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Biomedical subjects

E Christophers

Publications and source records attributed to E Christophers.

At least 199 records · Page 11Linked to original sources
PubMed1990

[Sweet's syndrome after T-lymphoblastic lymphoma and before the manifestation of a secondary acute myeloid leukemia].

Thirty-two months after the diagnosis and treatment of a T-lymphoblastic lymphoma with bone marrow involvement had been made in a 30-year-old patient, he developed fever up to 40 degrees C during maintenance treatment with methotrexate and 6-mercaptopurine. Later there were tender, blue-red skin eruptions, leukocytopenia (1.4 x 10(9)/l) and thrombocytopenia (29 x 10(9)/l). Histological examination of a skin biopsy revealed acute febrile neutrophilic dermatosis (Sweet's syndrome). Bone marrow biopsy revealed hyperplastic myelopoiesis. There was no evidence for acute myeloid leukaemia or lymphoma recurrence. After the maintenance treatment had been discontinued, treatment with methylprednisolone, 60 mg, was begun. The signs of Sweet's syndrome regressed, but thrombocytopenia and mild leukocytopenia remained. Six months later it was found by morphological and immunological tests that he had acute myeloid leukaemia without any chromosomal abnormalities. There was still no evidence for a recurrent T-lymphoblastic lymphoma.

Adult↗
PubMed1990

Elafin: an elastase-specific inhibitor of human skin. Purification, characterization, and complete amino acid sequence.

A potent inhibitor of human leukocyte elastase (EC 3.4.21.37) and porcine pancreatic elastase (EC 3.4.21.36) was purified to homogeneity from human horny layers. It inhibits human leukocyte elastase and porcine pancreatic elastase in a 1:1 molar ratio and shows equilibrium dissociation constants of 6 x 10(-10) M and 1 x 10(-9) M, respectively. Inhibition of plasmin, trypsin, alpha-chymotrypsin, and cathepsin G was not observed. This inhibitor proved to be an acid stable basic peptide with an isoelectric point of 9.7. The complete amino acid sequence appears to be unique with 38% homology to the C-terminal half of antileukoprotease. The sequence shows that the inhibitor is composed of 57 amino acids and predicts a Mr of 7017. The high affinity as well as the apparent specificity for elastases suggests a functional role in preventing elastase-mediated tissue proteolysis. It is suggested that the term "elafin" be used to designate this inhibitor.

Amino Acid Sequence↗
PubMed1990

Lipopolysaccharide-stimulated human monocytes secrete, apart from neutrophil-activating peptide 1/interleukin 8, a second neutrophil-activating protein. NH2-terminal amino acid sequence identity with melanoma growth stimulatory activity.

Purification of monocyte-derived NAP-1/IL-8 by preparative reversed-phase (RP)-HPLC led to the detection of a second peak with polymorphonuclear leukocyte (PMNL)-activating (degranulation, chemotaxis) properties. The monokine responsible for this biological activity, which we tentatively termed NAP-3, could be purified to homogeneity by three different RP-HPLC steps. Tricine-SDS-PAGE analysis gave a single line at Mr 5.3 kD (NAP-1/IL-8 = 5.8 kD). NH2-terminal amino acid sequence analysis read as a major sequence (ASVATELRXCXLQT. .), which shows greater than 40% homology to that of NAP-1/IL-8. The sequence is identical to that found for the 13-kD moiety of melanoma growth stimulating activity (MGSA) and the product of the oncogene gro. Determination of neutrophil chemotactic activity of NAP-3 revealed a typical bell-shaped dose-response curve (ED50 = 2 ng/ml) with no significant neutrophil chemotactic activity at doses greater than 200 ng/ml. Also, in cytochalasin B-pretreated PMNL, NAP-3 elicited release of myeloperoxidase and beta-glucuronidase. Crossdesensitization studies in PMNL enzyme release revealed crossreactivities with the NAP-1/IL-8-R on PMNL. NAP-3 (MGSA/gro) appears to represent the first member of the novel supergene family of beta-thromboglobulin-like host defense cytokines, which expresses both mitogenic as well as proinflammatory properties at the nanogram level.

Amino Acid Sequence↗
PubMed1990

IL-1 alpha or tumor necrosis factor-alpha stimulate release of three NAP-1/IL-8-related neutrophil chemotactic proteins in human dermal fibroblasts.

Human dermal fibroblasts in culture secrete three protein-like neutrophil chemotactic factors, when stimulated either with human rIL-1 alpha or IL-1 beta; not, however, after incubation with LPS. These three fibroblast-derived neutrophil-activating proteins (FINAP) could be purified by subsequently performed reversed phase and size exclusion HPLC. By high resolution SDS-PAGE, all the proteins were shown to migrate with an Mr of 6,700 (alpha-FINAP), 3,600 (beta-FINAP), and 5,300 (gamma-FINAP). All purified cytokine preparations were found to be chemotactic for human neutrophils. In addition, all FINAP induced release of lysosomal enzymes in neutrophils. Deactivation of chemotaxin-elicitable enzyme release showed cross-desensitization of all FINAP with NAP-1/IL-8. Western blot analysis of alpha-FINAP by using mAb against neutrophil-activating protein (NAP)-1/IL-8 reveals immunologic cross-reactivity with NAP-1/IL-8. By amino-terminal amino acid sequence analysis alpha-FINAP could be identified as the 77-residue extended form of NAP-1/IL-8 containing the 79-residue form as a minor contaminant. Whereas beta-FINAP has been found to be a truncation product of alpha-FINAP, gamma-FINAP shows identity with authentic melanoma growth stimulatory activity with respect to retention time upon reversed phase HPLC, high resolution SDS-PAGE, and biologic properties, as well as amino-terminal amino acid sequence. These data show that human dermal fibroblasts may actively participate in inflammatory reactions by secretion of proinflammatory cytokines.

Amino Acid Sequence↗
PubMed1990

Identification of a novel and highly potent eosinophil chemotactic lipid in human eosinophils treated with arachidonic acid.

Purified human eosinophils generate eosinophil chemotactic lipids (ECL), when incubated with arachidonic acid without any stimulus. Reversed phase HPLC of incubation supernatants revealed major lipid-like eosinophil chemotactic activity eluting in a peak containing 5(S), 15(S)dihydroxy-6,13-trans-8,11-cis-eicosatetraenoic acid (5,15-DiHETE) as well as a 8,15-dihydroxyeicosatetraenoic acid. For further characterization of the ECL, some authentic dihydroxyeicosatetraenoic acids were tested for eosinophil chemotactic activity. Only 5,15-DiHETE as well as 8(S), 15(S)-dihydroxy-5,11-cis-9,13-trans-eicosatetraenoic acid were found to be chemotaxins for human eosinophils, however, with an ED50 near 0.3 microM and 1.5 microM, respectively. The presence of high titer eosinophil chemotactic activity in ECL preparations let us look for a contaminating ECL with higher specific activity. By using a different reversed phase HPLC-system 5,15-DiHETE as well as 8(R), 15(S)-dihydroxy-5,11-cis-9,13-trans-eicosatetraenoic acid could be separated from a highly potent ECL. Final purification of this ECL by the use of straight phase HPLC resulted in a single at 260 nm absorbing peak giving an UV spectrum different from that known for eosinophil chemotactic factors indicating a novel type of eosinophil chemotactic lipid. Eosinophil chemotactic activity of purified ECL has been found to be similar to that seen for platelet-activating factor, the most potent chemotaxin so far known, either in the number of migrating cells or the ED50. Cross-desensitization experiments with ECL, leukotriene B4, and platelet-activating factor revealed the existence of a separate ECL receptor on eosinophils. The production of potent ECL by the responder cells themselves supports the idea that there exists a self-sustaining mechanism of eosinophil accumulation.

Arachidonate 15-Lipoxygenase↗
PubMed1990

Detection of neutrophil-activating peptide NAP/IL-8 and NAP/IL-8 mRNA in human recombinant IL-1 alpha- and human recombinant tumor necrosis factor-alpha-stimulated human dermal fibroblasts. An immunocytochemical and fluorescent in situ hybridization study.

A neutrophil-activating peptide (NAP)/IL-8 produced by LPS-stimulated human peripheral blood monocytes was biochemically purified and functionally characterized by different investigators. Work conducted in our laboratory showed that NAP/IL-8 as well as variants of this peptide are produced by a variety of cells (e.g., monocytes, T lymphocytes, endothelial cells) and that lesional psoriatic scales contain large amounts of biologically active NAP/IL-8. We now investigated human dermal fibroblasts for production of NAP/IL-8. The peptide was detected by immunocytochemistry by using the mAb 46E5. NAP/IL-8 mRNA was visualized by high resolutive fluorescent in situ hybridization with biotinylated antisense/sense RNA probes. Among the various stimuli used [human (h)rIL-1 alpha, hrTNF-alpha, hrIL-3, hr-granulocyte-macrophage-CSF, LPS, FMLP, and platelet-activating factor (PAF)] only hrIL-1 alpha (100 U/ml) and hrTNF-alpha (100 ng/ml) induced the transcription and translation of NAP/IL-8. In contrast to monocytes, LPS was without effect in cultured human dermal fibroblasts. Both NAP/IL-8 and NAP/IL-8 mRNA were found in the cytoplasm adjacent to the nucleus, but interestingly NAP/IL-8 mRNA was not restricted to the cytoplasm. In positive cells only two small bright spots were randomly distributed in the nucleus. Most likely these spots represent transcription sites where NAP/IL-8 mRNA is accumulated during gene expression. Our observations show that stimulation of dermal fibroblasts with the cytokines hrIL-1 alpha and hrTNF-alpha results in expression of IL-8.

Cell Compartmentation↗
PubMed1990

A family study of atopic dermatitis. Clinical and genetic characteristics of 188 patients and 2,151 family members.

The history of 188 caucasian patients with atopic dermatitis (AD) and of 2,151 family members has been analyzed. Of the AD patients 48% suffered from respiratory atopy (36% rhinitis, 28% asthma, and 15% both). AD showed by far the earliest onset of all atopic diseases: 50% of our patients had skin lesions before the age of 2 years and 60% before the age of 5 years. In contrast, symptoms of allergic asthma developed in 40% of AD patients before the age of 5 years in comparison with only 25% who had allergic rhinitis. AD affects females more frequently than males (male to female ratio 1:1.5), regardless of whether additional respiratory atopies are present or not. In contrast, respiratory atopies develop more frequently in males than in females (male to female ratio 1.5:1). Mothers with respiratory atopy more often have atopic children (26%) than do fathers with respiratory atopy (13%). Finally, risk figures for genetic counselling are given. In short, the general risk of developing AD (3%) and atopy (7%) increases by a factor of two with each first-degree family member already suffering from atopy.

Adolescent↗
PubMed1990

Production of 15-hydroxyeicosatetraenoic acid by purified human eosinophils and neutrophils.

In the presence of high concentrations of exogenous arachidonic acid (greater than or equal to 10 microM), eosinophils produced 15-hydroxyeicosatetraenoic acid (15-HETE) in the absence of stimuli. The calcium ionophore A23187, as well as the chemotaxins used in this study--complement split product C5a, platelet-activating factor (PAF), and N-formyl-methionyl-leucyl-phenylalanine (FMLP)--failed to increase 15-HETE production, indicating that eosinophil 15-lipoxygenase is already active. Production of 15-HETE from eosinophils increased with increasing concentrations of arachidonic acid, exogenously added. Maximal 15-HETE production was observed to be 1111 +/- 380 ng per 10(6) eosinophils at the concentration of 100 microM of arachidonic acid. With low concentrations of exogenous arachidonic acid (below 2 microM), eosinophils were considered to incorporate exogenous arachidonic acid into their cell membrane, and did not produce 15-HETE. In contrast, 15-HETE formation in highly purified neutrophils (eosinophils less than 1%) was negligible compared with that in eosinophils (300-fold less), suggesting that 15-HETE-forming activity in granulocytes is derived from the eosinophil 15-lipoxygenase pathway and that neutrophils may lack 15-lipoxygenase activity.

Arachidonic Acid↗
PubMed1989

Production and characterization of monoclonal antibodies against the novel neutrophil activating peptide NAP/IL-8.

LPS-stimulated human mononuclear cells have recently been shown to produce large amounts of a novel neutrophil-activating cytokine termed neutrophil-activating peptide NAP/IL-8. This chemotactic factor has in the meantime been biochemically and functionally well characterized. We now report on four distinct murine mAb directed against this peptide. All four mAb are different in respect to isotype and IEF pattern. The cross-reactivity with partially homologous peptides like beta-thromboglobulin and platelet factor 4 showed defined differences. With the use of these antibodies we were able to detect solid phase as well as soluble NAP/IL-8 as tested in a sandwich-ELISA. Also dose-dependent neutralization of NAP/IL-8 chemotactic activity in the Boyden chamber chemotaxis assay was observed. Immunoaffinity columns prepared with these four mAb bound NAP/IL-8 from supernatants of LPS-stimulated mononuclear cells. Furthermore, Western immunoblots showed a single protein band in the expected region of Mr of 10 kDa with all four mAb presented.

Animals↗
PubMed1989

Secretion of novel and homologous neutrophil-activating peptides by LPS-stimulated human endothelial cells.

Human umbilical vein endothelial cells in culture produce two chemotactic polypeptides when stimulated with LPS. The chemotactic factors could be purified to apparent homogeneity by HPLC techniques and were identified as 7.5-kDa and 15-kDa polypeptides by SDS-PAGE under nonreducing conditions. Both factors are potent chemotaxins for human neutrophils demonstrating half-maximal chemotaxis at 2 ng/ml and g ng/ml, respectively. In addition both peptides elicited release of azurophilic granule constituents when neutrophils were pretreated with cytochalasin B. Cross-desensitization experiments by using human neutrophils revealed cross-reactivities between both chemotaxins, not, however, with C5a or FMLP, indicating that both endothelial cell-derived neutrophil activating peptides (ENAP) are homologous. In addition, the 7.5-kDa factor (beta-ENAP) proved to be the quantitatively dominating and more potent chemotaxin as compared to the 15 kDa factor (alpha-ENAP). beta-ENAP shows biochemical and biologic similarities to monocyte- and lymphocyte-derived neutrophil-activating peptides MONAP and LYNAP, which recently were purified and sequenced.

Chemotactic Factors↗
PubMed1989

Transendothelial cell diapedesis of neutrophils in inflamed human skin.

The mode of extravasation of neutrophils (PMNs) in cutaneous inflammation was studied in sequential biopsy specimens taken from human skin. Inflammatory skin reactions were produced by intracutaneous injection of endogeneous mediators of inflammation--C5ades arg, LTB4, neutrophil-activating peptide (NAP) and interleukin-1 (IL-1). Within 30 min after injection neutrophils were observed in close contact with endothelial cells of postcapillary venules and, following cytoplasmic engulfment, the cells were found to be transported transcellulary through the endothelial layer. In a total of 20 biopsy specimens taken at various times, cell migration via interendothelial gaps was absent. Instead, the transcellular pathway appeared to be the first and foremost mode of diapedesis. During this migratory process PMNs lacked signs of degranulation and numerous electron-lucent vesicles and secondary lysosomes were found. In addition, coated pits present on leukocyte as well as endothelial-cell membranes were indicative of receptor-mediated endocytotic processes.

Cell Movement↗
PubMed1989

Chemotactic responsiveness of eosinophils isolated from patients with inflammatory skin diseases.

We determined the chemotactic responsiveness of peripheral eosinophilic granulocytes (eosinophils) isolated from patients with inflammatory dermatoses and healthy volunteers. Ten patients with atopic dermatitis, five patients with drug reactions, ten patients with psoriasis, and fourteen healthy volunteers were studied. Well characterized chemotaxins, the complement split product C5a, leukotriene B4 (LTB4), platelet activating factor (PAF), and N-formyl-methionyl-leucyl-phenylalanine (FMLP), were used as chemoattractants. Eosinophils from healthy volunteers showed strong migratory responses towards C5a and PAF but responded poorly to LTB4 and FMLP. When patients were grouped by disease severity, eosinophil chemotactic responses to PAF were significantly enhanced in severely affected patients (p less than 0.05); this was not true with C5a, LTB4 or FMLP. This enhanced eosinophil chemotaxis to PAF was not related to a specific disease. No correlation between eosinophil chemotactic activity and peripheral blood eosinophil count was observed. The increased responsiveness of circulating eosinophils towards PAF may be related to altered receptor expression during cutaneous inflammation.

Chemotactic Factors, Eosinophil↗
PubMed1989

Differential sensitivities of purified human eosinophils and neutrophils to defined chemotaxins.

Functions of eosinophils and neutrophils isolated from normal human blood were determined by measuring chemotactic migration and release of beta-glucuronidase. Four well-characterized chemotaxins, the complement fragment C5a, formyl-methionyl-leucyl-phenylalanine (FMLP), platelet-activating factor (PAF), and leukotriene B4 (LTB4) were used as stimuli. Neutrophils showed remarkable chemotactic responses to all four chemotaxins. In contrast, eosinophils showed a significant chemotactic response to C5a and PAF, but only weak responses to FMLP and LTB4. Using these chemotaxins we found the following order of chemotactic potency (maximal number of migrated cells): C5a = LTB4 greater than FMLP greater than PAF for neutrophils and PAF = C5a greater than LTB4 = FMLP for eosinophils. Neutrophils elicited a significant beta-glucuronidase release when stimulated by C5a and FMLP, whereas only small amounts were released with PAF and LTB4. On the other hand, an amount of beta-glucuronidase released from eosinophils comparable to that from neutrophils was elicited only with C5a. FMLP, LTB4, and PAF caused the release of small percentages of beta-glucuronidase. The important cellular functions of eosinophils and neutrophils, chemotaxis and enzyme release, are thought to be controlled by differential responsiveness to stimuli.

Adult↗
PubMed1989

Recombinant human tumour necrosis factor beta (lymphotoxin) lacks chemotactic activity for human peripheral blood neutrophils, monocytes, and T cells.

Human recombinant tumour necrosis factor beta (rhuTNF beta)/lymphotoxin was tested for human neutrophil granulocyte (PMN), monocytes (MO), and T-cell chemotactic activity by means of a modified Boyden chamber system. Over a wide range of concentrations (10(-7)-10(-14)M)rhuTNF beta showed no chemotactic activity for PMN, MO, or T cells. In contrast, strong chemotactic migration was elicited in PMN and MO with the tripeptide N-formyl-methionyl-leucyl-phenylalanine (FMLP) and in T cells when complement split product C5a and leukotriene B4 (LTB4) were used as chemotaxins. The results of this study indicate that rhuTNF beta/lymphotoxin is not a chemotaxin for human PMN, MO, or T lymphocytes in vitro.

Chemotactic Factors↗
PubMed1989

Decreased releasability of basophils from patients with cold urticaria after cold exposure.

Histamine release from peripheral blood basophils challenged with C5a, f-met-peptides and calcium ionophore was studied in patients with cold urticaria before and after exposure to low environmental temperatures. Compared to healthy controls, stimulated mediator release before cold exposure was increased in 7 of 11 patients. When challenged by cold exposure mediator release from in vitro-stimulated basophils was decreased. This decrease was more pronounced after stimulation with receptor-mediated stimuli (e.g. C5a) as compared to receptor-unrelated stimuli, e.g. calcium ionophore. In 4 of 11 patients stimulated mediator release before cold exposure was moderately increased. Also after cold exposure only a weak decrease of stimulated histamine release was seen. Levels of activated complement components (C3a) before and after cold exposure failed to provide evidence for complement activation in vivo. Also the number of circulating basophils as well as their cellular histamine content remained normal after cold exposure. The results show that in these patients release of histamine is altered before and after cold exposure. These changes in basophil responsiveness are not due to complement activation in vivo.

Basophils↗
PubMed1989

Identification of two endogenous neutrophil-activating peptides in psoriatic skin and inflammatory cells: C5ades arg and NAP.

Migration of polymorphonuclear leukocytes (PMN) into the upper layers of involved epidermis represents a characteristic feature of psoriasis. By analysis of psoriatic scale material we were able to identify two potent proinflammatory peptides, which are present in the upper epidermis from psoriatic lesions. Both factors (C5ades arg and NAP) show strong chemotactic activity for human neutrophils in vitro as well as in vivo. Whereas C5ades arg is a known mediator activated by either alternative or classical activation of the complement cascade, NAP represents a newly detected peptide with a molecular weight of 8,000 daltons which is produced by a variety of cells participating in the psoriatic tissue reaction.

Cell Movement↗
PubMed1989

Acute inflammatory effects of a monocyte-derived neutrophil-activating peptide in rabbit skin.

The inflammatory effects of a monocyte-derived neutrophil-activating peptide (MONAP), purified to homogeneity from lipopolysaccharide-stimulated human peripheral blood monocytes, have been evaluated in rabbit skin. Intradermal injection of MONAP alone caused a mild infiltration of polymorphonuclear leucocytes (PMNL) but did not induce any change in plasma extravasation. When combined with prostaglandin E2(PGE2), MONAP caused a marked and synergistic increase in PMNL infiltration and plasma extravasation into the injected skin sites. The increase in vascular permeability induced by MONAP depended on the presence of circulating PMNL. MONAP is a novel cytokine with pro-inflammatory properties and may have physiological and pathophysiological roles in health and disease.

Animals↗