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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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In vitro generation of effector cells in the presence of anti-IFN-γ antibody. CD4+ memory effector T cells obtained from the peripheral blood of a healthy donor (normal) or from RASF (patient) were stimulated as described in Fig. 3 with immobilized anti-CD3 mAb (OKT3, 0.01 μg/ml), anti-CD28, and cytokines. In addition, cultures were carried out in the presence of increasing concentrations of a neutralizing anti-IFN-γ antibody as indicated. After priming, the cells were washed, rested for 2.5 days and restimulated with mitogen to assess intracellular cytokines; 104 cells were collected for each analysis. The data are representative of at least two experiments. No significant difference was observed in the presence of anti-IFN-γ antibody (P > 0.05).
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Figure 4: In vitro generation of effector cells in the presence of anti-IFN-γ antibody. CD4+ memory effector T cells obtained from the peripheral blood of a healthy donor (normal) or from RASF (patient) were stimulated as described in Fig. 3 with immobilized anti-CD3 mAb (OKT3, 0.01 μg/ml), anti-CD28, and cytokines. In addition, cultures were carried out in the presence of increasing concentrations of a neutralizing anti-IFN-γ antibody as indicated. After priming, the cells were washed, rested for 2.5 days and restimulated with mitogen to assess intracellular cytokines; 104 cells were collected for each analysis. The data are representative of at least two experiments. No significant difference was observed in the presence of anti-IFN-γ antibody (P > 0.05).

Mentions: In order to determine whether cytokine production by rheumatoid T cells was resistant to modulation in the in vitro priming cultures because of the secretion of IFN-γ, the in vitro priming was also carried out in the presence of anti-IFN-γ antibody. As shown in Fig. 4, the addition of increasing concentrations of a neutralizing antibody to IFN-γ had little effect on the generation of cytokine-producing cells from either normal synovial fluid or RASF CD4+ T cells. Then we examined whether the combined effects of anti-IFN-γ antibody and IL-4 could influence the effector phenotype of cells isolated from the synovium. As shown in Table 1, Tm were isolated from matching RAPB and RASF, and were cultured under conditions that optimized the generation of IL-4-secreting effector cells [17]. RAPB Tm generated a marked increase in the percentage of IL-4-producing cells when primed with the combination of IL-4 and anti-IFN-γ antibody. In contrast, T cells from the synovial fluid of the same patient generated virtually no IL-4-producing cells and few IL-2-producing cells, despite supplemental IL-4 and anti-IFN-γ antibody.


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)

In vitro generation of effector cells in the presence of anti-IFN-γ antibody. CD4+ memory effector T cells obtained from the peripheral blood of a healthy donor (normal) or from RASF (patient) were stimulated as described in Fig. 3 with immobilized anti-CD3 mAb (OKT3, 0.01 μg/ml), anti-CD28, and cytokines. In addition, cultures were carried out in the presence of increasing concentrations of a neutralizing anti-IFN-γ antibody as indicated. After priming, the cells were washed, rested for 2.5 days and restimulated with mitogen to assess intracellular cytokines; 104 cells were collected for each analysis. The data are representative of at least two experiments. No significant difference was observed in the presence of anti-IFN-γ antibody (P > 0.05).
© Copyright Policy
Related In: Results  -  Collection

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Figure 4: In vitro generation of effector cells in the presence of anti-IFN-γ antibody. CD4+ memory effector T cells obtained from the peripheral blood of a healthy donor (normal) or from RASF (patient) were stimulated as described in Fig. 3 with immobilized anti-CD3 mAb (OKT3, 0.01 μg/ml), anti-CD28, and cytokines. In addition, cultures were carried out in the presence of increasing concentrations of a neutralizing anti-IFN-γ antibody as indicated. After priming, the cells were washed, rested for 2.5 days and restimulated with mitogen to assess intracellular cytokines; 104 cells were collected for each analysis. The data are representative of at least two experiments. No significant difference was observed in the presence of anti-IFN-γ antibody (P > 0.05).
Mentions: In order to determine whether cytokine production by rheumatoid T cells was resistant to modulation in the in vitro priming cultures because of the secretion of IFN-γ, the in vitro priming was also carried out in the presence of anti-IFN-γ antibody. As shown in Fig. 4, the addition of increasing concentrations of a neutralizing antibody to IFN-γ had little effect on the generation of cytokine-producing cells from either normal synovial fluid or RASF CD4+ T cells. Then we examined whether the combined effects of anti-IFN-γ antibody and IL-4 could influence the effector phenotype of cells isolated from the synovium. As shown in Table 1, Tm were isolated from matching RAPB and RASF, and were cultured under conditions that optimized the generation of IL-4-secreting effector cells [17]. RAPB Tm generated a marked increase in the percentage of IL-4-producing cells when primed with the combination of IL-4 and anti-IFN-γ antibody. In contrast, T cells from the synovial fluid of the same patient generated virtually no IL-4-producing cells and few IL-2-producing cells, despite supplemental IL-4 and anti-IFN-γ antibody.

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