scholarly journals LOCALIZATION OF THE ANTIGEN IN POPLITEAL LYMPH NODES OF RABBITS DURING THE FORMATION OF ANTIBODIES TO HORSERADISH PEROXIDASE

1970 ◽  
Vol 18 (2) ◽  
pp. 131-142 ◽  
Author(s):  
WERNER STRAUS
Keyword(s):  
Horseradish Peroxidase ◽  
Lymph Nodes ◽  
Specific Antibody ◽  
Plasma Cells ◽  
Repeated Injection ◽  
Before And After ◽  
Reticular Cells ◽  
Popliteal Lymph Nodes ◽  
Tissue Material

The localization of an antigen (horseradish peroxidase) in popliteal lymph nodes of rabbits was investigated in order to detect the possible interrelationship with the location of the specific antibody in the same tissue material. Staining procedures for peroxidase with benzidine, diaminobenzidine and 3-amino-9-ethyl-carbazole, as well as double staining procedures for the antigen and the antibody and for the antigen (or antibody) and acid phosphatase, were applied before and after adsorption of the antigen to sites of antibody in vitro. The appearance of the antigen in the cells lining the lymph sinuses, in reticular cells of medullary cords, in macrophages and in the "intercellular web" of lymphoid follicles was studied after a single and repeated injection of peroxidase, and the persistence of the antigen at these sites was observed. It was found that the localization of the antigen in the cortex and medulla of the lymph node was different depending on whether or not specific antibodies were present in the blood at the time of injection, and that at certain periods a considerable number of plasma cells and lymphoblasts contained the antigen together with the specific antibody.

scholarly journals LOCATION OF ANTIBODY TO HORSERADISH PEROXIDASE IN POPLITEAL LYMPH NODES OF RABBITS DURING THE PRIMARY AND EARLY SECONDARY RESPONSE

1970 ◽  
Vol 18 (2) ◽  
pp. 120-130 ◽  
Author(s):  
WERNER STRAUS
Keyword(s):  
Horseradish Peroxidase ◽  
Lymph Nodes ◽  
Specific Antibody ◽  
Plasma Cells ◽  
Secondary Response ◽  
Double Staining ◽  
Intercellular Spaces ◽  
Reticular Cells ◽  
Popliteal Lymph Nodes ◽  
Medullary Cords

After a primary injection of horseradish peroxidase into the footpads of rabbits, the specific antibody reaction in popliteal lymph nodes was first seen in plasma cells of medullary cords and, later, in lymphoblasts of germinal centers in the cortex. During the late primary or early secondary response, the reaction also became positive in reticular cells and lymphocytes. Specific antibodies were also present in the intercellular spaces between lymphocytes (reticulum) in the lymphoid follicles and in globules distributed throughout the lymph node. Two weeks following a primary injection of the antigen, many plasma cells containing the specific antibody were located in proximity of macrophages and reticular cells. The phagolysosomes of many macrophages and reticular cells became antibody-positive at about the same time. Double staining procedures were developed by which the antigen and antibody and the antigen or antibody and acid phosphatase (lysosomes) could be visualized in the same tissue section.

scholarly journals CYTOCHEMICAL DETECTION OF SITES OF ANTIBODY TO HORSERADISH PEROXIDASE IN SPLEEN AND LYMPH NODES

1968 ◽  
Vol 16 (4) ◽  
pp. 237-248 ◽  
Author(s):  
WERNER STRAUS
Keyword(s):  
Horseradish Peroxidase ◽  
Lymph Nodes ◽  
Plasma Cells ◽  
Specific Binding ◽  
Repeated Injection ◽  
Similar Reaction ◽  
Fixed Tissue ◽  
Tissue Sections ◽  
Spleen And Lymph Nodes

After repeated injection of small amounts of horseradish peroxidase into the perivertebral muscle and footpads of rabbits, certain sites in spleen and popliteal lymph nodes showed intense adsorption of antigen (horseradish peroxidase) during treatment of fixed tissue sections in vitro. The reaction occurred in certain cells, probably plasma cells, and was also positive in the reticulum extending between lymphocytes. These sites were considered to contain antibodies against horseradish peroxidase, since sections from the same block, not treated with antigen in vitro, and corresponding sections from nonimmunized rabbits did not give a similar reaction. The blood sera of rabbits which showed specific binding of antigen in the spleen caused approximately 60% inhibition of horseradish peroxidase activity, but no inhibition was caused by the blood sera of nonimmunized control animals.

scholarly journals Staining patterns for the anti-horseradish peroxidase antibody reaction in proplasma cells developing in the medulla of rat popliteal lymph nodes during the secondary response.

1981 ◽  
Vol 29 (4) ◽  
pp. 525-530 ◽  
Author(s):  
W Straus
Keyword(s):  
Horseradish Peroxidase ◽  
Lymph Nodes ◽  
Plasma Cells ◽  
Small Cells ◽  
Secondary Response ◽  
Microscopic Appearance ◽  
Stage Of Development ◽  
Antibody Reaction ◽  
Popliteal Lymph Nodes ◽  
The One

The development of plasma cells from lymphocytes was studied in the medulla of popliteal lymph nodes of rats during the secondary response to horseradish peroxidase (HRP). Changes in the microscopic appearance of proplasma cells were compared with changes in the intensity of the anti-HRP antibody reaction in these cells. Early proplasma cells, appearing 2 to 3 days after the injection of HRP into the footpads, were relatively small cells similar in size to lymphocytes. Their small nuclei were eccentrically located due to the one-sided enlargement of the pyroninophilic cytoplasm. The reaction for the anti HRP antibody in these cells was weak or negative. Other proplasma cells located in the same medullary cord regions showed a more intense antibody reaction. This change was correlated, in many cases, with an enlargement of the nucleus, giving the cells a blast-like appearance. Three to 6 days after the reinjection of the antigen, the medullary cords contained many mature plasma cells characterized by an intense antibody reaction. The mature plasma cells were always accompanied by proplasma cells, the latter varying in microscopic appearance (stage of development) asd staining intensities (antibody contents). The staining intensities and the microscopic appearance of proplasma cells, and the proportion of proplasma cells to plasma cells, varied in different medullary cord regions of the same lymph nodes. The staining patterns, together with the microscopic appearance of the cells, seemed to show whether antibody formation was inhibited or stimulated.

scholarly journals HETEROGENEITY OF ANTIBODY-FORMING CELLS

1969 ◽  
Vol 129 (5) ◽  
pp. 1029-1044 ◽  
Author(s):  
Cesare Bosman ◽  
Joseph D. Feldman ◽  
Edgar Pick
Keyword(s):  
Lymph Nodes ◽  
Plasma Cells ◽  
Specific Antigen ◽  
Cell Suspensions ◽  
Electron Microscopic ◽  
Cytoplasmic Vacuoles ◽  
Draining Lymph Nodes

Cell suspensions from draining lymph nodes of immune and nonimmune rats were reacted in vitro with 125I-labeled antigens. In light microscopic radioautographs of smears, 17% of the immunized cells were tagged by specific antigen; 2.0% of control cells were positive. In electron microscopic radioautographs, 90% of the labeled elements from immune donors were lymphocytes, blast and plasma cells; 10% were monocytes-macrophages or other elements, including naked nuclei. 15% of the labeled cells from control materials were lymphocytes and plasma cells, while 85% were monocytes-macrophages and naked nuclei. Within cell suspensions derived from immunized animals there were almost twice as many lymphocytes marked by isotope as plasma cells, and the lymphocytes ranged in morphology from mature monoribosomal elements to immature polyribosomal cells. Antibody-forming cells fixed labeled antigen at their surfaces. The monocyte-macrophage class was distinguished by a high mean grain count and by distribution of grains within cytoplasmic vacuoles and lysosomes.

scholarly journals Production of Macroglobulins in Vitro and a Study of Their Cellular Origin

Blood ◽  
1962 ◽  
Vol 20 (1) ◽  
pp. 56-64 ◽  
Author(s):  
DOROTHEA ZUCKER-FRANKLIN ◽  
EDWARD C. FRANKLIN ◽  
NORMAN S. COOPER
Keyword(s):  
Bone Marrow ◽  
Tissue Culture ◽  
Lymph Node ◽  
Lymph Nodes ◽  
Plasma Cells ◽  
Buffy Coat ◽  
Cellular Origin

Abstract Lymph nodes of three patients with macroglobulinemia of Waldenström were studied in tissue culture and shown to synthesize 19S γ-globulin in vitro. Lymph node imprints, bone marrow, and buffy coat smears of the same patients consisted almost entirely of lymphocytes. When these were stained with fluorescein-conjugated antiserum to macroglobulin, large and medium-sized lymphocytes and lymphoblasts rather than mature lymphocytes or plasma cells were shown to contain the protein. It is suggested that 19S γ-globulin may also be synthesized by cells belonging to the lymphoid series under normal circumstances.

The route of re-circulation of lymphocytes in the rat

1964 ◽  
Vol 159 (975) ◽  
pp. 257-282 ◽  
Keyword(s):  
Lymph Nodes ◽  
Thoracic Duct ◽  
Large Scale ◽  
Plasma Cells ◽  
Tritiated Thymidine ◽  
Autoradiographic Study ◽  
Total Output ◽  
White Pulp ◽  
Radioactive Label

The experiments presented in this paper support the idea that the output of small lymphocytes from the thoracic duct of the rat (about 10 9 /day) is normally maintained by a large-scale re-circulation of cells from the blood to the lymph. It has been shown that the main channel from blood to lymph lies with in the lymph nodes and that small lymphocytes enter the nodes by crossing the walls of a specialized set of blood vessels, the post-capillary venules. In order to trace the fate of small lymphocytes, cells from the thoracic duct of rats were incubated for 1 h in vitro with tritiated adenosine. This labelled the RNA of about 65% of the small lymphocytes and more than 95% of the large lymphocytes; it also labelled the DNA of a proportion of the large lymphocytes. The mixture of small and large labelled lymphocytes was transfused into the blood of two groups of rats which belonged to the same highly inbred strain as the cell donors. At various times after the transfusions the thoracic ducts in one group of rats were cannulated to determine the proportion of labelled cells which could be recovered in the lymph; at corresponding times, the rats in the other group were killed and autoradiographs prepared from their tissues to determine the location of the labelled cells. The radioactive label in the RNA of small lymphocytes was stable enough to ensure that the labelled small lymphocytes which were recovered in the lymph several days after a transfusion were those which had originally been transfused into the blood. When the thoracic duct was cannulated 20 to 27 h after a transfusion, about 70% of the labelled small lymphocytes which had been transfused into the blood could be recovered from the thoracic duct over a 5-day period of lymph collection. During the first 36 to 48 h after cannulation, while the total output of small lymphocytes was falling rapidly, the proportion of labelled cells in the lymph remained approximately constant. The pool of the animal’s own cells with which the labelled cells had mixed contained between 1·5 and 2 × 10 9 small lymphocytes; this was identified as the re-circulating pool. An autoradiographic study showed that after their transfusion into the blood the labelled small lymphocytes ‘homed’ rapidly and in large numbers into the lymph nodes, the white pulp of the spleen and the Peyer’s patches of the intestine. The concentration of labelled cells in other tissues was trivial in comparison. Labelled small lymphocytes were seen penetrating the endothelium of the post-capillary venules in the lymph nodes within 15 min of the start of a transfusion; they were traced into the cortex of the nodes and finally into the medullary lymph sinuses. Labelled small lymphocytes did not migrate into the adult thymus but a few entered the thymus of newborn rats. It was concluded that the re-circulating pool of small lymphocytes was located in the lymphoid tissue, the thymus excepted, and that the rapid ‘homing’ of cells into the lymph nodes had its basis in the special affinity of small lymphocytes for the endothelium of the post-capillary venules. The interpretation of these experiments was not complicated by the presence of large, as well as of small lymphocytes in the suspensions of labelled cells which were transfused. Other experiments, in which the large lymphocytes alone were labelled with tritiated thymidine, showed that most of them migrated from the blood into the wall of the gut where they assumed the appearance of primitive plasma cells; very few divided to form small lymphocytes.

scholarly journals AN ELECTRON MICROSCOPE STUDY OF LYMPHATIC TISSUE IN RUNT DISEASE

1965 ◽  
Vol 25 (3) ◽  
pp. 149-177 ◽  
Author(s):  
Leon Weiss ◽  
Alan C. Aisenberg
Keyword(s):  
Lymph Nodes ◽  
Plasma Cells ◽  
Cell Types ◽  
Sprague Dawley ◽  
Connective Tissues ◽  
Splenic Cells ◽  
Reticular Cells ◽  
Spleen And Lymph Nodes ◽  
Myelin Figure ◽  
Rats And Mice

The thymus, spleen, and lymph nodes were studied in runt disease induced by a graft of intravenously injected homologous splenic cells into newborn rats and mice. Adult Long-Evans cells (70 x 106) were injected into Sprague-Dawley rats. Adult DBA cells (7 x 106) were injected into C57BL/6 mice. Runted rats were sacrificed at 14 to 28 days of age; mice at 10 to 20 days. The thymic cortex is depleted of small lymphocytes. Those remaining are severely damaged and phagocytized. Evidence of damage includes swelling of mitochondria, myelin figure formation, margination of chromatin, and sharp angulation in nuclear contour. Large numbers of macrophages are present. Epithelial-reticular cells which envelop small cortical blood vessels are often retracted, with the result that the most peripheral layer in the thymic-blood barrier suffers abnormally large gaps. Lymphocytes of the periarterial lymphatic sheaths of spleen and of the cortex of lymph nodes are reduced in number and damaged. Vast numbers of plasma cells and many lymphocytes are evident throughout lymph nodes, in the periarterial lymphatic sheaths, and in the marginal zone and red pulp of the spleen. Plasma cells are of different sizes, the larger having dilated sacs of endoplasmic reticulum. Lymphocytes are small to medium in size. They contain, in varying quantity, ribosomes and smooth membrane-bounded cytoplasmic vesicles approximately 350 to 500 A in diameter. Most plasma cells and lymphocytes are damaged and many of these are phagocytized. Many lymphocytes in lymph nodes, however, show no evidence of damage. Reticular cells and other fixed cells of the connective tissues seldom appear affected. Thus, the major cell types reacting in runt disease are lymphocytes, plasma cells, and histiocytes or macrophages. It appears, therefore, that both the delayed and immediate types of sensitivity play a part in this disease.

scholarly journals A novel cancer immunotherapy using tumor-infiltrating B cells in the APCmin/+ mouse model

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0245608
Author(s):  
Xinying Wang ◽  
Shohei Asami ◽  
Daisuke Kitamura
Keyword(s):  
B Cells ◽  
Cancer Immunotherapy ◽  
Tumor Antigens ◽  
Plasma Cells ◽  
Somatic Hypermutation ◽  
Serum Antibody ◽  
Good Prognosis ◽  
Repeated Injection ◽  
Lymphoid Structures

Accumulating evidence has suggested a correlation of tumor infiltrating B cells (TiBcs) and a good prognosis of cancer diseases. In some cases, TiBcs appear to have experienced antigen stimulation since they have undergone class-switching and somatic hypermutation and formed tertiary lymphoid structures around tumors together with T cells. Assuming TiBcs include those that recognize some tumor antigens, we sought to investigate their possible usefulness for cell-mediated immunotherapies. To expand usually small number of TiBcs in vitro, we modified our B cell culture system: we transduced B cells with ERT2-Bach2 so that they grow unlimitedly provided with tamoxifen, IL-21 and our original feeder cells. Such cells differentiate into plasma cells and produce antibodies upon withdrawal of tamoxifen, and further by addition of a Bach2-inhibitor in vitro. As a preliminary experiment, thus expanded splenic B cells expressing a transgenic antigen receptor/antibody against hen egg lysozyme were intravenously injected into mice pre-implanted with B16 melanoma cells expressing membrane-bound HEL in the skin, which resulted in suppression of the growth of B16 tumors and prolonged survival of the recipient mice. To test the usefulness of TiBcs for the immunotherapy, we next used APCmin/+ mice as a model that spontaneously develop intestinal tumors. We cultured TiBcs separated from the tumors of APCmin/+ mice as above and confirmed that the antibodies they produce recognize the APCmin/+ tumor. Repeated injection of such TiBcs into adult APCmin/+ mice resulted in suppression of intestinal tumor growth and elongation of the survival of the recipient mice. Serum antibody from the TiBc-recipient mice selectively bound to an antigen expressed in the tumor of APCmin/+ mice. These data suggest a possibility of the novel individualized cancer immunotherapy, in which TiBcs from surgically excised tumor tissues are expanded and infused into the donor patients.

scholarly journals RESISTANCE OF LONG-LIVED LYMPHOCYTES AND PLASMA CELLS IN RAT LYMPH NODES TO TREATMENT WITH PREDNISONE, CYCLOPHOSPHAMIDE, 6-MERCAPTOPURINE, AND ACTINOMYCIN D

1967 ◽  
Vol 126 (1) ◽  
pp. 109-125 ◽  
Author(s):  
John J. Miller ◽  
Leonard J. Cole
Keyword(s):  
Lymph Nodes ◽  
Autoimmune Diseases ◽  
Actinomycin D ◽  
Plasma Cells ◽  
Tritiated Thymidine ◽  
Lymphoid Tissues ◽  
Organ Transplants ◽  
Appreciable Effect ◽  
Low Doses ◽  
Popliteal Lymph Nodes

The cells of the popliteal lymph nodes of rats were labeled for 4 days after a secondary immunological stimulus. 31 days after the last dose of tritiated thymidine, groups of rats were started on courses of daily, intraperitoneal injections of prednisone, cyclophosphamide, 6-mercaptopurine, or actinomycin D. The initially low doses of these agents were doubled in successive weeks until either lymphoid hypoplasia or death occurred. Rats from each group were killed weekly, and the percentages of persisting, labeled small lymphocytes in the popliteal nodes were determined. Sections of these nodes were examined for persisting, labeled plasma cells. The per cent of lymphocytes labeled increased while the total number of lymphocytes decreased during treatment with prednisone and cyclophosphamide. Prednisone decreased the numbers of long-lived plasma cells, but these cells were preferentially resistant to cyclophosphamide. Neither 6-mercaptopurine nor actinomycin D had an appreciable effect on lymphoid tissues histologically nor on the proportions of labeled, long-lived lymphocytes and plasma cells before causing the deaths of the rats receiving them. These results indicate that long-lived lymphocytes and plasma cells survive treatment with the immunolytic drugs studied, and that long-lived lymphocytes are specifically resistant to prednisone and cyclophosphamide. We believe these results have an application to the attempts to find drugs useful in the treatment of immunologic rejections of organ transplants, and for therapy of autoimmune diseases.

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