Fig. 1

Astrocyte-T cell co-culture enhances immune cell vitality but inhibits ConA-mediated and antigen-specific T cell proliferation. A Experimental scheme: T cells were seeded with antigen presenting cells in absence or presence of astrocytes, and received media containing ConA or MOG_{35 − 55} peptide 24 h later. After 72 h incubation cells were harvested and T cell vitality and proliferation were addressed by flow cytometry and/or thymidine incorporation. B Percentage of living CD3 + T cells as assessed by 7AAD incorporation in unstimulated (ns) or ConA-supplemented cultures without (black bars) or with (grey bars) primary neonatal astrocytes. C ConA-induced T cell proliferation assessed by [3H] thymidine incorporation in cultures without (black bars) or with (grey bars) primary neonatal astrocytes. Data are reported as counts per minute (c.p.m.). D CFSE levels (x axis) in mouse CD3 + T cells in ConA-activated cultures without (black bars) or with (grey bars) neonatal astrocytes. E Percentage of T lymphocytes that underwent 0, 1 or > 2 cell cycle divisions. (F, G) CFSE levels in mouse 2D2 CD4 + T cells after exposure to MOG_35 −55 peptide in cultures without (black bars) or with (grey bars) neonatal astrocytes (F), and relative quantification (G). In D and F dotted lines represent mean CFSE levels in unstimulated cells (ns). H–J TNF-α (H), IL-4 (I) and IL-17 (J) released by 2D2 CD4 + Th1 (white dots), Th2 (grey dots) and Th17 (black dots) under resting conditions or after 24 h ConA stimulation. K–M MOG_35 −55 peptide induced proliferation of 2D2 CD4 + Th1 (K), Th2 (L), or Th17 (M) lymphocytes cultured without (black bars) or with (grey bars) neonatal astrocytes. Representative data of one out of 2–5 independent experiments are shown. In (B–C, E, G, H–M) data are reported as mean ± SD. Statistical analysis was performed using T test. *p-value < 0.05, **p-value < 0.01, ***p-value < 0.001