Demonstration of <i>Listeria Monocytogenes</i> with the PAP Technique in Formalin Fixed and Paraffin Embedded Tissues of Experimentally Infected Mice

Domingo, Mariano; Ramos, Jonathan Hernández; Domı́nguez, Lucas; Ferrer, L.; Marco, Alberto

doi:10.1111/j.1439-0450.1986.tb00065.x

Cited by 23 publications

(8 citation statements)

References 15 publications

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“…Reaction was developed with the avidin-biotin horseradish peroxidase complex using as chromogen 0.05% solution of 3,3Јdiamenobenzidine (Sigma Chemical Co., St. Louis, MO) with 0.03% H 2 O 2 in 0.1 mol/L imidazole buffer (pH 7.1). L. monocytogenes was immunohistochemically detected by a modified technique from Domingo et al 13 using the avidin-biotin complex system (Dako). Sections incubated with isotype-matched antibodies served as negative controls in each technique.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Expression of E-Selectin, P-Selectin, and Intercellular Adhesion Molecule-1 during Experimental Murine Listeriosis

López¹,

Prats²,

Marco³

1999

The American Journal of Pathology

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The expression of adhesion molecules E-selectin, P-selectin, and intercellular adhesion molecule-1 (ICAM-1) was immunohistochemically investigated during the course of experimental murine listeriosis. Infection was monitored by microbiological count of blood, liver, and spleen. After an early generalized expression of P-selectin and ICAM-1, a later regulation occurred specifically to areas of inflammation. Expression of E-selectin was faint and inconstantly detected in all of the studied organs. In the liver, typical lesions of murine listeriosis were related to the expression of ICAM-1 on sinusoidal endothelial cells and the biliary system and to the de novo expression of P-selectin in hepatic portal vessels. Inflammation in the spleen was related to the expression of ICAM-1 on red pulp sinusoidal cells, especially in the marginal sinus. High endothelial venules of inflamed lymph nodes also expressed P-selectin and ICAM-1. Lesions in the central nervous system appeared on day 3 after infection as a pyogranulomatous leptomeningitis associated with an intense expression of P-selectin and ICAM-1 in meningeal vessels, especially those in the hippocampal sulcus, suggesting a way through which inflammation initially reach the central nervous system during experimental murine listeriosis. Leptomeningitis was followed by the presence of ventriculitis, which was related to the up-regulation of ICAM-1 on choroid plexus epithelial cells, periventricular vessels and ependymal cells. Up-regulation of P-selectin and ICAM-1 during experimental murine listeriosis could play an important role in the recruitment of leukocytes, especially to the liver, lymphoid organs, and central nervous system.

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Section: Immunohistochemistrymentioning

confidence: 99%

Expression of E-Selectin, P-Selectin, and Intercellular Adhesion Molecule-1 during Experimental Murine Listeriosis

López¹,

Prats²,

Marco³

1999

The American Journal of Pathology

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“…This immunoperoxidase technique also presents the advantage of using purified sera instead of whole sera, thus reducing background staining. Moreover, the use of an only sort of biotin-labeled purified antibody permits to remove two steps of conventional immunochemical tests: the incubation with a labeled secondary antibody and with a normal serum of the same species in which secondary antibody is produced (DOMINGO et al, 1986). This fact simplifies, shortens and makes cheaper this methodology, also diminishing inespecific reactions.…”

Section: Discussionmentioning

confidence: 99%

“…A possible disadvantage of the diagnosis based on formalin-fixed and paraffinembedded tissues could be the reduction of the antigenicity of bacteria, because the high temperature and lipid-solvent reagents could remove or alter outer antigenic material of the bacterial structure (DOMINGO et al, 1986). However, the great amount of positive results obtained seems to indicate that the histological processing of tissues do not decrease the antigenicity of A .…”

Section: Discussionmentioning

confidence: 99%

Evaluation of an Immunoperoxidase Technique Using an Only Biotin‐Labeled Antibody for the Demonstration of Actinobacillus pleuropneumoniae in Tissue Sections

Gutiérrez¹,

Rodríguez-Barbosa²,

Tascon³

et al. 1993

Journal of Veterinary Medicine, Series B

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Summary In this report an immunoperoxidase technique (IPB) using an only biotin‐labeled antibody is compared to culture isolation method (CIM) for the demonstration of Actinobacillus pleuropneumoniae serotype 1 in tissues of mice inoculated intravenously. The organism was isolated in 86.7% of mice and detected by IPB in 100% of cases. A. pleuropneumoniae antigen was mainly demonstrated in the liver, spleen and lungs, but also in the kidney and brain, being especially located in the cytoplasm of macrophages. Significant differences were observed between the results obtained by both tests (P< 0.001). Cross‐reactions occurred by IPB when a rabbit anti‐A. pleuropneumoniae serotype 11 serum was tested but never when an anti‐serotype 9 serum was used. The immunoperoxidase test here described yielded much better results than CIM and it could be useful in routine diagnosis of A. pleuropneumoniae infections.

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“…Auf3er durch den kulturellen Erregernachweis konnen histologische Verdachtsdiagnosen durch den immunhistologischen Nachweis von Listerien-Antigen abgesichert werden. I n dieser Arbeit sol1 die beim Listerien-Antigen-Nachweis in formalinfixierten Paraffinschnitten bereits erprobte ungekoppelte Peroxidase-Antiperoxidase-(PAP)-Technik (2,3,8,11) rnit der Avidin-Biotin-Peroxidase-Komplex-(ABC)-Technik bei der Diagnose der Listerienenzephalitis verglichen werden, d a erste Untersuchungen (4) unter Verwendung der ABC-Methode auf eine qualitative Verbesserung beim Listerien-Antigen-Nachweis hindeuten.…”

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Untersuchungen zur Diagnostik der Listerienenzephalitis bei Wiederkäuern unter Anwendung kultureller und immunhistologischer Verfahren

Peters

Hewicker‐Trautwein

Amtsberg

1992

Journal of Veterinary Medicine, Series B

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Summary Studies on the diagnosis of listeric encephalitis in ruminants using cultural and immunohistological techniques II. Communication: Immunohistological investigations on formalin‐fixed paraffin sections The unlabelled peroxidase‐antiperoxidase (PAP)‐technique was compared with the avidin‐biotin‐peroxidase‐complex (ABC)‐technique in the identification of Listeria antigen in formalin‐fixed paraffin sections of 58 ruminant brains, 44 of which showed histopathological lesions typical for listeric encephalitis. Rabbit hyperimmune serum, obtained by immunization with a Listeria monocytogenes strain of serotype 1/2 a, served as primary serum in the immunohistochemical investigations. The antiserum was tested for specificity using formalin‐fixed smears of various bacterial species. Listeria antigen was demonstrated in 40 of the 44 brains with histopathological CNS alterations, as seen in listeriosis, using the ABC method, whereas their identification with the PAP method only succeeded in 34 of the brains. Despite the higher dilution of the primary antibody, the ABC method distinguished itself further, in comparison with the PAP method, through more intense immunohis‐tochemical staining of Listeria antigen. Listeria spp. could be isolated from 46 of 52 brains, which were also examined bacteriologically. They could be isolated from 14 brains, which showed no histopathological lesions indicative of listeriosis. In contrast to this, Listeria antigen was only detected immunohistologically in brains with typical histological listeric CNS alterations. In three cases the presumptive histological diagnosis could not be confirmed immunohistologically. Zusammenfassung An formalinfixierten Paraffinschnitten von 58 Wiederkäuergehirnen, von denen 44 histopathologische Veränderungen wie bei Listeriose zeigten, wurde die ungekoppelte Peroxidase‐Antiperoxidase‐(PAP)‐Methode mit der Avidin‐Biotin‐Peroxidase‐Komplex‐(ABC)‐Methode beim Listerien‐Antigen‐Nachweis verglichen. Als Primärserum für die immunhistologischen Untersuchungen diente ein durch Immunisierung mit einem L. m.‐Stamm der Serovar 1/2a hergestelltes Kaninchenhyperimmunserum, welches hinsichtlich seiner Spezifität an formalinfixierten Objektträgerausstrichen verschiedener Bakterienspezies überprüft wurde. In 40 von 44 Gehirnen mit auf Listeriose hinweisenden histopathologischen Veränderungen konnte mit Hilfe der ABC‐Methode Listerien‐Antigen darge‐stellt werden, während der Nachweis mittels PAP‐Methode nur in 34 der 44 Gehirne gelang. Trotz einer höheren Gebrauchsverdünnung des primären Antiserums zeichnete sich die ABC‐Methode gegenüber der PAP‐Methode zudem durch eine höhere Farbintensität des immunhistochemisch dargestellten Listerien‐Antigens aus. Listeria spp. wurden aus 46 der 52 auch bakteriologisch untersuchten Gehirne isoliert. In 14 Fällen gelang der kulturelle Nachweis, ohne daß sich histologische Veränderungen wie bei Listeriose fanden. Demgegenüber ließ sich Listerien‐Antigen immunhistologisch ausschließlich in Gehirnen mit auf Listeriose hinweisenden histop...

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Demonstration of Listeria Monocytogenes with the PAP Technique in Formalin Fixed and Paraffin Embedded Tissues of Experimentally Infected Mice

Cited by 23 publications

References 15 publications

Expression of E-Selectin, P-Selectin, and Intercellular Adhesion Molecule-1 during Experimental Murine Listeriosis

Expression of E-Selectin, P-Selectin, and Intercellular Adhesion Molecule-1 during Experimental Murine Listeriosis

Evaluation of an Immunoperoxidase Technique Using an Only Biotin‐Labeled Antibody for the Demonstration of Actinobacillus pleuropneumoniae in Tissue Sections

Untersuchungen zur Diagnostik der Listerienenzephalitis bei Wiederkäuern unter Anwendung kultureller und immunhistologischer Verfahren

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