Detection of antibody-forming cells directed against Newcastle disease virus and their immunoglobulin class by double immunoenzyme histochemistry.
Antibody-forming cells (AFCs) against Newcastle disease virus (NDV) and their immunoglobulin (Ig) class were demonstrated by a double immuno-enzyme histochemical technique. The AFCs were stained and quantified in spleen sections of chickens euthanatized at day 7 postexposure to the Roakin strain of NDV. The sections were incubated with NDV to determine the specificity of the AFCs. Bound virus was subsequently visualized with a primary monoclonal antibody (MAb), a secondary horseradish peroxidase-conjugated MAb, and 3-amino-9-ethylcarbazole as substrate. IgM and IgA were stained with MAbs and an alkaline phosphatase (AP)-conjugated secondary antibody. IgG class antibodies were demonstrated with an AP-conjugated rabbit serum. The final substrate for the three Igs was naphthol AS-MX-phosphate and fast blue BB. About 64-159/mm2 AFCs against NDV were detected. Of these virus-binding cells, about 55% produced IgM, 37% produced IgG, and the remaining 8% produced IgA.
Al-Garib
S O
SO
Central Institute for Disease Control (CIDC-Lelystad), P.O. Box 2004, 8203 AA Lelystad, The Netherlands.
Gruys
E
E
Gielkens
A L J
AL
Koch
G
G
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis* / Q000276:immunology; D002645:Chickens; D007124:Immunoenzyme Techniques / Q000379:methods*; D007136:Immunoglobulins / Q000145:classification* / Q000276:immunology*; D009521:Newcastle Disease / Q000276:immunology*; D009522:Newcastle disease virus / Q000276:immunology*; D010950:Plasma Cells / Q000276:immunology*; D013154:Spleen / Q000166:cytology / Q000276:immunology
D000914:Antibodies, Viral; D007136:Immunoglobulins