Skin reactive factor and lymph node permeability factor

We have compared the properties of lymph node extracts from rat tissue with the properties of the supernatant obtained from the lectin-transformed rabbit peritoneal exudate lymphocytes in culture. Both extracts possess large amounts of lysosomal hydrolases, including particularly a cathepsin D-like protease. Both extracts are capable of increasing cutaneous permeability in the rat and causing a significant amount of cellular infiltration into the site of injection in the skin of these animals. This permeability-increasing activity and the cellular infiltration response to injection of these materials is completely inhibited by pepstatin and not by a variety of other inhibitors. Both extracts' permeability-increasing activity has a molecular weight range between 50,000 and 100,000 daltons and an isoelectric point of approximately 4.2. The molecular weight, the isoelectric point, and the inhibition by pepstatin are characteristic of cathepsin D from liver or spleen or granulocytes. Like cathepsin D, the permeability factor from LNPF and SRF extracts will release acid kinins from substrates which have been shown by Greenbaum and Houck to exist in the ground substance of rat skin. Finally, the breakdown products of the hydrolytic action of cathepsin D-like enzymes found in both lymph node extracts and supernatant from transformed lymphocytes are chemotactic for white blood cells. Thus, the two properties of cathepsin D described above, namely (a) the release of acid kinins which are capable of increasing the permeability of the microcirculation, and (b) the generation of chemotactic breakdown products via proteolysis could explain the two primary biological effects of both lymph node permeability factor and skin reactive factor.     

Nature of immune lymph node factors stimulating humoral and cellular immune responses

Institute of Immunology, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. D. Ado.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 107, No. 4, pp. 460–462, April, 1989.     

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.     

Studies on lymph humoral factor. Biological characteristics of a lymphocytopoietic factor in rat thoracic duct lymph.

The biological characteristics of a lymphocytopoietic factor obtained from rat thoracic duct lymph (Yamashita, Fukumoto & Miyamoto, 1976) were further investigated. The lymph extract from normal rats failed to stimulate both large pyroninophilic cell-proliferation and mitotic response in the spleen and lymph node of the thymectomized, irradiated and marrow reconstituted rat (B rat). This suggests that target or responsive cells for the factor are not marrow-derived (B) cells, but thymus-derived (T) cells. On the other hand, the lymph extract from the lymphopenic lymph-drained B rats showed similar high lymphopoietic activity to those of normal rats, indicating that the existence of a thymus is not essential for the production or secretion of the factor. The fact that lymphocytotic rats produced by syngeneic lymphocyte transfusion are most sensitive to the lymph extract suggests that liver endogenous level of this factor and the responsiveness of target cells are regulated by the number of circulating T lymphocytes.     

The effects of field stimulation on bovine mesenteric lymph node contractility

In these studies isometric tension was recorded from isolated strips of bovine mesenteric lymph node capsule. The possible influence of intrinsic nerves on their contractile activity was examined using field stimulation and the pharmacology of the response was investigated. Over 75% of the tissues studied demonstrated regular spontaneous activity at a frequency of 3.9±0.2 contractions/5 min under control conditions. Field stimulation at 2, 8 and 32 Hz (pulse width = 0.3 ms, nominal voltage 60 V) produced frequency-dependent increases in the rate of this spontaneous activity associated with a raised baseline tension. These responses were not diminished on repeat stimulation under control conditions. The contraction frequency response to stimulation at 8 Hz was blocked completely in the presence of 1 M tetrodotoxin, while the increase in baseline tension was reduced by over 80%, suggesting that field stimulation was activating intrinsic nerves. The responses to stimulation (8 Hz) were also greatly reduced by 10 M phentolamine but were unaffected by 10 M propranolol. These experiments suggest that intrinsic nerves can modulate the contractile activity of lymph node capsule acting, at least in part, through excitatory -adrenoceptors. The possible functional significance of this is discussed.Part of this work has been communicated to the Physiological Society of Great Britain and Ireland (Gallagher and McGeown 1990)     

Purification of serum proteins from lymph node cell extracts with permeability activity characteristic of the lymph node permeability factor

The biologically active aqueous extract of rat lymph node cells, previously shown to contain the lymph node permeability factor LNPF, was fractionated by gel filtration and ion exchange chromatography. The fractions obtained were characterized by immunoelectrophoretic analysis and found to contain a wide range of serum proteins. The ability of the fractions to increase vascular permeability to plasma protein was measured. Activity was found to be distributed among several serum proteins and was mainly associated with albumin, though minor quantities of γ-globulin were also found to have high specific activity. Comparative analysis of γ-globulin and albumin prepared from lymph node cell extract and serum, respectively, suggested that the active factor was most likely to be released from the lymph node cells to form complexes with these proteins.     

Regulatory role of humoral factors released by macrophages in rat lymphocyte reactivity in vitro. II. Influence of humoral factors released by normal peritoneal macrophages on spontaneous lymphocyte stimulation

Supernatants harvested from peritoneal macrophage cultures, derived from normal rats, exhibit in vitro two antagonistic activities: stimulating and suppressing spontaneous stimulation of syngeneic lymphocytes. Stimulation of DNA synthesis by macrophage supernatants occurs in lymph node lymphocytes, which show low values of spontaneous stimulation. Suppression of DNA synthesis by macrophage supernatants is revealed in thymocytes and cortisone-resistant thymocytes, which demonstrate high values of spontaneous stimulation.     

A low molecular weight ES-20 protein released in vivo and in vitro with diagnostic potential in lymph node tuberculosis

PURPOSE: To determine role of antigens released in vivo and in vitro in immunodiagnosis of tuberculosis (TB). METHODS: In vivo released circulating tuberculosis antigen (CTA) was obtained from TB sera by ammonium sulphate precipitation and in vitro released excretory-secretory (ES) antigens from Mycobacterium tuberculosis culture filtrate. CTA and ES antigens were fractionated by SDS-PAGE and electro-eluted gel fractions were analysed for antigen by ELISA. RESULTS: Low molecular weight proteins CTA-9 and ES-9 showed high titre of antigen activity. To explore the diagnostic potential of low molecular weight ES antigen, M. tuberculosis ES antigen was further fractionated by gel filtration chromatography followed by purification on anion exchange column using fast protein liquid chromatography and a highly seroreactive ESG-5D (ES-20) antigen was obtained. Competitive inhibition showed that CTA-9 and ES-9 antigens inhibit the binding of ES-20 antigen to its antibody. Seroanalysis showed sensitivity of 83 and 80% for ES-20 antigen and antibody detection, respectively, in pulmonary TB and 90% in lymph node TB. CONCLUSIONS: Seroreactivity studies using M. tuberculosis ES-20 antigen showed usefulness in detection of TB; in particular, lymph node TB.     

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.     

The effect of transfer factor on lymph node morphology in murine toxoplasmosis

Mice were infected intraperitoneally with a low virulence strain of Toxoplasma gondii (TO) and transfer factor (TF) was prepared from the spleens of infected (TFT) and uninfected control mice (TFC). Three experimental groups of 12 mice were given either saline, TFC or TFT, by intraperitoneal injection. After 24 h half of each group of these animals were infected by intraperitoneal injection of TO cysts. In three separate experiments animals were killed at 11, 28 and 35 days and the flank and axillary nodes removed for histological examination. There was generalized lymph node enlargement with cortical and paracortical expansion. In most animals there was diffuse infiltration of the nodes by clusters of histiocytes. Administration of TFC alone led to a mild increase in node size at 11 and 28 days. Administration of TFT alone had a moderate stimulatory effect on the mouse lymph nodes with a significant increase in size at 11 days due predominantly to expansion of the paracortex. Administration of TFT and TFC followed by inoculation of TO led to an increased and more consistent histiocyte response and an increased number of paracortical T blasts compared with animals given TO alone. TFT and TFC had no demonstrable protective effect in experimental murine toxoplasmosis as assessed by quantitation of toxoplasma brain cysts. The effect of transfer factor was not antigen specific in this system.     

The lymph node neutrophil

Secondary lymphoid organs provide a specialized microenvironment tailored to foster communication between cells of the innate and adaptive immune systems. These interactions allow immune cells to coordinate multilayered defense against pathogens. Until recently dendritic cells and macrophages were thought to comprise the main innate immune cell subsets responsible for delivering signals that drive the adaptive immune response, while the function of neutrophils was largely confined to the innate immune system. However, the discovery of neutrophils in lymph nodes has raised the question of whether neutrophils might play a more extensive role not only in innate immunity per se, but also in coordinating the interactions between innate and adaptive immune responses. In this review we discuss the mechanisms and consequences of neutrophil recruitment to lymph nodes and how this recruitment influences subsequent immune responses both in situ and at distant sites.     

Deflating the lymph node

Sensing pathogens is an essential first step in the initiation of a host response to infection. In this issue of Immunity, Kane et?al. (2011) used mouse models to demonstrate that Toll-like receptor 7 is required for the generation of an antibody response to infection by retroviruses.     

The artificial lymph node

Adsorption of lymphocytes on the sorbents in correction of inflammatory lesions has significant clinical perspectives as the cells and active biological substances they secrete function actively in inflamed tissues acting in the site of its application: every cell adsorbs and inactivates foreign substances, every cytokin affects certain target cells. The products of these cells functioning and dissociation as well as the products of immune system reaction to them remain on the sorbent and are eliminated together with granules during bandaging or by the end of the treatment thus do not entering the organism. Lymphocytic monolayer on the sorbent does not prevent its specific action in pathologic nidus. In this case sorbent does not only drain the tissue duplicating certain primitive non specific functions of regional lymph node (barrier, filtrating, transport, draining and protective functions) but also performs certain highly specialized functions of immunocompetent organs--selective adsorption and inactivation of antigenic substances. It is reasonable to use sorbents for adsorbing lymphocytes in kapron containers which exclude the preparation leak into wounds and cavities and act as a filter preventing fast inactivation of the preparation by large fragments of tissue detritus.     

A case of heterotopic pancreas in lymph node

Summary A case of pancreatic heterotopia in a lymph node is described. Small pieces of pancreatic tissue were found incidentally in four lymph nodes located around the common hepatic artery in a 65-year-old man. Both exo- and endocrine elements of the pancreas, together with ductal structures were found to constitute the heterotopic tissue. No authentic case of heterotopic pancreas in lymph node has been previously reported in the literature.     

A local subset of mesenchymal cells expressing the transcription factor Osr1 orchestrates lymph node initiation

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