Journal:

Article Title: Cultured basophils but not cultured mast cells induce human IgE synthesis in B cells after immunologic stimulation

doi: 10.1046/j.1365-2249.1998.00474.x

Figure Lengend Snippet: Production of IgE, IgG4 and soluble CD23 by normal B cells mixed with cultured human mast cells or basophils. Mast cells or basophils were stimulated for 2h with antigen after sensitization with murine IgE, washed, added to highly purified B cells, and then cultured for 12 days. Stimulated mast cells were incubated with B cells in the absence or presence of recombinant IL-4. Stimulated basophils were incubated with B cells in the absence or presence of two neutralizing anti-IL-4 and anti-IL-13 MoAbs or a soluble form of CD40. Supernatants were collected and measured for concentrations of IgE, IgG4 and soluble CD23. The net synthesis of IgE and IgG4 was determined, as described in Materials and Methods. Results represent the mean ± s.e.m. of triplicate cultures. One of two comparable experiments is shown. IgE production by B cells was neither induced by unstimulated basophils, nor significantly affected in the cultures containing B cells plus stimulated basophils by the addition of control antibody or IgM (data not shown).

Article Snippet: Reagents, antigen, and antibodies Recombinant human stem cell factor (SCF) and IL-6 were generous gifts from Kirin Brewery Co. (Maebashi, Japan).

Techniques: Cell Culture, Purification, Incubation, Recombinant, Control

Journal:

Article Title: Cultured basophils but not cultured mast cells induce human IgE synthesis in B cells after immunologic stimulation

doi: 10.1046/j.1365-2249.1998.00474.x

Figure Lengend Snippet: Analysis of germ-line and mature Cε mRNA expression in normal B cells mixed with cultured human mast cells or basophils. The cell pellets obtained in Fig. 5 were used for the analysis of germ-line and mature Cε transcription. Highly purified B cells were incubated with medium alone (lane 1) or with stimulated mast cells in the absence (lane 2) or presence (lane 3) of recombinant IL-4. Stimulated basophils were incubated with B cells in the absence (lane 4) or presence of two neutralizing anti-IL-4 and anti-IL-3 MoAbs (lane 5) or a soluble form of CD40 (lane 6). After extraction of total cellular RNA, mRNA for germ-line Cε, mature Cε, and G3PDH was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR), subjected to electrophoresis, and then visualized by ethidium bromide. One of two comparable experiments is shown. Neither germ-line nor mature transcripts were detected in the cultures containing B cells plus unstimulated basophils (data not shown).

Article Snippet: Reagents, antigen, and antibodies Recombinant human stem cell factor (SCF) and IL-6 were generous gifts from Kirin Brewery Co. (Maebashi, Japan).

Techniques: Expressing, Cell Culture, Purification, Incubation, Recombinant, Extraction, Amplification, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Electrophoresis