scholarly journals RECENT ADVANCEMENT ON ISOLATION, ACTIVATION AND CRYOPRESERVATION OF LYMPH NODE CELLS IN MICE.

Author(s):  
Seema Gupta ◽  
Madhuri Grover ◽  
Vasundhara Saxena

ABSTRACT                                                                                                                                      Lymph nodes are found within the body has B, T and other immune cells and help to filter and trap foreign particles. Like any other primary culture lymph node culture would retain many of differentiated characteristics of cells in vivo thus they have potential for acting as alternative method to mammalian model. For setting up primary lymph node culture in mice different types of lymph nodes were collected from mice followed with isolation, activation and cryopreservation of cells from lymph node. The present review emphasize on various procedures used for isolation, activation and cryopreservation of lymph node cells. Isolation of cells was performed by collagenase digestion, teasing apart of lymph node using dissecting needle or lymph nodes were disrupted between two frosted slides. Concanavalin A have been widely used to stimulate mice lymph node cells. Low dose of Con A have stimulatory effect on T cells but high dose have inhibitory action and caused suppression of proliferation of T cell. Balb/c mice and C57Bl/6 mice were used for different dose of Con A. The addition of cryoprotective agents, e.g.dimethylsulphoxide and careful control of cooling rates affords protection from cell damage during freezing.

Author(s):  
O. Faroon ◽  
F. Al-Bagdadi ◽  
T. G. Snider ◽  
C. Titkemeyer

The lymphatic system is very important in the immunological activities of the body. Clinicians confirm the diagnosis of infectious diseases by palpating the involved cutaneous lymph node for changes in size, heat, and consistency. Clinical pathologists diagnose systemic diseases through biopsies of superficial lymph nodes. In many parts of the world the goat is considered as an important source of milk and meat products.The lymphatic system has been studied extensively. These studies lack precise information on the natural morphology of the lymph nodes and their vascular and cellular constituent. This is due to using improper technique for such studies. A few studies used the SEM, conducted by cutting the lymph node with a blade. The morphological data collected by this method are artificial and do not reflect the normal three dimensional surface of the examined area of the lymph node. SEM has been used to study the lymph vessels and lymph nodes of different animals. No information on the cutaneous lymph nodes of the goat has ever been collected using the scanning electron microscope.


1959 ◽  
Vol 110 (2) ◽  
pp. 207-219 ◽  
Author(s):  
Milton Kern ◽  
Herman N. Eisen

Isolated lymph node cells incorporate inorganic orthophosphate into a protein fraction. The phosphorylated product is a phosphoprotein. The rate of phosphate incorporation into phosphoprotein was determined in cells isolated from regional lymph nodes at varying times after antigen injection. The rate was unaltered on the 3rd day, but was enhanced on the 4th day after injection. Parallel results were obtained with L-methionine incorporation into the same gross protein fraction. Possible relationships between antibody formation and the observed enhancement in phosphate incorporation into phosphoprotein are discussed.


1980 ◽  
Vol 30 (2) ◽  
pp. 578-587
Author(s):  
Carole Davis-Scibienski ◽  
Blaine L. Beaman

Normal and specifically activated rabbit alveolar macrophages were infected in vitro with Nocardia asteroides GUH-2. In the presence of serum from normal rabbits, no significant differences were noted between normal and activated alveolar macrophages with respect to phagocytosis, incidence of phagosomelysosome fusion, or nocardicidal activity. However, all of these macrophage functions were enhanced by various immunological components. Serum from immunized rabbits enhanced phagocytosis of nocardial cells by activated macrophages, and there was an additional increase in phagocytosis observed when alveolar lining material was present. Complement had no effect on the ability of the macrophages to phagocytize nocardial cells. The greatest percentage of organisms phagocytized was observed when specifically primed lymph node cells, alveolar lining material, and serum from immunized rabbits were present in the incubation medium. N. asteroides GUH-2 inhibited phagosome-lysosome fusion in normal macrophages in the presence of serum from normal rabbits. However, addition of serum from immunized rabbits or the addition of specifically primed lymphocytes increased the amount of phagosome-lysosome fusion, whereas complement had no effect on this fusion process. Nocardial viability was not reduced when either normal or activated macrophages were infected with bacteria in the presence of normal serum, immune serum, or alveolar lining material. However, specifically activated macrophages incubated with primed lymph node cells obtained from immunized rabbits were able to both decrease the number of viable organisms recovered and to increase the incidence and extent of bacterial cell damage. The greatest number of organisms were killed by specifically activated macrophages when the bacterial cells were incubated with primed lymph node cells suspended in immune serum and alveolar lining material. These results indicate that activated macrophages alone are not sufficient to kill ingested N. asteroides GUH-2 and that specifically primed lymphocytes are important in host resistance to nocardial infections.


2001 ◽  
Vol 114 (19) ◽  
pp. 3463-3477
Author(s):  
Shulamit B. Wallach-Dayan ◽  
Valentin Grabovsky ◽  
Jürgen Moll ◽  
Jonathan Sleeman ◽  
Peter Herrlich ◽  
...  

Cell motility is an essential element of tumor dissemination, allowing organ infiltration by cancer cells. Using mouse LB lymphoma cells transfected with standard CD44 (CD44s) cDNA (LB-TRs cells) or with the alternatively spliced CD44 variant CD44v4-v10 (CD44v) cDNA (LB-TRv cells), we explored their CD44-dependent cell migration. LB-TRv cells, but not LB-TRs or parental LB cells, bound soluble hyaluronic acid (HA) and other glycosaminoglycans (GAGs), and exclusively formed, under physiological shear force, rolling attachments on HA substrate. Furthermore, LB-TRv cells, but not LB-TRs cells or their parental LB cells, displayed accelerated local tumor formation and enhanced accumulation in the peripheral lymph nodes after s.c. inoculation. The aggressive metastatic behavior of i.v.-injected LB-TRV cells, when compared with that of other LB-transfectants, is attributed to more efficient migration to the lymph nodes, rather than to local growth in the lymph node. Injection of anti-CD44 monoclonal antibody or of the enzyme hyaluronidase also prevented tumor growth in lymph nodes of BALB/c mice inoculated with LB-TRv cells. The enhanced in vitro rolling and enhanced in vivo local tumor growth and lymph node invasion disappeared in LB cells transfected with CD44v cDNA bearing a point mutation at the HA binding site, located at the distal end of the molecule constant region. These findings show that the interaction of cell surface CD44v with HA promotes cell migration both in vitro and in vivo, and they contribute to our understanding of the mechanism of cell trafficking, including tumor spread.


2014 ◽  
Vol 80 (3) ◽  
pp. 295-300 ◽  
Author(s):  
Paul Trottman ◽  
Katrina Swett ◽  
Perry Shen ◽  
Joseph Sirintrapun

Radical antegrade modular pancreatosplenectomy (RAMPS) has been reported to provide improved margin resection and lymph node retrieval for tumors of the body and tail of the pancreas compared with standard resection. We examined our experience with RAMPS and standard resection to determine differences in clinicopathologic outcomes. A comparison of RAMPS procedures was made to standard distal pancreatectomy and splenectomy examining various clinicopathologic variables through retrospective chart review. Twenty-six patients underwent distal pancreatectomy with or without splenectomy between November 2004 and June 2011. Twenty patients underwent standard resection and six patients underwent RAMPS procedures for a variety of histologies. As a result of the heterogeneity of diseases, which included benign lesions, margin status was not applicable in some cases and therefore was not assessed overall. Fisher's exact test and Wilcoxon rank sum tests demonstrated a significant difference in number of lymph nodes removed with mean of 4.3 and 11.2 lymph nodes obtained for standard resection and RAMPS, respectively ( P = 0.03). The RAMPS procedure for lesions of the body and tail of the pancreas retrieved significantly more lymph nodes than standard distal pancreatectomy and splenectomy. It should be the preferred surgical approach when lymph node count is important for tumor staging.


Blood ◽  
1989 ◽  
Vol 73 (8) ◽  
pp. 2171-2181 ◽  
Author(s):  
L Ostlund ◽  
P Biberfeld ◽  
KH Robert ◽  
B Christensson ◽  
S Einhorn

Abstract The influence of interferon (IFN) on cellular proliferation, blast transformation, and differentiation was studied in lymph node cells from 17 patients with B-cell lymphomas, one patient with T-cell lymphoma, and eight patients with enlarged, non-malignant lymph nodes. The effects of IFN on lymph node cells were compared with effects on mononuclear blood cells from chronic lymphocytic leukemia (CLL) patients and healthy donors. Natural IFN-alpha (nIFN-alpha) induced a proliferative response in cells from seven of 17 of the B-cell lymphomas, in two of eight of the non-malignant lymph nodes, and in lymphoid blood cells from two of 32 CLL patients. With few exceptions, the proliferating cells were B cells and the data suggest that IFN acts directly on the B cells. Proliferation was not induced with IFN in cells from the T-cell lymphoma or in mononuclear blood cells from 13 healthy donors. nIFN-alpha induced blast transformation in cells from ten of 14 of the B-cell lymphomas and in four of seven of the non- malignant lymph nodes. Also beta- and gamma-IFN were shown to induce proliferation and blast transformation in lymph node cells from some patients. No major effect on the expression of various differentiation markers could be observed following culture in the presence of nIFN- alpha. We conclude that IFNs can induce proliferation and blast transformation in malignant and non-malignant B cells from lymph nodes.


1968 ◽  
Vol 128 (6) ◽  
pp. 1437-1449 ◽  
Author(s):  
David M. Lubaroff ◽  
Byron H. Waksman

The precise origin of cells infiltrating tuberculin skin reactions was studied with the technique of immunofluorescence. Thymectomized, irradiated Lewis rats were restored with bone marrow from allogeneic or F1 donors. They were passively sensitized to tuberculin by a subsequent transfer of Lewis lymph node cells and were given intradermal skin tests with tuberculoprotein. In 24 hr reactions the majority of cells were shown to be derived from the infused marrow. These results were the same regardless whether the lymphocyte transfer was performed on the day of irradiation and marrow injection or 7 days later. The cells in the tuberculin reactions, marrow, spleen, and lymph nodes not derived from the bone marrow were found to originate in the transferred lymph node cells. The relative percentages of marrow-derived and lymph node-derived cells in the tuberculin reactions remained the same during the 9–24 hr period following skin test.


1968 ◽  
Vol 127 (2) ◽  
pp. 307-325 ◽  
Author(s):  
Vera S. Byers ◽  
Eli E. Sercarz

A set of conditions has been described under which primed rabbit lymph nodes produce a secondary antibody response upon in vivo stimulation with a large dose of antigen, but are subsequently "exhausted;" that is, lymph node cultures prepared at intervals following the booster injection cannot be re-stimulated to display tertiary responses. Rabbits given 100-fold less antigen in the booster inoculum were able to give a tertiary response upon in vitro challenge. The system used permits neither induction nor continuation of a primary response to BSA in vitro. Since it could be demonstrated that no memory cells were generated by the booster injection within the intervals between in vivo injection and culture, the tertiary response in nonexhausted nodes must have been due to residual memory cells which remained untriggered by the in vivo booster injection. The unresponsive state was not caused by antibody feedback. These results are interpreted to mean that a population of memory cells can be exhausted by a supraoptimal dose of antigen, rendering the node temporarily incapable of further response. This implies that long-lived memory is not due to asymmetric division of memory cells. The source and fate of memory cells is discussed with regard to this evidence.


1970 ◽  
Vol 131 (5) ◽  
pp. 936-969 ◽  
Author(s):  
Niels C. Pedersen ◽  
Bede Morris

The rejection of renal homografts has been studied in sheep by transplanting kidneys into the neck and preserving the renal lymphatic drainage intact. Chronic fistulae were established in the transplanted renal lymphatics and lymph collected throughout the life of the graft. The changes that occurred in homografts during the process of rejection were reflected in changes in the lymph. Large numbers of basophilic, blast, lymphoid cells appeared in the lymph, and lymph production in the grafted kidney increased 20–50 fold. Over a period of about 10 days, up to 60 g wet weight of lymphoid cells and up to 10 liters of lymph were collected from the graft. Within 24 hr of grafting, the host cells present in the renal lymph had become sensitized to the graft and transformed into blast cells when cultivated in Millipore chambers in vitro. When the cells leaving the graft during the first 18–48 hr were injected into distant nonstimulated lymph nodes of the host sheep, they evoked significant cellular and antibody responses in the nodes. Within the graft, the main pathological changes were found in the vascular endothelium and many of the peritubular capillaries become plugged with emboli comprised of blast cells. There was extensive infiltration of the renal parenchyma with lymphoid cells and evidence of their transformation and proliferation within the renal blood capillaries. When all the lymph and cells leaving the homograft were diverted from the body, there was a greatly decreased reaction in the regional prescapular lymph node, and no reaction in lymph nodes distant from the graft. In these circumstances, the survival of the graft was not prolonged, and it was rejected without involvement of the lymph nodes of the host. Humoral antibody was produced in the lymph node regional to the homograft within 48–60 hr of grafting. Antibody was not detected in the blood or in the renal lymph until near to the time the graft was rejected. It was thought that this was due to the binding of antibody by the kidney graft tissue. We conclude that all the events which lead to the recognition and rejection of renal homografts can occur centrally within the graft itself.


Immuno ◽  
2021 ◽  
Vol 1 (3) ◽  
pp. 223-230
Author(s):  
Yoshihiro Komohara ◽  
Toshiki Anami ◽  
Kenichi Asano ◽  
Yukio Fujiwara ◽  
Junji Yatsuda ◽  
...  

Lymph nodes are secondary lymphoid organs that appear as bean-like nodules usually <1 cm in size, and they are localized throughout the body. Many antigen-presenting cells such as dendritic cells and macrophages reside in lymph nodes, where they mediate host defense responses against pathogens such as viruses and bacteria. In cancers, antigen-presenting cells induce cytotoxic T lymphocytes (CTLs) to react to cancer cell-derived antigens. Macrophages located in the lymph node sinus are of particular interest in relation to anti-cancer immune responses because many studies using both human specimens and animal models have suggested that lymph node macrophages expressing CD169 play a key role in activating anti-cancer CTLs. The regulation of lymph node macrophages therefore represents a potentially promising novel approach in anti-cancer therapy.


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