Article Figures & Data
Figures
- FIGURE 1.
Expression of trkB gp95 on PBMC. a, FACS analysis of unstimulated lymphocytes. PBL were stained with anti-trkB gp95 polyclonal rabbit Ab and CD4 Ab. Twelve percent of the Th cells carried the receptor. b, Expression of trkB gp95 mRNA in CD4-positive (+) and CD4-depleted (−) PBMC. Cells were either unstimulated or stimulated with the following: PHA (1%).or anti-CD3 (20 μg) in combination with 20 U/ml IL-2 or 100 U/ml IL-4. Only unstimulated CD4+ cells expressed the receptor mRNA, whereas after stimulation CD4+ as well as CD4− cells were positive for trkB gp95 mRNA. PC*, positive control (cDNA of neuroblastoma cell line SMS-KCN); NC§, negative control (no cDNA added). The constitutively expressed GAPDH served as a PCR and gel-loading control.
- FIGURE 2.
Amplification of trkB, trkC, and NT-3 mRNA in various cell lines and clones by RT-PCR. trkB gp95 mRNA (a),trkB gp145 mRNA (b), trkC mRNA (c), and NT-3 mRNA (d) expression in B-LCL, the monocyte cell line MM1, the γδ− clone EM-17, and CD4+ T cell clones 200, 234, 305, 401, 403, 407, 411, and 425. The CD4+ clones 305 and 401 were also stimulated with anti-CD3 (20 μg) and 20 U/ml IL-2 for 3 days. The constitutively expressed GAPDH served as a PCR and gel-loading control.
- FIGURE 3.
Influence of different cytokines on trkB gp145 expression. trkB gp145 mRNA was amplified by RT-PCR. Th0 clone 234, Th2 clone 305, and PBMC were treated with IL-2 (20 U/ml), IL-4 (100 U/ml), BDNF (100 ng/ml), NT-4 (100 ng/ml), IFN-γ (500 U/ml), or with medium alone (-) for 24 h. PC*, positive control (cDNA of the neuroblastoma cell line SMS-KCN was added), NC§, negative control (cDNA of PC-12 cells, not expressing the trkB mRNA, was added). The constitutively expressed GAPDH served as a PCR and gel-loading control.
- FIGURE 4.
Phenotyping of CD4+ Th clones according to their secretion of IL-4 and IFN-γ determined by ELISA of cell culture supernatants after 24 h. a, IFN-γ production as measured in nanograms per ml. b, IL-4 production in pg/ml. Data represent the mean ± SEM of eight experiments. c, Clones producing mainly IL-4 were assigned a Th2 phenotype; clones producing mainly IFN-γ as Th1 and clones secreting both cytokines in higher amounts were assigned a Th0 phenotype.
- FIGURE 5.
Effect of irradiation on BDNF secretion. BDNF levels in pg/ml were determined by ELISA for three untreated and 20-Gy-irradiated B-LCL (A.N., B.E., R.E.), the monocyte cell line MM1, and four Th clones (234, 305, 401, 411). The cell culture supernatants were collected 3 days after irradiation. Data represent the mean ± SEM of three experiments.
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