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Rheumatoid synovial CD4+ T cells exhibit a reduced capacity to differentiate into IL-4-producing T-helper-2 effector cells.

Davis LS, Cush JJ, Schulze-Koops H, Lipsky PE - Arthritis Res. (2000)

Bottom Line: RAPB Tm contained significantly more IFN-gamma producers than normal cells.Normal and RAPB Tm differentiated into both IFN-gamma- and IL-4-producing effectors.RA synovial fluid (RASF) Tm demonstrated defective responsiveness, exhibiting diminished differentiation of IL-4 effectors, whereas RA synovial tissue (RAST) Tm exhibited defective generation of IFN-gamma and IL-4 producers.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Harold C Simmons Arthritis Research Center and Rheumatic Diseases Division, Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, Texas 75390-8884, USA. laurie.davis@UTSouthwestern.edu

ABSTRACT
CD4+ memory T cells (Tm) from rheumatoid arthritis peripheral blood (RAPB) or peripheral blood from normal donors produced IL-2, whereas fewer cells secreted IFN-gamma or IL-4 after a brief stimulation. RAPB Tm contained significantly more IFN-gamma producers than normal cells. Many rheumatoid arthritis (RA) synovial Tm produced IFN-gamma alone (40%) and fewer cells produced IL-2 or IL-4. An in vitro model was employed to generate polarized T-helper (Th) effectors. Normal and RAPB Tm differentiated into both IFN-gamma- and IL-4-producing effectors. RA synovial fluid (RASF) Tm demonstrated defective responsiveness, exhibiting diminished differentiation of IL-4 effectors, whereas RA synovial tissue (RAST) Tm exhibited defective generation of IFN-gamma and IL-4 producers.

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RA synovial memory T cells are deficient in IL-4-producing cells after in vitro priming. CD4+ memory effector T cells were placed into priming cultures to generate effector cells. Effector T cells were generated by priming in the presence of anti-CD28 mAb and cytokines, as described in Materials and methods, or with additional immobilized anti-CD3 mAb, OKT3, at the indicated concentrations (T3lo, 0.01 μg/ml; and T3hi, 1 μg/ml). After priming, the cells were harvested and rested in cytokines before restimulation to detect intracellular cytokines as described in Materials and methods; 104 cells were collected for each analysis. The data shown were derived from independent experiments; for normal donors (n = 4), RAPB (n = 4), RASF (n = 4), and RAST (n = 3), low-dose OKT3 was omitted in one experiment. The frequency of each subset of cytokine-producing cells is shown. The total number of cytokine-producing cells in each sample is shown in the upper right corner. *P < 0.05, versus freshly isolated samples shown in Fig. 2 (two-tailed Student's t-test).
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Figure 3: RA synovial memory T cells are deficient in IL-4-producing cells after in vitro priming. CD4+ memory effector T cells were placed into priming cultures to generate effector cells. Effector T cells were generated by priming in the presence of anti-CD28 mAb and cytokines, as described in Materials and methods, or with additional immobilized anti-CD3 mAb, OKT3, at the indicated concentrations (T3lo, 0.01 μg/ml; and T3hi, 1 μg/ml). After priming, the cells were harvested and rested in cytokines before restimulation to detect intracellular cytokines as described in Materials and methods; 104 cells were collected for each analysis. The data shown were derived from independent experiments; for normal donors (n = 4), RAPB (n = 4), RASF (n = 4), and RAST (n = 3), low-dose OKT3 was omitted in one experiment. The frequency of each subset of cytokine-producing cells is shown. The total number of cytokine-producing cells in each sample is shown in the upper right corner. *P < 0.05, versus freshly isolated samples shown in Fig. 2 (two-tailed Student's t-test).

Mentions: Peripheral blood and synovial Tm were than examined in an in vitro culture system in order to determine the relative capacity of differentiation signals to modulate cytokine production. Our previous studies with normal T cells [17] demonstrated that this system effectively generates polarized IFN-γ-producing or IL-4-producing effector cells. Fig. 3 shows the composite cytokine profiles of CD4+, CD45RO+ T cells obtained from cultures primed with anti-CD28 and cytokines in the absence or presence of anti-CD3 mAb. As seen in Fig. 3, stimulation of normal T cells after priming with anti-CD28 alone resulted in a significant decrease in the percentage of IL-2 producers (35% versus 75%) compared with cells immediately stimulated after isolation, and a significant increase in the percentages of IFN-γ (31% versus 6%) and IL-4 producers (17% versus 4%). After priming with a low dose of anti-CD3 (Fig. 3; middle panel), the percentage of IL-2 producers decreased further and the percentage of IFN-γ-secreting cells increased. As previously shown, the generation of IL-4-secreting cells was optimally induced by priming with anti-CD28 alone and was inhibited by costimulation with anti-CD3 mAb [17].


Rheumatoid synovial CD4+ T cells exhibit a reduced capacity to differentiate into IL-4-producing T-helper-2 effector cells.

Davis LS, Cush JJ, Schulze-Koops H, Lipsky PE - Arthritis Res. (2000)

RA synovial memory T cells are deficient in IL-4-producing cells after in vitro priming. CD4+ memory effector T cells were placed into priming cultures to generate effector cells. Effector T cells were generated by priming in the presence of anti-CD28 mAb and cytokines, as described in Materials and methods, or with additional immobilized anti-CD3 mAb, OKT3, at the indicated concentrations (T3lo, 0.01 μg/ml; and T3hi, 1 μg/ml). After priming, the cells were harvested and rested in cytokines before restimulation to detect intracellular cytokines as described in Materials and methods; 104 cells were collected for each analysis. The data shown were derived from independent experiments; for normal donors (n = 4), RAPB (n = 4), RASF (n = 4), and RAST (n = 3), low-dose OKT3 was omitted in one experiment. The frequency of each subset of cytokine-producing cells is shown. The total number of cytokine-producing cells in each sample is shown in the upper right corner. *P < 0.05, versus freshly isolated samples shown in Fig. 2 (two-tailed Student's t-test).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC17825&req=5

Figure 3: RA synovial memory T cells are deficient in IL-4-producing cells after in vitro priming. CD4+ memory effector T cells were placed into priming cultures to generate effector cells. Effector T cells were generated by priming in the presence of anti-CD28 mAb and cytokines, as described in Materials and methods, or with additional immobilized anti-CD3 mAb, OKT3, at the indicated concentrations (T3lo, 0.01 μg/ml; and T3hi, 1 μg/ml). After priming, the cells were harvested and rested in cytokines before restimulation to detect intracellular cytokines as described in Materials and methods; 104 cells were collected for each analysis. The data shown were derived from independent experiments; for normal donors (n = 4), RAPB (n = 4), RASF (n = 4), and RAST (n = 3), low-dose OKT3 was omitted in one experiment. The frequency of each subset of cytokine-producing cells is shown. The total number of cytokine-producing cells in each sample is shown in the upper right corner. *P < 0.05, versus freshly isolated samples shown in Fig. 2 (two-tailed Student's t-test).
Mentions: Peripheral blood and synovial Tm were than examined in an in vitro culture system in order to determine the relative capacity of differentiation signals to modulate cytokine production. Our previous studies with normal T cells [17] demonstrated that this system effectively generates polarized IFN-γ-producing or IL-4-producing effector cells. Fig. 3 shows the composite cytokine profiles of CD4+, CD45RO+ T cells obtained from cultures primed with anti-CD28 and cytokines in the absence or presence of anti-CD3 mAb. As seen in Fig. 3, stimulation of normal T cells after priming with anti-CD28 alone resulted in a significant decrease in the percentage of IL-2 producers (35% versus 75%) compared with cells immediately stimulated after isolation, and a significant increase in the percentages of IFN-γ (31% versus 6%) and IL-4 producers (17% versus 4%). After priming with a low dose of anti-CD3 (Fig. 3; middle panel), the percentage of IL-2 producers decreased further and the percentage of IFN-γ-secreting cells increased. As previously shown, the generation of IL-4-secreting cells was optimally induced by priming with anti-CD28 alone and was inhibited by costimulation with anti-CD3 mAb [17].

Bottom Line: RAPB Tm contained significantly more IFN-gamma producers than normal cells.Normal and RAPB Tm differentiated into both IFN-gamma- and IL-4-producing effectors.RA synovial fluid (RASF) Tm demonstrated defective responsiveness, exhibiting diminished differentiation of IL-4 effectors, whereas RA synovial tissue (RAST) Tm exhibited defective generation of IFN-gamma and IL-4 producers.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Harold C Simmons Arthritis Research Center and Rheumatic Diseases Division, Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, Texas 75390-8884, USA. laurie.davis@UTSouthwestern.edu

ABSTRACT
CD4+ memory T cells (Tm) from rheumatoid arthritis peripheral blood (RAPB) or peripheral blood from normal donors produced IL-2, whereas fewer cells secreted IFN-gamma or IL-4 after a brief stimulation. RAPB Tm contained significantly more IFN-gamma producers than normal cells. Many rheumatoid arthritis (RA) synovial Tm produced IFN-gamma alone (40%) and fewer cells produced IL-2 or IL-4. An in vitro model was employed to generate polarized T-helper (Th) effectors. Normal and RAPB Tm differentiated into both IFN-gamma- and IL-4-producing effectors. RA synovial fluid (RASF) Tm demonstrated defective responsiveness, exhibiting diminished differentiation of IL-4 effectors, whereas RA synovial tissue (RAST) Tm exhibited defective generation of IFN-gamma and IL-4 producers.

Show MeSH
Related in: MedlinePlus