Foamy cells associated with phagocytosis of glutaraldehyde-treated red blood cells and red cell membranes

In order to clarify the mechanism for the formation of foamy cells (macrophages with foamy appearance) associated with increased erythrophagocytosis, we tried to reproduce these cells in mice by subcutaneous injection of intact red blood cells (RBCs), OsO4-treated RBCs (Os-RBCs), glutaraldehyde-treated RBCs (G-RBCs), or isolated red cell membranes, and time-course observation was done by light and electron microscopy. Foamy cells were induced by the latter two methods. Within the macrophages, G-RBCs were fragmented into spherules by newly formed small vacuoles, and with time these spherules lost their hemoglobin content transforming into small vacuoles with translucent matrix. In most of these vacuoles, red cell membrane structure was discernible adjacent to the phagocytic vacuole. Such macrophages containing abundant small vacuoles appear foamy in light microscopy. Foamy cells induced by injection of red blood cell membranes were positive for lipid stains and contained abundant laminated membrane structures in electron microscopy. These results suggest that the foamy cells related with increased erythrophagocytosis are heterogeneous with respect to their pathogenesis and cellular inclusions, and proteinaceous constituents resistant to intracellular digestion are also responsible for the occurrence of foamy cells.     

FOAMY CELLS ASSOCIATED WITH PHAGOCYTOSIS OF GLUTARALDEHYDE-TREATED RED BLOOD CELLS AND RED CELL MEMBRANES

In order to clarify the mechanism for the formation of foamy cells (macrophages with foamy appearance) associated with increased erythrophagocytosis, we tried to reproduce these cells in mice by subcutaneous injection of intact red blood cells (RBCs), OsO, -treated RBCs (Os-RBCs), glutaraldehy de-treated RBCs (G-RBCs), or isolated red cell membranes, and time-course observation was done by light and electron microscopy. Foamy cells were induced by the latter two methods. Within the macrophages, G-RBCs were fragmented into spherules by newly formed small vacuoles, and with time these spherules lost their hemoglobin content transforming into small vacuoles with translucent matrix. In most of these vacuoles, red cell membrane structure was discernible adjacent to the phagocytic vacuole. Such macrophages containing abundant small vacuoles appear foamy in light microscopy. Foamy cells induced by injection of red blood cell membranes were positive for lipid stains and contained abundant laminated membrane structures in electron microscopy. These results suggest that the foamy cells related with increased erythrophagocytosis are heterogeneous with respect to their pathogenesis and cellular inclusions, and proteinaceous constituents resistant to intracellular digestion are also responsible for the occurrence of foamy cells.     

ELECTRON MICROSCOPIC STUDIES OF THE SPLEEN AND LIVER IN HEREDITARY SPHEROCYTOSIS

Electron microscopic findings of the red pulp of the spleen and liver from three patients with hereditary spherocytosis have been reported.
The sinus lumen of the spleen contained various amounts of red cell with reduced deformability. Pronounced engorgement of erythrocytes in the cordal space and various stages of erythrophagocytosis by the cordal macrophages were characteristic findings. The sinus lining cells also showed erythrophagocytosis and contained numerous dense bodies. Cordal macrophages and fibrous elements seemed to be more increased in the older patient.
Acid phosphatase activity of varied intensity was demonstrated in the erythrophagocytic vacuoles of cordal macrophages. As the intracellular degradation progressed, more intense activity of this enzyme was noted, indicating their lysosomal origin.
Splenic conditioning and enhanced destruction of defective red cells in the spleen were the main cause of hemolysis in hereditary spherocytosis.
In the liver, erythrophagocytosis by the Kupffer cells was also not infrequently noted, but the liver was estimated to play a minor role in HS.
The mechanism of hemolysis and the role of the spleen and liver in elimination of hereditary spherocytes have been discussed.     

Erythrophagocytosis and recycling of heme iron in normal and pathological conditions; regulation by hepcidin]

Most of the iron required for erythropoiesis is provided by heme iron recycling following degradation of senescent erythrocytes by tissue macrophages. Accumulation of biochemical modifications at the red blood cell membrane during ageing (externalisation of phosphatidyl-serine, peroxydation of membrane-bound lipoproteins, loss of sialic acid residues and formation of senescence neoantigens) constitute a series of signals that will allow the macrophage to identify the red blood cells to be eliminated, through interaction with specific receptors. After this initial recognition step, the red blood cell is internalised by phagocytosis, and phagosome maturation, which can comprise recruitment of the endoplasmic reticulum, will favour degradation of red blood cell constituents. Heme is catabolised by heme oxygenase 1, anchored in the endoplasmic reticulum membrane. A fraction of the released iron will be recycled back to the plasma through ferroportin, a membrane-bound Fe (II) export molecule, and a fraction will retained within the ferritin molecules, to be released at later stages. Multiple evidence coming from human diseases (type 4 hemochromatosis) and animal models indicate that ferroportin is essential for heme iron recycling by macrophages. Furthermore, ferroportin seems to be the molecular target of hepcidin, this circulating peptide synthesized by the liver and acting as a negative regulator of intestinal iron absorption and iron recycling by macrophages. Perturbations in erythrophagocytosis play a physiopathological role in several diseases, including hemochromatosis, anemia of chronic disorders and thalassemia.     

Effect of erythrocyte ingestion on macrophage antibacterial function.

Individuals with sickle cell anemia are subject to serious infections caused by a number of bacteria, including Salmonella species and Staphylococcus aureus. It has been suggested that in sickle cell anemia, extensive erythrophagocytosis by macrophages may interfere with their antibacterial function and thereby predispose to infection. As a means of investigating this possibility, we evaluated the effects of erythrocyte ingestion on the Killing of Salmonella typhimurium by peritoneal macrophages and of S. aureus by alveolar macrophages. Monolayers of rabbit macrophages were exposed to erythrocytes or latex particles immediately before and during bacterial challenge. Erythrophagocytosis markedly inhibited intracellular killing of S. typhimurium by peritoneal macrophages (bacterial survival was 181% of control) and of staphylococci by alveolar macrophages (bacterial survival was greater than 200% of control). Exposure to latex particles depressed the bactericidal activity of alveolar macrophages to a lesser degree. Next we investigated the possibility that erythrophagocytosis inhibits oxidative bactericidal mechanisms in macrophages. Hexose monophosphate shunt activity was stimulated by erythrocyte ingestion. However, zymosan-induced superoxide generation and chemiluminescence were suppressed by erythrocytes. Furthermore, a cell-free (hypoxanthine-xanthine oxidase) system for chemiluminescence generation was also depressed in the presence of erythrocytes (intact or lysate) or by purified hemoglobin. These studies reveal that erythrophagocytosis inhibits macrophage antibacterial function, probably because of interactions between erythrocyte components and reactive products of phagocyte oxygen metabolism. This host defense abnormality may predispose to bacterial infection in certain hemolytic anemias.     

Electron microscopy of the red pulp of the dog spleen including vascular arrangements,periarterial macrophage sheaths (Ellipsoids), and the contractile,innervated reticular meshwork

The vascular and stromal arrangements of the red pulp in congested and contracted dog spleens were studied by transmission electron microscopy. Each dog had been injected intravenously with Thorotrast to label actively endocytizing cells. Only macrophages ingested Thorotrast. The proximal portion of each arterial capillary was surrounded by a “periarterial macrophage sheath” (PAMS), a term we introduce to replace the term “ellipsoid”. PAMS were composed of a fine meshwork of reticular cells and reticular fibers which held tightly-packed macrophages and interspersed blood cells. These macrophages, as well as those in the reticular meshwork of red pulp, contained Thorotrast, cell debris, and deposits of hemosiderin. The arterial capillary at the center of each PAMS was formed by parallel, rod-shaped endothelial cells and discontinuous layers of basement membrane and reticular-cell cytoplasm. PAMS were tapered at their distal ends; the terminal portion of the arterial capillary continued beyond the PAMS to end in the reticular meshwork of red pulp. Endothelial cells in the terminal arterial capillaries were separated by gaps through which blood cells passed into the spaces of the reticular meshwork of red pulp. The reticular meshwork was formed by reticular cells which appeared to be specialized for contraction. These cells were filled with thin filaments and possessed plasmalemmal dense bodies as found in smooth muscle cells. Furthermore, the reticular meshwork was innervated by unmyelinated adrenergic axons which probably were derived from nerves that followed arterioles. Axons were enclosed in surface invaginations of cells which were similar to reticular cells in shape and cytologic detail and which we called “axon-bearing reticular cells”. Axon-bearing reticular cells were inserted between the branches of the reticular cells that formed the meshwork. Venous sinuses formed an anastomosing system of vessels draining into pulp veins which then joined trabecular veins. Sinuses were formed by parallel, rodshaped endothelial cells encircled by strands of basement membrane and reticular-cell branches. Endothelial cells lay closely side by side except where inter-endothelial slits were opened by blood cells passing into the lumen or by pseudopodia of macrophages which lay outside the sinus. Cell traffic across the sinus wall was greatest in areas where blood cells were mixed with plasma. Congested spleens stored concentrated red cells in both sinuses and the reticular meshwork; contracted spleens were emptied of blood. The reticular meshwork may contract to assist trabecular and capsular smooth muscle in expelling stored red cells and effecting hemoconcentration.     

Effect of membrane-bound IgG and desialysation in the interaction of monocytes with senescent erythrocytes

Abstract Determination of the erythrocyte lifespan is a complex process affected by many cellular parameters. In the present study we measured and characterised the red blood cell (RBC) membrane proteins, mainly band 3, and quantified membrane-bound IgG in senescent RBC (SeRBC) and young RBC (YRBC). We also investigated, through a functional assay, the interaction between SeRBC and peripheral blood monocytes. We applied this erythrophagocytosis assay to study the phagocytosis of desialysed RBC. The results obtained showed no changes in the protein content between SeRBC and YRBC and no differences when examining membrane proteins by SDSPAGE. Then, considering that the accumulation of autologous IgG on RBC membrane provides a direct mechanism for the removal of SeRBC, we measured the IgG content of intact RBC using an enzyme-linked antiimmunoglobulin test finding that the number of IgG molecules bound to SeRBC was significantly higher than that observed for YRBC. The increase observed in the percentage of erythrophagocytosis with SeRBC and sensitised RBC (SRBC) confirmed the involvement of autologous IgG in the selective removal of erythrocytes. We also observed a higher percentage of monocytes with phagocytosed and adherent RBC (AM) obtained with neuraminidase-treated RBC than those obtained with YRBC. This finding suggests that a decrease in sialic acid content of SeRBC may be involved in physiological erythrophagocytosis     

Vascular pathways in nonsinusal red pulp—an electron microscope study of the cat spleen

The red pulp of the cat spleen, including terminal segments of arterial capillaries, pulp venules, and the reticular meshwork, was studied by transmission electron microscopy. Splenic congestion and contraction were produced by barbiturate anesthetic and norepinephrine. Terminal segments of arterial capillaries were ampullary and flared. Blood escaped into surrounding pulp spaces through interendothelial gaps. Pulp venules originated as open-ended vessels in the reticular meshwork near trabeculae and drained into trabecular veins. Venule walls were thin and composed of squamous endothelial cells, a continuous basement membrane, and reticular cells. Venules in congested spleens had many mural apertures, but venules in contracted spleens had few. The interstices of the reticular meshwork in congested spleens contained large amounts of blood, which often was concentrated, many macrophages, lymphocytes, and plasma cells. Fewer blood cells and scant plasma were present in contracted spleens. The vascular arrangements are anatomically open. Blood takes pathways through the reticular meshwork from arterial terminations to pulp venules. Some pathways through the reticular meshwork probably function as closed vascular channels conveying rapidly flowing blood. Other pathways are functionally open and probably contain slowly moving blood that constitutes a reservoir of red cells. Macrophages formed associations with mature red cells and with reticulocytes. Mature red cells were attached to macrophages in a manner indicating erythrophagocytosis. Reticulocyte attachment had a different appearance and likely resulted in reticulocyte sequestration. Platelets bore pseudopodia which would impede their passage through irregular and cell-filled pulp spaces. The change in platelet shape probably is responsible for the formation of the splenic pool of platelets.     

Interaction between macrophage and parasite cells in lobomycosis. The thickened cell wall of Paracoccidioides loboi exhibits apertures to the extracellular milieu

Lesioned, cutaneous, tissue fragments from five indians of the Caiabi tribe with lobomycosis, living in the Xingu National Park (Central Brazil), are analyzed by light and electron microscopy. Clusters of macrophages filled with parasite and/or cell wall debris, separated by collagen fiber bundles, characterize the morphological pattern seen in thick and thin sections. Paracoccidioides loboi within the phagocytic cells are multinucleate organisms whose cytoplasm contains mitochondria with few cristae, ribosomes and vacuoles; a large, dense, globular body and peculiarly curved mitochondrial profiles are described. From the outer portion of the double layered parasite cell wall, radial projections commonly emerge, rendering the structure conspicuously thicker and more irregularly surfaced than that seen in many other phagocytized yeast cell species. The cell wall layers from fungi possessing distinct subcellular organization show a weak or no reaction for acid phosphatase. Most of the foamy cells commonly seen by light microscopy are macrophages filled with fungal cell wall remnants which exhibit marked acid phosphatase activity. Occasionally, microchannels extending from the outer layer of the parasite cell wall to the macrophage surface and exocytic-like openings, possibly derived from the fusion of the macrophage membrane covering the parasite cell wall and the macrophage plasmalemma can be seen. Through such routes, material of texture and density similar to that of the outermost cell wall layer appears to be deposited extracellularly.     

Myospherulosis: further ultrastructural observations

The changes of myospherulosis were observed in masses from the buttocks of three patients. Ultrastructural examination of parent bodies and spherules from two patients revealed some spherules to contain dense bodies and filaments. Spherules had a double electron dense wall: the outer layer had a thickness corresponding to that of a cell membrane. Incubation of packed red blood cells with tetracycline ointment produced a similar electron dense deposit on the inner aspect of the cell membrane of the erythrocytes. No filaments or parent bodies were seen. Our findings support the hypothesis that myospherulosis represents altered red blood cells. The filaments seen in our patients may represent polymers of haemoglobin. It is suggested that parent bodies may be derived from histiocytes.     

The effects of malnutrition on murine peritoneal macrophages

Differences were detected between peritoneal macrophages (both resident and elicited) from mice on a low protein diet and from normal animals. The concentration of resident peritoneal macrophages was lower in animals on low protein diets than in normal controls. Although total protein (and therefore cell mass) of resident macrophages from malnourished mice was increased, their contents of thiamine pyrophosphatase, succinate dehydrogenase, and non-specific esterase were disproportionately reduced. In addition they did not ingest as many glutaraldehyde-fixed sheep erythrocytes or attach to as many adherent C3b sensitized sheep red blood cells as those from normal animals, although reduction of nitroblue tetrazolium was unaffected. Initially (24 hr after thioglycollate), elicited macrophages from malnourished mice did not divide as frequently as those from normal mice but by 48 hr the differences were insignificant. The elicited macrophage possessed lower levels of total protein (indicating a reduced cell mass); the levels of acid phosphatase, thiamine pyrophosphatase, succinate dehydrogenase, and nonspecific esterase and nitroblue reducing activity were also proportionately reduced. They ingested fewer glutaraldehyde-fixed erythrocytes and reacted with fewer C3b sensitised sheep red blood cells than those from normal mice; ingestion of IgG-coated sheep erythrocytes, on the other hand, was somewhat increased. These abnormalities may influence adversely the efficiency of early phlogistic responses and favor the establishment of infection in malnourished animals.     

脂多糖对巨噬细胞铁代谢的影响

目的检测二价金属离子转运体(DMT1)和金属转运蛋白(FPN1)在原代培养巨噬细胞中的分布及脂多糖(LPS)对巨噬细胞铁代谢的影响。方法采用原代细胞培养、MTT显色、细胞化学染色和细胞免疫荧光等方法检测LPS对原代培养的巨噬细胞活性的作用及对DMT1和FPN1分布的影响。结果不带铁反应元件的二价金属离子转运体(DMT1-IRE)主要在细胞核中表达,在细胞质中分布较少。带铁反应元件的二价金属离子转运体(DMT1+IRE)主要分布于细胞质中,可以将吞噬小体中的铁向细胞质中转运。FPN1在巨噬细胞的细胞质和细胞膜均有表达,但主要分布于细胞质。结论巨噬细胞吞噬衰老的红细胞以后,其中的亚铁血红素在细胞质中分解,FPN1可能介导了亚铁血红素分解释放出来的铁在巨噬细胞质中的转运过程。LPS在低浓度时有促进巨噬细胞生长的作用,LPS浓度为10-5μg/L时达到高峰,随着浓度的增加,又开始抑制细胞生长。     

FINE STRUCTURES OF THE SPLEEN IN HEREDITARY ELLIPTOCYTOSIS

Light and electron microscopic studies of the spleen and liver from a patient with hereditary elliptocytosis have been reported. Characteristic findings are congestion of the cordal space, diminished variation of the erythrocyte shape In the cord, relatively empty sinuses and erythrophagocytosis by the cordal macrophages. The same basic defect of the red cell membrane as has been documented in hereditary spherocytosis might be a possible explanation on the pathogenesis of this rare hemolytic anemia.     

AM-3K,A NOVEL MONOCLONAL ANTIBODY SPECIFIC FOR TISSUE MACROPHAGES AND ITS APPLICATION TO PATHOLOGICAL INVESTIGATION

An anti-human macrophage monoclonal antibody, AM-3K, was produced using human alveolar macrophages as antigen. The molecular weights of the antigen recognized by AM-3K were 120 and 70 kD. Immunohistochemically, AM-3K reacted intensely with most macrophages in lymphoreticular organs and in many other organs and tissues. In the spleen, AM-3K reacted with red pulp macrophages, some white pulp macrophages, and tingible body macrophages in lymphoid follicles. In the lymph nodes, many macrophages distributed in the outer cortex, paracortical area, medulla, capsule, or within lymphoid follicles showed an intense reaction for AM-3K. Kupffer cells of the liver, macrophages in the connective tissues, and interstitial macrophages of the kidneys, pancreas, testis, and many other organs were also strongly labelled. AM-3K also reacted with macrophages in many pathological conditions. This antibody, however, did not react with dendritic cell populations, such as epidermal Langerhans cells, interdigitating cells in the paracortex of the lymph nodes, and follicular dendritic cells within the lymphoid follicles, nor with cells other than macrophages, including epithelial cells, vascular endothelial cells, lymphocytes, and granulocytes. Reaction products for AM-3K were found on the cytoplasmic membrane of macrophages by immunoelectron microscopy. In both cryostat sections and formalin-fixed paraffin sections, this monoclonal antibody recognized the antigen present on the cell surface membrane of tissue macrophages, but not monocytes or dendritic cells.     

Neutral lipid accumulation in macrophages during lipid-induced macrophage growth

We previously reported that lipids such as cholesterol esters, triglycerides, and some phospholipids that are components of cell membranes or serum lipoproteins induced the growth of mouse peritoneal macrophages in vitro. Here, we report that macrophage growth was directly induced by lipids without any soluble factors and that the continuous presence of lipids was required for their growth during culture. When macrophages were cultured with several phospholipids that stimulated macrophage growth, their triglyceride content was dramatically increased during culture, whereas their contents of phospholipids, cholesterol, and esterified cholesterols were scarcely changed. On the other hand, phosphatidylcholine and sphingomyelin, which did not stimulate macrophage growth, caused less accumulation of triglycerides in the cells. Macrophages in which triglycerides accumulated resembled foam cells, since they contained many particles that stained with oil red O. Macrophages that cultured with cholesteryl linoleate also contained many oil red O-stainable particles. These data suggest that accumulation of lipid droplets is correlated with macrophage growth induced by lipids and that foam cells in pathological states such as those in atherosclerotic or xanthoma lesions might proliferate in situ.     

BACH1 regulates erythrophagocytosis and iron-recycling in β-thalassemia

Macrophages play essential roles in erythrophagocytosis and iron recycling. β-thalassemia is characterized by a genetic defect in hemoglobin synthesis, which increases the rate of iron recycling. We previously showed that reduced expression of the BTB and CNC homolog 1 (BACH1) gene leads to increased phagocytosis of abnormal RBCs by activated monocytes. However, the mechanisms underlying this abnormal RBC clearance remained unclear. Herein, the spleen and bone marrow cells of β-thalassemic mice were examined for erythrophagocytosis CD markers and iron-recycling genes. Higher expression levels of CD47 and CD163 on RBCs and macrophages, respectively, were observed in β-thalassemic mice than in wild-type cells. The decreased expression of BACH1 caused an increase in Nrf2, Spic, Slc40a1, and HMOX1 expression in splenic red pulp macrophages of thalassemic mice. To investigate BACH1 regulation, a macrophage cell line was transfected with BACH1-siRNA. Decreased BACH1 expression caused an increase in CD163 expression; however, the expression levels were lower when the cells were cultured in media supplemented with β-thalassemia/HbE patient plasma. Additionally, the iron recycling-related genes SPIC, SLC40A1, and HMOX1 were significantly upregulated in BACH1-suppressed macrophages. Our findings provide insights into BACH1 regulation, which plays an important role in erythrophagocytosis and iron recycling in thalassemic macrophages.     

Effects of orchidectomy on the adrenal macrophage system

Macrophages of the adrenal cortex were studied in normal and orchidectomized rats. In normal rats, few macrophages with numerous cytoplasmic granules were observed, mainly in the zona reticularis. Granules were limited by a single membrane and contained either a finely granular dense matrix or heterogeneous materials made up of electron-lucent parts, dark granular and membranous areas. An aminotriazole-resistant peroxidatic activity was confined to the dense granules. In orchidectomized rats, the number of macrophages was markedly increased, and the cells were concentrated at the border between the zonae fasciculata and reticularis and disseminated throughout the zona reticularis. Lysosomes were more numerous in each macrophage, and those of heterogeneous matrix were larger and their contents were more complex than in normal rats. These results show that orchidectomy stimulates the adrenal macrophage system.     

Immune complex receptors on cell surfaces. II. Cytochemical evaluation of their abundance on different immune cells: distribution, uptake, and regeneration.

A recently developed method for ultrastructural demonstration of cell surface receptors for immune complexes is applied to evaluation of these receptors on various cell types. The method entailing incubation with a complex of horesradish peroxidase (HRP) and antibody to HRP (anti-HRP) disclosed dense foci indicative of immune complex receptors distributed at 30- to 120-mmu intervals over macrophage surfaces. Invaginations, loop-like evaginations, and pinocytotic vasicles stained prominently. The number of stained immune complex receptors averaged 200,000 per oil-induced macrophage and 120,000 per noninduced macrophage, as determined from counts of focal deposits in electron micrographs. Receptor periodicity on giant cells present in oil-induced exudates resembled that on macrophages, but the larger giant cells contained an estimated 1.5 million sites. Although receptor periodicity on eosinophils and neutrophils equaled that on macrophages, the staining was lighter and was interrupted by intervals of unstained membrane. Neutrophils averaged 28,000 and eosinophils 35,000 receptors per cell, whereas those lymphocytes with receptors averaged 3,500 per cell. Viable cells incubated with anti-HRP sequentially exhibited about half as many reactive sites as did cells incubated with immune complex. When warmed to 37 C, viable macrophages and eosinophils pinocytosed soluble immune complexes almost completely within 30 minutes and phagocytosed insoluble complexes more slowly. The endocytosed soluble immune complexes were sequestered within tubulovesicular structures in addition to the expected phagocytic vacuoles. Receptors appeared fully active on macrophages that were restained with soluble, cold immune complex after they had endocytosed immune complex in the course of a 30-minute warming interval.     

Origin of myofibroblasts in the avascular capsule around free-floating intraperitoneal blood clots

The origin of cells comprising the capsule which forms around free-floating intraperitoneal blood clots has been examined in rats. One day after implantation the surface of the clot was covered by 1-2 layers of rounded nucleated cells. The majority of these cells were monocyte/macrophage in type with many cytoplasmic folds. Over the next 2 wk a thick capsule composed of dense connective tissue and myofibroblasts developed, completely covering the clot. The surface became covered by a contiguous layer of mesothelial cells, apparently derived from displaced cells of the damaged peritoneum. Detailed examination of the formation of the capsule suggested that a proportion of cells with the characteristic features of peritoneal macrophages gradually develop over a 3-4 d period the characteristic features of fibroblasts. These cells with time then develop filament bundles and a basal lamina to become myofibroblasts.     

Tonsillar lymphangiomatous polyp in an adult dog

A female spayed cocker spaniel dog, aged 8 years, developed a large, smooth pedunculated mass arising from the right palatine tonsil. Histology revealed that the mass was composed of many, variably dilated, thin-walled lymphatic channels filled with pale eosinophilic fluid lacking red blood cells and embedded in a dense, fibrovascular stroma. The dilated lymphatic channels were lined by a single layer of flattened, discontinuous endothelium with scattered intraluminal valves. Periodic acid-Schiff staining highlighted the discontinuous basement membrane and immunohistochemistry revealed strong cell membrane and cytoplasmic immunoreactivity for CD31 and von Willebrand factor, respectively. The clinical and histological findings were consistent with a tonsillar lymphangiomatous polyp - an uncommon, benign tumour of the tonsil in man that has previously been unrecognized in veterinary medicine.