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D D Metcalfe

Publications and source records attributed to D D Metcalfe.

At least 163 records · Page 9Linked to original sources
PubMed1990

Demonstration of IgE antibodies to nucleic acid antigens in patients with SLE.

Using a combination of avidin-biotin microELISA and solid phase radioimmunoassay, we examined sera from 23 patients with systemic lupus erythematosus (SLE), two patients with established sensitivity to ingested shrimp, and 15 healthy normal subjects. In addition to IgG antibodies, varying amounts of IgE antibodies specific for native DNA (nDNA), denatured or single-stranded DNA (dnDNA), RNA, and tRNA were demonstrable in the sera of SLE patients, but not in the sera of normal subjects. A comparison of the specificity of nucleic acid-specific IgE antibodies present in the sera of shrimp-sensitive patients with those present in the sera of seven SLE patients revealed that the IgE antibodies in the sera of shrimp-sensitive patients specifically recognized shrimp tRNA but not yeast tRNA, calf thymus RNA, or calf thymus DNA, while those present in the sera of patients with SLE recognized all these nucleic acid antigens. The IgE antibodies directed against nDNA, dnDNA, RNA, and tRNA may mediate mast cell and basophil degranulation and thus contribute both to immediate-type hypersensitivity phenomena including hives seen in patients with SLE and to the localization of IgE-nucleic acid complexes in target tissues.

Antibodies, Antinuclear↗
PubMed1989

Mast cells chemotax to laminin with enhancement after IgE-mediated activation.

The increase of mast cells at sites of tissue inflammation suggests the production of local factors chemotactic for mast cells. In this report, we demonstrate that the murine mast cell line PT18 and primary mouse bone marrow-derived mast cells chemotax to the basement membrane glycoprotein laminin, and that the synthetic laminin A chain-derived peptide, PA22-2, represents a region of laminin that contains a major chemoattractant site. Mast cell chemotaxis to laminin is enhanced after activation of mast cells by the calcium ionophore, A23187, or PMA and by sensitization of the cells with IgE followed by exposure to antigen. Chemotaxis is not increased in the presence of IL-3 and is independent of mast cell degranulation, as histamine release did not occur when cells were activated with PMA. Mast cell chemotaxis to laminin and its enhancement by IgE-dependent mast cell activation provides a mechanism by which these cells may be attracted to sites of tissue injury. Such activity may be particularly relevant in the response of host tissues to inflammation accompanying parasitic infestations, allergic reactions, and wound healing.

Adjuvants, Immunologic↗
PubMed1989

Laminin promotes mast cell attachment.

Tissue mast cells often localize in close proximity to the basement membrane of endothelial cells and increase at sites of inflammation. The reason for this unique tissue distribution is unknown. We report here that both the murine mast cell line PT18 and mouse bone marrow-derived mast cells possess functional receptors for laminin, and exhibit adhesion, spreading and redistribution of histamine-containing granules on a laminin substratum. This adherence is enhanced in the presence of purified IL-3 and can be inhibited by antibodies to laminin and by antibodies to laminin receptors. Northern analysis showed a high level of mRNA for a 32-kDa laminin receptor in PT18 mast cells. Mouse bone marrow-derived cultures initially exhibited a low level of the mRNA expression. However, the expression of the laminin receptor mRNA is induced rapidly within 1 wk of culture with IL-3. Thus, mast cells exhibit functional laminin receptors that may explain the tissue distribution of mast cells and their accumulation at sites of tissue injury.

Animals↗
PubMed1989

A staphylococcal protein A rosetting assay for the demonstration of high affinity IgE receptors on rIL-3-dependent human basophil-like cells grown in mixed cell cultures.

Mature basophils can be differentiated from other blood born cells by their morphology, metachromatically staining granules, histamine content, and the presence on their surfaces of high-affinity IgE receptors. The identification of basophil-like cells arising in mixed human bone marrow cultures, however, is made more difficult because of their immaturity and the morphologic alterations due to in vitro culture. Identification of IgE receptors on individual cells in cell preparations which could simultaneously be examined for their histochemical properties would facilitate the study of basophil growth and differentiation. Because this was difficult using existing techniques, we developed an IgE staphylococcal protein A rosetting assay which allows the identification of cells bearing high-affinity IgE receptors and permits the same cells to be examined by a variety of staining techniques. We then examined the appearance of basophil-like cells bearing IgE receptors in cultures of human bone marrow and correlated this data with measurements of IgE receptor number and affinity. The percentage of total cells grown in the presence of human recombinant IL-3 (rIL-3) that rosetted with Staphylococcus aureus increased from 2 +/- 0.5% at 1 week of culture to 7 +/- 2% at 2 weeks, 14 +/- 5% at 3 weeks, and 18 +/- 5% at 4 weeks. Using 125I-labeled IgEPS, the number of IgE receptors per rosetting basophil-like cell was calculated to be 7.3 x 10(4) at 2 weeks, 6.8 x 10(4) at 3 weeks, and 3.9 x 10(4) receptors per rosette positive basophil-like cell at 4 weeks. Finally, the rate of dissociation of IgE from these cultured cells was found to be 4.0 +/- 1.3 x 10(-5) s-1, indicating that IgE bound to IgE receptors on cultured basophil-like cells with high affinity.

Antigens, Differentiation, B-Lymphocyte↗
PubMed1989

Inhibition of the growth of IL-3-dependent mast cells from murine bone marrow by recombinant granulocyte macrophage-colony-stimulating factor.

The mouse mast cell line PT-18 demonstrates [3H] thymidine uptake in the presence of either mouse IL-3 or mouse recombinant granulocyte-macrophage CSF (rGM-CSF). Experiments were thus undertaken to determine whether rGM-CSF would affect IL-3-dependent growth of mast cells from mouse bone marrow cells (BMC). BMC placed in liquid culture containing 50 U/ml of IL-3 gave rise to cultures containing up to 95% mast cells by 2 to 3 wk. The rise in percentage of mast cells was accompanied by an increase in total cell-associated histamine. In contrast, BMC grown in the presence of 50 U/ml of rGM-CSF gave rise to cultures containing primarily macrophages and granulocytes with less than 1% mast cells. The addition of increasing amounts of rGM-CSF to BMC cultures grown in the presence of IL-3 resulted in a decrease in the number of mast cells present in culture at 2 to 3 wk. Cells other than mast cells in these cultures consisted principally of granulocytes and macrophages. The rGM-CSF-related inhibition of mast cell growth was not abrogated by the addition of indomethacin to cultures. Granulocyte-macrophage cell populations added to IL-3-containing cultures did not inhibit mast cell growth. The suppressive effect of rGM-CSF on IL-3-dependent mast cell growth may indicate an important role for GM-CSF in the down-regulation of mast cell proliferation in tissues.

Animals↗
PubMed1989

Induction of allergen-specific IgE and IgG responses by anti-idiotypic antibodies.

In order to explore idiotypic, anti-idiotypic, and anti-anti-idiotypic responses to allergens, BALB/c mice were immunized with affinity-purified human idiotypic antibodies directed against a highly purified shrimp allergen. This resulted in the production of anti-idiotypic antibodies which were quantitated by using rabbit idiotypic antibodies raised against the same purified allergen. The mouse anti-idiotypic antibodies recognized shrimp-specific human idiotypic antibodies of the IgE isotype from 18 of 20 individuals, and IgG antibodies from 14 of 20 shrimp-sensitive patients. Immunization of BALB/c mice with affinity-purified, allergen-specific anti-idiotypic antibodies induced anti-allergen IgE and IgG responses in the absence of the allergen. This paper thus presents evidence that anti-idiotypic antibodies raised against allergen-specific idiotypic antibodies may substitute for the original allergen in the induction of allergen-specific idiotypic antibodies. The demonstration of shared idiotopes on IgG and IgE antibodies in the sera of shrimp-sensitive patients supports the use of allergen-specific anti-idiotypic antibodies as surrogate allergens.

Adult↗
PubMed1989

IL-3-dependent growth of basophil-like cells and mastlike cells from human bone marrow.

Human bone marrow cultured in the presence of human rIL-3 has been reported to give rise to basophils. In contrast, mouse bone marrow, cultured in the presence of mouse IL-3, leads to the growth of mast cells. To determine if human rIL-3 might also stimulate the growth of human mast cells, we cultured human bone marrow in the presence of human rIL-3 in suspension cultures, methylcellulose, and in "interphase" cultures where cells are layered over agar. The presence of mast cells was determined using a variety of histochemical techniques. In agreement with previous reports, basophil-like cells were identified in all culture systems. Mastlike cells were identified only in interphase cultures. By 3 wk, such cultures consisted of basophil-like cells (20 to 50%) and mastlike cells (1 to 5%). Cultures supplemented with rIL-4 showed no additional increase in basophil-like and mastlike cells. Both basophil-like and mastlike cells fluoresced with o-phthaldialdehyde and exhibited IgE receptors. Unlike basophil-like cells, mastlike cells were chloroacetate esterase, amidase, and human mast cell tryptase positive. We conclude that human rIL-3 can support the growth of human mastlike cells under selected culture conditions.

Basophils↗
PubMed1989

Analysis of the wheal-and-flare reactions that follow the intradermal injection of histamine and morphine in adults with recurrent, unexplained anaphylaxis and systemic mastocytosis.

It has been suggested that patients with recurrent, unexplained anaphylaxis may be more responsive, and patients with systemic mastocytosis, less responsive, to mast cell-derived mediators, including histamine, compared to normal subjects. This would help explain why patients with recurrent, unexplained anaphylaxis have an anaphylactic response and, conversely, why patients with systemic mastocytosis can tolerate high levels of plasma histamine. To test this hypothesis, intradermal titrations (0.02 ml of solution from 1 ng/ml to 2 micrograms/ml) of histamine and morphine sulfate (MS) (10 ng/ml to 10 micrograms/ml) were administered to normal volunteers (N = 15), patients with recurrent, unexplained anaphylaxis (N = 10), and patients with systemic mastocytosis (N = 18). Antihistamines were stopped at least 72 hours before the study. Resultant areas of wheal and flare were determined with a computerized morphometric system. Comparison of any two given means at each dose of histamine or morphine with the two-sample Student's t test with Bonferroni inequality demonstrated no significant differences (p greater than 0.05) among the three groups. The median amount of MS or histamine required to produce a half-maximal response was compared for equality. None of the differences observed reached statistical significance, in agreement with the similarity of the dose-response curves. An analysis of the correlation between response to MS and to histamine in individual subjects revealed the responses to be significantly correlated in all cases, with the exception of wheal in patients with recurrent, unexplained anaphylaxis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1989

A double-blind, placebo-controlled, crossover trial of ketotifen versus hydroxyzine in the treatment of pediatric mastocytosis.

To asses the efficacy of ketotifen (Zaditen; Sandoz Pharmaceuticals, Basel, Switzerland) for the treatment of pediatric mastocytosis, eight children who exhibited symptoms as a result of mastocytosis were enrolled in a 12-week, double-blind, placebo-controlled, crossover trial of ketotifen versus hydroxyzine (Atarax; Roerig, New York, N.Y.). Efficacy of each drug was assessed by daily symptom scores and plasma- and 24-hour urine-histamine levels. After completion of the study, symptom scores revealed that seven of the eight children exhibited a greater reduction in symptoms while they were receiving hydroxyzine (p less than 0.05). The symptoms most likely to improve with treatment with hydroxyzine were flushing and abdominal pain. Analysis of plasma- and 24-hour urine-histamine levels at the beginning and end of each trial period of each drug revealed no significant differences (p greater than 0.20). Changes in 24-hour urine-histamine levels, but not plasma-histamine levels, correlated with changes in symptom scores. We conclude that ketotifen offers no advantage over hydroxyzine in the treatment of pediatric mastocytosis.

Child↗
PubMed1989

Isolation and characterization of heat-stable allergens from shrimp (Penaeus indicus).

Shrimp are among the more common causes of immediate hypersensitivity reactions to food. To characterize better the allergenic substances within shrimp, extracts from heated shrimp were systematically examined with solid-phase radioimmunoassay and sera from patients clinically sensitive to shrimp. Two heat-stable protein allergens, designated as Sa-I and Sa-II, were identified from boiled shrimp (Penaeus indicus) extracts. Sa-I was isolated by ultrafiltration, Sephadex G-25, and diethylaminoethyl-Sephacel chromatography, whereas, Sa-II, the major allergen, was purified by successive chromatography on diethylaminoethyl-Sephacel, Bio-Gel P-200, and Sepharose 4B columns. Sa-I, was homogeneous by polyacrylamide gel electrophoresis (PAGE), elicited a single band on sodium dodecyl sulfate-PAGE corresponding to a molecular weight of 8.2 kd. Sa-II was also found to be homogeneous by PAGE, crossed immunoelectrophoresis, and immunoblotting. On sodium dodecyl sulfate- PAGE, it elicited a single band with a molecular weight of 34 kd. Sa-II was found to contain 301 amino acid residues and was particularly rich in glutamate/glutamate and aspartate/asparagine. Solid-phase radioimmunoassay-inhibition studies revealed that Sa-I and Sa-II share 54% of the allergenic epitopes, suggesting that Sa-I may be a fragment of Sa-II.

Adult↗
PubMed1989

The histamine content of oriental foods.

Several of the symptoms of scombroid poisoning (i.e. histamine toxicity) resemble those observed in people suffering from Chinese restaurant syndrome. Therefore, the histamine content of representative Chinese cuisine, which included 31 common dishes, 12 condiments and 12 basic ingredients from several sources, was measured using a sensitive and specific radioenzymatic assay. A further enzymatic procedure involving diamine oxidase was used to verify that the substance measured was histamine. A total of 184 assays were performed on 57 samples in the study. High levels of histamine were found in the cheeses, which were used as positive controls (863.6 micrograms histamine/g blue cheese and 107.4 micrograms histamine/g Parmesan cheese), and in some common condiments, including tamari (2392.2 micrograms histamine/g sample) and one brand of soy sauce (220.4 micrograms histamine/g sample). The histamine content of four condiments and three common dishes was over 10 micrograms histamine/g sample, while four condiments and 16 common dishes contained less than 1 microgram histamine/g sample. Calculations involving representative amounts of food that can be consumed at a typical oriental meal suggest that, in some cases, histamine intake may approach toxic levels. The results are discussed with regard to the possible role of histamine in reactions associated with restaurant meals.

Animals↗
PubMed1989

Analysis of plasma histamine levels in patients with mast cell disorders.

PURPOSE: The use of plasma histamine determinations as a screening tool to distinguish patients with recurrent unexplained anaphylaxis, flushing, or both from those with mastocytosis has never been evaluated. This retrospective study was designed to determine if plasma histamine levels can be used as a screening test. PATIENTS AND METHODS: Values of plasma histamine levels, measured using a sensitive radioenzymatic assay, from 41 patients with mastocytosis, 26 patients with recurrent unexplained anaphylaxis, and 76 normal subjects were statistically analyzed to determine diagnostic usefulness and accuracy. Patients with mastocytosis were subdivided into four smaller groups on the basis of clinical and histopathologic findings: (1) isolated urticaria pigmentosa, (2) indolent systemic mastocytosis, (3) mastocytosis with dysmyelopoiesis, and (4) lymphadenopathic mastocytosis with eosinophilia. RESULTS: The distribution of plasma histamine values among patients with unexplained anaphylaxis strongly resembled that among the normal subjects (p greater than 0.50, Smirnov test), whereas patients with mastocytosis tended to show moderate to marked elevations above the upper limit of normal (617 pg/mL). The geometric mean plasma histamine levels in mastocytosis subgroups 2, 3, and 4 were found to be quite similar (1,085, 1,976, and 1,433 pg/mL; p greater than 0.50, F-test); moreover, each mean level was significantly greater than those of the normal subjects and of patients with unexplained anaphylaxis (p less than 0.01, Scheffé multiple comparison test). Analysis of the 27 sets of plasma histamine values collected on patients with indolent systemic mastocytosis revealed that the earliest value observed fell below 617 pg/mL in eight patients (30%). A similar analysis applied to the two earlier values indicated that both values would fall below 617 pg/mL in 9% of the patients. Data in four patients with mastocytosis demonstrated a diurnal variation in plasma histamine, with the highest values observed in the early morning (approximately 2:00 A.M.) and the lowest values in the afternoon (approximately 2:00 P.M.). CONCLUSIONS: We conclude that, on average, patients with mastocytosis have elevated plasma histamine levels, whereas patients with unexplained anaphylaxis have plasma histamine levels within the normal range during asymptomatic periods; that plasma histamine levels in patients with mastocytosis exhibit a diurnal variation; and that plasma histamine determinations alone are not useful to screen patients for mastocytosis.

Adult↗
PubMed1989

Hematopathology of the bone marrow in pediatric cutaneous mastocytosis. A study of 17 patients.

Sixteen bone marrow aspirates and 15 trephine core biopsies from 17 children with cutaneous mastocytosis, of which 15 exhibited urticaria pigmentosa and 2 exhibited diffuse cutaneous mastocytosis, were evaluated for the presence of bone marrow-associated pathologic conditions. Eight bone marrow aspirates from 8 children and 23 trephine core bone marrow biopsies from 16 children who had had evaluation for hematologic abnormalities not associated with cutaneous mastocytosis served as a control population. Eosinophilia was a prominent finding in bone marrows of 9 of 17 patients who had cutaneous mastocytosis. Increased mast cell numbers in bone marrow aspirates were observed in 5 children with cutaneous mastocytosis (5 of 16) and in 2 of the control children (2 of 8). Examination of the trephine core bone marrow biopsies obtained from patients with cutaneous mastocytosis demonstrated focal perivascular and paratrabecular aggregates consisting of mast cells, eosinophils, and early myeloid cells in 10 of 15 individuals. Similar lesions were observed in trephine core bone marrow biopsies of 3 of 16 control patients. The focal mast cell lesions characteristic of adult systemic mastocytosis were not observed. The authors conclude that cutaneous mastocytosis in the pediatric age group is rarely associated with definitive bone marrow findings suggestive of systemic mast cell disease and that this observation is consistent with previous reports that cutaneous mastocytosis in the majority of pediatric cases resolves by adulthood.

Bone Marrow↗
PubMed1989

Mastocytosis.

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Humans↗
PubMed1989

Human macrophages cultured on agar but not agarose resemble mast cells.

It has been reported that rare colonies of mast cells may be identified when human bone marrow cells are cultured in agar. Based on this observation, we attempted to culture mast cells from human bone marrow using a modified agar interphase culture. Metachromatically staining cells appearing in culture peaked in number by 2-3 weeks and consisted of basophil-like cells, mast-like cells and larger, granulated cells superficially resembling mast cells. The large cells, which constituted the majority of metachromatic cells, were berberine sulphate-positive but negative for histamine, chloroacetate esterase, and mast cell tryptase. These larger granulated cells were subsequently identified as macrophages by FACS analysis with Leu M3 and by electron microscopy. Berberine sulphate-staining of macrophages was not due to the de novo synthesis of heparin, as shown by analysis of radiolabelled proteoglycans from cultured cells. The macrophage metachromasia was eliminated with the use of agarose. The metachromatic and berberine sulphate-staining of cultured human macrophages was therefore considered to be due to phagocytosed agar. Recognition of this phenomenon will aid in the proper identification of human mast cells and basophils in cultures where agar or similar substances are present, and demonstrates that berberine sulphate-staining is not specific for heparin.

Agar↗
PubMed1988

Functional and biochemical characterization of rat bone marrow derived mast cells.

The functional and biochemical characterization of rat bone marrow derived mast cells (RBMMC) confirms both species-related differences between rat and mouse bone marrow-derived mast cells (MBMMC) as well as mast cell heterogeneity in a single species. Such RBMMC have the staining characteristics of mucosal mast cells and contain the mucosal mast cell protease. The RBMMC release the preformed granule mediator beta-hexosaminidase both in response to immunologic stimulation with 200 ng Ag (net release 15.8 +/- 3.8%) and in response to 1 microM calcium ionophore A23187 (net release 21.8 +/- 6.8%). However, compound 48/80, substance P, and somatostatin did not induce mast cell degranulation. In experiments with optimal beta-hexosaminidase release, the RBMMC generated similar quantities of the newly formed arachidonic acid metabolites leukotriene C4 and PGD2 when stimulated with either Ag or calcium ionophore A23187. The RBMMC incorporate [35S]sulfate into proteoglycans consisting of 90% chondroitin sulfates and 10% heparin. The chondroitin sulfates were comprised of chondroitin 4 sulfate and chondroitin sulfate diB sulfated disaccharides in a ratio of 4/1. Although we show that RBMMC and MBMMC share a low histamine content, functional IgE receptors and unresponsiveness to cromolyn and selective secretagogues (compound 48/80, substance P, and somatostatin), we also provide evidence that RBMMC differ from MBMMC in their profile of newly generated mediators, preformed granule proteoglycan, and lack of proliferative response to mouse IL-3.

Animals↗
PubMed1988

Identification of chondroitin sulfate E in human lung mast cells.

Human lung mast cells (HLMC) enriched up to 99% purity by counter current elutriation and density gradient centrifugation were labeled with 35S-sulfate to determine cell-associated proteoglycans. The 35S-labeled proteoglycans were extracted by the addition of detergent and 4 M guanidine-HCl, and separated from unincorporated precursor by Sephadex G-50 chromatography. 35S-Proteoglycans chromatographed over Sepharose 4B with a Kav of 0.48. 35S-Glycosaminoglycans separated from the parent 35S-proteoglycans by beta-elimination and chromatographed over Sepharose 4B with a Kav of 0.63. Characterization of 35S-proteoglycans by chondroitin ABC lyase treatment revealed approximately 36% of the proteoglycan to be composed of chondroitin sulfates. Analysis by HPLC of component disaccharides liberated by chondroitin ABC lyase using an amino-cyano-substituted silica column indicated that the chondroitin sulfates consisted of the monosulfated A disaccharide (GlcUA----GaINAc4SO4) (75%) and the over-sulfated E disaccharide (GlcUA----GaINAc4,6-diSO4) (25%). Nitrous acid/heparinase-susceptible heparin proteoglycans accounted for approximately 62% of the total 35S-proteoglycans present in the HLMC. Proteoglycans remaining after exposure of the original proteoglycan extract to either heparinase or chondroitin ABC lyase were of similar size, suggesting that the majority of heparin and chondroitin sulfate glycosaminoglycans were on separate protein cores. Proteoglycans extracted from HLMC were protease insensitive. Hence, in addition to heparin proteoglycans, HLMC synthesize a hitherto unrecognized quantity of chondroitin sulfate E proteoglycans.

Chondroitin Lyases↗