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 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 CELLULAR SITES OF FORMATION OF GAMMA GLOBULIN

1957 ◽  
Vol 106 (5) ◽  
pp. 627-640 ◽  
Author(s):  
L. G. Ortega ◽  
R. C. Mellors
Keyword(s):  
Germinal Center ◽  
Gamma Globulin ◽  
Plasma Cells ◽  
Serum Proteins ◽  
Fluorescent Antibody ◽  
Germinal Centers ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Reticular Cells ◽  
Parenchymal Cells

The cellular sites of formation of γglobulin in lymphatic tissues of man and in a representative human lymphoid infiltrate have been studied by fluorescent antibody technique. The findings indicate that γ-globulin is formed in the germinal centers of lymphatic nodules and in the cytoplasm of mature and immature plasma cells of two types—those with and those without Russell bodies. The germinal center cells that synthesize γ-globulin have been designated "intrinsic" cells to distinguish them from the medium and large lymphocytes, and the primitive reticular cells that occur elsewhere and do not produce γ-globulin. Unlike the plasma cells, which function as individual units, the intrinsic cells apparently form γ-globulin only when they are arranged in discrete aggregations. The function, the blood supply, and the systematic cellular arrangement of germinal centers justifies the postulate that they are miniature organs of internal secretion of γ-globulin. The release of γ-globulin from its sites of formation appears to be accomplished by holocrine and apocrine secretion. Presumably, these secretory mechanisms are adaptations required for the production of antibody since they have not been described in parenchymal cells that form the other serum proteins. The cells found to form γ-globulin appear to be identical with those previously shown to form specific antibody in response to a variety of antigens in the experimental animal. This evidence indicates that normal γ-globulin, if it exists, originates in the same cells that produce antibody. It is suggested, also, that each of the 3 morphologically distinct categories of cells that synthesize γ-globulin represents a response to a particular form of antigenic stimulation. Nuclear participation in the process of γ-globulin synthesis was not detected by the technique employed.

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 Use of anti-horseradish peroxidase antibody-gold complex in the ABC technique.

1991 ◽  
Vol 39 (6) ◽  
pp. 863-869 ◽  
Author(s):  
B Gee ◽  
M J Warhol ◽  
J Roth
Keyword(s):  
Horseradish Peroxidase ◽  
Polyclonal Antibodies ◽  
Light And Electron Microscopy ◽  
Gold Complex ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Endogenous Peroxidase ◽  
Pre Treatment ◽  
Lowicryl K4m ◽  
Antigen Antibody

We report a modification of the avidin-biotin-peroxidase complex (ABC) technique for the light and electron microscopic detection of antigens in tissue sections. An immunological approach was used instead of the DAB reaction to reveal ABC bound to antigen-antibody complexes. Affinity-purified polyclonal antibodies against horseradish peroxidase were complexed to particles of colloidal gold and applied for reaction with the horseradish peroxidase molecules of the ABC. For light microscopic immunolabeling, the signal produced by the anti-horseradish peroxidase antibody-gold complex required silver intensification. The ABC immunogold reaction as compared with the standard ABC technique, in particular with silver intensification of the DAB reaction product, provided superior resolution in paraffin sections. Furthermore, section pre-treatment to block endogenous peroxidase activity could be omitted and no potentially hazardous substrate was used. The ABC immunogold reaction was successfully applied for electron microscopic immunolabeling on Lowicryl K4M thin sections. We propose that the ABC immunogold reaction is a useful alternative to the standard ABC technique and can be equally well applied to light and electron microscopy.

scholarly journals THE CELLULAR ORIGIN OF HUMAN IMMUNOGLOBULINS (γ2, γ1M, γ1A)

1963 ◽  
Vol 118 (3) ◽  
pp. 387-396 ◽  
Author(s):  
Robert C. Mellors ◽  
Leonhard Korngold
Keyword(s):  
Germinal Center ◽  
Plasma Cells ◽  
Individual Cell ◽  
Germinal Centers ◽  
Small Extent ◽  
Fetal Thymus ◽  
Cellular Origin ◽  
Human Immunoglobulins ◽  
Intermediate Size ◽  
Reticular Cells

A study was made of the cellular origin of human immunoglobulins (γ2, γ1M, γ1A). The results indicated that two closely related families of cells form immunoglobulins in human lymphoid tissue: germinal (reticular) centers and plasma cells. Thus their cellular origin in addition to their known antigenic relations further justifies placing the immunoglobulins in one family of proteins. Immunoglobulins were also formed to a small extent in primitive reticular cells which resembled those of germinal centers but were separated from them. Possibly such cells were undergoing transition to the much more numerous plasma cells with which they were commonly associated. The mantles of small lymphocytes which surrounded germinal centers did not contain detectable quantities of immunoglobulins. While in general only one type of immunoglobulin was present in an individual cell or germinal center, γ2- and γ1M-globulin were identified on occasion in the same plasma cell and germinal center. A peculiarity of the fetal thymus gland was the presence of immunoglobulin, mainly γ1M, in a small number of cells of small and intermediate size and primitive reticular appearance and in Hassall's corpuscles.

scholarly journals ELECTRON MICROSCOPIC AUTORADIOGRAPHY OF GERMINAL CENTER CELLS IN MOUSE SPLEEN

1965 ◽  
Vol 25 (3) ◽  
pp. 109-119 ◽  
Author(s):  
D. C. Swartzendruber ◽  
M. G. Hanna
Keyword(s):  
Tritiated Thymidine ◽  
Cell Types ◽  
Ultrastructural Level ◽  
Germinal Centers ◽  
Mouse Spleen ◽  
Cell Type ◽  
Electron Microscopic ◽  
Electron Microscopic Autoradiography ◽  
Golgi Region ◽  
Reticular Cells

The fine structure of tritiated thymidine-labeled cells in antigen-stimulated mouse spleen germinal centers is described. In studies on the ultrastructural level, two labeled cell types found in germinal centers are observed. Large lymphocytes are characterized by their very numerous free ribosomes, a paucity of endoplasmic reticulum, relatively few mitochondria, and a poorly developed Golgi region. The nuclei are large and vesicular, and large nucleoli are present. A second labeled cell type appears to contain more mitochondria and has a higher development of the Golgi area. The nucleus contains large, numerous blocks of chromatin, indicative of a more differentiated cell type. Reticular cells, both phagocytic and non-phagocytic, were not observed to be labeled in the germinal centers.

scholarly journals REQUIREMENT FOR CONTINUOUS ANTIGENIC STIMULATION IN THE DEVELOPMENT AND DIFFERENTIATION OF ANTIBODY-FORMING CELLS

1969 ◽  
Vol 129 (5) ◽  
pp. 953-971 ◽  
Author(s):  
M. G. Hanna ◽  
P. Nettesheim ◽  
Mary W. Francis
Keyword(s):  
Germinal Center ◽  
Immune Cell ◽  
Passive Immunization ◽  
Germinal Centers ◽  
Primary Response ◽  
Antigenic Stimulation ◽  
Transfer System ◽  
Cell Compartment ◽  
Cell Conversion ◽  
Development And Differentiation

The essential role of continuous antigenic stimulation in the development and differentiation of antibody-forming cells as defined in the X-Y-Z immune cell maturation scheme was examined in these studies. Mice were primed with sheep erythrocytes (SRBC) in an attempt to induce maximum immune progenitor cell conversion (X → Y). Subsequently antigen was depleted at 1 or 4 days after priming with isologous specific antibody in order to interrupt further immune cell differentiation (Y → Z). It was reasoned that this condition would result in depression of the functional antibody-producing cell compartment as measured in the intact mice and subsequently in enhancement of the sensitized (Y cell) compartment as measured in the spleen cell transfer system. These data were also correlated with systematic studies of the hyperplasia of the spleen germinal centers. The effect of passive antibody on the primary response to SRBC was a marked decrease indirect and indirect hemolysin-producing cells (DPFC and IPFC). However, there was a lack of correlation in the degree of antibody-mediated 19S and 7S immune cell suppression during the primary response, the DPFC being much less depressed than the IPFC. As measured in the transfer system there was an enhanced 19S sensitized cell compartment and a depressed 7S sensitized cell compartment in 1 day passively immunized mice. This was true whether or not transfers were performed 1, 2, or 4 wk after priming. Similarly, there was an enhanced 19S-sensitized cell compartment with little or no effect on the 7S-sensitized. cell compartment in 4 day passively immunized mice. These data suggest that progeny of the antigen-stimulated progenitor cells (X cell), as a consequence of lack of further antigenic stimulation, were forced into maturation arrest. These studies further demonstrate that isologous passive antibody suppresses germinal center growth regardless of whether the antibody is infused 1, 2, or 4 days after priming. In terms of formation of sensitized cells, the marked depression of 7S sensitized cell compartment after passive immunization at 24 hr in contrast to the enhancement of the 19S sensitized cell compartment corresponds to the suppressed growth of germinal centers during the primary response. Thus, if the germinal center is, as suggested, the site of proliferative expansion of immunocompetent cells, these data indicate that the germinal center growth is related to the 7S antibody response and in the formation of "7S memory."

mRNA localization by Electron microscopic in situ hybridization

1991 ◽  
Vol 49 ◽  
pp. 430-431
Author(s):  
J. A. Pollock ◽  
M. Martone ◽  
T. Deerinck ◽  
M. H. Ellisman
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Nucleic Acids ◽  
Recombinant Dna ◽  
Light And Electron Microscopy ◽  
Electron Microscopic ◽  
High Voltage Electron Microscopy ◽  
Functional Sites ◽  
Voltage Electron

Localization of specific proteins in cells by both light and electron microscopy has been facilitate by the availability of antibodies that recognize unique features of these proteins. High resolution localization studies conducted over the last 25 years have allowed biologists to study the synthesis, translocation and ultimate functional sites for many important classes of proteins. Recently, recombinant DNA techniques in molecular biology have allowed the production of specific probes for localization of nucleic acids by “in situ” hybridization. The availability of these probes potentially opens a new set of questions to experimental investigation regarding the subcellular distribution of specific DNA's and RNA's. Nucleic acids have a much lower “copy number” per cell than a typical protein, ranging from one copy to perhaps several thousand. Therefore, sensitive, high resolution techniques are required. There are several reasons why Intermediate Voltage Electron Microscopy (IVEM) and High Voltage Electron Microscopy (HVEM) are most useful for localization of nucleic acids in situ.

Electron Microscopic Characterization and Quantitation of Air Borne Particulate Matter with Special Emphasis on Asbestos

1972 ◽  
Vol 30 ◽  
pp. 356-357
Author(s):  
Peter K. Mueller ◽  
Glenn R. Smith ◽  
Leslie M Carpenter ◽  
Ronald L. Stanley
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Ambient Air ◽  
Quality Standard ◽  
Primary Objective ◽  
Cellulose Ester ◽  
Main Concept ◽  
Electron Microscopic ◽  
Air Samples ◽  
Diffraction Patterns

At the present time the primary objective of the electron microscopy group of the Air and Industrial Hygiene Laboratory is the development of a method suitable for use in establishing an air quality standard for asbestos in ambient air and for use in its surveillance. The main concept and thrust of our approach for the development of this method is to obtain a true picture of fiber occurrence as a function of particle size and asbestos type utilizing light and electron microscopy.We have now available an electron micrographic atlas of all asbestos types including selected area diffraction patterns and examples of fibers isolated from air samples. Several alternative approaches for measuring asbestos in ambient air have been developed and/or evaluated. Our experiences in this regard will be described. The most promising method involves: 1) taking air samples on cellulose ester membrane filters with a nominal pore size of 0.8 micron; 2) ashing in a low temperature oxygen plasma for several hours;

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