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Biomedical subjects

E J Goetzl

Publications and source records attributed to E J Goetzl.

At least 109 records · Page 6Linked to original sources
PubMed1989

Appearance of neuropeptides in ascitic fluid after peritoneal therapy with interleukin-2 and lymphokine-activated killer cells for intraabdominal malignancy.

Administration of intravenous interleukin-2 (IL-2), followed by intraperitoneal IL-2 and autologous lymphokine-activated killer (LAK) cells to six patients with colonic, ovarian, or endometrial carcinoma restricted to peritoneal spread increased significantly the ascitic fluid concentrations of the neuropeptides substance P (SP) and calcitonin-gene related peptide (CGRP). After intravenous IL-2 alone, the level of SP rose 10- to 140-fold, without a change in that of CGRP. Intraperitoneal IL-2 and LAK cells led to elevations in the concentrations of SP and CGRP to respective maximal means of 319 and 175 pM after 8 hr, which were maintained for 24-48 hr without alterations in the levels of vasoactive intestinal peptide or somatostatin. SP and CGRP from peritoneal fluid were chromatographically indistinguishable from synthetic neuropeptides. The increases in concentrations of SP and CGRP after IL-2 and LAK-cell therapy are the first demonstration of a neural response to a human cellular immunological reaction. The time course and magnitude of the neuropeptide response suggest a role in the vascular side effects of this form of treatment.

Ascitic Fluid↗
PubMed1989

Late appearance of phospholipid platelet-activating factor and leukotriene B4 in human skin after repeated antigen challenge.

Inflammatory mediators were assessed in supernatants of chamber fluids from eight ragweed- or grass-sensitive subjects during antigen-induced cutaneous inflammatory responses. Platelet activating factor (PAF) accumulated at concentrations of 1 pm to 90 mumol/L in six of eight subjects beginning at 3 hours and continuing for 9 hours after antigen challenge. Leukotriene B4 (LTB4) was detectable at cutaneous sites of antigen challenge in five of five subjects throughout the 9-hour period at levels from 1 to 36 nmol, a range of 38% to 80% of which were omega-oxidation metabolites. Histamine levels peaked in the first hour at 106 +/- 18 ng/ml and decreased to a plateau of 11 to 13 ng/ml at 3 to 9 hours after antigen challenge. No PAF and only very low levels of LTB4 (0.1 to 1.3 nmol) and of histamine (less than 2 ng/ml) were detected at buffer-control sites during the 9 hours of study. Continuous antigen exposure thus results in the persistent release of histamine and LTB4 and the late appearance of PAF, all of which may contribute to the chronicity of allergic disorders and may have a bearing on the IgE-mediated, late-phase cutaneous response.

Adult↗
PubMed1989

Identification of leukotriene B4 as the neutrophil chemotactic factor released by antigen challenge from passively sensitized guinea pig lungs.

Neutrophils are prominent in some IgE-mediated allergic reactions and may contribute to the pathophysiology of immediate hypersensitivity. Antigen challenge of fragments of guinea pig lung tissue that were passively sensitized with IgE or IgG antibody evoked the release of neutrophil chemotactic activity (NCA) in parallel with histamine. The NCA released from lung tissue by both IgG- and IgE-dependent stimulation coeluted from a column of Sephacryl S-300 with synthetic leukotriene B4 (LTB4). The NCA in eluates from the Sephacryl S-300 column contained LTB4, as determined by high-performance liquid chromatography and specific radioimmunoassay, in quantities that accounted for the observed chemoattractant activity in the eluates. Furthermore, the NCA of supernatants from antigen-challenged lung fragments was reduced by a mean of 80% after absorption with a monoclonal antibody to LTB4. LTB4 thus constitutes the major functional constituent of NCA released after anaphylactic challenge of IgE- and IgG-sensitized guinea pig lung tissue.

Anaphylaxis↗
PubMed1989

Mediation of primary afferent peripheral hyperalgesia by the cAMP second messenger system.

Cyclooxygenase (prostaglandin E2 and prostaglandin I2) and lipoxygenase [8(R), 15(S)-dihydroxyicosa-(5E-9,11,13Z)-tetraenoic acid] products of arachidonic acid metabolism are thought to produce peripheral hyperalgesia by a direct action on the primary afferent nociceptor. In this study we investigated the possibility that these eicosanoids generate hyperalgesia through a common second messenger in the rat. We report that 8-bromo cAMP, a membrane permeable analogue of cAMP, produces a dose-dependent hyperalgesia that is not affected by treatments that interrupt indirect routes of hyperalgesia production including sympathectomy with 6-hydroxydopamine, depletion of polymorphonuclear leukocytes (a source of hyperalgesic eicosanoids) with hydroxyurea, or blockade of the cyclooxygenase pathway of arachidonic acid metabolism with indomethacin. The phosphodiesterase inhibitor isobutyl-methylxanthine markedly prolongs the hyperalgesic effect of 8-bromo cAMP as well as those of the directly acting hyperalgesic agents prostaglandin E2, prostaglandin I2 and 8(R),15(S)-dihydroxyicosa-(5E-9,11,13Z)-tetraenoic acid. We conclude that the effect of all known hyperalgesic eicosanoids is mediated by the cAMP second messenger system and suggest, therefore, that cAMP mediates peripheral hyperalgesia in primary afferent nociceptors.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1989

Diverse responses of human fibroblasts to a highly purified fibroblast-activating factor from the U937 line of human monocytes.

The responses of human dermal fibroblasts to highly purified fibroblast-activating factor (FAF), derived from supernatants of U937 human monocytes stimulated with phorbol myristate acetate (PMA), were investigated with several in vitro assays of specific synthetic functions. The highly purified peptide was detected as a single 16,000-18,000 MW protein, by both silver staining and Western blot analysis, with an antiserum generated against a synthetic peptide representing the amino-terminal sequence of 17 amino acids. At concentrations that induced similar levels of fibroblast proliferation, transforming growth factor-beta (TGF-beta), interleukin-1 (IL-1), acidic fibroblast growth factor (aFGF) and FAF also stimulated fibroblasts to generate and release prostaglandin E2 (PGE2) and proteoglycans. TGF-beta had the least effect on proteoglycan production. In contrast, the production and secretion of collagen evoked by FAF was only minimal when compared to that observed with IL-1 and aFGF. FAF and aFGF promoted fibroblast-induced collagen gel contraction with similar potency. Thus, the profile of fibroblast effects is a specific property of each cytokine.

Antigens, Neoplasm↗
PubMed1989

Unique pattern of cleavage of vasoactive intestinal peptide by human lymphocytes.

Human cultured T lymphocytes of the Jurkat line and myeloma cells of the U266 line cleaved the 28 amino acid vasoactive intestinal peptide (VIP1-28) preferentially at three sites with time- and temperature-dependence. The fragments VIP4-28 and VIP23-28) from an endopeptidase activity, and VIP15-28 from a trypsin-like peptidase, together represented a range of 26-65% of the VIP1-28 recovered after 2 hr at 37 degrees C or 4 hr at 22 degrees C, based on the absorbance of purified peptides and the radioactivity of [125I]Tyr10 VIP1-28. The endopeptidase activity was associated with membranes recovered after disruption of U266 cells by nitrogen cavitation. Pretreatment of intact U266 and Jurkat cells with diisopropylfluorophosphate (DFP) and the subsequently isolated subcellular particles with phenylmethylsulphonylfluoride (PMSF) and leupeptin inhibited the trypsin-like enzyme by a mean of 80%, without suppressing endopeptidase activity. In contrast, 0.1 mM DL-thiorphan and phosphoramidon blocked selectively a range of 35-70% of the endopeptidase activity in membrane preparations and intact cells. The capacity of lymphocytes to degrade VIP1-28 may substantially alter the effects of this neuromediator on functions of some subsets of T and B cells.

B-Lymphocytes↗
PubMed1988

Isolation and partial characterization of a fibroblast-activating factor generated by U-937 human monocytic leukocytes.

Human cultured monocyte-like tumor cells of the U-937 line, that are incubated for 24 h at 37 degrees C with 100 ng/ml of PMA, generate and release a fibroblast-activating factor (FAF) capable of enhancing the uptake of [3H]TdR by human dermal fibroblasts in vitro. The predominant FAF activity in unpurified supernatants from U-937 cells was associated with one protein of 16 to 18 kDa, as assessed by SDS-PAGE and pI 4 to 5, as determined by IEF in gel. Isolation of the FAF in 15-liter batches of supernatant from cultures of 1.5 x 10(10) PMA-stimulated U-937 cells by ammonium sulfate precipitation and sequential filtration on Sephadex G-50, anion exchange chromatography, and reversed phase HPLC yielded microgram quantities of a homogeneous protein of sufficient purity for structural studies. Purified FAF was not absorbed by affinity columns bearing antisera to IL-1 beta, TNF or basic fibroblast growth factor die not share any tryptic peptides with IL-1 alpha or acidic fibroblast growth factor when analyzed by two-dimensional electrophoresis/chromatography on cellulose thin-layer plates, and has an amino-terminal sequence of amino acids that is different from any known fibroblast growth factor. FAF thus represents a unique human monocyte-derived protein that selectively stimulates human fibroblast proliferation and other functions.

Cell Line↗
PubMed1988

Structurally distinctive vasoactive intestinal peptides from rat basophilic leukemia cells.

Peptides recognized by rabbit antibodies to vasoactive intestinal peptide (VIP) were extracted from diisopropyl fluorophosphate-treated rat basophilic leukemia (RBL) cells and resolved by filtration on Sephadex G-25 in 50 mM acetic acid. The immunoreactive VIPs of RBL cells eluted from Sephadex G-25 at 35-41%, 53-60%, and 69-73% bed volume, but not at 63-68% as for the neuropeptide VIP1-28. The two forms of immunoreactive VIP larger than VIP1-28 reacted with antibodies to both VIP1-9 and VIP10-28, but the smallest was bound only by antibodies to VIP10-28. The smallest immunoreactive VIP was purified by ion-exchange and reverse-phase high-performance liquid chromatography, and the amino acid sequence was determined to be that of VIP10-28 with asparagine-free acid at the carboxyl terminus rather than the amide of VIP neuropeptide. Challenge of RBL cells with 1 microM ionophore A23187 at 37 degrees C released VIP10-28 rapidly to a mean of 75% at 5 min and 77% at 30 min. The VIP generated and released by mast cells thus consists of a mixture of peptides that all differ structurally from the neuropeptide VIP.

Amino Acid Sequence↗
PubMed1988

Selective modulation by guanine nucleotides of the high affinity subset of plasma membrane receptors for leukotriene B4 on human polymorphonuclear leukocytes.

Isolated human polymorphonuclear (PMN) leukocyte plasma membranes express high affinity (mean Kd = 0.12 nM) and low affinity (mean Kd = 50 nM) receptors for the chemotactic factor leukotriene B4 (5(S),12(R)-dihydroxy-eicosa-6,14 cis-8,10 trans-tetraenoic acid; LTB4) that are similar to those on intact PMN leukocytes. A portion of high affinity LTB4-R on PMN leukocyte membranes were converted to the low affinity state by GTP (mean +/- SE = 28.6 +/- 14.0%) and nonhydrolyzable GTP analogues, such as 5'-guanylylimidodiphosphate (GMP-PNP), in a concentration-dependent, nucleotide-specific, and reversible manner, without altering the intrinsic binding affinities of either class. [3H]GMP-PNP bound specifically to one class of receptors (mean Kd = 13 nM) on PMN leukocyte membranes. The interdependence of the LTB4-binding membrane protein and guanine nucleotide-binding protein was suggested by the capacity of LTB4 to enhance by a maximum of 150% the binding of [3H]GMP-PNP to PMN leukocyte membranes by increasing the number, but not altering the affinity, of receptors for GMP-PNP. Pertussis toxin, but not cholera toxin, reversed the enhancement of binding of [3H]GMP-PNP produced by LTB4. Guanine nucleotide-binding proteins and high affinity LTB4-R thus exhibit a mutual regulation that differs mechanistically from that of peptide chemotactic factor receptors on PMN leukocytes.

Cell Membrane↗
PubMed1988

Leukotriene D4-induced increases in the cytoplasmic pH of human myelocytic leukocytes.

The effects of leukotriene D4 on the intracellular pH of human myelocytes, derived from cultured HL-60 cells by dimethylsulfoxide-induced differentiation, were quantified with the fluorescent indicator 2',7'-bis-(2-carboxy-ethyl)-5,6-carboxyfluorescein. Leukotriene D4, but not C4 or E4, increased intracellular pH optimally by 3 min with a half-maximal effect at 1-2 nM. The increases in intracellular pH stimulated by leukotriene D4 were prevented by pretreatment of myelocytes with leukotriene D4 but not peptide chemotactic factors. Analogs of amiloride that inhibit selectively the Na+/H+ antiport also prevented the intracellular alkalinization induced by leukotriene D4. The rate of recovery of intracellular pH after an acid load with 30 mM sodium propionate was approximately 30% higher at each level of intracellular pH for myelocytes exposed to leukotriene D4 than for those challenged in buffer alone. The increase elicited by leukotriene D4 in the adherence of myelocytic leukocytes to surfaces thus is associated with an enhanced sensitivity of the Na+/H+ antiport to intracellular pH, that is, not coupled to an earlier rise in the cytosolic level of Ca+2.

Calcium↗
PubMed1988

Sulfidopeptide-leukotriene peptidases in pulmonary edema fluid from patients with the adult respiratory distress syndrome.

The human pulmonary edema fluid concentrations of LTC4 and of LTD4 and LTE4, derived peptidolytically from LTC4, were assessed by radioimmunoassays of the mediators resolved by reverse-phase high-performance liquid chromatography. The mean pulmonary edema fluid concentration (+/- SD) of LTD4 of 19.2 +/- 25.6 nM for 12 patients with the adult respiratory distress syndrome and of LTE4 of 192 +/- 309 nM for 10 of the patients were significantly higher (P less than 0.005 and P less than 0.05) than those of 2.2 +/- 2.4 and 11.0 +/- 18.2 nM, respectively, for 10 patients with cardiogenic pulmonary edema, whereas the lower mean concentrations of LTC4 were not significantly different for the two groups. Pulmonary edema fluid from five patients with adult respiratory distress syndrome, one with cardiogenic pulmonary edema, and one with an indeterminate syndrome contained similar concentrations of peptidoleukotriene peptidases. The LTC4 and LTD4 peptidolytic activities in ARDS fluids were 81 and 142 kD, respectively, by gel filtration. The extents of peptidolysis of [3]LTC4 and [3]LTD4 by 100 microliter of pulmonary edema fluid attained respective mean maximum levels of 74.5 +/- 2.9% (N = 5) and 37.7 +/- 10.2% (N = 4) after 30 min at 37 degrees C and were inhibited by serine-borate and by cysteine, respectively. The predominance of LTD4 and LTE4 over LTC4 in states of altered pulmonary vascular pressure and permeability thus is attributable to two distinct peptidases.

Adolescent↗
PubMed1988

Distinctive patterns of release of neuroendocrine peptides after nasal challenge of allergic subjects with ryegrass antigen.

The concentrations of the neuropeptides substance P, somatostatin, and calcitonin gene-related peptide in human nasal secretions were quantified by radioimmunoassays, concurrently with that of histamine, in the course of nasal challenge of allergic and control subjects with ryegrass antigen to examine contributions of neuromediation of the tissue response. Each of the neuropeptides and histamine were detected in nasal lavage fluid prior to challenge. In allergic patients, but not normal controls, antigen evoked significant increases of 3-fold in histamine at 15-60 min, 1.5- to 4-fold in calcitonin gene-related peptide at 15 min-24 hr, and more than 2-fold in somatostatin at 6 hr, without altering the concentration of substance P in nasal lavage fluid. The identity of the neuropeptides was confirmed chromatographically. Thus calcitonin gene-related peptide may mediate nasal congestion directly and somatostatin may be one of the factors regulating the late involvement of basophils and mast cells in allergic rhinitis.

Administration, Intranasal↗
PubMed1988

Generation and recognition of leukotriene mediators of hypersensitivity and inflammation.

The potent mediators generated by the 5- and 15-lipoxygenation of arachidonic acid have diverse effects on smooth muscles, blood vessels, leukocytes, epithelial cells and glands, and sensory neurons, which suggest possible roles in the initiation and regulation of physiological and biochemical events. The responses to leukotrienes and related mediators are attributable to binding by stereospecific cellular receptors and consequent activation of biochemical transductional sequences analogous to those characteristic of other receptor systems. The elevated concentrations of these mediators in lesional fluids and tissues of inflammatory bowel disease and other hypersensitivity and inflammatory states are, in some instances, clearly related to the time course of development of the disease process. Systematic application of specific inhibitors and antagonists that are becoming available will define more clearly the involvement of leukotrienes in health and disease and possibly lead to new therapeutic approaches.

Arachidonate 15-Lipoxygenase↗
PubMed1988

Enzymatic properties of the 15-lipoxygenase of human cultured keratinocytes.

The arachidonic acid 15-lipoxygenase or linoleic acid omega-6 lipoxygenase of human neonatal foreskin cultured keratinocytes converts arachidonic acid to 15-hydroxy-eicosatetraenoic acid and linoleic acid to 13-hydroxy-linoleic acid. A mean of 93% of the 15-lipoxygenase activity in sonicates of cultured keratinocytes was recovered in the 400,000 X g supernatant, attesting to the cytosolic localization of this enzyme. Optimal 15-lipoxygenase activity in the 400,000 X g supernatant was expressed at pH 6.7-7.3 and in the presence of calcium at a concentration of 2 mM or higher. Keratinocyte 15-lipoxygenase metabolized arachidonic acid (Km = 10.6 microM) and linoleic acid (Km = 9.5 microM) with similar efficiency. Nordihydroguaiaretic acid and 5,8,11,14-eicosatetraynoic acid both inhibited the conversion of arachidonic acid to 15-HETE with respective 50% inhibitory concentrations of 2.0 microM and 0.9 microM, while ATP, GTP, and cyclic AMP had no effect on activity at pH 6.8-7.2. The enzymatic properties of human keratinocyte 15-lipoxygenase thus resemble those of PMN leukocyte 15-lipoxygenase and the mediators generated may contribute to the regulation of cutaneous sensation and inflammation.

Arachidonate 15-Lipoxygenase↗
PubMed1988

Effect of eicosapentaenoic acid in asthma.

The role of arachidonic acid metabolites in the pathogenesis of airway inflammation and clinical asthma is currently unknown. The addition of eicosapentaenoic acid (EPA) to the diet of humans has been shown to generate metabolites that are less potent than their arachidonic acid counterparts. The substitution of EPA for arachidonic acid metabolites in patients might cause a decrease in airway inflammation and an improvement in clinical asthma. We studied the effect of addition of EPA to the diet of twelve asthmatic patients. Standard clinical evaluations and pulmonary function tests were done on weeks 0, 3, 6, 10, 12 and 14. Patients ingested either low-dose EPA (0.1 g/day) or high-dose EPA (4.0 g/day) from weeks 6-14 (total of 8 weeks). There was no difference in clinical status or pulmonary function between groups at the start of the study. There was no change in clinical status or pulmonary function between or within groups at the end of 8 weeks of EPA ingestion.

Adult↗
PubMed1988

Leukotriene B4 decreases the mechanical and thermal thresholds of C-fiber nociceptors in the hairy skin of the rat.

1. We have recently shown that leukotriene B4 (LTB4), a product of the 5-lipoxygenase pathway of arachidonic acid metabolism, sensitizes nociceptors to mechanical stimuli. The present study examined whether LTB4 also induces a heat sensitization of cutaneous C-fiber nociceptors. The C-fiber nociceptors studied had von Frey hair thresholds greater than 5 g and were characterized according to their responses to noxious heat and chemical stimuli, including glacial acetic acid, bradykinin, and capsaicin. Thirty-four of the C-fibers that were activated by intense thermal stimulation were also activated by topical application of glacial acetic acid. They were classified as C-polymodal nociceptors (2, 28). Those that were activated by intense mechanical and thermal stimulation, but were unresponsive to acid, were classified as C-mechanoheat nociceptors (27). 2. Ninety-four percent of C-polymodal nociceptors and 60% of C-mechanoheat nociceptors were sensitized by LTB4. All C-fiber nociceptors that showed a decrease of their heat threshold also had a decrease of their mechanical threshold. LTB4 (75 ng) lowered the average heat threshold from 45 degrees C to 35 degrees C and produced an average decrease in the mechanical threshold of 86%. 3. The magnitude of the LTB4-evoked decrease in thermal threshold was similar to that produced by 75 ng of prostaglandin E2 (PGE2). These data demonstrate that LTB4 sensitizes C-mechanoheat nociceptors to both mechanical and thermal stimuli. 4. We conclude that LTB4 may contribute to the component of hyperalgesia that is resistant to nonsteroidal anti-inflammatory agents.

Action Potentials↗
PubMed1988

Neuropeptide regulation of immediate and delayed hypersensitivity.

Peptide mediators of sensory nerves that are released in tissues by noxious stimuli or inflammatory reactions rapidly elicit local and systemic responses similar to those of immediate hypersensitivity. These sensory neuropeptides affect functions of smooth muscles, blood vessels, leukocytes, and epithelial glands both directly and indirectly, through the actions of mediators released from mast cells stimulated by the peptides. Stereospecific receptors transduce the effects of neuropeptides of the peripheral nervous system (PNS) and central nervous system (CNS) on diverse functions of human, murine and guinea pig mononuclear and polymorphonuclear leukocytes, mast cells, and basophils in vitro and in vivo. Stimulatory and inhibitory effects of neuropeptides on leukocytes are attained in vitro at concentrations which are similar to those in the circulation and in tissues. The dissociation constant (KD) for the binding of a neuropeptide to its leukocyte receptor is within the range of concentrations that evoke cellular responses critical to immunity and hypersensitivity. Neuropeptides exhibit both cellular and stimulus specificities, as exemplified by the greater potency of substance P in activating mucosal than connective tissue mast cells and the capacity of somatostatin to inhibit the release of mediators from basophils challenged by IgE-dependent mechanisms, but not by basic peptides or ionophores. The selective release of distinct neuropeptides from different subsets of sensory nerve endings, the specificity of neuropeptide recognition by mast cells, basophils, lymphocytes, and other target cells, and the diversity of relevant activities of the neuropeptides suggest that the nervous system may initiate and modulate immediate and delayed hypersensitivity by unique mechanisms.

Basophils↗