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Biomedical subjects

T Landsverk

Publications and source records attributed to T Landsverk.

At least 55 records · Page 3Linked to original sources

Ontogeny of reticular cells in the ileal Peyer's patch of sheep and goats.

The ontogeny of reticular cells in the ileal Peyer's patch of sheep from 70 days gestational age was studied by light and electron microscopy and by enzyme histochemistry. Small to medium-sized lymphocytes were seen in the lamina propria at 97 days, when the stroma was essentially still mesenchymal. By 110 days, the stromal cells in the dome/follicle primordia had differentiated into reticular fibroblasts, whose processes and fibers were seen to surround groups of lymphocytes. With advancing age the number and size of primordia increased, and proliferation was obvious among the lymphocytes. Processes of reticular cells increased in number and penetrated between individual lymphocytes of the groups. Coarser desmosome-like contacts were seen between the reticular cells from 115 days onwards. A central light area in the follicle was apparent from 130 days onwards. The fine structure of the stromal cells in this light follicle center developed towards but never became similar to that of follicular dendritic cells in a typical germinal center. The fine interdigitating end branches of the stromal cells were less numerous, and the dense homogeneous material present in between the end branches was not observed in the ileal Peyer's patch follicle. Instead, small particles and vesicles were seen between the various cell types of the light center and were not restricted to the intercellular spaces between the stromal cells. In the dark peripheral zone of the follicle, the stromal cells retained more immature features. The follicle became bordered by a capsule at an early stage. This capsule was formed by multiple layers of flattened fibroblasts separated by small amounts of intercellular material only. The alkaline phosphatase, Mg(2+)-dependent adenosine triphosphatase and 5' nucleotidase reactivities of the follicular dendritic cells in the ileal Peyer's patch were similar to those of early prenatal primary follicles of sheep lymph nodes. This study indicates that the stromal cells of the ileal Peyer's patch are mesenchymal in nature and different from those of germinal centers and the epithelial stromal cells of bursa Fabricii of birds.

Alkaline Phosphatase↗

Ontogeny, distribution and structure of aggregated lymphoid follicles in the large intestine of sheep.

Aggregates of lymphocytes were demonstrated from 70 days gestation (term 150 days in sheep) in the proximal colon and rectum. Immunoperoxidase staining for 5'-bromo-2'-deoxyuridine incorporation and IgM, indicated that the lymphocyte population of lymphoid follicles in fetal sheep colon was actively dividing and surface IgM positive. Enzyme histochemistry for 5'-nucleotidase showed that the lymphocytes developed in a meshwork of positive reticular cells, suggestive of developing follicles. Follicle aggregates were distributed in a characteristic pattern in lambs, with major accumulations in the ascending colon and in the rectum. In adult sheep a partial atrophy of follicle aggregates was observed. The microscopic structure of large intestinal aggregates showed similarities to the jejunal Peyer's patch (PP), with broad follicles containing a prominent corona and wide interfollicular areas in older lambs. The apparent deep penetration of crypts into the lymphoid tissue proper, which is a frequently reported phenomenon for colon follicles, was dependent on the contractile state of the mucosa, as judged from its absence in specimens where the intestinal wall had been stretched before fixation.

Age Factors↗

The intestinal habitat for organized lymphoid tissues in ruminants; comparative aspects of structure, function and development.

Unlike the Peyer's patches of rats and mice, which are considered to be secondary lymphoid organs, the ileal Peyer's patch of sheep is thought to be responsible for the primary generation of B cells, like the bursa of Fabricius of birds. The ileal Peyer's patch of sheep shows prenatal maturation, antigen-independent lymphopoiesis, a rate of lymphocyte production larger than that of the thymus, and involution at a young age. Follicles contain few T cells and have an IgM+, relatively immature B lymphocyte population, as judged by B-cell differentiation markers. The follicle-associated epithelium of the ileal Peyer's patch is of a special type that sheds carbonic anhydrase-rich, 50-nanometer membrane-bounded particles (carbonic anhydrase-reactive particles; CAP) into the intercellular spaces. The CAP filter into the follicle centre and are taken up by lymphocytes. They represent the epithelial (bursa-like) element in an otherwise mesenchymal stroma of reticular cells embedding the follicle lymphocytes. Transepithelial transport of macromolecules, with the formation of multivesicular body-like cytoplasmic vacuoles, appears to be the basis for CAP formation. The jejunal Peyer's patches are devoid of CAP, persist in the adult animal, contain M cells with clusters of B cells in the follicle-associated epithelium, and have many CD4+ lymphocytes in the follicles and in the interfollicular areas. Aggregates of lymphoid follicles in the large intestine resemble the jejunal Peyer's patches with respect to their lymphocyte population and the ileal Peyer's patch with respect to their follicle-associated epithelium.

Animals↗

A study of modified lymphatics in the deep cortex of ruminant lymph nodes.

Ruminant lymph nodes, except when very small, were found to have a system of smooth-walled channels in the periphery of the 'deep cortical units' defined by Bélisle & Sainte-Marie (1981 a,b). Each channel originated with many 'blind' branches in the subnodular layer of the cortex and ended by joining a medullary sinus. The wall consisted of a continuous endothelial lining, a sometimes thin or discontinuous basement membrane without a basal lamina, and at least one layer of flattened reticular fibroblasts. The endothelium was higher than in most typical lymphatics, with a cytoplasmic fine structure similar to that of sinus-lining cells in the medullary sinuses. The intercellular junctions were generally long and elaborate. The lumen often contained opaque material, especially in the branches, as for initial lymphatics, as well as a few lymphocytes and an occasional nonlymphoid cell, but sinus macrophages were never seen. In some lymph nodes the lumen was crowded with lymphocytes. When small ferripolymaltose particles arrived in the node with the afferent lymph, many of them rapidly passed through the outer cortex and reached the lumen of the smooth-walled channels by way of the intercellular junctions of the endothelium. When colloidal carbon was introduced the same way, some of it also reached the channels where it accumulated in the basement membrane and in vesicles and vacuoles of the endothelium. These channels are interpreted as initial lymphatics of the same type as in other lymphoid organs rather than lymph node sinuses. They seem to play an important role for the exit of lymphocytes from the nodes and also for the passage of particulate material, including antigens, through those areas where recirculating lymphocytes arrive in the cortex.

Animals↗

Computer-assisted morphometric analysis of absorptive and follicle-associated epithelia of Peyer's patches in sheep foetuses and lambs indicates the presence of distinct T- and B-cell components.

The phenotypes of lymphocytes infiltrating the epithelium of the jejunal and ileal Peyer's patches in foetal sheep at about 130 days gestation and 2-month-old lambs were examined using indirect immunoperoxidase histochemistry, a panel of monoclonal antibodies and enzyme histochemistry. Computer-assisted morphometric analysis enabled the relative size of reactive areas within epithelia to be estimated. The comparison of the intraepithelial lymphocyte populations associated with structurally developed Peyer's patches of foetal sheep and those of lambs allowed assessment of the impact of extrinsic factors from which the sheep foetus is shielded. The study confirmed the postnatal expansion in the villous intraepithelial lymphocyte population and showed that this expansion involved the CD8 and gamma delta phenotypes. CD4 lymphocytes did not appear in the follicle-associated epithelium until after birth. Unlike the villous epithelium, the follicle-associated epithelium had a high frequency of IgM+ and MHC II+ cells, which was dramatically reduced after birth. This postnatal reduction was particularly prominant in the follicle-associated epithelium of the jejunal Peyer's patch, where the frequency of IgM+ cells fell from 12.4% in foetal sheep to 0.7% in lambs. Double staining for alkaline phosphatase in the jejunal Peyer's patch suggested that clusters of IgM+ cells were associated with M cells.

Animals↗

Transfer of carbonic anhydrase-positive particles from the follicle-associated epithelium to lymphocytes of Peyer's patches in foetal sheep and lambs.

Carbonic anhydrase cytochemistry of the ileal Peyer's patch in foetal and neonatal lambs has indicated secretion from the follicle-associated epithelium to the follicles. Reaction for carbonic anhydrase in the follicle-associated epithelium was found in the luminal plasma membrane, in cytoplasmic vesicles, and in vacuoles containing 50-nm membrane-bounded particles that seemed to be shed to the intercellular space. The lateral plasma membrane was negative for carbonic anhydrase, indicating that formation of carbonic anhydrase-positive particles was restricted to vacuoles. Administration of ferritin to ileal loops of sheep foetuses showed ferritin localized in vesicles and vacuoles of the follicle-associated epithelium followed by exocytosis, together with carbonic anhydrase-positive particles, into the indentations of the lateral cell border. The carbonic anhydrase-positive particles seemed to be transported to the centres of lymphoid follicles where many were attached to the plasma membrane of lymphocytes. Carbonic anhydrase-positive particles were also seen in vesicles and sometimes free in the cytoplasm of the lymphocytes or attached to their nuclear envelope. Light microscopically, carbonic anhydrase reactivity of the follicle-associated epithelium was associated with the early formation of the ileal Peyer's patch at about 100 days gestation. At this time the follicle-associated epithelium showed a strong luminal but at most a weak lateral staining. With further foetal development there was a progressive increase in the amount of carbonic anhydrase-positive reaction product in extracellular particles, both along the lateral cell borders of the follicle-associated epithelium and among the lymphocytes of the follicle centres.

Animals↗

Organization of ruminant Peyer's patches as seen with enzyme histochemical markers of stromal and accessory cells.

Five enzyme histochemical reactions were used to characterize calf, goat kid and lamb Peyer's patches. 5'-nucleotidase and acid phosphatase gave a reticular pattern of staining in follicular and interfollicular regions, respectively. Different subpopulations of fibroblastic reticulum cells were suggested for the T cell area, the dome/corona region, and the follicle capsule. The T cell area and the neck portion of the follicle showed a positive reticular reaction with alkaline phosphatase and non-specific esterase. Follicular dendritic cells were positive for Mg2(+)-ATPase in the follicle centre, contrasting with a negative reaction in the periphery. Dendritic cells prevalent in the dome and T cell area showed a Mg2(+)-ATPase reactivity. Macrophages were stained with non-specific esterase and acid phosphatase. No principal differences were found in cell populations between the three species or between fetuses in late gestation and postnatal animals, although the size of the respective compartments varied.

5'-Nucleotidase↗

Morphology and immunoperoxidase studies of intestinal adenomatosis in the blue fox, Alopex lagopus.

In a blue fox farm about 300 of 400 weanling pups were affected with diarrhoea and prolapse of the rectum. About 40 pups died in an emaciated state after a few days of disease. Eight pups were submitted for examination. Pathological findings were restricted to the caecum, proximal colon and rectum, with occasional extension to the ileum. Affected areas revealed a thick and rigid intestinal wall, a narrow lumen and a thickened, wrinkled and sometimes ulcerated mucosa. Histologically, the mucosa showed long tortuous crypts outlined by a high pseudostratified epithelium which lacked goblet cells. Epithelial cells revealed increased basophilia, and mitoses were seen along the entire crypt length and in the surface epithelium. Silver impregnation showed curved organisms in the apical cytoplasm of the altered epithelial cells. Penetration of the muscularis mucosa by the altered epithelial cells was frequent. The avidin-biotin-immunoperoxidase complex (ABC) technique revealed positive organisms in the apical cytoplasm of altered epithelial cells, when rabbit antisera against different biotypes and serotypes of Campylobacter jejuni and Campylobacter coli were used. Antisera against Campylobacter sputorum subsp. muscosalis and Campylobacter hyointestinalis were negative.

Adenoma↗

Electron microscopy of intestinal adenomatosis in the blue fox, Alopex lagopus.

Scanning electron microscopy of adenomatous intestinal tissue in the blue fox revealed an irregular surface topography of the colon with increased diameter of the crypt openings and prominent ridge formations between crypts. The ileum showed villous atrophy and fusion. Microvilli were short and irregular. Small ulcerations of intestinal mucosa were seen. Freeze-fracture revealed curved intracellular organisms in the altered epithelial cells. Transmission electron microscopy showed features associated with immaturity and high protein synthesis. Filamentous extensions from the basolateral plasma membrane of altered epithelial cells sometimes penetrated the basal lamina. The cytoplasm contained numerous polyribosomes, nuclei had many indentations and large and irregular nucleoli. Intracellular bacteria, with morphology corresponding to Campylobacter spp. were found in the apical epithelial cytoplasm. No host-cell-derived membrane was seen to surround the bacteria.

Adenoma↗

Alterations of ultrastructure and of cytoplasmic filaments in remodelling rat jejunal epithelial cells during recovery from deoxycholate.

Structural features associated with reversibility of lesions induced by deoxycholic acid (DOC) were studied by electron microscopy and immunofluorescence techniques. Tied jejunal loops were incubated in vivo with 2.5-20 mmol/l DOC in isotonic solution. Immediately after this treatment, or after a recovery period of 15 or 150 minutes following washout of the bile acid, the loops were excised and processed. DOC produced epithelial lesions whose severity and reversibility were concentration-dependent. Ultrastructural features associated with the reversibility of the lesions were particularly apparent in specimens exposed to 10-20 mmol/l DOC. These features included cell flattening with the formation of thin, veil-like structures into the eroded area by cells at the edges of the erosions. Immunofluorescence studies showed a redistribution of actin and cytokeratin filaments to the margins and leading edges of the flattened cells. It is suggested that cell flattening and migration are responsible for the rapid morphological recovery of the injured epithelium. Actin and cytokeratin appear to be instrumental in the remodelling and migration.

Actins↗

Peyer's patches in experimental Salmonella dublin infection in calves. Microvascular and epithelial changes contributing to atrophy of lymphoid follicles.

Six calves were infected per os with Salmonella dublin and killed nine hours to seven days later. Early changes included occlusion of capillaries with a hyaline material, particularly in the ileal Peyer's patch (PP). Central areas of the follicles contained hemorrhages and edema. In later stages the follicle-associated epithelium (FAE) of both the jejunal and ileal PP was fused with the adjacent epithelium and the follicles were collapsed. As judged from 5'nucleotidase histochemistry, follicles were depleted of lymphocytes whereas reticular cells were retained. Carbonic anhydrase (CA) histochemistry showed a decreased reaction in the ileal FAE and a reduced amount of CA reactive material in the follicles of the ileal PP, indicating loss of FAE differentiation and function. Hyaline material and fibrinous thrombi were seen occluding the blood capillaries and the lymphatics, respectively. The villi were atrophied and covered with thick fibrin deposits. Using antifibrinogen antibodies, immunoperoxidase stained fibrin in the lymphatics and the lumenal deposits but not the hyaline material in the capillaries. Reaction for CA indicated that this hyaline material originated from erythrocytes. Factors contributing to the follicle atrophy may include anoxia due to stasis in the microcirculation with the formation of erythrocyte thrombi, and reduced lymphopoiesis due to a decrease in the stimulating factors provided by the FAE.

5'-Nucleotidase↗

Immuno-histochemical and -cytochemical evidence suggesting the presence of Campylobacter jejuni and Campylobacter coli in cases of porcine intestinal adenomatosis.

Antisera against a number of Campylobacter species were used in immuno-histochemical and -cytochemical studies on cases of porcine intestinal adenomatosis. Avidin-biotin-complex (ABC) and streptavidin immunoperoxidase methods were used on formalin-fixed, paraffin-embedded and frozen sections. Protein A gold method was used on formaldehyde fixed and frozen sections for immuno-cytochemistry. The antisera used were raised in rabbits by subcutaneous or intravenous injection of living or formalin treated organisms. Anti-sera against different serotypes of the thermotolerant, catalase positive campylobacters, Campylobacter jejuni and Campylobacter coli, gave positive reactions in the immuno-histochemical studies. The staining was found in intestinal epithelial cells both in the ileum and in the colon and was restricted to the apical cytoplasm of adenomatous epithelial cells. The staining had a granular pattern, the positive structures sometimes having the shape of Campylobacter. Epithelial cells in areas with normal differentiation of goblet cells did not stain. In contrast, no staining resulted with antisera against Campylobacter sputorum subsp. mucosalis and Campylobacter hyointestinalis. Immuno-cytochemistry, using antisera against Campylobacter jejuni, showed that the positive staining in altered epithelial cells were restricted to intracellular organisms having a structure resembling Campylobacter spp.

Animals↗

Cell differentiation in intestinal adenomatosis of pigs studied by histochemistry of laminin and enzymes of epithelial and subepithelial tissue.

The distribution of enzymes and laminin was examined in ileal tissue from pigs suffering from intestinal adenomatosis to reveal the nature of the lesion. A disruption of the normal and specific pattern of distribution was found. Thus, the normal ileal epithelium was characterised by brush border enzymes: alkaline phosphatase, magnesium-dependent adenosine triphosphatase (Mg-ATPase), fluoride resistant acid phosphatase and 5'-nucleotidase; enzymes of the basolateral border: Mg-ATPase; and cytoplasmic enzymes: beta-glucuronidase, non-specific esterase and acid phosphatase. Subepithelial fibroblasts seemed to be characterised by 5'-nucleotidase. Laminin was present as a continuous band under the surface and crypt epithelium, somewhat thicker in the former. In contrast, the branching proliferating crypts of intestinal adenomatosis largely lacked enzymes characteristic of both villus and crypt cells. Reactions for the subepithelial components, laminin and fibroblasts were also reduced. The deficient differentiation of the epithelial as well as subepithelial components in porcine intestinal adenomatosis distinguish the condition from crypt hyperplasia and indicate an adenoma-like character.

5'-Nucleotidase↗

Distribution of lymphocyte subsets in the large intestinal lymphoid follicles of lambs.

The phenotypes of lymphocytes in the large intestinal patches (LIP) of lambs were examined by flow cytometry and immunohistology, using a panel of monoclonal antibodies (mAb), and compared to those found in the jejunal (JPP) and ileal Peyer's patches (IPP). T-cell markers were detected on 25% of the LIP and JPP lymphocytes by cytofluorometry, and nearly all T cells expressed the CD4 molecule. In contrast, T cells were scarce in the IPP (less than 1%). The B-cell marker p220 was expressed by 74% of the LIP lymphocytes, whereas surface immunoglobulin-positive cells comprised 50-60% of the lymphocyte population. The adhesion molecule CD2 was expressed by a larger proportion of cells from the LIP and JPP than from the IPP, whereas the adhesion molecule CD44 was detected on more IPP lymphocytes. Major histocompatibility complex (MHC) class I antigens were expressed by nearly all lymphocytes from the LIP, JPP and IPP. The LIP contained 70-80% cells with MHC class II expression, whereas the majority of IPP cells (greater than 95%) were MHC class II positive. Immunohistology showed many CD4+ T lymphocytes in the follicles of the LIP and JPP, but none in the IPP follicles. CD8+ lymphocytes were found in the interfollicular areas and were absent from the follicles. The interfollicular areas of the rectal patch contained about 15% tau delta T cells. In contrast, the JPP, IPP and the colon patch at the beginning of spiral colon contained less than 3% tau delta T cells.

Animals↗

Morphological and functional recovery following exposure to deoxycholic acid. A study in the rat small intestine in vivo.

Whereas many studies deal with teh deleterious effects of unconjugated deoxycholic acid on epithelial morphology, few are concerned with the reversibility of these effects, the subject of the present study. Tied jejunal loops in the rat were incubated for 30 minutes with deoxycholic acid (2.5-20 mmol/litre) in isotonic solution. Immediately after this treatment, or after a subsequent recovery period of 15 or 150 minutes following wash out of the bile acid, the loops were excised, fixed and examined by light microscopy and scanning electron microscopy. Deoxycholic acid produced epithelial lesions whose severity and reversibility depended on the concentration applied. However, even the severely affected epithelium obtained by treatment at 10-20 mmol/litre was reverted to normal within 150 minutes, and a substantial normalisation was observed already after 15 minutes. An exception to this rapid restoration of epithelial morphology and integrity was noted in villi which had suffered necrosis of lamina propria. The revertion of epithelial pathology was paralleled with a normalisation of glucose absorption, of the potassium ion and protein content of the loop fluid, and of the paracellular epithelial permeability as measured with 3-H-poly-ethylenglycol. Morphometry showed that deoxycholic acid caused villous atrophy without affectin the crypt length. The extent and reversibility of this atrophy depended on dose and recovery time as above. It is suggested that the remarkably fast morphological restitution proceeds mainly by process involving cellular remodelling and migration.

Animals↗

Secretin empties bile duct cell cytoplasm of vesicles when it initiates ductular HCO3- secretion in the pig.

To determine whether secretin has any effect on bile duct cell ultrastructure, bile duct cells from liver biopsy specimens of pigs were analyzed morphometrically. During secretory rest, bile duct cell cytoplasmic vesicles totaled 96 (84-103) arbitrary units per cell volume (U). Secretin increased bile HCO3- secretion from 9 mumol/min (range 6-15) to 131 mumol/min (range 118-200) and lowered the bile duct cell vesicles to 5 U (range 3-9). Acute elevation of arterial PCO2 to 10.9 kPa (range 10.2-11.1) doubled vesicle number in resting duct cells and augmented the secretory response to secretin. At high arterial PCO2, secretin cleared the duct cell cytoplasm of vesicles and more than doubled the basolateral plasma membrane surface area. Taurocholate-induced canalicular choleresis, in contrast, did not alter duct cell morphology. It is concluded that secretin clears the bile duct cell cytoplasm of vesicles as it initiates ductular HCO3- secretion, possibly through causing exocytotic insertion of vesicle material into the basolateral plasma membrane.

Animals↗

Phagocytosis and transcytosis by the follicle-associated epithelium of the ileal Peyer's patch in calves.

Latex beads, 250 and 610 nm in diameter, and parapox virus isolated from ecthyma in sheep, were injected into intestinal loops containing either jejunal or ileal Peyer's patches (PP) of 3-4 week old calves. Uptake of latex and parapox virus was restricted to the ileal PP, 30-60 min after injection. The latex beads seemed to be embraced by thin surface protrusions extending from the concentric folds of the follicle-associated epithelial cells (FAE) of the ileal PP. Both latex and virus were internalized into cytoplasmic vacuoles. Some of the vacuoles containing virus showed reaction for acid phosphatase. The latex beads and virus were shed to the intercellular spaces of the FAE. The exocytosis appeared to occur through specialized indentations of the lateral plasma membrane where the production of 50 nm membrane-bounded particles by budding off from the lateral plasma membrane was a prominent phenomenon.

Animals↗

Effects of arterial pH and carbon dioxide on pancreatic exocrine H+/HCO3- secretion and secretin-dependent translocation of cytoplasmic vesicles in pancreatic duct cells.

To elucidate why arterial pH and carbon dioxide (PaCO2) modify the pancreatic H+/HCO3- secretory response to secretin stimulation, experiments were performed on anaesthetized pigs, recording the effects of arterial pH and PaCO2 on exocrine H+/HCO3- secretion and on morphology of pancreatic duct cells. Duct cells contained numerous cytoplasmic vesicles at secretory rest. Their number more than doubled during elevation of PaCO2 from 5.5 to 11.0 kPa. At arterial pH 7.40, maximal secretin stimulation cleared the cytoplasm of duct cells of more than 90% of the vesicles. At high PaCO2, this was accompanied by doubling the basolateral plasma membrane area and a 30% higher secretion rate than at PaCO2 5.5 kPa. Lowering arterial pH to 7.0 more than halved the secretin-induced vesicle clearance of duct-cell cytoplasm as well as exocrine H+/HCO3- secretion and abolished the secretin-dependent basolateral membrane area changes. Supramaximal secretin stimulation did not reverse the inhibitory effect of severe metabolic acidosis on secretion. It is concluded that PaCO2 and arterial pH may modify the secretory response to secretin through determining the incorporation of cytoplasmic vesicle material into the basolateral plasma membrane of duct cells.

Animals↗