M. Michael Glovsky

C3a-induced lysosomal enzyme secretion from human neutrophils: lack of inhibition by f met-leu-phe antagonists and inhibition by arachidonic acid antagonists.

C3a-induced lysosomal enzyme secretion from human peripheral neutrophils in a noncytolytic, dose-dependent (10-100 microgram/ml) process. Release of both primary and secondary granule constituents occurred when neutrophils were exposed to C3a plus cytochalasin B, however, C3 alone induced limited release of lysozyme. A competitive antagonist of the formyl-peptide receptor on neutrophils, t boc (phe-leu) 2-phe, did not block the release induced by C3a. Arachidonic acid antagonists, nordihydroguaiaretic acid and quercetin caused dose-dependent inhibition of release induced by C3a plus cytochalasin B, however, lysozyme release induced by C3a in the absence of cytochalasin B was minimally affected. Indomethacin at high concentration (greater than 10(-5) M) had similar inhibitory effects.

Identification of receptor regulatory proteins, membrane glycoproteins, and functional characteristics of adenylate cyclase in vesicles derived from the human neutrophil

Human neutrophils were disrupted by brief sonication under conditions which preserve the hormone sensitivity of adenylate cyclase and yield minimal granule lysis. Fractions enriched in adenylate cyclase were analysed for hormonal and guanine nucleotide regulation of the enzyme as well as structural proteins. Adenylate cyclase was activated by PGE1 and isoproterenol in a GTP-dependent fashion, while f-met-leu-phe and C5a gave no stimulation. Cholera toxin treatment, which specifically modifies cyclase-related GTP-binding proteins, caused a dose-dependent enhancement of GTP activation, in which GTP alone activated maximally and PGE1 was without further effect. The following proteins were detected in the cyclase-containing vesicles: a 42 K mol. wt protein labeled selectively by cholera toxin; protein subunits observed in SDS gels at 214, 165, 105 and 47 K, of which the 47 K band was the most prominent and comigrated with actin; prominent lectin-binding activities at 165 K (concanavalin A and wheat germ agglutinin) as well as at 100 K (wheat germ agglutinin); and a set of proteins and lectin-binding activities in fractions containing beta-glucuronidase activity distinct from adenylate cyclase containing vesicles. The identification of receptor-controlled cyclase, GTP-binding regulatory proteins, cytoskeletal elements and unique lectin-binding activities in a single vesicle preparation should contribute to an understanding of receptor-mediated control of neutrophil function.

Chronic chyluria: a clinical study of 3 patients.

Studies of patients with chyluria or chylothorax have demonstrated significant disruptions of protein, blood and fat metabolism that may result in iron deficiency anemia, hypoproteinemia, hypolipidemia and malnutrition. To document the sequential development of these complications we performed serial clinical and biochemical studies for 2 to 12 years in 3 patients with presumed filarial chyluria whose sole treatment had been diethylcarbamazine. Despite the chronic loss of chyle in the urine these 3 patients did not have significant complications during the period of observation. The weight and blood pressure remained stable. No persistent anemia, hypoproteinemia or hypolipidemia was noted. Except for 1 patient in whom a transient decrease of the creatinine clearance developed during pregnancy, no permanent renal function impairment occurred. These observations suggest that chronic chyluria may not always result in serious alterations of the physical status or body functions of these patients requiring surgical repair, and supports the hypothesis that untreated chyluria could be a relatively benign process in our milieu.

Morphological changes in human polymorphonuclear leukocytes induced by c3a in the presence and absence of cytochalasin b

Treatment of human peripheral polymorphonuclear leukocytes(PMNs) with C3a leads to profound changes in cell morphology, along with the limited secretion of lysozyme. Simultaneous addition of C3a and cytochalasin B enhances the release of lysosomal enzymes and renders the cell morphology similar to that seen when either C5a or n-formyl peptides are added to PMNs in the presence of cytochalasin B. The morphological response of PMNs treated with C3a is considerably more heterogeneous than that induced by either C5a or n-formyl-methionyl-leucyl-phenylalanine, indicating that some, but not all, PMNs have receptors for C3a.

Immunologic studies in patients with chyluria

Patients with chyluria, postoperative chylothorax, or surgical thoracic duct drainage are depleted of lymphocytes, IgA, and C3 as a result of the loss of lymphatic cellular and humoral elements, but the specific defects have not been well characterized. Therefore, we investigated the immunologic status of three patients with long-standing chyluria who were lymphopenic but did not have evidence of opportunistic infections. All patients had normal levels of immunoglobulins except for a moderate decrease in the IgA fraction, a normal antibody response to viral agents, and a normal hypersensitivity response to at least one of six recall antigens. Their complement levels were essentially normal. All were leukopenic because of decreased numbers of circulating lymphocytes. Two had depressed relative and absolute numbers of OKT4 (helper/inducer) T cells and normal or increased relative numbers of OKT8 (cytotoxic/suppressor) T cells, resulting in a marked reversal of the OKT4 to OKT8 ratio. The third was severely depleted of both OKT4 and OKT8 cells. The response of their peripheral blood mononuclear cells to mitogen stimulation was reduced, but they all had normal or increased absolute and relative numbers of natural killer (NK) cells as defined by HNK-1 or OKM-1. Their NK activity was less than that of normal controls unless stimulated by interferon, suggesting that many of the NK cells were immature. Normal delayed hypersensitivity, reduced but effective mitogen stimulation, and a normal NK response after interferon may be factors that protect these patients against opportunistic infections and neoplasms.

Gamma a Myeloma with Hyperviscosity and Obstructive Uropathy

A patient with gamma A myeloma, hyperviscosity and an obstructive uropathy is described. Operation revealed a proteinaceous mass obstructing the right renal pelvis. Immunoprecipitin and immunofluorescent analysis of this mass and concentrated urine demonstrated the presence of gamma A myeloma protein, kappa light chains and albumin. This is the first description of an obstructive uropathy in multiple myeloma owing to a proteinaceous matrix containing paraprotein.