Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes).
Jacqueline Cordell,Brunangelo Falini,Wendy N. Erber,A. K. Ghosh,Zainalabideen Abdulaziz,S M MacDonald,Karen Pulford,Harald Stein,D Y Mason +8 more
TLDR
The APAAP technique was found particularly suitable for labeling cell smears and for detecting low numbers of antigen-bearing cells in a specimen and could be used in conjunction with immunoperoxidase methods for double immunoenzymatic staining.Abstract:
A murine monoclonal antibody specific for calf intestinal alkaline phosphatase has been prepared and used in an unlabeled antibody bridge technique for labeling monoclonal antibodies. This procedure--the alkaline phosphatase monoclonal anti-alkaline phosphatase (APAAP) method--gives excellent immunocytochemical labeling of tissue sections and cell smears, comparable in clarity and intensity to that achieved with immunoperoxidase labeling. If the enzyme label is developed with a naphthol salt as a coupling agent and Fast Red or hexazotized new fuchsin as a capture agent, a vivid red reaction product is obtained which is very easily detected by the human eye. For this reason the APAAP technique was found particularly suitable for labeling cell smears (for both cytoplasmic and surface-membrane antigens) and for detecting low numbers of antigen-bearing cells in a specimen (e.g., carcinoma cells in a malignant effusion). It was found possible to enhance the intensity of the APAAP labeling reaction substantially by repeating the second and third incubation steps (i.e., the unlabelled "bridge" antibody and APAAP complexes). The APAAP technique was superior to immunoperoxidase labeling for staining tissues rich in endogenous peroxidase, and could be used in conjunction with immunoperoxidase methods for double immunoenzymatic staining. The method was also applicable to the detection of antigenic molecules following their electrophoretic transfer from SDS-polyacrylamide gels to nitrocellulose sheets ("immunoblotting").read more
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CD40 ligand on activated platelets triggers an inflammatory reaction of endothelial cells
Volker Henn,Joseph R. Slupsky,Michael Gräfe,Ioannis Anagnostopoulos,Reinhold Förster,Gert Müller-Berghaus,Richard A. Kroczek +6 more
TL;DR: In this paper, platelets express CD40L within seconds of activation in vitro and in the process of thrombus formation in vivo, indicating that platelets are not only involved in haemostasis but that they also directly initiate an inflammatory response of the vessel wall.
Journal ArticleDOI
Identification of tissue transglutaminase as the autoantigen of celiac disease
Walburga Dieterich,Tobias Ehnis,Michael Bauer,Peter Donner,Umberto Volta,Ernst Otto Riecken,Detlef Schuppan +6 more
TL;DR: Tissue transglutaminase is identified as the unknown endomysial autoantigen of celiac disease, and gliadin is a preferred substrate for this enzyme, giving rise to novel antigenic epitopes.
Journal ArticleDOI
The expression of the Hodgkin's disease associated antigen Ki-1 in reactive and neoplastic lymphoid tissue: evidence that Reed-Sternberg cells and histiocytic malignancies are derived from activated lymphoid cells.
Harald Stein,D Y Mason,Johannes Gerdes,N O'Connor,J. S. Wainscoat,Gorm Pallesen,K C Gatter,B Falini,Georges Delsol,Hilmar Lemke,R Schwarting,Karl Lennert +11 more
TL;DR: Results obtained indicate that Ki-1 antigen is an inducible lymphoid-associated molecule that identifies a group of hitherto poorly characterized normal and neoplastic large lymphoid cells in Hodgkin's disease and Disorders in which only a minority of cells express Ki- 1 antigen probably represent lesions in whichonly some of the abnormal cells have transformed into an "activation state.
Journal ArticleDOI
Cytoplasmic Nucleophosmin in Acute Myelogenous Leukemia with a Normal Karyotype
Brunangelo Falini,Christina Mecucci,Enrico Tiacci,Myriam Alcalay,Roberto Rosati,Laura Pasqualucci,Roberta La Starza,Daniela Diverio,Emanuela Colombo,Antonella Santucci,Barbara Bigerna,Roberta Pacini,Alessandra Pucciarini,Arcangelo Liso,Marco Vignetti,Paola Fazi,Natalia Meani,Valentina Pettirossi,Giuseppe Saglio,Franco Mandelli,Francesco Lo-Coco,Pier Giuseppe Pelicci,Massimo F. Martelli +22 more
TL;DR: Cytoplasmic NPM is a characteristic feature of a large subgroup of patients with AML who have a normal karyotype, NPM gene mutations, and responsiveness to induction chemotherapy.
Journal ArticleDOI
Activation-induced cytidine deaminase (AID) deficiency causes the autosomal recessive form of the Hyper-IgM syndrome (HIGM2).
Patrick Revy,Taro Muto,Yves Levy,Frederic Geissmann,Alessandro Plebani,Ozden Sanal,Nadia Catalan,Monique Forveille,Dufourcq-Lagelouse R,Andrew R. Gennery,Ilhan Tezcan,Fügen Ersoy,Hülya Kayserili,Alberto G. Ugazio,Nicole Brousse,Masamichi Muramatsu,Luigi D. Notarangelo,Kazuo Kinoshita,Tasuku Honjo,Alain Fischer,Anne Durandy +20 more
TL;DR: The phenotype observed in HIGM2 patients (and in AID-/- mice) demonstrates the absolute requirement for AID in several crucial steps of B cell terminal differentiation necessary for efficient antibody responses.
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Journal ArticleDOI
Alkaline phosphatase and peroxidase for double immunoenzymatic labelling of cellular constituents.
D Y Mason,R Sammons +1 more
TL;DR: It was found that the unlabelled antibody immunohistological procedure can be shortened without loss of sensitivity by carrying out two of the incubation steps simultaneously, and the total duration of this double immunostaining procedure is only a few minutes longer than that required for detection of a single antigen.