(A) The effect of anti-NGF MoAb on NGF detection in coculture supernatants of memory B cells cocultured with CD40L-transfected L cells. (B [donor 1] and C [donor 2]) The mean IgG production by memory B cells cocultured with CD40L-transfected L cells. B lymphocytes were purified from buffy coats by using anti-CD19 M450 Dynabeads (Dynal, Oslo, Norway), and memory CD19⁺ B cells were isolated by negative selection using a rabbit anti-human IgD antibody and anti-rabbit IgG antibody coated microbeads (Miltenyi Biotec, Bergisch, Germany). Memory B cells were cocultured with CD40L-expressing L cells (B cells: fibroblasts, 5:1) in IMDM containing IL-2 (20 U/mL) and IL-10 (10 ng/mL), in the presence of a neutralizing anti-NGF MoAb (15 μg/mL; R & D Systems, Minneapolis, MN) or negative mouse IgG (15 μg/mL). Cultures were performed in six parallel wells containing 1 × 10⁵ B cells. Cell-culture supernatants were collected after 5 days and tested for NGF or IgG content by capture ELISA.19 SEM were <10% for the NGF ELISA and <15% for the IgG ELISA. *P < .05 and **P< .005 by t-test.