Lung Expansion of Th1, Th2, Th17 and Tregs Table 1. Cont. Table 1. Fold change of gene expression in peribronchial, perivascular and alveolar lung tissue of OVA exposed mice compared to PBS control. RefSeq* Symbol Peribronchial Perivascular Alveolar tissue NM_008404 Itgb2 1,80 2,88 2,69 RefSeq* Symbol Peribronchial Perivascular Alveolar tissue NM_010735 Lta 1,33 1,12 21,21 NM_013854 Abcf1 2,17 2,03 2,22 NM_008518 Ltb 21,26 1,33 21,17 NM_009744 Bcl6 1,21 21,09 21,09 NM_010798 Mif 1,92 1,80 1,36 NM_009778 C3 1,65 1,34 1,22 NM_007926 Scye1 1,56 1,49 1,01 NM_009807 Casp1 4,96 6,05 7,36 NM_009263 Spp1 5,86 9,63 11,55 NM_011329 Ccl1 1,60 2,25 1,73 NM_011577 Tgfb1 22,38 21,62 21,30 NM_011330 Ccl11 2,95 4,34 2,73 NM_011609 Tnfrsf1a 23,07 23,64 23,61 NM_011331 Ccl12 3,86 3,94 3,63 NM_011610 Tnfrsf1b 22,28 22,33 22,64 NM_011332 Ccl17 2,08 3,41 4,08 NM_011616 Cd40lg 2,79 3,97 3,39 NM_011888 Ccl19 3,03 6,67 1,84 NM_023764 Tollip 21,68 21,81 21,58 NM_011333 Ccl2 4,86 5,73 12,13 NM_011798 Xcr1 1,20 1,31 1,45 NM_016960 Ccl20 5,13 9,11 6,23 Genes showed have fulfilled all quality criteria, including Ct value, melting curve NM_019577 Ccl24 3,92 13,07 3,63 and melting temperature. NM_013652 Ccl4 2,87 2,48 4,56 *RefSeq, reference sequence. doi:10.1371/journal.pone.0019889.t001 NM_013653 Ccl5 1,05 1,96 1,97 NM_009139 Ccl6 21,47 21,04 1,64 Discussion NM_013654 Ccl7 8,57 11,62 15,78 NM_021443 Ccl8 18,38 20,08 11,08 Our current study shows that airway exposure to allergen in NM_011338 Ccl9 2,53 3,03 4,23 sensitised mice expands all studied effector T cells in the lungs, including T-bet+, GATA-3+, RORct+ and Foxp3+ cells , both in NM_009912 Ccr1 1,51 2,69 3,39 those cell populations expressing the IL-2Ra (i.e. CD25) and those NM_009915 Ccr2 22,07 21,11 21,32 that do not. Among these, the GATA-3 effector Th2 cell is the one NM_009914 Ccr3 21,01 1,10 1,04 most prominently increased after allergen exposure, followed to a NM_009916 Ccr4 21,17 1,42 1,51 lesser extent by the T-bet+ Th1 and the RORct+ Th17 cells. In contrast, the Foxp3+ Treg cells increased in absolute cells numbers NM_009835 Ccr6 2,22 3,83 1,79 in the CD4+CD25+ and CD4+CD25- population but decreased NM_009913 Ccr9 2,38 2,29 2,89 compare to other T effector cells in the CD4+CD25+ fraction. All NM_009142 Cx3cl1 21,66 22,06 22,03 effector T cells are produced de novo and proliferate in situ in the NM_008176 Cxcl1 5,13 4,95 6,23 lungs, but with different profiles. Th1 and Th2 cells proliferate NM_021704 Cxcl12 21,57 21,22 1,09 more extensively than Th17 and Treg cells during allergic NM_011339 Cxcl15 23,14 22,81 21,48 inflammation. Allergen exposure subsequently results in an NM_019932 Pf4 24,29 22,62 23,53 accumulation of Th1, Th2 and Treg cells in the peribronchial tissue, as well as an increase in the alveolar lung tissue of Th17 NM_009141 Cxcl5 2,08 22,32 21,95 cells. In perivascular tissue, only Th2 and Treg cells increase. A NM_008599 Cxcl9 5,10 6,81 4,69 general inflammatory milieu in the lung was showed to involve NM_009910 Cxcr3 2,99 4,43 2,62 Th2 cytokines such as IL-4 as well as Th1 cytokines including NM_008348 Il10ra 21,65 21,37 21,21 IFN-c, and the Th17 cytokine IL-17 as well as several pro- NM_008349 Il10rb 1,01 1,35 1,46 inflammatory cytokines including IL-6. The low concentration of NM_008355 Il13 4,08 4,95 6,02 IL-10 argues for a relatively low activity of Treg cells. Finally, allergen exposure induced a wide range of inflammatory genes in NM_133990 Il13ra1 1,44 21,55 21,77 lung, whereas some of them only in specific tissues, arguing for the NM_008357 Il15 21,18 1,32 1,42 importance of the local microenvironment. Several chemokine NM_008360 Il18 28,11 25,99 26,68 genes supported the accumulation pattern of T effector cells. NM_010554 Il1a 21,06 21,19 1,11 As far as we know this is the first time that all mentioned T cells 22,50 21,95 21,21 NM_008361 Il1b are quantified simultaneously in allergic airway inflammation, especially as both their numbers and their topographical NM_008362 Il1r1 2,36 3,63 3,27 localizations are determined, as well as their proliferative status. NM_010555 Il1r2 3,05 3,63 3,46 Furthermore, this is also the first time that the local inflammatory NM_008368 Il2rb 22,46 21,59 22,03 milieu in three main lung tissue compartments, i.e. peribronchial, NM_013563 Il2rg 21,62 1,16 21,23 perivascular and alveolar tissue, is described in relation to the NM_008370 Il5ra 2,14 2,07 2,60 relative expression of a substantial number of relevant pro- and NM_010560 Il6st 23,18 22,74 23,20 anti-inflammatory genes. CD25 (IL-2Ra) expression is induced upon T cell activation to NM_009909 Il8rb 26,15 26,37 22,87 form part of the trimeric, high affinity IL-2R complex in NM_008401 Itgam 1,07 1,25 1,75 combination with CD122 (IL-2Rb) and CD132 (IL-2Rc) and PLoS ONE | www.plosone.org 12 May 2011 | Volume 6 | Issue 5 | e19889