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  • Title: Interleukin-2 secretion by human B lymphocytes occurs as a late event and requires additional stimulation after CD40 cross-linking.
    Author: Kindler V, Matthes T, Jeannin P, Zubler RH.
    Journal: Eur J Immunol; 1995 May; 25(5):1239-43. PubMed ID: 7539752.
    Abstract:
    While Epstein-Barr virus (EBV)-immortalized B cell lines have been shown to secrete interleukin (IL)-2 after stimulation with either teleocidin or phorbol myristate acetate (PMA) and ionomycin, experimental conditions leading to IL-2 production by normal human B cells have not been reported. In the present study we investigated various B cell activating conditions, including--by analogy to EBV-immortalized B lymphocytes--stimulation of B cells that are already proliferating (in cultures with IL-4 and immobilized anti-CD40 monoclonal antibody; the anti-CD40 system). This approach showed that B lymphocytes secreted IL-2 in the culture medium, but only if they were first activated for more than 24 h in the anti-CD40 system before exposure to PMA plus ionomycin. The production rate of IL-2 by B lymphocytes reached a maximum after 6 days of priming in such cultures followed by 48 h of stimulation with PMA plus ionomycin, corresponding to 7% or 15% of that of fresh CD4+ T cells activated, respectively, with phytohemagglutinin plus PMA, or with PMA plus ionomycin for 48 h. This IL-2 production could not be attributed to T cell contamination nor to EBV-infected B cells according to flow cytometric and reverse transcriptase-polymerase chain reaction analysis of cultured B cells. Lower IL-2 expression (detected only as mRNA synthesis) was also induced in the cultured B lymphocytes after incubation with cross-linking anti-IgM antibodies instead of PMA plus ionomycin. The appearance of IL-13 mRNA, but not IL-4 mRNA, was detected under the same stimulation conditions as for IL-2 mRNA. These results show that the production of IL-2 by normal B lymphocytes occurs as a late event relative to their activation and proliferation, and is in this respect subject to regulation different to that found in T lymphocytes.
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