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Biomedical subjects

L Enerbäck

Publications and source records attributed to L Enerbäck.

At least 19 recordsLinked to original sources
PubMed1992

Mast cells and eosinophils in the allergic mucosal response to allergen challenge: changes in distribution and signs of activation in relation to symptoms.

An allergen challenge was performed in 10 asymptomatic patients with strictly seasonal allergic rhinitis. For comparison; seven nonallergic subjects were challenged with allergen, and seven allergic patients were challenged with diluent. Cell samples, obtained with use of a brush technique to recover cells from within the epithelium and nasal lavage to collect cells from the epithelial surface, and symptom scores were taken before challenge and at 2-hour intervals during 12 hours. The cell suspensions were cytocentrifuged onto object slides for light microscopy. Histamine was determined in the cell pellets. In brush samples from the allergic patients challenged with allergen, eosinophils, expressed as a percentage of the total granulocytes, increased from 4.3% +/- 2.7% (mean +/- SEM) to 10.3% +/- 3.8% (p < 0.05) 4 hours after challenge. This level was maintained for up to 12 hours. A similar increase was noted in the lavage specimens 2, 6, and 8 hours after the challenge. In the brush samples the proportion of eosinophils containing two or more cytoplasmic vacuoles, taken as a sign of activation, increased from 20% to 72% (p < 0.05) 8 hours after provocation. In brush samples from the allergic patients challenged with allergen, the numbers of metachromatic cells increased to a maximum of eightfold at 10 hours. In the lavage specimens, no metachromatic cells were observed before provocation, but they progressively increased in number 2 to 12 hours after provocation. Cell pellet histamine content decreased temporarily 2 to 4 hours after challenge (p < 0.05) in brush samples from allergen-challenged allergic patients. The local metachromatic cell density before challenge, as reflected in the brush specimens, correlated with nasal congestion, sneezing, and the degree of eosinophilia.

Adult↗
PubMed1990

DNA content and nuclear size of megakaryocytes in thrombocythaemia.

Total nuclear DNA content and nuclear size of megakaryocytes were studied in biopsies of the iliac bone marrow of individuals with normal or increased platelet counts. The DNA content was determined using Feulgen cytophotometry of bone marrow smears and the nuclear area by morphometric analysis of megakaryocytes of bone marrow sections. The mean DNA content and the mean nuclear area were both significantly larger in megakaryocytes of patients with thrombocytosis as a result of myeloproliferative disease than in patients with secondary thrombocytosis as well as in two control groups of individuals with normal platelets counts, one comprising healthy volunteers, the other with various non-haematological disorders. There was a statistically significant correlation between the DNA content and nuclear area of the megakaryocytes (r = 0.92) in the entire group of bone marrows studied.

Adolescent↗
PubMed1990

Mast cells, tissue histamine and eosinophils in early- and late-phase skin reactions: effects of a single dose of prednisolone.

Skin prick tests with allergen and histamine were performed on the volar aspect of the forearms in a double-blind, cross-over study with 40 mg of prednisolone and placebo in 16 pollen-allergic subjects. Skin biopsies were taken before any treatment and 15 min (group 1; n = 8) and 6 h (group 2; n = 8) after local challenge with allergen, corresponding to the timing of an early- and late-phase reaction. The specimens were used for the histological evaluation of mast cell and eosinophil density as well as for the determination of the histamine and protein content. The size of the induced weal and flare area as well as of any late-phase reaction was determined using digitized planimetry. The single dose of prednisolone, given 2 h prior to challenge, did not affect the size of the weal and flare response. Only 4 of the individuals developed a visible late-phase response. Eosinophils were virtually absent before allergen exposure, but were already present 15 min after allergen challenge, largely associated with the blood vessels, and were numerous at 6 h. There was, however, no relationship between eosinophil density and the presence or extent of any visual late phase. The mast cells/basophils showed a tendency to increase at the 6-hour determination. The infiltration of eosinophils was blocked by the glucocorticoid. This treatment also induced a difference in the mast cell density at the 6-hour determinations, associated with a similar difference in the histamine content of the biopsy specimens.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1989

The mast cells.

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Animals↗
PubMed1989

Histamine and mucosal mast cells in interstitial cystitis.

Interstitial cystitis (IC) is a chronic inflammatory disorder of the urinary bladder of unknown aetiology and pathogenesis. The classic form (Hunner's ulcer) is characterized by high mast cell numbers in the detrusor muscle and an expansion of mucosal mast cells in the lamina propria and also in the epithelium. Such cells can be recovered in bladder washings and in the urine in single cell suspensions. We have counted the mast cells and measured the histamine in bladder washings from 16 patients with classic IC and from a control group of 15 patients with so-called early, non-ulcerative IC. The bladder washings from all patients with classic IC contained well preserved mast cells (median 2.16, range 0.5-8.6 x 10(3) cells/I) and histamine (median 14.3, range 6-66 ng/l), while only occasional mast cells and traces of histamine were found in washings from patients with non-ulcerative IC. The histamine content was strongly correlated to the number of mast cells (r = 0.87). The mean histamine content per mast cell was estimated at 7.6 +/- 0.65 (SEM) pg/cell. The high histamine content per mast cell in relation to previously published data (2.8-4.6 pg/cell) can be attributed to the mild and rapid handling of the specimens.

Adult↗
PubMed1989

Prolonged treatment with topical glucocorticoids results in an inhibition of the allergen-induced weal-and-flare response and a reduction in skin mast cell numbers and histamine content.

The inhibitory effects of topical glucocorticosteroid treatment on the immediate dermal allergic reaction were studied in 10 patients in a double, randomized, placebo-controlled fashion. The aim was to study whether a prolongation of the treatment time would enhance the inhibitory effect beyond the 30-40% reduction previously reported after 1 week of treatment, and whether any changes in skin reactivity were accompanied by changes in the level of mast cells or histamine at the challenge site. Allergen and histamine skin-prick tests were performed on both forearms before the start of the study and after 2 and 4 weeks of treatment with placebo cream on one forearm and with 0.05% clobetasol-17-propionate cream on the other. Punch biopsies from the skin treated actively and with placebo were taken after 4 weeks in eight of the patients. The specimens were used for the light-microscopic evaluation of mast cell density and for the measurement of histamine and protein content. After 4 weeks of treatment we found a reduction in the allergen-induced weal (72%; P less than 0.001) and flare (62%; P less than 0.05) response. There was also a minor reduction in the histamine-induced weal (38%; P less than 0.05) but not the flare response, suggesting that the glucocorticoid treatment induced a reduced mediator release at allergen challenge. This could be partially explained by the finding of a reduction in the number of detectable skin mast cells (85%; from 0.78 to 0.11 mast cells per unit area) and in the histamine content of the skin as related to the tissue wet weight (36%; P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1989

Nasal mast cell response to natural allergen exposure.

A redistribution of mast cells into the epithelium and towards the mucosal surface was previously observed during the allergy season in individuals allergic to birch pollen. We have therefore attempted to investigate in greater detail the role of mast cells in mucosal allergy by the study of the morphological and biochemical changes that occur in the nasal mucosa of allergic individuals during natural allergen exposure. An activation of the intraepithelial mast cells was indicated by the observation of ultrastructural signs of a secretory activity. We also found that the normal strong correlation between mast cell numbers and histamine content in the nasal mucosa was absent in specimens taken during allergen exposure, which was interpreted as a result of a release of histamine from the mast cells with the appearance of a transient, non-mast cell pool of tissue histamine. Furthermore, the histamine content of the nasal mucosa during the pollen season was strongly correlated to the severity of symptoms experienced by the patients. These observations provide additional evidence that mucosal mast cells have a pathogenetic role in continuous allergic airway disease.

Allergens↗
PubMed1989

Nasal mucosal response to repeated challenges with pollen allergen.

In order to explore the dynamics of the cellular response of the airway mucosa to allergen exposure, controlled daily allergen challenges were performed for 7 days during the pollen-free winter months in nine patients with strictly seasonal allergic rhinitis caused by birch pollen allergen. Symptoms obtained after the challenge were related to morphologic changes within and on the surface of the nasal mucosa. Cell samples were obtained daily prior to challenge from the nasal mucosa using mucosal imprints on plastic strips, brush samplings from the nasal mucosa, and nasal lavage. For light microscopy, the cellular material obtained by brushing and nasal lavage were cytocentrifuged onto object slides. Histamine was measured in the cell pellets that were obtained using the lavage and brush procedures as well as in the lavage supernatant fluid. Four symptoms (sneezes, itching, secretion, and blockage) were recorded and expressed individually as well as in the form of a composite symptom score. Nasal blockage was measured using a nasal peak flow meter. The patients displayed an increase in nasal symptoms after the allergen challenge, which was further increased between Days 2 to 7 (p less than 0.05). The light microscopic evaluation of the lavage revealed an immediate increase in eosinophils from 2.3 to 13.1% and was maintained on the same level throughout the period of challenges. A similar increase was noted in the brush specimens, and a strong correlation was obtained between these two methods of sampling.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Intranasal↗
PubMed1989

Effect of nedocromil sodium on the compound exocytosis of mast cells.

The secretory activity of peritoneal mast cells was studied in vitro after challenge with compound 48/80 of cells obtained from normal rats and with anti-rat IgE of cells obtained from rats hyperimmunised with Nippostrongylus brasiliensis. Peritoneal mast cells were isolated from 2 groups of rats aged 2 to 3 months and 5 to 6 months. After purification, the cells were preincubated with or without nedocromil sodium for 5 minutes followed by addition of secretagogue and incubation for an additional period of 5 minutes (compound 48/80) or 15 minutes (anti-IgE). The secretory response was analysed by measurement of the release of endogenous histamine and serotonin (5-hydroxytryptamine) into the medium using high performance liquid chromatography, and by the assay of the heparin content of individual mast cells by cytofluorometry, using the fluorescent dye berberine as a quantitative marker. Concentrations of secretagogues which gave 30 to 40% amine release in controls without inhibitor were selected. In agreement with previous reports, the degree of release of histamine was slightly higher than that of serotonin, but nedocromil sodium suppressed the release of both amines in a parallel manner, without any differential effect, after stimulation with both compound 48/80 and anti-IgE. The effect of nedocromil sodium was concentration-dependent, with the lowest effective concentration in the range of 10(-8) to 10(-7) mol/L, and reduced release to about 40% of control values. Mast cells from young rats responded with a significantly higher amine release than mast cells from old rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1988

A brush method to harvest cells from the nasal mucosa for microscopic and biochemical analysis.

A method is described for the sampling of epithelial cells and other effector cells from the human airway mucosa for structural and biochemical analysis. The cell samples are obtained from the nasal mucosa using a small nylon brush which is rotated over the epithelium and soaked and shaken in a small volume of a balanced salt solution. Morphological evaluation using light microscopy and transmission electron microscopy revealed excellently preserved cytological detail. In asymptomatic individuals the cells harvested were as follows: 45 +/- 5.9% (mean +/- SEM) epithelial cells, 38 +/- 7.1% granulocytes, 16 +/- 2.3% large mononuclear cells (monocytes), and 1.3 +/- 2.3% eosinophils. Repeated measurements in the same individual revealed a coefficient of variation of the order of 40% for the proportions of cells harvested. In comparison with nasal airway lavage, a higher proportion of epithelial cells and monocytes were obtained with the brush method. The cells harvested could also be used for biochemical analysis. The histamine content of the cell pellets was found to be strongly correlated with the mast cell count (r = 0.93) and was estimated to about 10 pg/cell, which is higher than previously reported for mast cells obtained from human lung tissue dispersed by an enzymatic method. The present method appears to be appropriate for the study of cellular events in the nasal mucosal epithelium.

Adult↗
PubMed1988

Identification of oversulphated galactosaminoglycans in intestinal-mucosal mast cells of rats infected with the nematode worm Nippostrongylus brasiliensis.

The oversulphated galactosaminoglycans synthesized by rat mucosal mast cells were isolated from the small intestine of animals infected with the nematode Nippostrongylus brasiliensis, which causes proliferation of these cells. The 35S-labelled polysaccharides were degraded by digestion with chondroitinase ABC, and the structures of the disaccharide products were determined by cleavage with mercuric acetate followed by electrophoretic characterization of the resultant sulphated monosaccharides. It was concluded that about half of the disulphated disaccharide units in the polysaccharide consisted of chondroitin sulphate E-type structures [GlcA-GalNAc(4,6-di-OSO3)], in which both sulphate groups were located on the N-acetylgalactosamine unit. The remainder consisted of isomeric structures with one sulphate group on the N-acetylgalactosamine residue and one on the hexuronic acid unit and presumably represented the dermatan sulphate-type sequence [IdoA(2-OSO3)-GalNAc(4-OSO3)].

Animals↗
PubMed1988

Phenotypic expression of proteoglycan in mast cells of the human nasal mucosa.

The phenotypic expression of the proteoglycan of human mast cells in the nasal mucosa and normal skin was analysed using histochemical techniques. Nasal mucosa was obtained from normal subjects, from patients with seasonal allergic rhinitis before and during the pollen season and from patients with nasal polyps. In the latter groups, specimens were taken from both polyp tissue and adjacent nasal mucosa. Formaldehyde treatment blocked the cationic dye binding in 75-84% of the mast cells located in the nasal mucosa, as compared to the optimum fixation with IFAA (iso-osmotic formaldehyde-acetic acid). A significantly lower degree of blocking of dye binding was obtained in the human skin where 45% of the mast cells were susceptible to formaldehyde treatment (P less than 0.01). The mast cells of the polyp tissue also showed a relatively low degree of blocking (54%), which was significantly lower than the blocking of mast cells of the nasal mucosa taken from the same individuals (P less than 0.05). Staining of serial tissue sections in Alcian Blue containing graded concentrations of MgCl2 was used to determine the critical electrolyte concentration (CEC) of the dye binding, defined as the salt concentration at which the staining of 50% of the mast cells is extinguished. The CEC of the skin mast cells was 0.64M MgCl2 which is significantly higher than that of the mast cells of the nasal mucosa of normal subjects [0.49M (P less than 0.05)], allergic subjects [0.52M (P less than 0.01)], patients with polyp disease [0.52M (P less than 0.01)] and the polyp tissue proper [0.57M (P less than 0.05)].(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1988

Histochemical heterogeneity of dermal mast cells in athymic and normal rats.

Mucosal mast cells (MMC) and connective tissue mast cells (CTMC) of the rat contain different proteoglycans, which can be distinguished using histochemical methods. The chondroitin sulphate proteoglycan of the MMC, unlike the heparin of the CTMC, does not show fluorescent berberine binding, is susceptible to aldehyde fixatives and stains preferentially with Alcian Blue in a staining sequence with Safranin. The majority of the dermal mast cells are typical CTMC and are located in the deep part of the dermis. Subepidermal mast cells are comparatively few in normal rats but numerous in athymic rats and mice. These cells differ from other dermal mast cells in that they stain preferentially with Alcian Blue and they appear to contain little histamine. We examined some of the histochemical properties of the skin mast cells of female PVG-rnu/rnu rats and their heterozygous littermates aged from 5 to 29 weeks. The thiazine dye-binding of the subepidermal mast cells was partially blocked by formaldehyde fixation and only about half of them showed a weakly fluorescent berberine binding. The critical electrolyte concentration of the Alcian Blue staining of the subepidermal mast cells was between that of CTMC and MMC. Deaminative cleavage with nitrous acid abolished the staining of all skin mast cells, while that of the MMC was unaffected. There were no statistically significant differences in the staining patterns of the dermal mast cells between different ages or groups of rat. These results indicate that the subepidermal mast cells contain a heparin proteoglycan which is, however, different from that of the typical CTMC of other sites. They thus appear to represent a second example of a mast cell within a defined anatomical location exhibiting a distinct proteoglycan expression.

Animals↗
PubMed1988

Secretory activity of nasal mucosal mast cells and histamine release in hay fever.

Although theoretical considerations and experimental evidence implicate the mast cells in the pathophysiology of the immediate type hypersensitivity reaction, the evidence of their active participation in human allergic disease is still fragmentary. We have therefore sought evidence of mast cell activation in allergic mucosal disease using strictly seasonal allergic rhinitis as a model. Twelve patients with birch pollen-induced hay fever were examined before and well into the birch pollen season. Allergen exposure was monitored by pollen counts and the degree of symptoms registered daily. Small surgical biopsies and mucosal imprints were obtained from each patient before and during the season. Mast cells were analysed by light and electron microscopy and mucosal histamine was measured using a sensitive HPLC assay. We found a reduction in the number of mast cells in the nasal mucosa during pollen exposure (p less than 0.05) but no significant reduction of the histamine content. There was a correlation between the nasal mucosal mast cell density and histamine content before the pollen season (r = 0.76; p less than 0.01), but no such correlation was found during the period of pollen exposure (r = 0.19; n.s.). This finding points to secretory activity by the mast cells during the pollen season and to the appearance of a non-mast cell pool of tissue histamine. Evidence for a secretory activity of the mast cells during the pollen season was also confirmed by electron microscopy. In addition, we found a strong correlation (r = 0.77; p less than 0.01) between the histamine content of the nasal mucosa during the pollen season and the degree of nasal symptoms. The number of epithelium-associated mast cells found on mucosal imprints prior to the pollen season showed a strong correlation with the symptoms experienced later during the period of pollen exposure (r = 0.83; p less than 0.01). Taken together these observations indicate that the mast cell has a pathogenetic role in continuous allergic airway disease and re-emphasizes the role of histamine in the induction of the symptoms of allergic rhinitis.

Adult↗
PubMed1987

Mucosal mast cells in the rat and in man.

The proteoglycan structure of mucosal mast cells (MMC) of the two species has been analyzed with histochemical in situ techniques. The findings indicate that human MMC, like human mast cells of several other sites, contain a heparin proteoglycan, unlike rat MMC which lack heparin but contain an oversulphated chondroitin sulphate. However, the dye-binding of the human MMC proteoglycan, like that of the rat, is highly susceptible to blocking by formaldehyde. Human MMC also exhibit a lower critical electrolyte concentration (CEC) of dye-binding than mast cells of other connective tissue sites, suggesting a relatively lower charge density and/or molecular weight of the glycosaminoglycan of the MMC. These findings thus suggest that the human MMC like those of the rat have a distinctive proteoglycan structure. Recent findings of another group indicate that the human MMC like those of the rat have also a distinctive proteinase composition. Finally, the mast cell response of the nasal mucosa during birch pollen allergy shows fundamental similarities to the nematode response of the rat intestinal mucosa. During both conditions mast cells are redistributed from the lamina propria into the epithelium, probably as a result of migration of mast cells or mast cell precursors. Taken together, these findings suggest the existence of a distinctive MMC phenotype also in man.

Animals↗