scholarly journals Light microscopic, immunohistochemical localization of the pia-glial basal lamina.

1980 ◽  
Vol 28 (4) ◽  
pp. 347-353 ◽  
Author(s):  
T F Kowalski ◽  
H L Vahlsing ◽  
E R Feringa
Keyword(s):  
Basal Lamina ◽  
Wistar Rats ◽  
Staining Technique ◽  
Immunohistochemical Localization ◽  
Basement Membrane Zone ◽  
Rabbit Serum ◽  
Antibody Technique ◽  
Antigenic Source ◽  
Hyperimmune Rabbit ◽  
Different Tissues

The current histologic methods for studying the pia-glial basal lamina (BL) are inappropriate for high contrast, permanent light microscopy preparation. We have developed a staining technique for epithelial BL which is highly specific, extremely sensitive, permanent, relatively inexpensive, and suitable for light or electron microscopy (EM). Central nervous system (CNS) basement membrane zone (BMZ) antigens were isolated by the technique of Meezan (1975) from female albino Wistar rats. Using this CNS BMZ preparation as an antigenic source, a hyperimmune rabbit serum was developed. This serum was exhaustively adsorbed with rat splenic pulp to remove undesirable antibodies to endothelial BL and collagen. The peroxidase-antiperoxidase indirect antibody technique was used to test the staining specificity of this splenic adsorbed serum on different tissues containing BLs of known origin and/or function. The results indicated that this BL staining technique was specific for epithelial BL of the rat and of some other species.

scholarly journals ENZYME-LABELED ANTIBODIES FOR THE LIGHT AND ELECTRON MICROSCOPIC LOCALIZATION OF TISSUE ANTIGENS

1967 ◽  
Vol 33 (2) ◽  
pp. 307-318 ◽  
Author(s):  
Paul K. Nakane ◽  
G. Barry Pierce
Keyword(s):  
Acid Phosphatase ◽  
Basement Membrane ◽  
Fluorescent Antibody ◽  
Enzymatic Reaction ◽  
Immunohistochemical Localization ◽  
Fluorescent Antibody Technique ◽  
Reaction Products ◽  
Electron Microscopic ◽  
Antibody Technique ◽  
Tissue Antigens

Enzymes, either acid phosphatase or horseradish peroxidase, were conjugated to antibodies with bifunctional reagents. The conjugates, enzymatically and immunologically active, were employed in the immunohistochemical localization of tissue antigens utilizing the reaction product of the enzymatic reaction as the marker. Tissues reacted with acid phosphatase-labeled antibodies directed against basement membrane were stained for the enzyme with Gomori's method, and those reacted with peroxidase-labeled antibody were stained with Karnovsky's method. The reaction products of the enzymes localized in the basement membrane. Unlike the preparations of the fluorescent antibody technique, enzyme-labeled antibody preparations were permanent, could be observed with an ordinary microscope, and could be examined with the electron microscope. In the latter, specific localization of antibody occurred in the basement membrane and in the endoplasmic reticulum of cells known to synthesize basement membrane antigens. The method is sensitive because of the amplifying effect of the enzymatic activity. The ultrastructural preservation and localization were better with acid phosphatase-labeled antibody than with peroxidase-labeled antibody, but acid phosphatase conjugated antibody was unstable and difficult to prepare. Peroxidase-antibody conjugates were stable and could be stored for several months at 4°C, or indefiniely in a frozen state.

scholarly journals STUDIES ON HERPETIC INFECTION IN MICE

1946 ◽  
Vol 84 (5) ◽  
pp. 429-447 ◽  
Author(s):  
Charles A. Evans ◽  
Howard B. Slavin ◽  
George Packer Berry
Keyword(s):  
Nervous System ◽  
Immune Serum ◽  
Rabbit Serum ◽  
Herpetic Infection ◽  
Focus Of Infection ◽  
System 2 ◽  
The Central Nervous System ◽  
Foot Pad ◽  
Old Mice ◽  
Hyperimmune Rabbit

Two-week-old mice inoculated with herpes virus on the pad of a hind foot regularly developed paralysis of the infected limb followed by paraplegia and encephalitis terminating fatally 5 or 6 days after inoculation. Hyperimmune rabbit serum given intraperitoneally at the time virus was inoculated on the foot pad prevented the formation of an herpetic lesion of the foot pad. When the antiserum was given 12 hours after inoculation of the virus, a typical infection of the epithelium of the foot pad developed, but the virus was prevented from causing obvious signs of infection of the nervous system in many of the animals. Amputation of the foot 2 hours after the inoculation of the virus prevented the paralysis of the hind leg. Some of the mice died of a delayed encephalitis. Amputation of the foot at 24 hours neither prevented nor delayed the sequence of paralysis of the hind leg, encephalitis, and death. In order to study immune serum therapy of an infection of the nervous system uncomplicated by a peripheral focus of infection or by traumatic disturbance of the central nervous system, 2-week-old mice were inoculated on the foot pad, the infected feet were amputated 24 hours later, and the immune serum was administered at varying intervals thereafter. Using litter mate controls and statistically significant numbers of mice, it was shown that hyperimmune rabbit serum, administered during the first one-third of the incubation period, retards and, in some cases, arrests the progress of herpetic infection within the nervous system.

scholarly journals Ultrastructural immunohistochemical localization of anti-invasion factor (AIF) in bovine cartilage matrix.

1982 ◽  
Vol 30 (6) ◽  
pp. 524-531 ◽  
Author(s):  
F H Wezeman ◽  
G V Childs
Keyword(s):  
Staining Technique ◽  
Ultrastructural Localization ◽  
Immunohistochemical Localization ◽  
Cartilage Matrix ◽  
Fixed Tissue ◽  
Antibody Complex ◽  
Protein Components ◽  
Antigen Antibody ◽  
Unlabeled Antibody ◽  
Bovine Cartilage

Rabbit antibodies prepared against bovine cartilage anti-invasion factor (AIF) were tested for their affinity toward antigenic sites in glutaraldehyde-fixed bovine hyaline cartilage matrix. Ultrastructural localization of the antigen-antibody complex was accomplished by the unlabeled antibody peroxidase-antiperoxidase staining technique. Unextracted and salt-extracted (1 M NaCl or 3 M GuHCl) cartilage slices were incubated with anti-AIF antibodies at a working dilution of 1:20,000. Staining occurred in unextracted matrix distributed throughout the tissue, but with regional variation in the lacunar matrix. Significantly less stain was noted in extracted tissues. The results suggest that at least certain protein components in AIF are morphologically associated with matrix complexes in aldehyde-fixed tissue.

scholarly journals Immunohistochemical localization of human immunoglobulins and lysozyme in epoxy-embedded lymph nodes: effect of different fixatives and of proteolytic digestion.

1982 ◽  
Vol 30 (7) ◽  
pp. 630-636 ◽  
Author(s):  
P Dell'Orto ◽  
G Viale ◽  
R Colombi ◽  
P Braidotti ◽  
G Coggi
Keyword(s):  
Lymph Nodes ◽  
Staining Technique ◽  
Immunohistochemical Localization ◽  
Immunoperoxidase Staining ◽  
Proteolytic Digestion ◽  
Enzymatic Pretreatment ◽  
Heavy Chains ◽  
Tissue Sections ◽  
Human Immunoglobulins ◽  
Immunohistochemical Studies

The postembedding immunoperoxidase staining technique for the localization of immunoglobulins (light and heavy chains) and of lysozyme has been successfully applied to epoxy-embedded human lymph nodes, after removal of the resin. Glutaraldehyde-containing fixatives appear to be suitable for the immunohistochemical localization of human immunoglobulins and lysozyme, provided that the masked antigenicity of these proteins is recovered by proteolytic digestion of the tissue sections using 0.4% pepsin or 0.1% trypsin. Nonglutaraldehyde-containing fixatives allow the immunolocalization of human immunoglobulins without any enzymatic pretreatment. This study shows that tissues routinely fixed in glutaraldehyde and embedded for ultrastructural investigations are actually suitable for immunohistochemical studies on human immunoglobulins and lysozyme.

scholarly journals ULTRASTRUCTURAL LOCALIZATION OF A SOLUBLE COLLAGEN ANTIGEN

1971 ◽  
Vol 19 (11) ◽  
pp. 663-669 ◽  
Author(s):  
LIVIA LUSTIG
Keyword(s):  
Chick Embryo ◽  
Light And Electron Microscopy ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Collagen Fibrils ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Neutral Salt ◽  
Antibody Technique ◽  
Soluble Collagen

Soluble collagen antigen was localized at the ultrastructural level using an enzyme-labeled antibody technique. Specific antiserum against neutral salt-soluble collagen was conjugated with horseradish peroxidase. Tissue culture of fibroblasts, sections of chick embryo connective tissue and skin and tendon of newborn and adult chickens were incubated with the conjugated antiserum and processed for light and electron microscopy. Light microscopic observations showed a localization of the collagen antigen in the cytoplasm of fibroblasts and in the extracellular collagen fibers. Ultrastructurally, the antigen was found on the ribosomes, on the membranes of the rough endoplasmic reticulum, in the intracisternal material of fibroblasts and extracellularly in the collagen fibrils. The collagen fibrils of chick embryo cartilage were intensely stained while the matrix itself remained unstained. Controls performed with absorbed antiserum, conjugated normal rabbit serum or different antigens failed to react with the tissues.

scholarly journals Phage Lytic Enzyme Cpl-1 as a Novel Antimicrobial for Pneumococcal Bacteremia

2003 ◽  
Vol 71 (11) ◽  
pp. 6199-6204 ◽  
Author(s):  
Jutta M. Loeffler ◽  
Svetolik Djurkovic ◽  
Vincent A. Fischetti
Keyword(s):  
Streptococcus Pneumoniae ◽  
Treatment Efficacy ◽  
Lytic Enzyme ◽  
Rabbit Serum ◽  
Resistant Bacteria ◽  
Intravenous Therapy ◽  
Antibiotic Resistant ◽  
Pneumococcal Bacteremia ◽  
Hyperimmune Rabbit

ABSTRACT Streptococcus pneumoniae is becoming increasingly antibiotic resistant worldwide, and thus new antimicrobials are badly needed. We report the use of Cpl-1, the lytic enzyme of a pneumococcal bacteriophage, as an intravenous therapy for pneumococcal bacteremia in a mouse model. A 2,000-μg dose of Cpl-1 reduced pneumococcal titers from a median of log10 4.70 CFU/ml to undetectable levels (<log10 2.00 CFU/ml) within 15 min. This dose given 1 h after intravenous infection led to 100% survival at 48 h, compared to the 20% survival of buffer-treated controls. In advanced bacteremia, treatment with two doses at 5 and 10 h still resulted in significantly longer survival (P < 0.0001) and a hazard ratio of 0.29 (95% confidence interval, 0.04 to 0.35). The enzyme is immunogenic, but the treatment efficacy was not significantly diminished after previous intravenous exposure of mice and hyperimmune rabbit serum did not neutralize the activity. Cpl-1 is also very effective as a topical nasal treatment against colonization by S. pneumoniae. In vitro, the enzyme is active against many serotypes of S. pneumoniae, independent of their penicillin resistance, and it is very specific for this species. Bacteriophage enzymes are unusual but extremely effective antimicrobials and represent a new weapon against infections with resistant bacteria.

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