Effects of Canavalia lineata lectin-bound "vaccine" on the proliferation of S-180 sarcoma cells in mice.
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Animals↗
Biomedical subjects
S M Hsu
Publications and source records attributed to S M Hsu.
Russell bodies: a light and electron microscopic immunoperoxidase study.
Russell bodies have been previously regarded as aggregates of immunoglobulin. Light and electron microscopic immunoperoxidase studies show no detectable immunoglobulin determinants in the Russell body cores. Denaturation of antigens during tissue preparation appears to be an unlikely explanation, since immunoglobulins in the cytoplasm of plasma cells are clearly demonstrated. The presence of immunoglobulins on the surface of small intracellular Russell bodies may represent the immunoglobulin determinants in the surrounding rough endoplasmic reticulum. It seems likely that Russell bodies contain non-immunoglobulin molecules, by-products of immunoglobulin synthesis, or some altered form of immunoglobulins that no longer can be recognized by the anti-immunoglobulin antibody. The non-uniform dye staining pattern of Russell bodies further suggests that Russell bodies may be heterogenous in nature.
Fluorescent Antibody Technique↗
Warthin's tumor--epithelial cell differences.
An immunohistochemical technic to localize IgA and a histochemical technic to demonstrate beta-D-galactose and N-acetyl-S-galactosamine by peanut agglutinin are applied to paraffin-embedded sections of Warthin's tumor. The luminal layer of epithelium is positive for IgA, while the basal layer is positive for peanut agglutinin. The luminal layer epithelium apparently serves as the route through which IgA (derived from plasma cells in the lymphoid stroma) is secreted into the cystic spaces. The apical cytoplasmic staining for peanut agglutinin in the basal layer is uniquely different from the peanut agglutinin staining pattern in the normal parotid gland. We conclude that the two-layer epithelium of Warthin's tumor is different in morphology, biochemistry, and function.
Adenolymphoma↗
Demonstration of myoglobin in formalin-fixed renal sections by immunoperoxidase technic.
The use of an immunohistochemical technic offers a very sensitive method to detect myoglobin in formalin-fixed, paraffin-embedded tissues. Thus, the technic is suitable for retrospective studies. Myoglobin can be demonstrated at the brush border and apical cytoplasm of the tubular epithelium of proximal tubules. Granular substances and casts in the lumens also react positively with antimyoglobin. Our findings confirm the results of a previous animal study, showing that myoglobin is filtered through glomeruli and reabsorbed by the proximal tubules.
Cytoplasm↗
Discovery of a high affinity of Phaseolus vulgaris agglutinin (PHA) with gastrin-secreting cells.
A lectin, Phaseolus vulgaris agglutinin (PHA), isolated from the red kidney bean, was found to have a very high affinity with gastrin-secreting cells (G cells). The unique PHA-G cell interaction is confirmed by (1) G cell reaction only with PHA, not with other lectins, and (2) PHA reaction with G cells but not with other endocrine cells. The authors concluded that gastrin-secreting cells are composed of a large quantity of galactose-N-acetyl-glucose-manose-containing glycoconjugates. The significance of this glyco-conjugate in G cells is not ascertained. The use of a PHA histochemical technic may well be suitable to study the carbohydrate (or pathophysiological) changes in G cells.
Endocrine Glands↗
Concanavalin A (CON A) reaction with plasma membrane.
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Cell Membrane↗
Color modification of diaminobenzidine (DAB) precipitation by metallic ions and its application for double immunohistochemistry.
Three metallic ions, NiCl2, CoCl2, and CuSO4, were found to modify the color of the normally brown diaminobenzidine (DAB) reaction. The colors ranged from purplish blue (NiCl2), dark blue/bluish black (CoCl2), to greyish blue (CuSO4). We have found that the CoCl2 + DAB is the ion of choice because: 1) it yields a distinct dark blue color that is easily distinguishable from brown DAB; 2) the blue reaction product is very stable throughout the entire staining procedure; and 3) background staining is minimal. These findings can be applied to the double staining technique of two different antigens in the same section. Among three staining procedures discussed, the avidin-biotin peroxidase complex (Co-DAB)-peroxidase-antiperoxidase (DAB) technique produced the best results because: 1) no antibody elution was needed following the avidin-biotin-peroxidase complex procedure when the CoCl2-DAB modification was used; and 2) no background staining occurred.
3,3'-Diaminobenzidine↗
Versatility of biotin-labeled lectins and avidin-biotin-peroxidase complex for localization of carbohydrate in tissue sections.
Carbohydrate residues in tissue can be localized by a very sensitive, specific, and simple technique, utilizing a biotin-labeled lectin followed by an avidin-biotin-peroxidase complex (ABC). Distribution of peanut receptors in benign and malignant tissues was found to be quite different. Normal colonic mucosa occasionally shows small dot staining in the supranuclear cytoplasm, while the tumor cells of colonic adenocarcinoma tend to show diffuse cytoplasmic or membranous staining. Normal breast ductal epithelium shows membranous staining at luminal borders, while medullary carcinoma of the breast shows distinct membranous and cytoplasmic staining. The use of this avidin-biotin-peroxidase complex technique may contribute to understanding the process of tumorigenesis as well as providing a sensitive method to determine early stages of malignant transformation.
Avidin↗
Medullary carcinoma of breast: an immunohistochemical study of its lymphoid stroma.
Immunoperoxidase staining for IgA, IgG, IgM, and secretory component (SC) was performed on ten cases each of medullary carcinoma and infiltrating duct carcinoma of breast. Plasma cell-rich stroma of medullary carcinoma was found to contain predominantly IgA plasma cells. Tumor cells also contained IgA and SC. In contrast, the few plasma cells of infiltrating duct carcinoma were found to be predominantly IgG type, and the tumor cells contained none or very small amounts of IgA and SC. Because the presence of IgA plasma cells and IgA and SC in lining epithelial cells are characteristic features of organs of the secretory Ig system, these findings in medullary carcinoma may suggest a good degree of functional differentiation of these tumor cells and correlate well with the well-known favorable prognosis associated with this tumor.
Breast Neoplasms↗
Warthin's tumor: an immunohistochemical study of its lymphoid stroma.
The lymphoid stroma of Warthin's tumor was studied using an immunoperoxidase method. We found that IgA plasma cells constituted approximately 80 per cent of the total number of plasma cells in this tumor. IgA was present also in the tumor epithelium and in the intraluminal sections. The predominance of IgA plasma cells in Warthin's tumor is in contrast to the lymphoid pattern observed in autoimmune diseases and usual host-tumor reactions. It seems likely that the lymphoid stroma of Warthin's tumor represents an exaggerated secretory immune response. The neoplastic tissue of Warthin's tumor may elaborate or contain an unknown factor that causes the accumulation of lymphoid tissue and the differentiation of IgA plasma cells.
Adenolymphoma↗
Spironolactone bodies. An immunoperoxidase study with biochemical correlation.
The present immunoperoxidase study attempted to localize aldosterone and cortisol in formalin-fixed paraffin-embedded adrenal glands or tumors. Apparently, steroid substances are solubilized during routine tissue processing, and the cytoplasm of adrenal cortical cells is not stained by either antialdosterone or anticortisol antibodies. However, spironolactone bodies react positively with antialdosterone antibody. The presence of aldosterone in the concentric laminations of spironolactone bodies supports the concept that spironolactone bodies are not artifacts. Furthermore, the fact that aldosterone does survive in spironolactone bodies suggests that it is bound in some unusual form. The spironolactone bodies are probably derived from endoplasmic reticulum, and the membranes of endoplasmic reticulum are considered capable of storing steroids. It is likely that during the formation of spironolactone bodies are probably derived from endoplasmic reticulum, and the membranes of endoplasmic reticulum are considered capable of storing steroids. It is likely that during the formation of spironolactone bodies, aldosterone or a few cortisol-like substances are trapped in these laminated concentric bodies.
Adrenal Glands↗
A comparative study of the peroxidase-antiperoxidase method and an avidin-biotin complex method for studying polypeptide hormones with radioimmunoassay antibodies.
A highly sensitive immunoenzymatic technic is presented. The method involves three sequential steps: (1) primary antibody, (2) biotin-labeled secondary antibody, and (3) avidin-biotin-peroxidase complex. Avidin, an egg white protein, has four binding sites for the low-molecular-weight vitamin biotin. Many moieties of biotin can be coupled to the peroxidase molecule. Thus, since a relatively large amount of avidin is incubated with biotin-labeled peroxidase, avidin serves as a link between biotin-peroxidase molecules; in turn, biotin-peroxidase serves as a link between avidin molecules. Consequently, this large lattice-like complex with biotin-binding capability can be attracted to the sites of biotin-labeled antibody, producing a superior staining sensitivity. Several commercially available radioimmunoassay antibodies (e.g., antiglucagon, prolactin, gastrin, growth hormone, and thyroid-stimulating hormone antibodies) were tested for immunohistochemical staining. The unlabeled antibody peroxidase-antiperoxidase method fails to stain gastrin or thyroid-stimulating secretory cells when using these antibodies, and a relatively high antibody concentration is required to produce a positive reaction for glucagon, prolactin, and growth hormone. In contrast, the avidin-biotin-peroxidase complex method successfully demonstrates polypeptide hormones even when antibodies are diluted 20 to 40 times.
Antibodies↗
The use of antiavidin antibody and avidin-biotin-peroxidase complex in immunoperoxidase technics.
Avidin has an extraordinary affinity for the small-molecule vitamin biotin. Covalently coupling biotin or avidin to peroxidase molecules does not interfere with their normal biochemical functions. The avidin or biotin molecules, either peroxidase conjugated or unconjugated, can be brought to the antigen sites by means of an antiavidin antibody. Several immunohistochemical staining technics based on this principal have been described. The method utilizing an avidin-biotin-peroxidase complex was found to be more sensitive than the unlabeled antibody (PAP) method. This method involved four sequential staining procedures: (1) primary antibody (goat anti-human antigen); (2) secondary antibody (rabbit antigoat IgG) added in relative excess; (3) goat antiavidin antibody; (4) avidin-biotin-peroxidase complex. The applications of this technic are discussed.
Animals↗
Protein A, avidin, and biotin in immunohistochemistry.
Conjugated and unlabeled peroxidase antibody methods have proven to be quite satisfactory in localizing sites of antigen-antibody reaction. The use of avidin-biotin-peroxidase complex (ABC), as well as protein A, can contribute significantly to the field of immunohistochemistry. The sensitivity and specificity of several immunohistochemical methods is compared. In general, the ABC method produced the most intense staining and the least background staining of any method tested. The unlabeled antibody (peroxidase-antiperoxidase: PAP) method also yielded satisfactory results, but it was less intense than the ABC method. In comparison to the PAP method, the indirect conjugated method presented slightly inferior staining intensities and significantly higher background staining. Protein A techniques produced a range of staining sensitivities similar to or inferior to the PAP technique. The main disadvantage in using protein A is that it reacts with intrinsic immunoglobulin (Ig) G, thus producing an intense background. Therefore, its use is not recommended on tissues that have either abundant immunoglobulins in their interstitium or numerous IgG-containing plasma cells.
Avidin↗
Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures.
The use of avidin-biotin interaction in immunoenzymatic techniques provides a simple and sensitive method to localize antigens in formalin-fixed tissues. Among the several staining procedures available, the ABC method, which involves an application of biotin-labeled secondary antibody followed by the addition of avidin-biotin-peroxidase complex, gives a superior result when compared to the unlabeled antibody method. The availability of biotin-binding sites in the complex is created by the incubation of a relative excess of avidin with biotin-labeled peroxidase. During formation of the complex, avidin acts as a bridge between biotin-labeled peroxidase molecules; and biotin-labeled peroxidase molecules, which contains several biotin moieties, serve as a link between the avidin molecules. Consequently, a "lattice" complex containing several peroxidase molecules is likely formed. Binding of this complex to the biotin moieties associated with secondary antibody results in a high staining intensity.
Avidin↗
Self-sandwich method. An improved immunoperoxidase technic for the detection of small amounts of antigens.
The unlabeled antibody peroxidase-antiperoxidase (PAP) sequence has been widely accepted as the most sensitive method for demonstrating antigens in paraffin sections. However, the results have been unpredictable in routine surgical specimens, with frequent false-negative stains. In order to amplify the staining reactions a self-sandwich method was used. The amplifier (antigen itself) was added after the application of specific antibody. The procedure may be repeated several times before the application of the bridge antiserum followed by PAP complexes. The method was designed to increase the number of antigen-antibody layers without increasing the number of heteroantisera in the system. Specimens derived from routine tonsillectomy were studied semi-quantitatively. Based on the number of positively stained Ig-containing cells in germinal centers, the sensitivity of the self-sandwich method was estimated to be 20 to 50 times that of the PAP method. In addition, extracellular Ig (reticular staining) and surface Ig were also stained positively in the germinal centers and lymphocytic mantles, respectively, but were not demonstrated by the PAP method. The enhancement of the sensitivity was achieved without compromising the specificity.
Adolescent↗
Demonstration of IgA and secretory component in human hepatocytes.
By the immunoperoxidase technique, immunoglobulin A(IgA) was demonstrated in 50% of human hepatocytes. The positively stained cells showed a tendency towards periportal clustering. Very few plasma cells were identified in the liver. The intrahepatocyte IgA is most probably derived from serum and on its way to the bile ducts. Secretory component (SC) had the same distribution as that of IgA. The presence of SC in the hepatocytes reflects either the synthesis of SC or the transport of pre-assembled SC-IgA by liver cells. The significance of transfer of IgA from serum into bile is as yet unknown. However, the transhepatic passage of IgA may represent a reinforcement of local intestinal immunity, as part of the gut-originated IgA antibodies, which fail to go through the intestinal mucosa, may still gain access to the lumen of the gut.
Bile Ducts, Intrahepatic↗
Clinical applications of immunoperoxidase technique.
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Fluorescent Antibody Technique↗