![Figure 2. Dendritic cells from CVID patients display impaired functional properties. (A) Dendritic cells from CVID patients are impaired in their allogeneic CD4+ T-cell stimulatory capacity in an MLR, as measured by [3H]-thymidine uptake. Graded doses of DCs were seeded with 1 × 105 responder T cells in RPMI 1640 medium supplemented with 10% human AB serum. After 4 days, the cells were pulsed for 16 hours with 1 μCi (0.037 MBq) [3H]thymidine. Radioactive incorporation was measured by standard liquid scintillation counting, and results were expressed as counts per minute (cpm, mean ± SD of triplicate values) after subtraction of values from culturing stimulator cells alone. The level of [3H]-thymidine uptake by T cells in medium alone was in the range of 1320 ± 160 cpm and was less than 1000 cpm in stimulator cells alone when 10 000 cells were tested. (B) Dendritic cells from CVID patients display altered capacities for maturation. DCs (6 days old) from CVID patients (thick lines) and selective antibody-deficient patients (thin lines) were stimulated with CD40L-transfected fibroblasts (10:1) for 48 hours. The mean fluorescence intensity for 1 of the 4 CVID patients (patient 5, bold numbers) and 1 of the 3 selective antibody-deficient patients (patient 3, light numbers) analyzed are indicated. (C) Dendritic cells from CVID patients (n = 4, patients 3, 4, 5, and 12) produce significantly lower amounts of IL-12 compared with that of DCs from healthy donors (n = 3) (▪) and selective antibody-deficient patients (n = 3, patients 2, 3 and 4) (□). Cytokine secretion was measured in a cell-free culture supernatant by Quantikine kit (R&D Systems, Abingdon, Oxon, United Kingdom) following stimulation of cells with CD40L-transfected fibroblasts for 48 hours. The level of IL-12 production by 4 CVID patients (patients 3, 4, 5, and 12) was in the range of 5 to 86 pg/mL per 0.5 × 106 cells. DCs of patient 3 produced 29 pg/mL, those of patients 4 and 5 produced 5 pg/mL or less, and those of patient 12 secreted 86 pg/mL IL-12 upon CD40 stimulation. Statistical significance as determined by the nonparametric Mann-Whitney test is indicated (*P < .01).](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/104/8/10.1182_blood-2004-04-1325/6/zh80200468110002.jpeg?Expires=1736062170&Signature=FJ1Ol4KXPKhVU6qubAAfLJKQH6TT7P58-eAnhoTcm1YHgYJfiXvyv1yW2APVNfd786sUcDM~rvwJIzjP-AQnFYXu1sudCs6HJP-iwdDPyUNhCiTxLBdF4wC~zOPcv-yHyXa2AlP13dtbQQhQfazYJv4t6l-ZpOU5ipqyQMaqr9w7AZgCgqEHqjP~bZ5e44WscNOjEKDsW9t3ZTk0x8RhYhifu3M7to8qVv2riniutWxzu-2byuRy4VaxRJ5rptF2qWAt09B3-63dzQtiowsJiNO72J7GnB8VFv5yhDq2QpwlQIOtzZS-o5R--d03n3cVShMsDD5gDMPFGiTlPahukw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Dendritic cells from CVID patients display impaired functional properties. (A) Dendritic cells from CVID patients are impaired in their allogeneic CD4+ T-cell stimulatory capacity in an MLR, as measured by [3H]-thymidine uptake. Graded doses of DCs were seeded with 1 × 105 responder T cells in RPMI 1640 medium supplemented with 10% human AB serum. After 4 days, the cells were pulsed for 16 hours with 1 μCi (0.037 MBq) [3H]thymidine. Radioactive incorporation was measured by standard liquid scintillation counting, and results were expressed as counts per minute (cpm, mean ± SD of triplicate values) after subtraction of values from culturing stimulator cells alone. The level of [3H]-thymidine uptake by T cells in medium alone was in the range of 1320 ± 160 cpm and was less than 1000 cpm in stimulator cells alone when 10 000 cells were tested. (B) Dendritic cells from CVID patients display altered capacities for maturation. DCs (6 days old) from CVID patients (thick lines) and selective antibody-deficient patients (thin lines) were stimulated with CD40L-transfected fibroblasts (10:1) for 48 hours. The mean fluorescence intensity for 1 of the 4 CVID patients (patient 5, bold numbers) and 1 of the 3 selective antibody-deficient patients (patient 3, light numbers) analyzed are indicated. (C) Dendritic cells from CVID patients (n = 4, patients 3, 4, 5, and 12) produce significantly lower amounts of IL-12 compared with that of DCs from healthy donors (n = 3) (▪) and selective antibody-deficient patients (n = 3, patients 2, 3 and 4) (□). Cytokine secretion was measured in a cell-free culture supernatant by Quantikine kit (R&D Systems, Abingdon, Oxon, United Kingdom) following stimulation of cells with CD40L-transfected fibroblasts for 48 hours. The level of IL-12 production by 4 CVID patients (patients 3, 4, 5, and 12) was in the range of 5 to 86 pg/mL per 0.5 × 106 cells. DCs of patient 3 produced 29 pg/mL, those of patients 4 and 5 produced 5 pg/mL or less, and those of patient 12 secreted 86 pg/mL IL-12 upon CD40 stimulation. Statistical significance as determined by the nonparametric Mann-Whitney test is indicated (*P < .01).
