Detection of antibody, idiotype, and anti-idiotype forming cells by in situ immunocytochemical staining.
Methods are described for the immunocytochemical staining of cryostat sections of lymphoid tissue with enzyme conjugates of antigen, idiotype (Id) and anti-idiotype. Results established this as a useful approach, for simultaneously detecting Id and anti-Id antibody forming cells (AFC) in situ. As a model, the 5AF6 Id family associated with the BALB/c mouse antibody response against the p-azophenyl-arsonate (Ar) epitope was examined by two-color immunocytochemical staining, allowing the simultaneous detection of both Id+ and Id- anti-Ar AFC. Spleens from mice secondarily immunized with Ar antigen but not normal mice contained anti-Id AFC stained with the 5AF6 Id but not with another immunoglobulin of the same isotype. A sequential staining method was developed which allowed the detection of both Id and anti-Id AFC in the same tissue, thus providing a means of examining Id and anti-Id antibody networks in intact lymphoid tissues.
Brown
A R
AR
Department of Immunology, St. Jude Children's Research Hospital, Memphis, TN 38101.
Claassen
E
E
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000921:Antibody-Producing Cells / Q000166:cytology*; D006241:Haptens; D007130:Immunoglobulin Idiotypes / Q000032:analysis*; D007150:Immunohistochemistry / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D010132:p-Azobenzenearsonate / Q000276:immunology
D000906:Antibodies; D006241:Haptens; D007130:Immunoglobulin Idiotypes; D010132:p-Azobenzenearsonate