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 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.

In vitro autoradiographic and in vivo scintigraphic localization of somatostatin receptors in human lymphatic tissue

Blood ◽  
1993 ◽  
Vol 82 (7) ◽  
pp. 2143-2151 ◽  
Author(s):  
JC Reubi ◽  
B Waser ◽  
U Horisberger ◽  
E Krenning ◽  
SW Lamberts ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Gamma Camera ◽  
Somatostatin Receptors ◽  
Immune Functions ◽  
Thymic Tissue ◽  
Tissue Sections ◽  
Spleen And Lymph Nodes ◽  
Neuroendocrine Functions

Abstract Receptors for the neuropeptide somatostatin (SS) were evaluated in vitro and in vivo in various human lymphatic tissues, ie, thymus, spleen, and lymph nodes; thymic carcinoids and thymomas were also tested. The receptors were measured in vitro using receptor autoradiography on tissue sections incubated with the SS analog 125I- [Tyr3]-octreotide or 125I-[Leu8,D-Trp22,Tyr25]-SS-28. All tissues were SS-receptor positive for either radioligand, except the thymomas. In thymic tissue, the receptors were diffusely located in the medulla, presumably on epithelial cells. In the spleen, the red pulp was strongly labeled. In the lymph nodes, the germinal centers were preferentially labeled. In all tissues, the receptors were of high affinity (kd thymus, 0.84 nmol/L; kd spleen, 1.6 nmol/L; kd lymph node, 0.62 nmol/L) and specific for SS. Displacement by nanomolar concentrations of SS-14, SS-28, and octreotide was observed, as was guanosine triphosphate dependency. The in vivo visualization of somatostatin receptors was performed after injection of 111In-DTPA- octreotide and gamma-camera scintigraphy. The spleen, but not thymus or lymph nodes, were visualized. These data suggest an important role for SS in regulating immune functions through SS receptors in thymus, spleen, and lymph nodes. Furthermore, SS may regulate neuroendocrine functions in the thymus.

scholarly journals STUDIES ON ANTIBODY PRODUCTION

1955 ◽  
Vol 102 (1) ◽  
pp. 49-60 ◽  
Author(s):  
Albert H. Coons ◽  
Elizabeth H. Leduc ◽  
Jeanne M. Connolly
Keyword(s):  
Lymph Nodes ◽  
Plasma Cells ◽  
Histochemical Demonstration ◽  
Green Fluorescence ◽  
Spleen And Lymph Nodes ◽  
Specific Reactions ◽  
A Minor ◽  
Hyperimmune Rabbit ◽  
In Cells

A method for the specific histochemical demonstration of antibody in cells and parts of cells is described. It consists of carrying out a two stage immunological reaction on frozen sections of tissues: (a) allowing reaction between antibody in the tissue and dilute antigen applied in vitro, and (b) the detection of those areas where this antigen has been specifically absorbed by means of a precipitin reaction carried out with fluorescein-labelled antibody. Examination under the fluorescence microscope reveals the yellow-green fluorescence of fluorescein over those areas where a precipitate has formed. A study of the hyperimmune rabbit on the first few days after the last of a series of intravenous antigen injections reveals that antibody against human γ-globulin or ovalbumin is present in groups of plasma cells in the red pulp of the spleen, the medullary areas of lymph nodes, the submucosa of the ileum, and the portal connective tissue of the liver. Because of extensive non-specific reactions, the bone marrow could not be examined. Small amounts of antibody were occasionally visible in cells in the lymphoid follicles of the spleen and lymph nodes, so that a minor contribution by lymphocytes to antibody synthesis cannot be excluded.

scholarly journals In vitro autoradiographic and in vivo scintigraphic localization of somatostatin receptors in human lymphatic tissue

Blood ◽  
1993 ◽  
Vol 82 (7) ◽  
pp. 2143-2151 ◽  
Author(s):  
JC Reubi ◽  
B Waser ◽  
U Horisberger ◽  
E Krenning ◽  
SW Lamberts ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Gamma Camera ◽  
Somatostatin Receptors ◽  
Immune Functions ◽  
Thymic Tissue ◽  
Tissue Sections ◽  
Spleen And Lymph Nodes ◽  
Neuroendocrine Functions

Receptors for the neuropeptide somatostatin (SS) were evaluated in vitro and in vivo in various human lymphatic tissues, ie, thymus, spleen, and lymph nodes; thymic carcinoids and thymomas were also tested. The receptors were measured in vitro using receptor autoradiography on tissue sections incubated with the SS analog 125I- [Tyr3]-octreotide or 125I-[Leu8,D-Trp22,Tyr25]-SS-28. All tissues were SS-receptor positive for either radioligand, except the thymomas. In thymic tissue, the receptors were diffusely located in the medulla, presumably on epithelial cells. In the spleen, the red pulp was strongly labeled. In the lymph nodes, the germinal centers were preferentially labeled. In all tissues, the receptors were of high affinity (kd thymus, 0.84 nmol/L; kd spleen, 1.6 nmol/L; kd lymph node, 0.62 nmol/L) and specific for SS. Displacement by nanomolar concentrations of SS-14, SS-28, and octreotide was observed, as was guanosine triphosphate dependency. The in vivo visualization of somatostatin receptors was performed after injection of 111In-DTPA- octreotide and gamma-camera scintigraphy. The spleen, but not thymus or lymph nodes, were visualized. These data suggest an important role for SS in regulating immune functions through SS receptors in thymus, spleen, and lymph nodes. Furthermore, SS may regulate neuroendocrine functions in the thymus.

scholarly journals Soufeng sanjie formula alleviates collagen-induced arthritis in mice by inhibiting Th17 cell differentiation

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Di Hua ◽  
Jie Yang ◽  
Qinghai Meng ◽  
Yuanyuan Ling ◽  
Qin Wei ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Lymph Nodes ◽  
Inflammatory Cytokines ◽  
Th17 Cell ◽  
Collagen Induced Arthritis ◽  
Expression Levels ◽  
Th17 Cell Differentiation ◽  
Spleen And Lymph Nodes ◽  
Seed Coats

Abstract Background Rheumatoid arthritis (RA) is a chronic autoimmune disease. Soufeng sanjie formula (SF), which is composed of scolopendra (dried body of Scolopendra subspinipes mutilans L. Koch), scorpion (dried body of Buthus martensii Karsch), astragali radix (dried root of Astragalus membranaceus (Fisch.) Bge), and black soybean seed coats (seed coats of Glycine max (L.) Merr), is a traditional Chinese prescription for treating RA. However, the mechanism of SF in treating RA remains unclear. This study was aim to investigate the anti-arthritic effects of SF in a collagen-induced arthritis (CIA) mouse model and explore the mechanism by which SF alleviates arthritis in CIA mice. Methods For in vivo studies, female DBA/1J mice were used to establish the CIA model, and either SF (183 or 550 mg/kg/day) or methotrexate (MTX, 920 mg/kg, twice/week) was orally administered to the mice from the day of arthritis onset. After administration for 30 days, degree of ankle joint destruction and serum levels of IgG and inflammatory cytokines were determined. The balance of Th17/Treg cells in the spleen and lymph nodes was analyzed using flow cytometry. Moreover, the expression levels of retinoic acid receptor-related orphan nuclear receptor (ROR) γt and phosphorylated STAT3 (pSTAT3, Tyr705) in the spleen were detected by immunohistochemistry. Furthermore, the effect of SF on Th17 cells differentiation in vitro was investigated in CD4+ T cells under Th17 polarization conditions. Results SF decreased the arthritis score, ameliorated paw swelling, and reduced cartilage loss in the joint of CIA mice. In addition, SF decreased the levels of bovine collagen-specific IgG in sera of CIA mice. SF decreased the levels of inflammatory cytokines (TNF-α, IL-6, and IL-17A) and increased the level of IL-10 both in the sera and the joint of CIA mice. Moreover, SF treatment rebalanced the Th17/Treg ratio in the spleen and lymph nodes of CIA mice. SF also reduced the expression levels of ROR γt and pSTAT3 (Tyr705) in the spleen of CIA mice. In vitro, SF treatment reduced Th17 cell generation and IL-17A production and inhibited the expression of RORγt, IRF4, IL-17A, and pSTAT3 (Tyr705) under Th17 polarization conditions. Conclusions Our results suggest that SF exhibits anti-arthritic effects and restores Th17/Treg homeostasis in CIA mice by inhibiting Th17 cell differentiation.

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 Factors affecting the sensitivity and specificity of the cytochemical reaction for the anti-horseradish peroxidase antibody in lymph tissue sections.

1980 ◽  
Vol 28 (7) ◽  
pp. 645-652 ◽  
Author(s):  
W Straus
Keyword(s):  
Horseradish Peroxidase ◽  
Lymph Nodes ◽  
Sensitivity And Specificity ◽  
Peroxidase Activity ◽  
Additional Advantage ◽  
Tissue Sections ◽  
Endogenous Peroxidase ◽  
Cytochemical Reaction ◽  
Cold Acetone ◽  
Antibody Reaction

Factors which increase the sensitivity and specificity of the cytochemical reaction for the antibody to horseradish peroxidase (HRP) in precursors of plasma cells and in lymphocytes were studied in sections of popliteal lymph nodes of rats. The lymph nodes were removed 3-5 days after a secondary injection of HRP into the footpads and were fixed for 5 hr in a 4% cold formaldehyde solution (Straus W: Histochemistry 53:273, 1977). Brief postfixation of the frozen sections with cold acetone improved the retention of the antigen at the sites of the antibody in the precursor cells, and it improved the quality of fixation without appreciably weakening the antigen-binding capacity of the antibody. The cytochemical reaction for the anti-HRP antibody was intensified by staining with diaminobenzidine (DAB) and H2O2 at pH 5-6, or by staining at pH 7.4 in the presence of imidazole. Imidazole partially inhibited endogenous peroxidase activity. Pretreatment with phenylhydrazine prevented nonspecific background adsorption of HRP. Phenylhydrazine had the additional advantage of inhibiting most of the endogenous peroxidase activity (Straus W: J Histochem Cytochem 20:949, 1972). The intensity of the antibody reaction in the proplasma cells developing in the medullary cords varied greatly depending on the stage of maturation from lymphocytes and blast cells. Many lymphocytes in the cortex of the lymph node showed a strong perinuclear antibody reaction when the tissue sections were postfixed with cold acetone, and the peroxidase complexed to the antibody was visualized by staining with DAB and H2O2 at pH 5-6. The antibody reaction also occurred at the surgace of many lymphocytes when the tissue sections, postfixed with cold acetone, were stained with DAB and H2O2 at pH 7.4 in the presence of imidazole. Other lymphocytes showed a strong surface, perinuclear, and cytoplasmic antibody reaction after staining at pH 5-6 as well as after staining at pH 7.4, while yet other lymphocytes remained unstained.

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.

A Basis for Distinguishing Cultured Dendritic Cells and Macrophages in Cytospins and Fixed Sections

2005 ◽  
Vol 8 (1) ◽  
pp. 43-51 ◽  
Author(s):  
Jukka Vakkila ◽  
Michael T. Lotze ◽  
Connie Riga ◽  
Ronald Jaffe
Keyword(s):  
Dendritic Cells ◽  
Cell Types ◽  
Fixed Tissue ◽  
Reliable Identification ◽  
Formalin Fixed Tissue ◽  
Tissue Sections ◽  
Hla Dr ◽  
Formalin Fixed

There is a burgeoning literature on the contrasting role of intratumoral dendritic cells (DCs) and tumor-associated macrophages, making reliable identification of both cell types in clinical and experimental tissue sections important. However, because these cell types are closely related and share several differentiation antigens, their absolute distinction in tissue sections is difficult. We differentiated DCs and macrophages from monocytes in vitro, prepared cytospins and paraffin-embedded sections of the various cell populations, and tested a variety of antibodies that purportedly recognize monocytes and DCs for their capacity to react and distinguish cells after conventional formalin fixation. Cultured DCs but not macrophages were detected by fascin, DC-LAMP, and CD83 with a predictable increase in staining that paralleled their maturation. Staining by CD1a was found on immature DCs but was weak and absent on mature DCs and macrophages, respectively. CD14 and CD163 were characteristic for macrophages and absent on DCs. CD68, HLA-DR, and S100 did not discriminate between DCs and macrophages. We conclude that antigens such as HLA-DR and S100 are not in themselves sufficient for identification of DCs in formalin-fixed tissue sections, but that additional macrophage-specific (CD14, CD163) and DC-specific (CD1a, CD83, fascin, DC-LAMP) antigens should be used to distinguish cell types from each other and to provide information on their state of maturation.

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.

Sign in / Sign up

Export Citation Format

Share Document