Production and preliminary characterization of monoclonal antibodies to chicken anemia agent.
Mice were immunized with partially purified preparations of the Cux-1 isolate of chicken anemia agent (CAA), and their splenocytes were fused with NSO myeloma cells. Three patterns of staining of CAA-infected cells were recognized when the resulting hybridomas were screened by indirect immunofluorescence (IIF). Hybridomas representative of each staining pattern were cloned, and the monoclonal antibodies (MAbs) were characterized. Type 1 staining was indistinguishable from that produced by polyclonal chicken antisera to CAA. Type 2 staining was confined to large nuclear inclusions. Type 3 staining was predominantly nuclear and granular, and differed from type 1 in being more intense and occurring in a higher proportion of nuclei. Three MAbs producing type 1 staining were predominantly Cux-1-specific by IIF; they also reacted to lower titers with the Gifu-1 isolate but not at all with three other CAA isolates. These MAbs had very slight neutralizing activity against Cux-1. Another MAb giving type 1 staining reacted with all CAA isolates tested to high titers in IIF and neutralization tests. MAbs with type 2 and type 3 staining reacted by IIF with all CAA isolates tested but possessed no neutralizing activity. The availability of MABs to CAA should facilitate development of diagnostic tests for the virus.
McNulty
M S
MS
Veterinary Research Laboratories, Stormont, Belfast, Northern Ireland.
Mackie
D P
DP
Pollock
D A
DA
McNair
J
J
Todd
D
D
Mawhinney
K A
KA
Connor
T J
TJ
McNeilly
F
F
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis*; D000914:Antibodies, Viral / Q000032:analysis / Q000096:biosynthesis; D002645:Chickens; D005455:Fluorescent Antibody Technique; D006825:Hybridomas; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D014780:Viruses / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral