Production of human antibodies to bee venom phospholipase A2 in vitro

Scand J Immunol. 1989 Feb;29(2):203-9. doi: 10.1111/j.1365-3083.1989.tb01117.x.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bee Venoms / immunology*
  • Blood Cell Count
  • Humans
  • Immunization
  • Immunoglobulin G / analysis
  • Immunoglobulin Isotypes / analysis
  • Immunoglobulin M / analysis
  • In Vitro Techniques
  • Phospholipases / immunology*
  • Phospholipases A / immunology*
  • Phospholipases A2
  • Pokeweed Mitogens / pharmacology


  • Bee Venoms
  • Immunoglobulin G
  • Immunoglobulin Isotypes
  • Immunoglobulin M
  • Pokeweed Mitogens
  • Phospholipases
  • Phospholipases A
  • Phospholipases A2