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T and B lymphocyte populations in human normal lymph node, regional tumour lymph node and inflammatory lymph node.

We have determined the T and B lymphocyte subpopulations in normal human lymph nodes. The lymphocyte profile was the same irrespective of the anatomical distribution and was similar to that found in peripheral blood with identical T and B cell values but with a lower Fc and a higher C3-receptor-bearing lymphocyte subpopulation. This pattern showed a marked change in the regional nodes of patients with mammary carcinoma and nodes draining a variety of other solid tumours, with a fall in T and pronounced elevation of B, Fc and C3 cells but with a persistence of C3 predominance. The lymphocyte profile found in tonsils and nodes draining inflammatory foci was a similar but further exaggeration of the tumour node pattern, with reversal of T and B cell ratios. The T and B lymphocyte percentages in the peripheral blood of patients with clinically localized breast cancer are identical to those of the healthy controls. Different Fc/C3 subsets exist in peripheral blood and lymphoid structures and probably represent a differential functional heterogeneity. Proximity of tumour to the draining node modified this profile.

B-Lymphocytes

Killer cell (K) activity in human normal lymph node, regional tumour lymph node and inflammatory lymph node.

Human normal lymph nodes, irrespective of their anatomical site of origin, have a low K cell activity, which may not be detected except with the appropriate target cell and at high lymphocyte to target cell ratios (100:1). This very low killer cell activity is also found in all the homolateral axillary nodes of patients with clinical stage I and II carcinoma of the breast and in the regional draining nodes of a variety of solid tumours, whether small and localized or large and with extensive spread. In all cases proximity to the tumour and obious hyperplastic changes in the nodes have no modifying effect. This pattern of minimal reactivity is similarly found with tonsillar lymphocytes and with nodes draining inflammatory foci. The Fc and C3 receptors on surface membranes are dectected with ease, and pretreatment of lymphocytes by incubation, washing and enzymatic treatment fail to alter their reactivity, thus excluding effector cell inhibition by immune complexes. The killer cell activity of lymphocytes from the blood of breast tumor patients is similar to the activity of lymphocytes from healthy controls.

Binding Sites

The influence of the lymph node on the protein concentration of efferent lymph leaving the node.

1. Experiments have been performed in sheep to determine the contribution of lymph formed within a lymph node to the total protein output in lymph leaving the node. 2. The lymphatic duct leaving the popliteal lymph node was cannulated and the protein and lymphocyte output in efferent lymph determined. The afferent lymph flow to the popliteal node was then diverted and lymph formed only within the lymph node collected from the efferent cannula. It appeared from the results that the popliteal lymph node forms lymph at the rate of approximately 1 ml. per hour and may contribute 30-50% of the protein output observed in efferent lymph. 3. The importance of lymph formation within the lymph node varied between nodes found in different regions of the body. This was due in part to the different protein concentrations in the afferent lymph to the different nodes. 4. A positive correlation was found between the protein and lymphocyte concentrations in efferent lymph from the popliteal lymph node in seven out of eleven sheep and in lymph formed within the popliteal lymph node in two out of three sheep. It is suggested that this relationship may be due to an increased transfer of plasma proteins through the post-capillary venules in the lymph node accompanying the continual traffic of lymphocytes across the wall of these vessels. The results indicated that the protein transfer across the post-capillary venules was not an indiscriminate transfer of plasma per se but a selective transport from the blood plasma compartment based on molecular size.

Animals

[Experimental study of lymph node metastasis in thoracic esophageal carcinoma--regarding lymph node metastasis and changes in lymphatic flow by ultrafine charcoal in rabbit esophageal carcinoma model].

Esophageal carcinoma models were created by transplanting VX2 cells to rabbit esophagus endoscopically. By injecting finely divided activated charcoal into normal rabbit esophagus and tumor sites of esophageal carcinoma model, lymph flow was observed directly. Existence of lymph node metastasis was studied in detailed pathology. In 30 rabbits with upper esophageal carcinoma, lymph node metastasis was seen in 77%. Metastasis to bilateral intrathoracic paratracheal lymph node was seen in 50%, and also concentration of lymphatic flows from tumor site was seen. However, there were no metastasis and no lymph flow to abdominal lymph nodes. While, metastasis to cervical lymph nodes showed around 13%. Esophageal lymphatic flows were also seen reaching the cervical area along the esophagus. In 40 models with mid lower esophageal carcinoma, lymph node metastasis were seen in 88%. Metastasis to right and left thoracic paratracheal lymph nodes was 75% and 53%, respectively, and 25% of metastasis went to cardia lymph nodes. The lymph flows were going up and down around these lymph nodes, and reaching to lymph nodes at upper highest mediastinum or left gastric artery. The metastatic rate to the cervical lymph nodes was about 5%. There were no significant differences in lymphatic metastasis between right and left mediastinum. These findings suggest the necessity of radical dissection for both sides of the mediastinum.

Animals

Pelvic lymph node metastases in cervical cancer: comparison of lymphography, inspection, radiography, and histologic examination of lymph nodes.

Lymphangiography is commonly performed in the pretreatment evaluation of patients with cervical carcinoma, but its value is controversial. The purpose of this report is to determine the reliability of lymphography in the indication of metastatic pelvic lymph nodes by comparing data from preoperative lymphangiography, inspection of lymph nodes during laparotomy, radiography of surgically removed lymph nodes, and postoperative histologic report. Twenty-one patients (mean age 51.1 years, SD 14.5) with cervical cancer FIGO stages I b to II b were enrolled in this study. They all underwent Wertheim's radical hysterectomy with pelvic lymphadenectomy. With reference to histologic report this series included 8 squamous carcinomas (38%), 10 adenocarcinomas (48%) and 3 sarcomas (14%). Seven patients (33%) had a positive preoperative lymphangiography, in 9 patients (43%) lymph nodes were considered positive at the inspection during laparotomy, postoperative radiography of the lymph nodes was considered probably positive in 7 patients (33%) and positive in 2 patients (10%), histologic report was positive for lymphonodal metastases in 4 patients (19%). A total of 335 lymph nodes were studied, and with reference to the evaluated methods (lymphangiography, inspection, radiography, histology), 1 positive method was found in 40 lymph nodes (12%), 2 positive methods in 6 lymph nodes (2%), 3 or 4 positive methods in none of the lymph, nodes, and 4 negative methods in 289 lymph nodes (86%). Histologic report was positive in 4 lymph nodes (1%). Lymphangiography in the pretreatment evaluation in cases of cervical carcinoma is not reliable in indicating possible metastatic lymph nodes. It remains a useful exam fixation to be routinely performed.

Adult

Lymph node involvement by direct extension in adenoid cystic carcinoma. Absence of classic embolic lymph node metastasis.

Thirty-four cases of adenoid cyctic carcinoma seen at the University of Virginia Hospital from 1946 to 1974 were reviewed, with special emphasis on lymph node involvement by tumor. Lymph node involvement was found in three cases of primary tumors of the submaxillary gland, and all of the affected lymph nodes were in the immediate vicinity of the primary tumor. Two lymph nodes were involved in two of the cases, and one node was involved in the third case. In all of these lymph nodes, adenoid cystic carcinoma was present in the soft tissue surrounding the node, and the tumor extended into the node. No metastatic tumors were observed in 46 lymph nodes removed incidentally at the time of local excision of the primary tumors in 10 additional cases or in 212 lymph nodes examined after unilateral radical neck dissections in six other cases. Five autopsies in this series showed no lymph node metastases. In this series of cases adenoid cystic carcinoma only invades lymph nodes in the immediate vicinity of the primary tumor. When lymph node involvement does occur, it does not result from embolic lymph node metastasis; rather, a direct invasion of the lymph node from tumor in the perinodal soft tissue occurs. Obviously, this small study does not completely exclude the possibility of embolic metastasis; however, if it does occur, it must be extremely rare.

Carcinoma, Adenoid Cystic

Lymph follicles and germinal centers in popliteal lymph nodes and other lymphoid tissues of germ-free and conventional rats.

A study was conducted to evaluate the influence of natural exogenous antigen stimulation on the development of lymph follicles in the peripheral lymphoid organs of the rat. The number of lymph follicles and germinal centers per popliteal node as well as the morphological features of popliteal nodes, mesenteric nodes and Peyer's patches were compared between 8-week-old male Sprague-Dawley rats reared in germ-free, specific pathogen-free (SPF) and conventional environments. In mesenteric nodes and Peyer's patches from conventional and SPF rats, almost every follicle contained a germinal center. In the mesenteric nodes from germ-free rats, only 9 of 195 lymph follicles examined contained a germinal center, but interestingly, in Peyer's patches all the lymph follicles examined showed a fairly well developed germinal center. The popliteal nodes from germ-free rats had no germinal centers and each node contained about 80 lymph follicles. In six conventional rats and two of five SPF rats used, the number of lymph follicles per popliteal node usually ranged from 100 to 130, and some lymph follicles contained a germinal center. However, in the other three SPF rats the popliteal node on both sides showed no germinal center, and each contained almost the same number of lymph follicles as the popliteal nodes of other SPF and conventional rats. The present results are consistent with the view that even in the absence of exogenous antigen stimulation, a regional lymph node of the germ-free rat develops a substantial number of primary lymph follicles by way of a non-immunological cellular activity, and that natural exogenous antigens may influence the development of lymph follicles by stimulating the cellular activity rather than evoking the humoral immune responses.

Animals

In vivo influences of phorbol ester and calcium ionophore on lymphocyte traffic, lymph flow and efferent lymph levels of thromboxane B2 in sheep.

In vitro systems have provided increasing evidence of significant lymphocyte transmembrane signalling by plasma membrane receptors which utilize antigen and other ligand activation of the inositol phosphate dual second messenger system of intracellular signalling. Elevation of intracellular Ca2+ and activation of protein kinase C are important products of these signals and appear to provide a complete set of mitogenic signals for both T and B cells. Calcium inophore and phorbol ester have been found to mimic these events in vitro and are here employed in vitro to study their effects on lymphocyte traffic and efferent lymph flow through primary peripheral lymph nodes of sheep and on the output into efferent lymph of the arachidonic acid metabolite, thromboxane B2. Calcium ionophore and phorbol ester were given alone or in combination to popliteal lymph nodes of sheep by drainage area injection or by acute infusion into cannulated afferent lympatics of study popliteal lymph nodes whose efferent lymphatic was chronically cannulated for study. The findings resembled those of drainage area immunization with an early increase in efferent lymph flow and prompt and marked depressions in the output into efferent lymph of both small recirculating and blast lymphocytes ('shutdown', 'recruitment'), followed by a marked increase in the output into efferent lymph of both small recirculating and blast lymphocytes. The greatest elevation in both small recirculating and blast lymphocyte outputs was at 24 and 48 h following phorbol ester and calcium ionophore administration. Acute phorbol ester and calcium ionophore administration was associated with a prompt and marked elevation in efferent lymph levels of thromboxane B2 which were of short duration. The findings observed here with lymph node drainage area infusion/injection of both phorbol ester and calcium ionophore are quite similar to those encountered in this sheep lymphocyte traffic model following popliteal lymph node drainage area immunization with killed Salmonella muenchen antigen.

Animals

Topography of the major superficial lymph nodes and their efferent lymph pathways in the koala (Phascolarctos cinereus).

The koala has an inguinoaxillary lymph trunk on either side of the ventral midline, and this carries efferent lymph from the superficial inguinal lymph node directly to the deep axillary lymph node. The superficial lymph nodes are large and soft compared with those of the domestic species, and each lymph centre usually contains only one or two large lymph nodes. Koalas have a rostral mandibular lymph node which has not been described in other species, but lack popliteal and subiliac lymph nodes. The superficial lymph nodes which are readily palpable in the live koala are the facial, rostral mandibular, mandibular, superficial axillary and superficial inguinal. All superficial lymph pathways terminate at the confluence of the common jugular and subclavian veins.

Animals

Draining lymph node cell activation in guinea pigs: comparisons with the murine local lymph node assay.

The local lymph node assay in the mouse is a novel predictive test for the identification of contact sensitizing chemicals. The purpose of the studies described was to determine whether a similar local lymph node assay could be performed successfully in guinea pigs; currently the species of choice for assessment of sensitizing potential for regulatory purposes. Ten sensitizing chemicals (oxazolone, picryl chloride, 2,4-dinitrofluorobenzene, benzocaine, cinnamic aldehyde, 2,4,-dinitrothiocyanobenzene, p-nitrosodimethylaniline, formaldehyde, p-phenylenediamine and cyanuric chloride) and equal concentrations of sodium lauryl sulphate were examined in a guinea pig local lymph node assay. Animals received three consecutive daily applications of various concentrations of the test chemical on the dorsum of both ears. Control animals were untreated. Five days following the initiation of exposure, draining auricular lymph nodes were excised and weighed. Suspensions of lymph node cells (LNC) were prepared and cultured for 24 or 48 h and proliferation measured by incorporation of [3H]thymidine. Exposure to at least one concentration of all sensitizing chemicals, other than benzocaine, induced proliferation by draining LNC. Responses were higher at 24 h rather than 48 h. Evidence is presented that guinea pig LNC proliferation may be enhanced or maintained by addition to culture of an exogenous source of the T cell growth factor interleukin 2 (IL-2). Draining lymph node weight was increased following exposure to some sensitizing chemicals but, compared with LNC proliferation, provided a less sensitive correlate of lymph node activation. Exposure to sodium lauryl sulphate failed to induce changes in either lymph node weight of LNC proliferation. Data are compared with three-day murine local lymph node assays performed concurrently. The available information indicates that the local lymph node assay may be performed in guinea pigs.

Allergens

Proportions of dog lung lymph in the thoracic and right lymph ducts.

We studied the external lymphatic drainage of the lung in anesthetized dogs, by simultaneously measuring lymph flows from the thoracic duct (TD) and right lymph duct (RLD) during base line and during pulmonary edema. We measured lymph flow for a 2-h base-line period, for 2 h after tying off the thoracic duct above the diaphragm to eliminate nonthoracic lymph contributions, and after giving alloxan. Following alloxan, all dogs developed moderately severe pulmonary edema. In eight dogs the average TD flows were 24.0, 0.9, and 8.2 ml/h and RLD flows were 1.1, 1.3, and 8.4 ml/h, respectively. If we assume that all increases in lymph flow after giving alloxan are due to increased lung lymph flow, then, on the average, 50% of lung lymph drains into the TD and 50% into the RLD. However, among the eight dogs, four had significant increases in TD flow after alloxan (8.9-24.6 ml/h), and four did not. RLD flow increased in all dogs following alloxan. It appears the fraction of lung lymph draining into the TD and RLD can vary greatly amone individual dogs but, on the average, the TD and RLD receive about equal fractions of the pulmonary lymph. In shamoperated control animals TD and RLD lymph flows did not change over a 5-h period.

Alloxan

The effect of anaesthesia and surgery on lymph flow, protein and leucocyte concentration in lymph of the sheep.

Anaesthesia and the trauma of surgery, associated with the cannulation of lymphatic ducts in various regions of the body of the sheep, had a profound effect on lymph flow, protein concentration and leucocyte concentration of lymph. In general lymph flow was depressed and the protein concentration elevated in lymph collected at the time of cannulation, or within the first 24 hours of recovery from surgery. The changes in protein concentration in lymph draining the peripheral regions of the body appeared to be due to surgical interference in the region of drainage. The greatest changes in lymph flow were observed in lymph draining peripheral regions (skin, tendon, muscular areas) while lymph draining soft tissues in central regions (kidney, liver) was less affected by the anaesthesia and surgical stress. A neutrophilia was observed in venous blood collected under anaesthesia while the overall numbers of lymphocytes in three sources of efferent lymph were depressed. It is suggested that corticosteroid hormones may play a role in the changes in leucocyte migration observed during anaesthesia and surgical stress. Changes observed in the cellular content of afferent lymph appeared to be due to a low grade inflammation associated with surgical interference in the region of lymphatic drainage.

Albumins

Pathways of lymph flow through intestinal lymph nodes in the horse.

In the horse, several thousand lymph nodes receive lymph from the intestine, part of which is very large and contains microorganisms that enable the animal to utilize refractory dietary constituents such as cellulose. The aim of this study was to describe the pathways by which lymph is delivered into, traverses, and is drained from these lymph nodes. These pathways were studied with either Microfil or methacrylate casting materials and with light and electron microscopy. The afferent lymphatic vessel delivering lymph into one of the nodes divides over the capsular surface and within trabeculae into terminal branches, and these are continuous with the subcapsular and trabecular sinuses through rounded holes up to 30 microns across. Lymph is conveyed from the subcapsular and trabecular sinuses through the cortex by four types of sinuses: trabecular sinuses, cortical tubular sinuses, tubule-like sinuses with a network of stellate cell processes, and sinuses between cortical cords. It is conveyed through the medulla by sinuses both within and between medullary cords. Lymph is drained from these sinuses by initial efferent lymphatics of three types: those between medullary cords, those within the subcapsular sinus overlying medullary or cortical cords, and those within trabeculae. All three types are continuous with surrounding sinuses through holes 5-30 microns across. These three alternative routes for lymph drainage may ensure adequate lymph flow during different intranodal conditions that may exist when the node is responding to microorganisms or other foreign materials.

Animals

[Study of para-aortic lymph node metastasis for advanced gastric cancer on consecutive lymph node slices].

Para-aortic lymph nodes (No. 16) from 21 patients with advanced gastric cancer who underwent extensive lymph node dissection, were sliced consecutively at intervals of 50 mu to study minute metastasis of No. 16 lymph nodes. Twenty-six of 30 metastatic lymph nodes had minute involvement, in which carcinoma cells were detected in marginal sinus of lymph node, microscopically. The rate of the metastases to No. 16 lymph nodes was 19% among N0-3 cases by conventional method (not consecutive slices). On the contrary, the percentage of metastasis to No. 16 lymph nodes by consecutive slices was 33% among N0-3 cases. Therefore, No. 16 nodes are involved in metastasis at high rate at operation and the extensive lymph node dissection including No. 16 nodes (R4) will be recommended to prevent lymph node recurrence caused by such a minute metastasis.

Aorta

[Cytokinetics of lymph nodes in lymph nodes in lymphatic system diseases (author's transl)].

Untreated malignant lymphatic system diseases are characterized by a preponderance of cell new formation (proliferation) against the destruction of lymphatic cells. If the lymph nodes are enlarged during these diseases, then cell new formation occurs largely or mostly in these lymph nodes. The proliferating cells of the lymph node are bigger than small lyphocytes and have, in general, a mean diameter of the nucleus of 10 mu and more. In normal lymph nodes they belong morphologically to the big lymphocytes, immunoblasts and plasmoblasts. In pathological lymph nodes they have to be looked for among the bigger cells of the disease-specific cell population. Whereas in healthy lymph nodes and in chronic lymphatic leukemia only about 1% of lymph node cells was found to proliferate, they amount on the average to 5% in lymphomas of lymphogranulomatosis and mostly to 30--50% in the lympho-reticulosarcoma (lymphoblast and immunoblast sarcoma, corresponding to large-cell, poorly differentiated lymphomas). The proliferating cells often appear as foci in the lymphomas. The generation times of the proliferating cells both in normal and pathological lymph nodes are about 24 hrs. or slightly longer. In lymphatic proliferation, apart from plasma cells big and smallymphocytes are produced in the normal lymph node; in CLL, big and small lymphocytes, in lymphogranulomatosis, big and small lymphocytes and Hodgkin-cells, and in poorly differentiated lymphomas, the corresponding lymphoma cells are produced. The clinicist is at the beginning of drawing conclusions from prevalent kinetic disturbances.

Cell Division

High concentration of bilirubin in post-nodal lymph associated with red blood cell catabolism in lymph nodes of the sheep.

Qualitative and quantitative analysis of post-nodal lymph of the sheep has shown that the distinct yellow colour of this fluid pool is due to the presence of relatively large amounts of bilirubin. In efferent lymph from thepopliteal, prefemoral, prescapular, renal and intestinal lymph nodes total bilirubin concentrations were 3-8 times higher than the corresponding concentrations in blood plasma. In contrast the total bilirubin concentrations in afferent lymph from the lower leg and kidney were less than the corresponding concentrations in blood plasma. Histological examination of several popliteal and mesenteric lymph nodes revealed the presence of free iron and bilirubin in the cytoplasm of cells located near the lymphatic sinuses of the node. In addition, the concentration of bilirubin in efferent lymph from the popliteal node was observed to increase following an induced rise in the number of red blood cells reaching the node by way of the afferent lymphatic duct. These latter observations suggest that the bilirubin in post-nodal lymph is associated with the catabolism of extravascular red cells by reticulo-endothelial cells within the lymph nodes.

Albumins

Studies on lymph humoral factor. Evidence for a lymphocytopoietic factor in rat thoracic duct lymph.

Normal rats were injected with the partially purified material extracted from the thoracic duct lymph which was collected from normal syngeneic rats. The cellular changes in the peripheral lymphoid tissues and thymus were examined cytologically and histologically. The intravenous injection of the lymph extract into normal syngeneic rats resulted in increase in weight of lymphoid tissues. Histologically, a massive proliferation of large pyroninophilic lymphoid cells and an increase in mitotic index was detected predominantly in the thymus-dependent areas and thymic cortex. In spleens from rats injected with lymph extract, the marginal zone bridging channel was shown as one route for the translocation or mobilization of newly borne lymphoid cells to the venous circulation. Similar lymphocytopoietic activity, but to a lesser extent than lymph extract, was also detected in lymph plasma, serum and serum extract. The lymph extract was shown to be non-immunogenic in syngeneic rats. It is suggested that the effects of the lymph extract on lymphoid cell proliferation are due to the presence of a lymphocytopoietic factor in body fluid, particularly in the lymph.

Animals