title: Production of human antibodies to bee venom phospholipase A2 in vitro.
abstract: Phospholipase A2 (PLA) is the major antigen of bee venom. Whereas individuals frequently stung by bees, such as bee keepers, show high levels of IgG4 anti-PLA antibodies in serum, most patients sensitive to bee venom possess increased IgE anti-PLA. We have established a culture system by which anti-PLA antibodies can be induced in vitro. Peripheral blood mononuclear cells were stimulated in a first step with PLA and/or pokeweed mitogen (PWM). After 3 days of culture the cells were washed and further incubated with fresh medium. Anti-PLA antibodies were estimated by an enzyme-linked immunosorbent assay (ELISA) and anti-PLA antibody secreting cells were counted by means of an ELISA plaque assay. Cells from bee keepers, but not those from normal donors, produced anti-PLA IgG in vitro. The isotype pattern of anti-PLA antibodies produced in vitro was identical to that found in serum of the respective donor. Anti-PLA IgM antibody-secreting cells developed at low frequencies in both bee keepers and normal donors whereas PLA-specific IgE could not be detected. The formation of anti-PLA IgG was suppressed by high amounts of antigen, while the IgM response was not affected. We conclude that PWM-induced antibodies reflect the in vivo situation of the donors and that anti-PLA IgG and IgM are regulated by different pathways and mechanisms.
authors.affiliation: Swiss Institute of Allergy and Asthma Research, Davos.
authors.forename: W
authors.initials: W
authors.lastname: Held
authors.forename: M
authors.initials: M
authors.lastname: Stucki
authors.forename: C
authors.initials: C
authors.lastname: Heusser
authors.forename: K
authors.initials: K
authors.lastname: Blaser
mesh_terms: D001514:Bee Venoms / Q000276:immunology*; D001772:Blood Cell Count; D006801:Humans; D007114:Immunization; D007074:Immunoglobulin G / Q000032:analysis; D007132:Immunoglobulin Isotypes / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D066298:In Vitro Techniques; D010740:Phospholipases / Q000276:immunology*; D010741:Phospholipases A / Q000276:immunology*; D054467:Phospholipases A2; D011043:Pokeweed Mitogens / Q000494:pharmacology
chemical_list: D001514:Bee Venoms; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D011043:Pokeweed Mitogens; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2