Detection of anticardiolipin antibodies in culture supernatants.
OBJECTIVE
We have attempted to devise a method for measuring the levels of anticardiolipin antibodies produced in in vitro culture of peripheral blood mononuclear cells isolated from patients with systemic lupus erythematosus (SLE), with or without circulating anticardiolipin antibodies, patients with the primary antiphospholipid syndrome (PAPS) and normal controls.


METHODS
Peripheral blood mononuclear cells were isolated and cultured for up to six days, in the presence or absence of added cytokines in the culture medium. Supernatants harvested after culture were tested by ELISA for the presence of anticardiolipin antibodies.


CONCLUSIONS
Despite variation of the culture conditions and of the cell numbers and populations used, it was found that accurately measurable levels of anticardiolipin antibodies could not be detected reliably in any of the culture supernatants. It is concluded that alternative methods of measurement of antibody production need to be explored.
Dean
G S
GS
Department of Immunology, University College London Medical School, London, UK. gillian.dean@ucl.ac.uk
Isenberg
D A
DA
D017153:Antibodies, Anticardiolipin / Q000032:analysis*; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D066298:In Vitro Techniques; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008180:Lupus Erythematosus, Systemic / Q000175:diagnosis* / Q000276:immunology*
D017153:Antibodies, Anticardiolipin; D007074:Immunoglobulin G; D007075:Immunoglobulin M