Mast Cell Heterogeneity in the Small Intestine of Normal, Gnotobiotic and Parasitized Pigs

Pabst, Reinhard; Beil, Waltraud J.

doi:10.1159/000234827

Cited by 10 publications

(9 citation statements)

References 11 publications

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“…Information concerning pig mast cells is sparse *To whom correspondence should be addressed at: PHLS Centre for Applied Microbiology and Research, Division of Pathology, Porton Down, Salisbury, Wiltshire SP4 0JG, UK. 0018-2214 9 1993 Chapman & Hall (Ashraf et al, 1988;Pabst & Bell, 1989) and the aim of this study was to characterize porcine mast cells histochemically and morphologically before investigating their role in immune responses in infectious enteropathies of pigs.…”

Section: Introductionmentioning

confidence: 99%

Histochemistry and morphology of porcine mast cells

Carr²,

Bland³

et al. 1993

Histochem J

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Mast cells have been described extensively in rodents and humans but not in pigs, and the objective of this study was to characterize porcine mast cells by histochemistry and electron microscopy. Carnoy's fluid proved to be a good fixative but fixation with neutral buffered formalin blocked staining of most mast cells. Alcian Blue stained more mast cells than did Toluidine Blue (pH 0.5), although Alcian Blue also stained goblet cells. In pigs, unlike rodents, the Alcian Blue method did not distinguish between mast cells in the intestinal mucosa and those in the connective tissue of the intestinal submucosa, tongue and skin. Mast cells were significantly larger in adult pigs than in piglets; in adult pigs and piglets, mast cells in the intestinal mucosa were significantly larger than those in submucosal connective tissue, and they were more varied in shape in piglets and adults. Granules in mast cells in the intestinal mucosa stained less intensely than those in mast cells in connective tissue of tongue, skin and intestinal submucosa. Mast cells in the connective tissue of the tongue, skin and intestinal submucosa fluoresced strongly when stained with berberine sulphate or with a mixture of berberine sulphate and Acridine Orange, but mast cells in the intestinal mucosa did not. All mast cells reacted positively in an enzyme-histochemical method previously used to detect human tryptase but not in a method previously used to detect human chymase. Mast cells in the medulla of thymus stained similarly to mast cells in the intestinal mucosa. Ultrastructural differences between mast cells were not detected.

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Section: Introductionmentioning

confidence: 99%

Histochemistry and morphology of porcine mast cells

Carr²,

Bland³

et al. 1993

Histochem J

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“…Three Göttingen minipigs were orally infected with larvae of A. suum (the porcine equivalent to human ascariasis) shortly after birth [32]. After 18 weeks, when mature nematodes were present in the intestine, the animals were killed together with 3 control minipigs to be used for a different study, which was conducted at the Medizinische Hochschule Hannover and corresponded to the ethical guidelines of that institution.…”

Section: Experimental Ascariasismentioning

confidence: 99%

“…Intestinal ascariasis was studied in 2 human cases and a porcine Ascaris suum infection model. Porcine and human intestines have been shown to bear a close resemblance to each other [32], to the extent that porcine grafts have proved suitable for use in transplantation surgery [see, for example, ref . 33].…”

Section: Introductionmentioning

confidence: 99%

Selective Alterations in Mast Cell Subsets and Eosinophil Infiltration in Two Complementary Types of Intestinal Inflammation: Ascariasis and Crohn’s Disease

et al. 2002

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Objective: Numbers of mast cells (MCs) of different subpopulations and the extent of eosinophil infiltration were compared in Crohn’s disease and ascariasis. These two types of intestinal inflammation are complementary with regard to T cell response (TH1 versus TH2), prevalence and environmental factors. Methods: Histochemical, immunohistochemical and ultrastructural tools were applied to biopsies of morphologically uninvolved colon, ileum and duodenum from Crohn’s and ascariasis patients, as well as resection margins and tissues from an experimental porcine ascariasis model. MC subsets were defined by their dye-binding properties, and their chymase content was analysed using biochemical tools. Results: The TH2 (IgE-mediated) response in ascariasis was characterised by a dramatic increase in mucosal- type MCs (MMCs) and eosinophils in both the mucosa and the deeper layers of the intestinal wall and a simultaneous decrease of connective tissue-type MCs (CTMCs). Uninvolved intestine of Crohn’s patients showed moderate proliferation of CTMCs in the deeper layers of the intestinal wall, but a significant decrease of the MMCs, associated with moderate eosinophilia in all layers of the gut. Similar changes were present in the uninvolved duodenum of Crohn’s patients. Comparable amounts of chymase could be extracted from mucosal and submucosal duodenum, with similar proportions of its two principal isoforms in each. Conclusions: Our results indicate that T cell responses (TH1 or TH2) are associated with different MC subsets in intestinal inflammation. Changes remote from the focus of inflammation point to the systemic nature of the different MC responses.

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“…in the skin 7-10 . In pigs, mucosal-type MCs and connective tissue-type MCs have likewise been described due to histochemical properties 11 . However, it is postulated that heterogeneity of MCs in tissues is much more diverse than merely two polarized phenotypes and is probably dynamically changing in accordance with micro-environmental conditions 3 …”

Section: Introductionmentioning

confidence: 99%

“…Several studies have shown that staining patterns of MCs, including the porcine ones, are critically dependent on the respective fixation technique employed. 11,15-17 Apparently, the detection of MCs in formalin fixed specimens is dependent on the species and the localization. 16,18-21 In the pig, mucosal and submucosal MCs did not stain or stained only faintly after formalin fixation 11,17 and a similar phenomenon of loss of metachromatic staining of mucosal MCs is seen in rats and humans.…”

Section: Introductionmentioning

confidence: 99%

Porcine intestinal mast cells. Evaluation of different fixatives for histochemical staining techniques considering tissue shrinkage

et al. 2013

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Staining of mast cells (MCs), including porcine ones, is critically dependent upon the fixation and staining technique. In the pig, mucosal and submucosal MCs do not stain or stain only faintly after formalin fixation. Some fixation methods are particularly recommended for MC staining, for example the fixation with Carnoy or lead salts. Zinc salt fixation (ZSF) has been reported to work excellently for the preservation of fixation-sensitive antigens. The aim of this study was to establish a reliable histological method for counting of MCs in the porcine intestinum. For this purpose, different tissue fixation and staining methods that also allow potential subsequent immunohistochemical investigations were evaluated in the porcine mucosa, as well as submucosa of small and large intestine. Tissues were fixed in Carnoy, lead acetate, lead nitrate, Zamboni and ZSF and stained subsequently with either polychromatic methylene blue, alcian blue or toluidine blue. For the first time our study reveals that ZSF, a heavy metal fixative, preserves metachromatic staining of porcine MCs. Zamboni fixation was not suitable for histochemical visualization of MCs in the pig intestine. All other tested fixatives were suitable. Alcian blue and toluidine blue co-stained intestinal goblet cells which made a prima facie identification of MCs difficult. The polychromatic methylene blue proved to be the optimal staining. In order to compare MC counting results of the different fixation methods, tissue shrinkage was taken into account. As even the same fixation caused shrinkagedifferences between tissue from small and large intestine, different factors for each single fixation and intestinal localization had to be calculated. Tissue shrinkage varied between 19% and 57%, the highest tissue shrinkage was found after fixation with ZSF in the large intestine, the lowest one in the small intestine after lead acetate fixation. Our study emphasizes that MC counting results from data using different fixation techniques can only be compared if the respective studyimmanent shrinkage factor has been determined and quantification results are adjusted accordingly.

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Mast Cell Heterogeneity in the Small Intestine of Normal, Gnotobiotic and Parasitized Pigs

Cited by 10 publications

References 11 publications

Histochemistry and morphology of porcine mast cells

Histochemistry and morphology of porcine mast cells

Selective Alterations in Mast Cell Subsets and Eosinophil Infiltration in Two Complementary Types of Intestinal Inflammation: Ascariasis and Crohn’s Disease

Porcine intestinal mast cells. Evaluation of different fixatives for histochemical staining techniques considering tissue shrinkage

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