ELISA-plaque assay for the detection of platelet antibody-secreting cells. I. Studies on optimization of reagent concentrations and culture conditions for enumeration of spot-forming cells.
This report describes a modified ELISA-plaque assay that we have succeeded in developing and utilizing for the detection of platelet antibody-secreting cells (PASC). We investigated the effects of various culture conditions on the formation and specificity of plaques obtained from spleen cells and peripheral blood leukocytes (PBL) of mice immunized with human platelets. The values of PASC using this plaque assay were compared with the corresponding serum platelet-binding IgG (PBIgG) levels. The results revealed that plaque formation from spleen cells was well correlated with the number of inoculations, whereas that from PBL was not. The rise of PBIgG levels in sera was shown to be delayed, as compared with the plaque formation from spleen cells.
Uchiyama
H
H
Department of Pediatrics, Jikei University School of Medicine, Tokyo, Japan.
Akatsuka
J
J
D000818:Animals; D000921:Antibody-Producing Cells / Q000276:immunology*; D001792:Blood Platelets / Q000276:immunology*; D002452:Cell Count; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006462:Hemolytic Plaque Technique; D007074:Immunoglobulin G / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D013997:Time Factors
D007074:Immunoglobulin G