 1.5 to 7 ng/ml for IL-6, and 100 to 3000 ng/ml for IL-8. IL-1 alpha was predominantly cell-associated, whereas most of the IL-8 and all of the IL-6 were detected in the supernatant. Little or no IL-1 beta was detected by ELISA in the supernatants or cell lysates. Whole tissue turbinates and isolated epithelium were also examined for IL-1 beta, IL-6, and IL-8 mRNA expression by Northern blot analysis. IL-6 and IL-8 mRNAs were detected, whereas IL-1 beta mRNA was not. Furthermore, IL-6 and IL-8 release from human nasal epithelial cell cultures was enhanced by addition to the cultures of lipopolysaccharide, and IL-6 release was inhibited by polymyxin B. Thus human nasal epithelium may be a major source of IL-1 alpha, IL-6, and IL-8 in allergic and nonallergic rhinitis. Production of those proinflammatory cytokines by epithelial cells of the nasal and sinus mucosa may contribute to the pathologic and clinical events that occur in these diseases.

Defective Mononuclear Leukocyte Chemotaxis: A Previously Unrecognized Immune Dysfunction: Studies in a Patient with Chronic Mucocutaneous Candidiasis

Abstract Immunologic defects in patients with chronic mucocutaneous candidiasis vary but generally include impaired cell-mediated immunity. Mononuclear leukocyte accumulation at cellular-immune inf...

Polyinosine-polycytidylic acid stimulates versican accumulation in the extracellular matrix promoting monocyte adhesion.

Viral infections are known to exacerbate asthma and other lung diseases in which chronic inflammatory processes are implicated, but the mechanism is not well understood. The viral mimetic, polyinosine-polycytidylic acid, causes accumulation of a versican- and hyaluronan-enriched extracellular matrix (ECM) by human lung fibroblasts with increased capacity for monocyte adhesion. The fivefold increase in versican retention in this ECM is due to altered compartmentalization, with decreased degradation of cell layer-associated versican, rather than an increase in total accumulation in the culture. This is consistent with decreased mRNA levels for all of the versican splice variants. Reduced versican degradation is further supported by low levels of the epitope, DPEAAE, a product of versican digestion by a disintegrin-like and metallopeptidase with thrombospondin type 1 motif enzymes, in the ECM. The distribution of hyaluronan is similarly altered with a 3.5-fold increase in the cell layer. Pulse-chase studies of radiolabeled hyaluronan show a 50% reduction in the rate of loss from the cell layer over 24 hours. Formation of monocyte-retaining, hyaluronidase-sensitive ECMs can be blocked by the presence of anti-versican antibodies. In comparison, human lung fibroblasts treated with the cytokines, IL-1beta plus TNF-alpha, synthesize increased amounts of hyaluronan, but do not retain it or versican in the ECM, which, in turn, does not retain monocytes. These results highlight an important role for versican in the hyaluronan-dependent binding of monocytes to the ECM of lung fibroblasts stimulated with polyinosine-polycytidylic acid.

The toxicity of constituents of cedar and pine woods to pulmonary epithelium

Occupational exposure to cedar and pine woods and pine resin (colophony) can cause asthma and chronic lung disease. Prior studies suggest that plicatic and abietic acids are responsible for the asthmatic reactions that occur in cedar-wood and colophony workers; however, the etiologic mechanism(s) of the chronic lung disease is unknown. To determine if plicatic acid from cedar wood and abietic acid from pine resin could directly damage lung cells, we exposed monolayers of rat type II and human A549 alveolar epithelial cells, intact rat lungs, and rat tracheal explants to solutions of plicatic and abietic acids. As indices of injury, we measured lysis of alveolar epithelial cells with a 51Cr technique, quantitative desquamation of epithelial cells from tracheal explants, and histologic alterations in tracheal explants and intact lungs. Plicatic and abietic acids both caused dose- and time-dependent lysis of alveolar epithelial cells. Instillation of plicatic and abietic acids into rat lungs produced bronchial epithelial sloughing. Abietic acid also caused destruction of the alveolar epithelium. The addition of either acid to rat tracheal explants caused epithelial desquamation that was dose- and time-dependent. Our results suggest that plicatic acid, a unique constituent of cedar wood, and abietic acid, the major constituent in pine resin, can produce lytic damage to alveolar, tracheal, and bronchial epithelial cells. We hypothesize that repeated occupational exposure to these substances might promote the chronic lung damage observed in some cedar- and pine-wood workers and in electronic workers exposed to colophony.

ABNORMALITIES OF LEUKOCYTE CHEMOTAXIS IN HUMAN DISEASE

Dysfunctions of the immune system are associated not only with an increased susceptibility to infection and neoplasia but also with a propensity for the development of systemic inflammatory diseases. The ability of the immune system to function normally is predicated upon its ability to discriminate self from nonself, then to efficiently localize and eliminate material recognized as nonself. The process of localization and elimination of foreign material can be termed an immune-effector function and is largely mediated by an immunologically initiated inflammatory response. The local accumulation of immune effector cells, such as polymorphonuclear leukocytes (PMNs) , macrophages, or lymphocytes, is instrumental in the localization and destruction of “nonself.” PMNs (neutrophils, eosinophils, basophils) and macrophages are wandering phagocytic cells that can migrate unidirectionally along chemical gradients. During the past decade, it has clearly been shown that the interaction of the immune system with antigenic material results in the production of chemotactic factors that attract PMNs and macrophages to sites of immunologic reactions. The development of chemotactic gradients and the ability of wandering cells to respond normally to such gradients appear to be critical to host defense. Dysfunctions of leukocyte migration may therefore render an individual more susceptible to infectious, inflammatory, and perhaps neoplastic diseases. The localization of leukocytes at sites of immune reactions is accomplished by a series of interrelated processes initiated by thc recognition of foreign or neoplastically transformed materials as nonself. The combination of host recognition factors, such as serum immunoglobulins or lymphocytes, with antigens can result in the production or release of biologically active products that enhance vascular permeability, cause local vascular stasis, and attract circulating tissue leukocytes (TABLE 1 ) . The factors that control the magnitude of the inflammatory response are complex and poorly understood but depend in part on the efficiency of the mobilization of inflammatory cells and on the amount of antigenic material present and its “digestability” by PMNs or macrophages. It should be empha-

Basidiomycete allergy: measurement of spore-specific IgE antibodies

RAST with 12 basidiospore extracts demonstrated that a significant percent of 42 individuals with symptoms of perennial rhinitis and/or asthma and skin reactivity to at least one spore extract had spore-specific IgE antibodies. These antibodies were not demonstrable in 14 atopic, symptomatic, skin test negative control subjects nor in five nonatopic, asymptomatic control subjects. There was a statistically significant association between RAST and skin test results for all but three of the spore extracts. A statistically significant association was also observed between RAST results obtained with most of the different spore extracts. A similar association was present for skin test results with different spore extracts. These results provide evidence that basidiospore extracts are suitable allergens for use in diagnostic studies of respiratory disease associated with exposure to basidiomycetes.

Glucocorticoid receptors in normal human eosinophils: comparison with neutrophils

Saturation analysis with [3H]-dexamethasone was employed to measure glucocorticoid binding in purified preparations of human eosinophils and neutrophils. Eosinophils contained 10.8 +/- 1.3 x 10(3) high-affinity receptor sites per cell, with a dissociation constant (Kd) of 15.3 +/- 0.6 nM dexamethasone. Cortisol was capable of competing with [3H]-dexamethasone in the binding reaction, whereas progesterone, estradiol, estriol, and testosterone were less effective. Saturable glucocorticoid binding in neutrophils had a Kd of 17.7 +/- 0.8 nM dexamethasone with 11.1 +/- 0.8 x 10(3) sites per cell and displayed similar steroid specificity. These data indicate that normal human eosinophils have glucocorticoid receptors with characteristics similar to those in neutrophils and that in these cells ligand-receptor interaction can occur at physiologic glucocorticoid concentrations. Furthermore, these results suggest that certain glucocorticoid effects on eosinophils and neutrophils may be mediated through specific receptors.

Comparison of the in vivo autologous skin test with in vitro diagnostic tests for diagnosis of chronic autoimmune urticaria.

Previous studies indicate that 30-50% of chronic urticaria patients have an autoimmune etiology. Clinical diagnosis of autoimmune urticaria is supported with the autologous serum skin test. The purpose of this study was to compare two laboratory tests for measurement of IgG autoantibodies to IgE or IgE receptors and compare the results with the autologous serum and plasma skin tests. We performed skin tests and two functional in vitro tests, basophil histamine release, and CD63 up-regulation to detect autoantibodies relevant to autoimmune urticaria. Both sera and citrated plasma were evaluated in the autologous skin test and histamine release assay. Thyroid autoantibodies were also measured. Basophils were incubated with patient plasma, sera, buffer, or anti-IgE. The cells were analyzed for CD63 expression and the supernatants were recovered for histamine analysis. There was high correlation between CD63 up-regulation and histamine release assays, but histamine release was more sensitive. There was a high concordance between sera and citrated plasma for the skin test. Sera from chronic urticaria patients produced higher mean histamine release (23%) compared with citrated plasma (12%). Thirty-one percent of patients positive in the histamine release assay were also positive for thyroid autoantibodies. This compares with 12% who were negative in the histamine release assay. These data show that in vitro basophil histamine release can be used to measure antibodies to FceRI, FceRII/CD23, or IgE and identify patients with autoimmune urticaria.