Pathways of lymph flow from superficial tissues in the legs of horses

Pathways of peripheral lymph flow from the legs in horses were studied with casts, and with light and electron microscopic techniques. Although lymph nodes in horses occur in large groups, each lymph vessel draining from the periphery appeared to terminate on a single node within a group. The larger branches of each vessel divided either on the node surface or after penetrating into the node, and 25 to 60 terminal afferent vessels entered either the subcapsular, medullary or trabecular sinuses. Numerous initial efferent lymphatics arose either within the medulla, or at its surface, and they often coalesced to form an anastomosing network on the node surface. Almost all of the one to four efferent lymphatics that left the vicinity of the node terminated on other nodes, usually within more centrally placed groups. This arrangement may aid in the amplification and propagation of immune responses initiated in primary nodes.     

Dynamics of mast cells in lymph node following antigenic stimulation

Dynamics of mast cells in rat cervical lymph nodes were examined using conventional histological techniques after injection of Salmonella paratyphi B-H antigen. There was no significant change in the number of mast cells at sixth hour and on the first day of stimulation compared with the controls. The number of mast cells was increased in all lymph node compartments on the second day of stimulation, which continued in the following 3 days. On the eighth day of stimulation, although the mast cell number decreased in the subcapsular area, it was still high in the paracortical area and medullary sinuses of the lymph nodes. On the second day of stimulation, the mast cell number was apparently increased in the subcapsular area than those of the other compartments. In the following days of stimulation, the highest number of mast cells was seen in the medullary sinuses. The highest paracortical mast cell number was determined on the third day of stimulation and some mast cells were observed near the high endothelial venules (HEVs). The changes of mast cell number among the lymph node compartments after antigenic stimulation support the hypothesis that the migration of mast cells occurred. This migration pattern indicates that mast cells enter the lymph node via afferent lymphatics and migrate to the lymph node compartments following antigenic stimulation.     

The parotid, mandibular and lateral retropharyngeal lymph nodes of the camel (Camelus dromedarius)

The parotid, mandibular and lateral retropharyngeal lymph nodes of the dromedary camel were examined using both light and electron microscopy. All three lymph nodes were lobulated. They did not show the characteristic medulla, cortex and paracortex of typical lymph nodes. Instead, they contained lymphatic nodules, dense anodular lymphoid tissue and diffuse lymphoid tissue dispersed throughout the lymph node. Networks of sinuses were present in the diffuse lymphoid tissue. The diffuse lymphoid tissue in the periphery of all lymph nodes examined was characterized by numerous erythrocytes within and around its network of sinuses. The nodal sinuses were contiguous with the septal vessels, which are considered the possible source of erythrocytes seen in this study. The lymph nodes that were seen in this study resembled the haemolymph nodes of other mammals with regard to their content of erythrocytes but were unique in being located in sites that were typical of ordinary lymph nodes. Morphometric analysis has shown that the percentage volume densities of the stroma and the various parenchymal components were similar in the three lymph nodes.     

Blood vessels of lymph nodes in the pig

Pig lymph nodes have an unusual arrangement of cortical and medullary tissue, and lack a hilus. The aim of the present study was to describe the pathways by which blood reaches these lymph nodes. Natural or synthetic latex casts were made of blood vessels of superficial inguinal, popliteal and jejunal lymph nodes. Major nodal arteries approach these lymph nodes and give rise to about five branches, most of which remain on the node surface. These branches often wrap around part or all of the node in a claw-like manner and they divide to form a network on its surface. Arteries from this network penetrate the capsule directly to supply the lymphoid parenchyma. Anastomoses are common at all levels between the arteries supplying the nodes and arteriovenous anastomoses occur within the nodes. These may help to maintain and regulate blood flow within the lymph node under different conditions of stimulation.     

Drainage of lymph from the foreleg to the superficial cervical lymph node in sheep

The superficial cervical (prescapular) node in sheep is large and readily accessible. It lies medial to the omotransverse muscle and 40 to 60 mm cranial to the ventral end of the scapular spine. Lymph vessels reaching this node from the foreleg all enter its ventral half. These lymphatics can be approached surgically opposite the elbow joint where they lie adjacent to the cephalic vein. Carbon particles infused into a single afferent lymphatic were restricted to a small segment of the ventrocranial quadrant of the node. When Evans' Blue dye, or a suspension of carbon particles, was injected subcutaneously just above the hoof they were carried to the node in two to five afferent lymphatics and were found mainly in the ventrocranial quadrant. A knowledge of the anatomy of the superficial cervical node and its afferent lymphatics may be of value in immunological studies, especially where it is desired to introduce antigen into a known part of a node and to leave the remainder for control observations.     

Immunophysiological activity of supramammary lymph nodes of the ewe during the very early phase of staphylococcal mastitis

Efferent mammary lymph was collected from lactating ewes which were unimmunised (controls) or immunised during pregnancy with two doses of an attenuated live Staphylococcus aureus vaccine either in the hindlimb ("directly primed' supramammary nodes) or in the brisket ("indirectly primed' supramammary nodes). Mammary lymph was also collected from unimmunised animals in which the supramammary nodes had been extirpated several months before. Ewes in which the supramammary nodes had been directly primed by staphylococcal vaccination before challenge had a significantly greater output of IgM- and IgG2-containing cells in lymph and higher concentrations of IgG1 and IgG2 antibody against S aureus surface antigens than did other groups. Lymphadenectomised ewes had fewer leucocytes in mammary lymph but a much higher proportion of neutrophils than other ewes, indicating that afferent mammary lymph has an unusually high number of neutrophils and most of these cells are filtered out in the supramammary lymph nodes under normal circumstances. The results indicated that most of the leucocytes in efferent lymph were derived from the supramammary nodes. After induction of experimental staphylococcal mastitis there was a rapid drop in leucocyte output in lymph within one to four hours after infection; the data indicated that events within the supramammary nodes were responsible for this phenomenon. The output of immunoglobulin-containing cells was reduced during this phase. No significant increases in output of lymphoblasts, immunoglobulin-containing cells or specific antibody occurred during the six hours immediately following infection.     

Lymph Drainage from the Ovine Tonsils: An Anatomical Study of the Tonsillar Lymph Vessels

Although the tonsils of sheep have gained much attention during the last decade, only few data are available on their lymph vessel architecture. Tonsillar lymph vessels are immunologically important as they form the efferent routes for locally activated immune cells to reach the draining lymph nodes. To gain insight into the tonsillar lymph drainage in the sheep, Indian ink and a casting polymer were injected into the interstitium of the five tonsils present in the heads of slaughtered sheep. This enabled us to determine the draining lymph node and to examine the microscopic organization of lymph vessels using light and scanning electron microscopy. No lymph vessels were observed within the tonsillar lymphoid follicles. The corrosion casts demonstrated that the lymphoid follicles are surrounded by numerous sacculated lymph sinuses that drain into a dense interfollicular lymph vessel network. From here, the lymph flows into single small lymph vessels that in turn drain into larger lymph vessels extending towards the medial retropharyngeal lymph node. The presented results can be valuable for immunological studies, for example during oral or intranasal vaccine development.     

The immune reactivity of the regional and distant lymph nodes to autochthonous ovine squamous cell carcinomata

Anti-tumor immune reactivity of lymphocytes derived from lymph nodes regional to and distant from tumor growth, as well as that of peripheral blood leucocytes, against autochthonous tumor cells, was investigated. Experiments were carried out in vitro using a 51Cr cytotoxic assay and in vivo by cannulating the afferent and efferent lymphatics of regional and distant lymph nodes and challenging via the afferent lymphatics with 10(7) cultivated autochthonous tumor cells. No anti-tumor cytotoxic reactivity was detected in vitro using lymphocytes derived from any of the sources studied. In vivo, while challenge with autochthonous tumor cells produced no response in the regional lymph node, significant blast cell response was obtained in the distant node. The response at the distant node was associated with the production of antibodies that could bind to tumor cells without causing their demise. The anergy observed at the regional lymph node, and the possibility of a relation between the events occurring at that node and those observed at the distant node, are discussed.     

Morphologic features of lymph vessels in caprine hemal nodes

The distribution and characteristics of lymph vessels in caprine hemal nodes were studied after glutaraldehyde fixation and epoxy resin embedding. Histologically, the lymph vessels were characterized by thin walls and wide lumens containing inspissated lymph in which a few cells were suspended. The lymph vessels contrasted sharply with adjacent blood sinuses that were filled with elements of circulating blood. A circumferential lymph vessel in the cortex joined radial branches in the medulla that met at the hilum to drain through a large efferent lymph channel. Ultrastructurally, the lymph vessel wall comprised endothelial cells supported by a continuous basal lamina, collagen fibrils, and adventitial reticular cells. The cytoplasm of endothelial cells had fenestrations, plasmalemma-associated vesicles, vacuoles, and focal splits that enclosed large compartments. Many compartments contained erythrocytes and lymphocytes. Modifications of the endothelial cells signified their endowment with features that favored transendothelial transport. The distribution of lymph vessels and the finding of only efferent lymph channels are related to the roles of hemal nodes in blood storage by hemoconcentration and in immune defense mechanisms.     

Morphology of the lymph nodes in bottlenose dolphin (Tursiops truncatus) and striped dolphin (Stenella coeruleoalba) from the Adriatic Sea

Morphology of the lymph nodes was examined in six bottlenose dolphins (Tursiops truncatus) and three striped dolphins (Stenella coeruleoalba) from the Adriatic Sea. All animals had been found dead in nature. One group of the nodes was taken from the tracheal branching area and was marked as bifurcational lymph node, and the other group was taken from the mesenteric root and was marked as mesenteric lymph node. Microscopic analysis showed that the lymph nodes in both dolphin species were surrounded by a connective tissue capsule comprising smooth muscle cells. The parenchyma of the mesenteric and bifurcational lymph nodes in bottlenose dolphin was divided into the peripherally situated cortex with the lymphatic nodules and diffuse lymphatic tissue, and the centrally situated medulla structured of the medullary cords separated by the medullary sinuses. These lymph nodes structurally correspond to the lymph nodes in the majority of terrestrial mammals. The mesenteric lymph node of striped dolphin also had a peripherally situated cortex and a centrally positioned medulla as the majority of terrestrial mammals. In the bifurcational lymph nodes of striped dolphin, there was a central dense lymphatic tissue with the lymphatic nodules and a peripheral less dense lymphatic tissue structured of the cell cords and sinuses. The bifurcational lymph node in striped dolphin resembled porcine lymph nodes and belonged to the inverse lymph nodes.     

Immunological events in the popliteal lymph node of sheep following injection of liver or killed Corynebacterium ovis into an afferent popliteal lymphatic duct

Efferent popliteal lymph and blood were collected daily for three weeks from sheep following injection of live or killed Corynebacterium ovis organisms into an afferent lymphatic duct. The total lymphocyte output, proportion of blast cells and class specificity of immunoglobulin-containing groups of animals and antibody titres and concentrations of IgG1, IgG2, IMg and IgA were measured in both lymph and blood...     

Comparison of B-mode and Doppler ultrasonographic findings with histologic features of benign and malignant superficial lymph nodes in dogs

OBJECTIVE: To compare and correlate B-mode and color Doppler ultrasonographic characteristics with histopathologic findings of benign and malignant superficial lymph nodes in dogs. STUDY POPULATION: 50 superficial lymph nodes that were normal, abnormally large on physical examination, or represented regional lymph nodes draining an area of suspected primary malignancy in 30 dogs. PROCEDURES: Before excision, lymph nodes were evaluated via B-mode and color Doppler ultrasonography to assess size, echogenicity, presence of a hilus, acoustic transmission, and vascular flow. Formalin-fixed, paraffin-embedded tissue sections of excised lymph nodes were stained with H&E and examined for the presence and extent of necrosis, fibrosis, fat, metastases, and tissue heterogeneity. To assess vascularity, the number and distribution of vessels stained by the Verhoeff van Gieson technique were recorded. RESULTS: In superficial lymph nodes, a varied echogenicity corresponded to tissue heterogeneity. The ultrasonographic detection of a hilus was associated with the presence of fibrous tissue, fat, or both in the hilar region. Acoustic enhancement corresponded to presence of areas of intranodal necrosis. There was significant correlation between both the distribution and the number of vessels detected via ultrasonography and that detected by histopathology. The amount of flow estimated via ultrasonography was typically higher than that estimated via histologic examination. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that histopathologic changes in canine lymph nodes have associated ultrasonographic changes and suggest that lymph node ultrasonography has an important role in the evaluation of lymph nodes in dogs in general and in dogs with neoplastic disease in particular.     

Cytokines in mammary lymph and milk during endotoxin-induced bovine mastitis

Cytokine kinetics were examined in milk and in afferent and efferent lymph of the supramammary lymph node after intramammary infusion of endotoxin from Escherichia coli. Cows were sampled 0, 2 and 4h after infusion (p.i.). Neutrophils appeared in afferent lymph 2h p.i., and in efferent lymph and milk 4h p.i. The milk contained high concentrations of interleukin-8 (IL-8) at 2 and 4h p.i. IL-8 was also found in lymph, but at lower concentrations. The tumor necrosis factor-alpha (TNF-alpha) concentration tended to increase in afferent lymph at 2h p.i., and increased in milk at 4h p.i. The level of IL-1beta increased at 4h p.i. in milk, but was not detected in lymph. Interferon-gamma was not detected in any sample, at any time. The results indicate a primary role for IL-8 in the recruitment of neutrophils into the gland, and suggest that IL-1beta and TNF-alpha are not necessary for IL-8 production and release in response to endotoxin.     

Dendritic cells, implications on function from studies of the afferent lymph veiled cell

Studies of afferent lymph veiled cells (ALVC) show that the full biological function of dendritic cells in peripheral tissue is not explained by a simple model in which immature dendritic cells at the body surface take up antigen, migrate via the afferent lymph ducts, mature and then effectively present antigens to T-cells in the draining lymph node. Furthermore, it is evident from various investigations that the dendritic cells in afferent lymph draining from the body surfaces are not a homogeneous population of cells. They comprise a mixture of cell phenotypes defined by staining with monoclonal antibodies, and the different sub-populations have distinct biological functions and roles in vivo. The molecular basis for differences between the function of afferent lymph dendritic cell subsets is only now being explored and defined but some progress has been made in understanding the role of co-stimulatory molecules. It should be possible to exploit knowledge of the functions of these cells and aid future vaccination strategies in domesticated animals thereby improving animal health and reducing economic loss, and, as a consequence, improving human health. By deliberately targeting functionally distinct subsets of either precursor or mature dendritic cells in vivo, it should become feasible to achieve an appropriately biased immune response.     

Total and Differential Leucocyte Counts and Lymphocyte Subpopulations in Lymph, Afferent and Efferent to the Supramammary Lymph Node, During Endotoxin-Induced Bovine Mastitis

Leucocyte trafficking in afferent and efferent mammary lymph and the supramammary lymph node in cows was examined during 4 h after intramammary infusion of endotoxin from Escherichia coli. Total and differential leucocyte counts were measured in milk, blood and lymph. The proportions of CD4(+), CD8(+), major histocompatibility complex (MHC) class II(+) and IgM(+) lymphocytes were examined in the lymph and lymph node. At post-infusion hour (PIH) 4, the flow rates of both lymph fluids had increased approximately eightfold. Total leucocyte concentration increased in afferent lymph, but decreased in efferent lymph. Neutrophils increased in afferent lymph at PIH 2 and in efferent lymph and milk at PIH 4. The predominant cell type in afferent lymph shifted from lymphocyte to neutrophil while lymphocyte was still at PIH 4 the predominant type in efferent lymph. Among the lymphocytes, B cells were predominant in afferent lymph and lymph node at PIH 4 while T cells, mainly CD4(+) cells, were predominant in efferent lymph both at PIH 0 and PIH 4. The CD4 : CD8 ratio was higher in efferent lymph and the challenged lymph node than in afferent lymph and the control node, respectively. There was a significant difference in proportions of each lymphocyte subpopulation except for IgM(+) cells, between afferent and efferent lymph after infusion. According to the results, there was already during the first hours of the immune response, a non-random trafficking of neutrophils and lymphocyte subpopulations resulting in a changed distribution of cells in afferent and efferent lymph and a difference in lymphocyte reactivity between the two lymph fluids.     

Total and nematode-specific IgE responses in intestinal lymph of genetically resistant and susceptible sheep during infection with Trichostrongylus colubriformis

Total and antigen-specific IgE responses in afferent (AIL) and efferent (EIL) intestinal lymph of sheep with a nematode resistant (R) or susceptible (S) genotype during challenge infection with the intestinal nematode parasite Trichostrongylus colubriformis were examined. Within each sheep line, lambs with a nematode naive or nematode field-primed pre-challenge status were used. Total IgE level in AIL and EIL was dependent on nematode infection and was further influenced by genotype or the immune phenotype (nematode immune mean FEC+/-SDM=77+/-179 or non-immune mean FEC+/-SDM=4016+/-4318) of the animal. During T. colubriformis challenge immune animals had higher levels of total IgE in lymph than non-immune sheep, R line sheep had higher concentrations of total IgE than S line sheep, and field-primed animals had higher total IgE levels than nematode naive animals. Concentrations of total IgE were consistently higher in AIL than EIL or serum and were higher in lymph draining the proximal than the distal jejunum demonstrating that polyclonal IgE in AIL was largely derived from the intestinal mucosa of the anatomical compartment where the nematodes reside. The consistently higher concentration of total IgE in AIL was dependent on phenotype or genotype and in S genotype sheep also on the pre-challenge status. Concentrations of nematode specific IgE were significantly higher in EIL than AIL indicating a preference for the production of IgE reacting with excretory secretory products of the infective T. colubriformis larvae in the regional lymph node.     

Indirect computed tomography lymphangiography with aqueous contrast for evaluation of sentinel lymph nodes in dogs with tumors of the head

Sentinel lymph node evaluation is widely used in human medicine to evaluate the first lymph node(s) to which a tumor drains. Sentinel lymph node biopsy allows avoidance of extensive lymphadenectomies in cases where the sentinel lymph node is negative for metastasis, thereby reducing patient morbidity. It has been shown that regional lymph nodes are not always the sentinel lymph node, thus identification and sampling of sentinel lymph nodes allows for more accurate staging, which is critical for treatment and prognostication in dogs with cancer. The objective of this prospective, pilot study was to determine if indirect computed tomography (CT) lymphangiography with aqueous contrast agent would successfully allow identification of sentinel lymph nodes in dogs with masses on the head. Eighteen dogs underwent CT lymphangiography. The sentinel lymph node was successfully identified within 3 min of contrast injection in 16 dogs (89%). Compression of lymphatic vessels from endotracheal tube ties and/or the patient's own body weight delayed or prevented identification of sentinel lymph nodes in two dogs (11%). Computed tomography lymphangiography with aqueous contrast can be used successfully to rapidly identify sentinel lymph nodes in dogs with masses on the head.     

Distinctive peripheral lymph node hyperplasia of young cats

Peripheral lymph node enlargement was found in 14 of a series of 132 feline lymph node biopsy specimens. Six of nine cats tested had antibodies for feline leukemia virus (FeLV). Half of the cats were clinically normal while the remainder had fever, lethargy, anorexia, and hepatosplenomegaly. There was severe distortion of lymph nodal architecture with variable loss of discernible follicles and sinuses. Histiocytes, lymphocytes, immunoblasts, and plasma cells were present in expanded paracortical regions which encroached on, and occasionally effaced, lymphoid follicles. Postcapillary venules were numerous and prominent throughout the paracortex. The lymphadenopathy was most commonly transient (86% of cases) with subsequent development of lymphoma in one cat. Lymph nodes from seven kittens with experimental FeLV infection were compared with spontaneously enlarged lymph nodes; four of seven had B and T lymphocyte hyperplasia with normal nodal architecture. Three had partial loss of nodal architecture as a result of expanded paracortical regions populated largely by histiocytes and lymphocytes. Proliferation of postcapillary venules was not prominent in nodes from FeLV-infected cats. The cause of spontaneous lymph node hyperplasia of young cats was not determined. However, the similarity of lesions to those of kittens with experimental FeLV infection and the association with FeLV by serologic tests in six of nine cats suggest that this retrovirus may be involved in the pathogenesis of the lesion.     

The sonographic evaluation of lymph nodes

Ultrasound can be used to detect and evaluate both normal and abnormal lymph nodes, as well as aid in biopsy sampling procedures, an important part of staging procedures in cancer patients. Several parameters can be evaluated using ultrasound; lymph node size, margins, echogenicity, echopattern (echotexture), acoustic transmission, presence and distribution of vascular flow, and vascular flow indices. The most diagnostically helpful include the short/long axis ratio of the lymph node, the pattern of distribution of the blood vessels within the lymph node, and to some extent the resistive and pulsatility indices. This review discusses the use of ultrasound for detecting, evaluating, and sampling peripheral, abdominal and thoracic lymph nodes in small animals.