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Functional anatomy of lymph nodes. II. Peripheral lymph-borne mononuclear cells.

In the rabbit a number of large mononuclear cells with ruffled surface membranes travel from the skin and superficial tissues of the leg, via the lymphatics, to the popliteal lymph node: they constitute 40-50% of the total cell population in the afferent lymph. About 10% of these cells are actively phagocytic when tested in vitro and about 3% are found to contain Langerhans granules. After isotopic labelling the majority of lymph-borne mononuclear cells can be detected within the regional node for at least 24 hours; most being located in the paracortex and a few in the interfollicular cortex. It is proposed that these cells, including those containing Langerhans granules, belong to the "mononuclear phagocyte system." Possible functions of these lymph-borne cells are discussed with particular reference to antigen transport.

Adenosine

Eosinophil chemotaxis of supernatants from cultured Hodgkin's lymph node cells.

Cultured lymph node cell supernatants from five out of six cases of Hodgkin's disease were relatively more chemotactic for peripheral blood eosinophils than neutrophils. In contrast, supernatants from two cases of lymphocytic lymphoma and four nodes showing reactive hyperplasia were chemotactic for neutrophils but had little eosinophil chemotactic activity. In most instances the dgree of eosinophil infiltration observed histologically in the Hodgkin's lymph node correlated with elaboration of eosinophil chemotactic activity from the cultured cells. Following gel-filtration of three of the Hodgkin's lymphnode supernatants, four peaks of eosinophil chemotactic activity were demonstrated in each case. One of these corresponded in molecular size to th previously described eosinophil chemotactic factor of anaphylaxis (ECF-A). It is suggested that the eosinophil chemotactic activity of cultured lymph node cell supernatants may be on value in the diagnosis and classification of Hodgkin's disease.

Anaphylaxis

Development of IgE-forming cells in vitro from rat mesenteric lymph node cells.

Mesenteric lymph node cells from normal rats and rats infected with Nippostrongylus brasiliensis (Nb) were cultured with pokeweed mitogen (PWM) or Nb antigen, and the development of IgM-, IgG2a-, or IgE-containing cells was assessed by immunofluorescence. Normal lymph node cells stimulated with PWM developed into both IgM- and IgE-containing cells, whereas similar stimulation of cells from Nb-infected rats resulted in the development of IgM-, IgG2a-, and IgE-containing cells. The in vitro plasma cell response to PWM was dependent on the presence of T lymphocytes. Lymph node cells from Nb-infected rats responsed to Nb antigen and developed into plasma cells of IgM, IgG, and IgE classes. The response was antigen specific and required antigen-primed T cells. Depletion of IgE-bearing cells or IgM-bearing cells before stimulation with either PWM or Nb antigen diminished the level of IgE forming cell development, suggesting that IgE-IgM double bearing cells are precursors of IgE-forming cells. The distribution of the three isotypes among the If-forming cells that developed in response to PWM was influenced by the source of both B and T cells. When B cells from Nb-infected rats were employed as a source of precursors, T cells from infected animals were more effective than normal T cells for the development of IgE-forming cells, whereas the latter cells were more effective for the development of IgG2a-forming cells than T cells from infected animals.

Animals

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

Doppler ultrasound examination of pathologically enlarged lymph nodes.

Pathologically enlarged lymph nodes have been examined with a commercially available 10 MHz continuous-wave Doppler flowmeter. Many enlarged lymph nodes gave rise to significant Doppler-shift signals indicating increased blood flow. The signals have been spectrum analysed and the large diastolic flow components suggest that there is considerable arterio-venous shunting within lymph glands involved in leukemia, lymphoma and carcinoma. It also seems that the signals tend to diminish in response to treatment. The Doppler signals have been used in an imaging system to produce a vascular map of the region of the enlarged gland. It is suggested that these findings might be applicable to the detection of neoplastic tissues in less accessible sites.

Doppler Effect

Mucocutaneous lymph node syndrome.

Mucocutaneous lymph node syndrome represents a series of clinical findings that has been observed primarily in Japanese children. The disease now appears to be migrating to this country. It involves the cervical lymph nodes, the skin, and mucus membranes. Although the course is usually benign and self-limiting, a number of deaths have resulted from coronary artery disease.

Acute Disease

Electrophoretic fractionation of guniea pig lymphocytes: evidence for different subsets of T and B cells in spleen and lymph node.

Guinea pig lymph node and blood lymphocytes have been physically fractionated in preparative cell electrophoresis into two functionally viable populations, the high mobility cell population (HMC) and the low mobility cell population (LMC). By using cell surface markers and functional tests known to be specific for T and B lymphocytes, respectively, it is shown that the T lymphocytes localize in the HMC population and the B lymphocytes in the LMC population. The spleen lymphocytes do not separate into the two populations. They move into one single broad peak containing both T and B cells. This finding indicates the presence of electrokinetically different subsets of T and B lymphocytes in the spleen on one hand and in the lymph node and blood on the other hand.

Animals

T and B cell populations in blood and lymph node in lymphoproliferative disease.

Lymph node and peripheral blood lymphocytes were studied simultaneously for surface markers of T and B cells in 22 patients with lymphoproliferative diseases and 8 patients with non-neoplastic lymphadenopathy. This resulted in the classification of the malignancy from involved lymph nodes into 4 groups. Six patients had B cell lymphomata with normal or strong immunofluorescent staining for surface membrane immunoglobulin; 8 patients had B cell chronic lymphocytic leukaemia with pale staining for surface membrane immunoglobulin; 5 patients had T cell lymphomata and 3 patients were not definitely classifiable. In 6 out of 8 patients with B cell CLL, histopathology of lymph nodes showed infiltration with well differentiated lymphocytes and in all T cell lymphomata, the infiltrating cells were poorly differentiated. By the use of these markers, malignant lymphocytes were identified in the circulation in only 3 out of 6 patients with B cell lymphoma, in all patients with B cell CLL but in none of those with T cell lymphoma or unclassifiable lymphoma. Therefore a more conclusive characterization of the malignant lymphocyte in lymphoproliferative diseases must include an examination of involved lymph nodes.

B-Lymphocytes

The appearance of non-specific antibody-forming cells in the efferent lymph draining antigen-stimulated single lymph nodes.

Immunization of single lymph nodes with various antigens led to the appearance of cells in the efferent lymph that secreted antibody specific for the antigen which induced their formation and for a number of unrelated, non-crossreacting antigens. Immunization of single lymph nodes with mitogens led to the appearance of cells secreting antibodies specific for an even greater number of antigens, including one (TNP) that in all probability is not present in the animals' natural environment. When the node was primed with one antigen, a subsequent challenge with an unrelated antigen 12 weeks later led to the appearance of greater numbers of cells containing and secreting antibody against the previously experienced antigen, than was the case in unprimed lymph nodes. These findings indicate that the immune response to antigen provokes the maturation of lymphocytes of specificities unrelated to that of the injected immunogen. Such a mechanism may be important in maintaining immunological memory. Mitogens may directly activate lymphocytes into maturation and expression as antibody-secreting cells, whereas antigens appear to act indirectly.

Animals

Lymphocyte locomotion. II. The lymphocyte traffic over the post-capillary venules analysed by phase contrast microscopy of thin sections of rat lymph nodes.

Thin sections of lymph nodes from 14 rats were examined by phase contrast microscopy as regards direction of lymphoctes with amoeboid movement configuration (AMC) relative to the basement membrane of post-capillary high-endothelium venules (HE-cenules). Out of 118 lymphocytes with AMC, 82 appeared to be on theyr way into the venule from the lymph node parenchyma. This observation suggests that the lymphocyte traffic over the HE-venules is bi-directional, with the main migratory stream of lymphocytes from the lymph node parenchyma into the post-capillary venules.

Animals

Lymph-node biopsy during simple mastectomy.

The distribution of pectoral (external mammary) nodes identified during the operation and removed with the axillary tail of the breast was studied in 45 patients treated by simple (total) mastectomy. Up to 13 nodes may lie within the axillary tail, and these are continuous with the pectoral nodes. Lymph-nodes were identified in 90 percent of patients treated by simple (total) mastectomy without dissection of the axilla.

Axilla

Surface immunoglobulin positive lymphocytes in human breast cancer tissue and homolateral axillary lymph nodes.

Surface immunoglobulins were determined on human lymph node lymphocytes by the use of immunofluorescence technique in 59 breast cancer patients undergoing radical mastectomy. In 10 of these cases, lymphocyte surface immunoglobulins were also studied on lymphocytes infiltrating the primary cancer mass. The most outstanding finding was a difference between the IgM lymphocyte populations in the lymph nodes of patients with and without lymph node metastases. When cancer tissue was present in one or more lymph nodes, the tumor-free as well as the tumor-positive nodes showed a higher percentage of IgM positive lymphocytes than did lymph nodes from patients without nodal metastases. The greatest difference was found when IgM lymphocytes from tumor-bearing lymph nodes were compared with those from the lymph nodes of patients without nodal metastases (p is less than .005). The lymphocyte populations infiltrating 5 of the 10 primary cancer masses studied showed no surface immunoglobulins; in the remainder, both IgG and IgM positive lymphocytes were found but in variable proportions. While the findings are not definitive, this is the first study dealing with the quantitation of immunoglobulin specific lymphocytes in the lymph nodes and tumor tissue of patients with breast cancer.

Breast Neoplasms