Journal:

Article Title: Expression of the ?1?1 integrin, VLA-1, marks a distinct subset of human CD4 + memory T cells

doi: 10.1172/JCI200319607

Figure Lengend Snippet: VLA-1 expression is associated with Th1 polarization. (a) CD4+ PBLs were purified into VLA-1+ and VLA-1– fractions, activated with PMA/ionomycin for 5 hours, and analyzed for intracellular IFN-γ production. (b) CD4+ PBLs were stimulated with TT or anti-CD3; 10 days later the cells were harvested and purified into VLA-1+ and VLA-1– fractions, activated with PMA/ionomycin, and analyzed for intracellular IFN-γ and IL-4. (c) CD4+ PBLs were labeled with CFSE, stimulated with TT, and 10 days later purified into VLA-1+ and VLA-1– fractions. Subsequently, the two subsets were activated and analyzed for divisions and intracellular IFN-γ. (d) CD4+ PBLs were stimulated with TSST-1 in two different environments: Th1 (IL-12+) or TH2 (IL-4+ and anti–IFN-γ). Ten days later cells were harvested and analyzed for VLA-1 and Vβ2 surface expression (left) or activated and analyzed for intracellular IFN-γ (right). (e) Fresh SFLs from RA (n = 4) and PsA (n = 2) patients were immediately activated and analyzed for surface expression of CD4, VLA-1, and intracellular IFN-γ. The dot plots were obtained by pregating on CD4+ events, and a representative patient sample is shown.

Article Snippet: To induce Th1 polarization, the CD4 + cells were stimulated with 2.5 μg/ml of anti-CD3 mAb in a medium containing 5 ng/ml recombinant human IL-12 (R&D Systems Inc., Minneapolis, Minnesota, USA).

Techniques: Expressing, Purification, Labeling