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 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 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 IMPROVED STAINING FOR PEROXIDASE WITH BENZIDINE AND IMPROVED DOUBLE STAINING IMMUNOPEROXIDASE PROCEDURES

1972 ◽  
Vol 20 (4) ◽  
pp. 272-278 ◽  
Author(s):  
WERNER STRAUS
Keyword(s):  
Acid Phosphatase ◽  
Horseradish Peroxidase ◽  
Lymph Nodes ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Plasma Cells ◽  
Double Staining ◽  
Technical Problems ◽  
Cytochemical Reaction ◽  
Antigen Antibody

The sensitivity of the cytochemical reaction for peroxidase with benzidine and H2O2 could be much enhanced and a noncrystalline, blue-brown reaction product could be obtained by decreasing the concentration of ethanol (for dissolving benzidine) and by increasing the time and temperature of incubation. This method, together with a new method for the inactivation of residual (injected) peroxidase, were incorporated in double staining procedures for horseradish peroxidase (HRP) and its antibody (antigen-antibody complexes) and in double staining procedures for the antibody to HRP and acid phosphatase activity. Double staining in contrasting colors was also applied to detect rabbit antibodies against two antigens (HRP and rat anti-HRP γ-globulin) in the same section of popliteal lymph nodes. It was found that the antibody against each antigen appeared in different plasma cells whether the rabbits were immunized against the two antigens separately or against both antigens together as antigen-antibody complexes. Certain technical problems arising in double staining procedures are discussed.

scholarly journals SPECIFIC ANTIBODY WITHIN LYMPHOID GERMINAL CENTER CELLS OF MICE AFTER PRIMARY IMMUNIZATION WITH HORSERADISH PEROXIDASE: A LIGHT AND ELECTRON MICROSCOPIC STUDY

1970 ◽  
Vol 131 (1) ◽  
pp. 77-91 ◽  
Author(s):  
Bernard Sordat ◽  
Martine Sordat ◽  
Max W. Hess ◽  
Richard D. Stoner ◽  
Hans Cottier
Keyword(s):  
Horseradish Peroxidase ◽  
Specific Antibody ◽  
Germinal Center ◽  
Light And Electron Microscopy ◽  
Germinal Centers ◽  
Antigenic Stimulation ◽  
Intercellular Spaces ◽  
Electron Microscopic ◽  
Perinuclear Space ◽  
Reticular Cells

The appearance in mice of specific antibody within newly formed germinal centers in lymph nodes was demonstrated by light and electron microscopy after regional primary antigenic stimulation with horseradish peroxidase (HRP). Lymphoid germinal center cells containing anti-HRP antibody in the perinuclear space and in the cytoplasm were detected from 17 to 26 days after antigenic stimulation. Extracellular anti-HRP antibody within germinal centers, localized between dendritic reticular cells and lymphoid elements, could not be found before the appearance of intracellular antibody. These findings strongly suggest antibody formation by lymphoid germinal center cells. Both antigen and corresponding antibody persisted in intercellular spaces up to 35 days after primary stimulation. The concomitant presence in a given lymph node of germinal centers which are positive or negative with regard to specific antibody provide evidence in favor of monospecificity of individual centers. The mechanisms of antigen-trapping within germinal centers are discussed in the light of the present observations.

scholarly journals STUDIES ON ANTIBODY PRODUCTION

1964 ◽  
Vol 119 (4) ◽  
pp. 685-696 ◽  
Author(s):  
Hans-Jobst Wellensiek ◽  
Albert H. Coons
Keyword(s):  
Antibody Response ◽  
Plasma Cells ◽  
Fluorescent Antibody ◽  
Secondary Response ◽  
Phagocytic Cells ◽  
Physical Presence ◽  
Primary Antibody Response ◽  
Test Cells ◽  
Blue Reaction ◽  
Medullary Cords

The physical presence of the antigen used to stimulate a secondary antibody response was demonstrated in the cells of popliteal lymph nodes. Rabbits previously injected with apoferritin (containing no iron), which was prepared from recrystallized horse ferritin, were given an injection of ferritin 5 weeks later. The antigen was traced by means of the Prussian blue reaction, by specific fluorescent antibody, and by electron microscopy. Antiferritin antibody was localized by immunofluorescence, though it was not possible to test cells simultaneously for antigen and antibody. Horse ferritin induces a rather weak primary antibody response, but a brisk secondary response characterized by the appearance in the medullary cords of numerous plasma cells containing antiferritin. Many intact ferritin molecules were found in the nucleus and cytoplasm of numerous reticular and other phagocytic cells in the sinuses. In decreasing amount, ferritin molecules were also clearly demonstrated in hemacytoblasts (plasmoblasts), and immature and mature plasmocytes.

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 Relationship between lymph concentration of cytokines and structural transformations in mesenteric lymph nodes in chemotherapy, surgical treatment, and chemotherapy of experimental breast cancer

2020 ◽  
pp. 37-45
Author(s):  
О.В. Казаков ◽  
А.Ф. Повещенко ◽  
Н.Б. Орлов ◽  
Т.В. Райтер ◽  
О.В. Повещенко ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Immune Response ◽  
Surgical Treatment ◽  
Lymph Nodes ◽  
Thoracic Duct ◽  
Plasma Cells ◽  
Local Immune Response ◽  
Mesenteric Lymph Nodes ◽  
Mesenteric Lymph ◽  
Medullary Cords

Цель исследования - анализ корреляции морфометрии брыжеечных лимфатических узлов и концентрации цитокинов в лимфе грудного протока при химиотерапии рака молочной железы, хирургическом лечении и последующей химиотерапии. Методика. Рак молочной железы индуцировали введением N-метил-N-нитрозомочевины 5 раз с интервалом 7 сут подкожно в область 2-й молочной железы справа. Курс химиотерапии проходил по схеме CMF. Корреляцию между концентрациями 24 цитокинов лимфы и числом клеток структурных зон лимфатических узлов оценивали по коэффициенту ранговой корреляции Спирмена. Результаты. После химиотерапии РМЖ, по сравнению с РМЖ без лечения, морфологические преобразования в лимфатических узлах свидетельствуют о снижении активности местного иммунного ответа. Исследование корреляции концентрации цитокинов в лимфе со структурными изменениями в лимфатических узлах выявило зависимости направленные на повышение иммуномодулирующего и противоопухолевого действия цитокинов. После оперативного лечения РМЖ и последующей химиотерапии, по сравнению только с химиотерапией РМЖ, выявлены положительные связи иммунобластов с цитокином GRO/KC в герминативных центрах, цитокина IL-6 - с митотически делящимися клетками в герминативных центрах и мозговых тяжах, IL-5 - с иммунобластами в мозговых тяжах, хемокина MIP-1a - со зрелыми плазматическими клетками в мозговых синусах. Увеличено количество иммунобластов, средних и малых лимфоцитов в герминативных центрах, возросло количество малых лимфоцитов, незрелых и зрелых плазматических клеток в мозговых синусах. Увеличены площади мозговых тяжей и паракортикальной зоны. Выявлена корреляция: цитокина IL-1α с малыми лимфоцитами, IL-6 с иммунобластами, IL-7 и IL-18 - со средними лимфоцитами, GRO/KC - с иммунобластами, IL-17 - с макрофагами в Т-зависимой зоне; IL-7 и IL-18 - с иммунобластами, IL-12 - с макрофагами, MIP-1a и MCP-1 со зрелыми плазматическими клетками в мозговых синусах. Заключение. После оперативного лечения РМЖ c последующей химиотерапией, по сравнению только с химиотерапией РМЖ, выявлены взаимозависимости концентрации цитокинов в лимфе грудного протока с морфологическими изменениями в брыжеечных лимфатических узлах, которые могут указывать на повышение активности местного звена иммунного ответа. The aim of this study was to analyze correlations of the morphometry of mesenteric lymph nodes with cytokine concentrations in thoracic duct lymph in chemotherapy and surgical treatment with subsequent chemotherapy of breast cancer. Methods. Breast cancer was induced by subcutaneous injection of N-methyl-N-nitrosourea 5 times with 7-day intervals, into the region of the 2nd breast on the right. The course of chemotherapy was performed according to the CMF scheme. Correlations between concentrations of 24 cytokines of the lymph and cells of lymph node structural regions were estimated by the Spearman rank correlation coefficient. Results. After chemotherapy for breast cancer compared to untreated breast cancer, morphological transformations in lymph nodes indicated decreased activity of the local immune response. Analysis of correlations between lymph concentrations of cytokines and structural changes in lymph nodes identified relationships aimed at increasing the immunomodulatory and antitumor effects of cytokines. After surgical treatment of breast cancer and subsequent chemotherapy compared to chemotherapy alone, positive correlations were found for immunoblasts with cytokine GRO/KC in germinative centers, for cytokine IL-6 with mitotically dividing cells in germinative centers and medullary cords, for IL-5 with immunoblasts in medullary cords, and for chemokine MIP-1a with mature plasma cells in medullary sinuses. Numbers of immunoblasts and medium and small lymphocytes were increased in germinative centers whereas numbers of small lymphocytes and immature and mature plasma cells were increased in medullary sinuses. Areas of medullary cords and the paracortical zone were increased. Correlations were found for cytokine IL-1α with small lymphocytes, for IL-6 with immunoblasts, for IL-7 and IL-18 with medium lymphocytes, for GRO/KC with immunoblasts, for IL-17 with macrophages in the T-dependent zone, for IL-7 and IL-18 with immunoblasts, for IL-12 with macrophages, and for MIP-1a and MCP-1 with mature plasma cells in medullary sinuses. Conclusion. After surgical treatment of breast cancer and subsequent chemotherapy compared to chemotherapy alone, cytokine concentrations in lymph of the thoracic duct were observed to correlate with morphological changes in mesenteric lymph nodes, which may indicate increased activity of the local immune response.

scholarly journals The compartments of the parenchyma of the lymph nodes in newborn bull calves of domestic cattle (Bos taurus)

2017 ◽  
Vol 8 (2) ◽  
pp. 169-178
Author(s):  
P. N. Gavrilin ◽  
О. G. Gavrilina ◽  
M. V. Kravtsovа
Keyword(s):  
Lymph Nodes ◽  
Plasma Cells ◽  
Bos Taurus ◽  
Relative Volume ◽  
Structural Components ◽  
Lymphocyte Migration ◽  
Clonal Proliferation ◽  
Bull Calves ◽  
High Level ◽  
Medullary Cords

The article analyzes the features of the structure of the lymphoid lobules of the parenchyma of the superficial somatic (Limphonodi subiliaci, L. cervicales superficiales), profund somatic (L. axillares proprii L. poplitei), somatovisceral (L. iliaci mediales, L. retropharyngei mediales) and visceral (L. mediastinales caudales, L. ileocolici) lymph nodes of newborn bull calves of domestic cattle. To visualize clearly the boundaries of the structural components of lymphoid lobules we used the author’s modification of the impregnation of total median frozen histological sections with silver nitrate. We have established a high level of tissue differentiation of the lymph nodes, a significant development of the lymphoid parenchyma, the division of the parenchyma into lymphoid lobules, the presence in the lobules of all the main structural components that are represented by two morphotypes. The first morphotype is ribbon-like perisinusoidal cords (interfollicular zone, paracortical and medullary cords). The second morphotype is rounded lymphoid formations (central zones of deep cortex units, lymphatic nodules). Lymphoid lobules are located along the marginal sinus in one row, they are better developed and differentiated in the visceral lymph nodes. In all the lymph nodes, the lymphoid lobules have a similar histoarchitectonic, and each structural component of the lymphoid lobules has a specific architectonic of the reticular meshwork and the density of the location of the fibroblastic reticulocytes. We determined that the structures of the first morphotype which provide the migration of lymphocytes, the detection of antigens and the accumulation of plasmocytes are more developed. We have established that the relative volume of structures of the first morphotype is 4.5–8.0 times larger than the volume of the structures of the second morphotype, which provide clonal proliferation of T and B lymphocytes, especially in deep somatic lymph nodes. Among the zones of the second morphotype, predominate T-dependent zones, the relative volume of which considerably exceeds the volume of B-dependent zones (lymphoid nodules): in the superficial somatic lymph nodes by 14–30 times, profound somatic by 12–14 times, somatovisceral by 6–7 times and visceral by 4.5–5.5 times. We determined that lymphatic nodules can form in different parts of compartments: in the interfollicular zone and paracortical cords of all lymph nodes and in the medullary cords of the visceral lymph nodes. The study shows that the parenchyma of the lymph nodes of newborn bull calves has a high degree of maturity, contains a full set of structural markers of immunocompetence, among which predominate the components that support lymphocyte migration, antigen detection and accumulation of plasma cells.

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