_:b403341660 . _:b403341404 . _:b403341724 . _:b5177938 "In a context of immunocompetent mice, TARC and MDC could lead to the recruitment of Th2 CCR4+ cells at the site of inflammation (>>51<<). In addition, IL-1\u03B2 and TNF-\u03B1 overexpression in Spink5\u2212/\u2212 epidermis may induce TSLP expression in keratinocytes (19, 54) and attract innate immune cells. These processes could explain worsening of the cutaneous phenotype observed in" . _:b403341407 . . . _:b403341406 . . _:b403341650 . _:b403341574 . . _:b403341401 . . . _:b403341624 . . _:b403341400 . . . _:b403341793 . _:b403341620 . . _:b403341831 . _:b403341403 . _:b403341411 . _:b403341402 . . _:b403341733 . _:b403341691 . _:b403341690 . . _:b403341413 . _:b5177950 "In addition, in an immune-competent system, TSLP has the potential to activate dendritic cells and epidermal LCs in particular, triggering the generation of proallergic CD4+ Th2 cells (19, 20, >>64<<)." . _:b403341639 . _:b403341492 . . _:b403341412 . _:b403341570 . _:b403341556 . _:b403341415 . . _:b403341414 . . _:b403341404 . _:b403341718 . _:b403341409 . . _:b403341409 . _:b403341408 . _:b403341411 . . . _:b403341839 . _:b403341555 . _:b403341829 . _:b403341410 . _:b403341562 . . _:b403341434 . . _:b5177916 "of the proinflammatory cytokine TNF-\u03B1, which is also produced in response to cutaneous barrier disruption (17) and tightly associated with inflammatory deviation of the immune response, such as asthma and probably allergic inflammation (>>48<<, 49). Despite unsuccessful immunostaining of TNF-\u03B1 on skin cross sections, we analyzed its transcriptional expression and showed that Tnf-\u03B1 mRNA was strongly overexpressed in the KO epidermis and also slightly induced in the dermis of KO" . _:b403341828 . _:b403341421 . _:b403341862 . . _:b403341655 . _:b403341831 . _:b403341420 . _:b403341838 . . _:b403341642 . "PMC0" . _:b5177908 "S5), which is in agreement with its degradation as previously described in newborn KO mice and NS skin (32, >>43<<). In summary, the abnormal differentiation pattern observed in KO graft, together with the desmosomal cleavage, confirmed that the KO grafted skin reproduces all major histological and ultrastructural anomalies seen in NS patients skin" . . _:b5177944 "Massive production of active \u03B21-tryptase by activated mast cells in Spink5\u2212/\u2212 grafted skin could also directly participate in the inflammatory response and favor a Th2-permissive microenvironment through PAR2 activation on mast cells (>>60<<), dendritic cells (61), and eosinophils (62, 63)." . _:b403341830 . _:b403341595 . _:b403341423 . _:b403341880 . . _:b403341825 . _:b403341422 . . _:b403341401 . _:b403341824 . _:b403341417 . _:b403341438 . _:b403341827 . _:b403341416 . . _:b403341386 . . _:b403341826 . _:b403341419 . . _:b403341837 . _:b403341848 . _:b403341418 . . _:b403341439 . _:b403341836 . _:b403341429 . _:b403341640 . _:b5177956 . _:b403341839 . _:b403341428 . . _:b403341838 . _:b5177934 "Nonetheless, IL-1\u03B2 overexpression in Spink5\u2212/\u2212 embryos epidermis is likely to be a consequence of SC detachment that is already present before birth (>>17<<). IL-1\u03B2 can be proteolytically activated by KLK7 (57), which is hyperactivated in Spink5\u2212/\u2212 skin (32). Thus, this protease could participate in the inflammatory process in our model as observed in KLK7 transgenic mice (58)." . _:b403341431 . _:b403341833 . _:b403341430 . _:b403341745 . _:b403341847 . _:b403341832 . _:b403341425 . . . . _:b403341835 . _:b403341424 . _:b5177911 "The proinflammatory cytokine IL-1\u03B2 is known to be secreted by keratinocytes after SC cohesion disruption (>>18<<). IL-1\u03B2 immunostaining was scant and localized at the GR-SC interface in WT epidermal grafts." . _:b403341786 . _:b403341834 . _:b403341427 . _:b5177922 "We analyzed two target genes of PAR2 activation in cultured keratinocytes, ICAM1 (>>47<<) and the chemokine IL8 (53)." . _:b403341608 . _:b403341845 . _:b403341426 . . _:b403341844 . _:b403341437 . _:b403341847 . _:b403341436 . . _:b403341545 . _:b403341846 . _:b403341439 . _:b403341746 . . . _:b403341794 . _:b403341423 . _:b403341841 . _:b403341438 . . _:b403341840 . _:b403341433 . _:b5177926 "Moreover, a functional NF-\u03BAB binding site was previously identified in the TSLP promoter in airway epithelial cells (>>54<<, 55). We therefore assessed the involvement of NF-\u03BAB signaling in TSLP, IL8, and TNF-\u03B1 up-regulation by KLK5 using BAY11-7082, a specific inhibitor of the NF-\u03BAB pathway. Pretreatment of NHK with BAY11-7082 decreased the effect of KLK5 on" . _:b403341843 . _:b403341432 . . _:b403341842 . _:b403341435 . . _:b403341725 . _:b403341616 . _:b403341853 . _:b403341434 . _:b5177937 . . _:b403341852 . _:b403341445 . . _:b403341855 . _:b403341734 . _:b403341444 . _:b403341402 . _:b403341566 . _:b403341822 . _:b403341854 . _:b403341447 . _:b403341717 . _:b403341849 . _:b403341446 . _:b403341678 . _:b403341848 . _:b403341441 . . . _:b403341753 . _:b403341851 . _:b403341440 . . _:b403341850 . _:b403341443 . _:b403341818 . _:b403341861 . _:b403341442 . _:b403341730 . _:b403341454 . _:b403341412 . _:b403341860 . _:b5177933 . _:b403341453 . _:b403341863 . _:b403341452 . . _:b403341862 . _:b403341455 . . . . . _:b5177952 . _:b403341857 . _:b403341541 . _:b403341454 . . _:b403341856 . _:b403341449 . . . _:b5177930 . _:b403341859 . _:b403341448 . _:b403341547 . . _:b403341502 . _:b403341858 . _:b403341451 . . . _:b403341869 . _:b403341450 . _:b403341618 . _:b403341868 . _:b5177931 . _:b403341461 . _:b5177929 . _:b403341476 . _:b403341871 . _:b403341460 . _:b403341782 . . _:b403341462 . _:b403341870 . _:b403341463 . _:b403341702 . . _:b403341865 . _:b403341462 . _:b403341864 . . _:b5177915 "In addition, we investigated the induction of the proinflammatory cytokine TNF-\u03B1, which is also produced in response to cutaneous barrier disruption (>>17<<) and tightly associated with inflammatory deviation of the immune response, such as asthma and probably allergic inflammation (48, 49)." . _:b403341457 . _:b403341433 . _:b403341867 . _:b403341456 . . _:b403341866 . _:b403341459 . _:b403341410 . _:b403341877 . _:b403341458 . . _:b403341876 . . _:b403341481 . _:b403341469 . _:b5177921 . _:b403341879 . _:b403341468 . _:b403341878 . _:b5177951 . _:b403341471 . _:b5177927 "Moreover, a functional NF-\u03BAB binding site was previously identified in the TSLP promoter in airway epithelial cells (54, >>55<<). We therefore assessed the involvement of NF-\u03BAB signaling in TSLP, IL8, and TNF-\u03B1 up-regulation by KLK5 using BAY11-7082, a specific inhibitor of the NF-\u03BAB pathway. Pretreatment of NHK with BAY11-7082 decreased the effect of KLK5 on" . . _:b403341873 . _:b403341470 . _:b403341534 . _:b403341599 . _:b403341872 . _:b403341465 . . _:b403341875 . _:b403341464 . _:b403341874 . _:b5177949 "In addition, in an immune-competent system, TSLP has the potential to activate dendritic cells and epidermal LCs in particular, triggering the generation of proallergic CD4+ Th2 cells (19, >>20<<, 64)." . _:b403341467 . _:b403341644 . _:b403341885 . . _:b403341829 . _:b403341466 . . . _:b5177901 . _:b403341884 . _:b403341477 . _:b403341781 . _:b403341476 . _:b403341664 . _:b5177935 . _:b403341886 . _:b403341479 . _:b403341881 . _:b403341478 . . _:b403341880 . _:b403341473 . . _:b403341761 . _:b403341883 . _:b5177899 . _:b403341472 . . _:b403341882 . _:b403341475 . _:b5177904 . . _:b403341679 . _:b403341474 . . . _:b403341485 . . . _:b403341484 . _:b403341646 . _:b403341487 . _:b403341677 . _:b403341490 . _:b403341816 . _:b403341486 . . . _:b403341470 . _:b403341481 . . _:b403341480 . . _:b403341483 . . . _:b403341482 . _:b403341673 . _:b403341493 . _:b403341563 . . _:b403341492 . _:b403341844 . _:b403341472 . _:b403341495 . _:b403341471 . _:b403341494 . _:b403341709 . _:b403341498 . _:b403341489 . _:b403341601 . _:b403341488 . _:b403341687 . _:b403341863 . _:b403341791 . _:b403341491 . _:b403341672 . _:b403341859 . . _:b403341490 . _:b403341864 . _:b403341496 . _:b403341501 . _:b403341500 . _:b403341503 . . _:b403341502 . _:b403341546 . _:b403341497 . _:b403341565 . _:b5177942 "These data support the notion that, in our model, TSLP-mediated skin inflammation mainly involves the innate immune system and that this cytokine may directly recruit and activate mast cells that express TSLP receptor (>>59<<). TSLP, synergistically with IL-1\u03B2 and TNF-\u03B1, stimulates the production of pro-Th2 cytokines by activated mast cells (59). Massive production of active \u03B21-tryptase by activated mast cells in Spink5\u2212/\u2212 grafted skin could also directly" . _:b403341598 . . _:b403341496 . . _:b403341499 . _:b5177952 . . . _:b403341498 . _:b403341375 "11"^^ . . _:b5177909 . _:b403341509 . _:b403341374 "11"^^ . _:b403341759 . _:b403341754 . _:b5177908 . _:b403341508 . _:b403341373 "11"^^ . _:b403341370 "16"^^ . _:b403341511 . _:b403341371 "15"^^ . _:b5177931 "That supports a major and direct role of KO keratinocytes in sensing and promoting inflammatory signal rather than the absorption of allergen by epidermal LC precursors (>>56<<), which in this condition will probably not encounter foreign particles." . _:b403341833 . _:b5177912 "Several studies have identified the proteinase-activated receptor 2 (PAR2) as a major effector of the cutaneous inflammatory response through its proteolytic activation by serine proteases (>>44<<\u201346). PAR2 is expressed in keratinocytes, and we therefore investigated its possible role in the development of the KO graft phenotype by immunohistochemistry analysis." . _:b403341372 "13"^^ . _:b403341510 . _:b5177904 "results" . _:b5177923 "We analyzed two target genes of PAR2 activation in cultured keratinocytes, ICAM1 (47) and the chemokine IL8 (>>53<<). In addition, we assessed whether IL-1\u03B2, TNF-\u03B1, and the pro-Th2 cytokine TSLP and chemokines TARC and MDC, which we showed to be overexpressed in LEKTI-deficient epidermis, could be directly up-regulated by hyperactive KLK5." . _:b403341390 . . _:b403341505 . . _:b5177913 . _:b403341504 . . . _:b403341489 . _:b403341507 . _:b403341430 . _:b5177914 "Recently, the major role of the epithelial cell\u2013derived cytokine TSLP was highlighted in the induction of cutaneous allergic inflammation and in AD epidermis (>>19<<), a key feature of NS. We therefore analyzed the expression of this cytokine in the epidermis of grafted skin." . _:b403341506 . _:b403341760 . _:b403341382 "8"^^ . _:b403341517 . . _:b403341383 "8"^^ . _:b403341884 . _:b403341516 . _:b403341380 "8"^^ . _:b403341821 . . _:b403341519 . . _:b403341381 "8"^^ . _:b403341518 . . . _:b5177918 "This subcellular localization is consistent with the presence of intracytoplasmic vesicles and/or Golgi pool\u2013containing PAR2 because of its internalization and/or neosynthesis after its activation by epidermal proteases (>>50<<) (Fig. 4, E and F; and Fig. S5). RT-PCR analysis showed that Par2 mRNA was slightly overexpressed in KO epidermis compared with WT, whereas no variation was detected between WT and KO dermis (Fig. 4 M)." . _:b403341378 "10"^^ . _:b403341513 . _:b403341398 . _:b403341379 "9"^^ . _:b403341377 "10"^^ . _:b403341512 . . _:b403341376 "10"^^ . _:b403341515 . _:b403341568 . _:b403341445 . . _:b403341514 . _:b403341390 "6"^^ . _:b403341391 "6"^^ . _:b403341525 . _:b403341453 . _:b403341524 . _:b403341388 "7"^^ . . _:b403341527 . . _:b403341389 "7"^^ . _:b403341444 . _:b403341526 . _:b403341386 "7"^^ . _:b403341521 . _:b403341387 "7"^^ . _:b403341700 . _:b403341590 . . . _:b403341755 . _:b403341520 . _:b403341384 "7"^^ . . _:b403341511 . . _:b403341408 . _:b403341523 . _:b403341385 "7"^^ . _:b403341398 "6"^^ . _:b403341522 . . . _:b403341779 . _:b403341533 . _:b403341399 "6"^^ . _:b403341763 . . _:b5177938 . _:b403341743 . _:b403341396 "6"^^ . _:b403341674 . _:b403341532 . . _:b403341397 "6"^^ . _:b403341701 . _:b403341535 . _:b403341510 . _:b403341394 "6"^^ . _:b403341879 . _:b403341534 . . _:b403341395 "6"^^ . _:b403341529 . . _:b403341525 . _:b403341392 "6"^^ . _:b403341528 . . _:b403341832 . . _:b403341393 "6"^^ . _:b403341531 . . _:b403341513 . _:b403341530 . _:b5177941 "It is of interest to note that elimination of T lymphocytes in K5-Tslp mice by crossing with TCR-\u03B2\u2013deficient mice does not affect eosinophilic and mast cell skin infiltration (>>21<<), as observed in Spink5 KO grafted skin." . . . _:b403341621 . _:b403341405 "5"^^ . _:b403341541 . . _:b403341741 . _:b403341750 . _:b403341404 "5"^^ . _:b403341540 . . _:b403341407 "5"^^ . _:b403341543 . _:b403341395 . _:b403341406 "5"^^ . _:b5177946 "activated mast cells in Spink5\u2212/\u2212 grafted skin could also directly participate in the inflammatory response and favor a Th2-permissive microenvironment through PAR2 activation on mast cells (60), dendritic cells (61), and eosinophils (>>62<<, 63). In addition, in an immune-competent system, TSLP has the potential to activate dendritic cells and epidermal LCs in particular, triggering the generation of proallergic CD4+ Th2 cells (19, 20, 64)." . . _:b403341542 . _:b403341817 . _:b403341537 . _:b403341738 . _:b403341400 "6"^^ . _:b403341536 . . . _:b403341749 . _:b403341403 "5"^^ . _:b403341643 . _:b403341401 "6"^^ . _:b403341539 . _:b403341402 "5"^^ . _:b5177920 . _:b403341860 . _:b403341538 . . . _:b403341413 "5"^^ . _:b403341610 . _:b403341549 . . _:b403341412 "5"^^ . . _:b403341548 . _:b403341415 "5"^^ . _:b403341551 . _:b403341841 . . _:b403341414 "5"^^ . _:b403341550 . _:b403341871 . _:b403341409 "5"^^ . _:b403341545 . _:b403341540 . . _:b403341408 "5"^^ . . . _:b403341544 . _:b403341411 "5"^^ . _:b403341829 "2"^^ . . . _:b403341484 . _:b403341735 . _:b403341547 . . _:b403341410 "5"^^ . _:b403341828 "2"^^ . . _:b403341546 . _:b403341494 . . . _:b403341421 "5"^^ . _:b403341831 "2"^^ . . _:b403341557 . _:b403341420 "5"^^ . _:b403341830 "2"^^ . . _:b403341556 . . _:b403341423 "5"^^ . _:b403341825 "2"^^ . . _:b403341559 . _:b403341422 "5"^^ . _:b403341824 "2"^^ . _:b403341558 . _:b403341634 . _:b403341417 "5"^^ . _:b5177912 . _:b403341827 "2"^^ . . _:b403341553 . _:b403341416 "5"^^ . _:b403341826 "2"^^ . _:b403341552 . _:b5177913 "The intercellular adhesion molecule 1 (ICAM1), which has been described as a direct target gene of PAR2 activation (>>47<<), was restricted to basal cells in the WT epidermis." . _:b403341419 "5"^^ . _:b403341837 "2"^^ . _:b403341555 . _:b403341652 . _:b403341418 "5"^^ . _:b403341836 "2"^^ . _:b403341532 . _:b403341554 . _:b403341429 "4"^^ . _:b403341839 "2"^^ . _:b403341565 . _:b403341428 "4"^^ . . _:b403341838 "2"^^ . . _:b403341564 . . _:b403341431 "4"^^ . _:b403341833 "2"^^ . _:b403341567 . _:b403341649 . _:b403341430 "4"^^ . . _:b403341832 "2"^^ . _:b403341566 . _:b403341425 "4"^^ . _:b403341835 "2"^^ . _:b403341561 . _:b403341424 "4"^^ . _:b403341796 . _:b403341834 "2"^^ . _:b403341560 . _:b403341427 "4"^^ . . _:b403341845 "2"^^ . _:b403341563 . _:b403341583 . _:b403341426 "4"^^ . _:b403341428 . _:b403341844 "2"^^ . _:b403341562 . _:b403341437 "4"^^ . _:b403341847 "2"^^ . _:b403341426 . _:b403341458 . _:b403341436 "4"^^ . _:b403341597 . _:b403341846 "2"^^ . _:b403341576 . . _:b403341439 "4"^^ . . _:b403341841 "2"^^ . _:b403341438 "4"^^ . _:b403341464 . _:b403341840 "2"^^ . _:b403341459 . _:b403341433 "4"^^ . _:b403341539 . . _:b403341843 "2"^^ . . _:b403341419 . _:b403341432 "4"^^ . . _:b403341842 "2"^^ . _:b403341435 "4"^^ . _:b403341526 . _:b403341853 "2"^^ . _:b403341434 "4"^^ . _:b403341852 "2"^^ . . . . . _:b403341499 . . _:b403341445 "4"^^ . _:b403341855 "2"^^ . . _:b403341444 "4"^^ . _:b403341854 "2"^^ . _:b403341447 "4"^^ . _:b403341617 . _:b403341849 "2"^^ . . _:b403341440 . _:b403341446 "4"^^ . _:b403341837 . _:b403341848 "2"^^ . . _:b403341627 . _:b403341441 "4"^^ . _:b403341509 . . _:b403341851 "2"^^ . . _:b403341440 "4"^^ . . _:b5177934 . _:b403341850 "2"^^ . _:b403341647 . _:b403341443 "4"^^ . . _:b403341861 "2"^^ . . _:b403341442 "4"^^ . _:b5177906 "We previously demonstrated in newborn KO mice that premature desmosome cleavage leading to SC detachment was a key event of NS (>>32<<). As SC cohesion was also affected in KO graft, desmosomes were examined by electron microscopy. Well-structured desmosomes were seen at the interface between the GR and the SC (GR-SC) of the WT grafted epidermis, whereas asymmetrical" . . _:b403341860 "2"^^ . _:b403341453 "4"^^ . _:b403341863 "2"^^ . _:b403341480 . . _:b403341452 "4"^^ . _:b403341862 "2"^^ . _:b403341463 . _:b403341852 . _:b403341455 "4"^^ . _:b403341857 "2"^^ . _:b403341424 . _:b403341454 "4"^^ . . _:b403341856 "2"^^ . _:b403341723 . . _:b403341449 "4"^^ . . _:b403341859 "2"^^ . _:b5177928 . _:b403341448 "4"^^ . _:b403341785 . _:b403341549 . _:b403341858 "2"^^ . _:b403341393 . _:b403341451 "4"^^ . _:b403341869 "2"^^ . _:b403341450 "4"^^ . _:b403341868 "2"^^ . _:b403341552 . _:b403341461 "4"^^ . _:b403341871 "2"^^ . _:b403341460 "4"^^ . _:b5177900 "Total dorsal skin from E19.5 mice was transplanted onto nude mice using the skin flap technique (>>67<<) and analyzed after 8 wk." . _:b403341824 . _:b403341870 "2"^^ . _:b403341463 "3"^^ . _:b403341865 "2"^^ . . _:b403341462 "3"^^ . . _:b403341589 . _:b403341587 . _:b403341864 "2"^^ . . _:b403341457 "4"^^ . _:b403341381 . _:b403341856 . _:b403341867 "2"^^ . _:b403341456 "4"^^ . _:b5177920 "These two chemokines are known to preferentially attract CCR4+ Th2 cell (>>51<<). In the epidermis and the dermis of E19.5 KO mice, Tarc and Mdc were overexpressed at the transcriptional level (Fig." . . _:b403341866 "2"^^ . . . _:b403341459 "4"^^ . _:b403341559 . _:b403341877 "2"^^ . _:b5177922 . . _:b403341458 "4"^^ . _:b403341697 . _:b403341876 "2"^^ . _:b403341469 "3"^^ . _:b403341876 . _:b403341879 "2"^^ . _:b5177924 . _:b403341626 . _:b403341684 . . _:b403341468 "3"^^ . . _:b5177925 . _:b403341878 "2"^^ . . _:b403341471 "3"^^ . _:b403341873 "2"^^ . . _:b403341470 "3"^^ . _:b403341720 . _:b403341872 "2"^^ . . _:b403341465 "3"^^ . _:b403341485 . _:b403341875 "2"^^ . _:b403341464 "3"^^ . _:b403341792 . _:b403341372 . _:b403341771 . _:b403341373 . _:b403341874 "2"^^ . . _:b403341374 . _:b403341375 . _:b403341858 . _:b403341467 "3"^^ . _:b403341885 "2"^^ . _:b403341370 . _:b403341371 . _:b403341466 "3"^^ . _:b403341836 . _:b403341884 "2"^^ . . _:b403341722 . . _:b403341477 "3"^^ . . _:b403341665 . _:b403341476 "3"^^ . . _:b403341886 "2"^^ . _:b403341479 "3"^^ . _:b403341881 "2"^^ . _:b403341478 "3"^^ . _:b403341396 . _:b403341397 . _:b403341880 "2"^^ . _:b403341398 . _:b403341399 . _:b403341473 "3"^^ . _:b403341392 . _:b403341393 . _:b403341883 "2"^^ . _:b403341394 . _:b403341395 . _:b5177945 . _:b403341472 "3"^^ . _:b403341404 . _:b403341405 . _:b403341882 "2"^^ . _:b403341406 . _:b403341604 . _:b403341407 . _:b403341475 "3"^^ . . _:b403341400 . _:b403341401 . _:b403341688 . _:b403341402 . _:b403341403 . _:b403341530 . _:b403341474 "3"^^ . _:b403341380 . _:b403341381 . _:b403341382 . _:b403341637 . _:b403341383 . _:b403341485 "3"^^ . _:b403341376 . _:b403341377 . _:b403341505 . . _:b403341538 . _:b403341378 . _:b403341379 . _:b403341484 "3"^^ . _:b403341388 . _:b403341392 . _:b403341389 . _:b403341390 . _:b403341391 . _:b403341575 . _:b403341487 "3"^^ . . _:b403341384 . _:b403341385 . _:b403341386 . . _:b403341442 . _:b403341387 . _:b403341486 "3"^^ . _:b403341428 . _:b403341429 . _:b403341430 . _:b403341431 . _:b403341481 "3"^^ . _:b403341424 . _:b403341425 . _:b403341815 . _:b403341426 . _:b403341427 . _:b403341480 "3"^^ . _:b403341436 . _:b403341751 . _:b403341437 . _:b403341438 . _:b403341439 . _:b403341483 "3"^^ . . _:b403341432 . . _:b403341433 . _:b403341434 . _:b403341435 . _:b5177926 . _:b403341482 "3"^^ . . _:b403341412 . _:b403341413 . _:b403341414 . _:b403341415 . _:b403341882 . _:b403341493 "3"^^ . _:b403341408 . . _:b403341409 . _:b5177918 . _:b403341410 . _:b403341878 . _:b403341641 . . _:b403341411 . _:b403341492 "3"^^ . . _:b403341420 . _:b403341421 . _:b403341504 . _:b403341422 . . . _:b403341423 . _:b5177927 . _:b403341495 "3"^^ . _:b403341416 . _:b403341417 . _:b403341418 . _:b403341419 . _:b403341494 "3"^^ . . _:b403341460 . _:b403341461 . _:b403341462 . _:b403341463 . _:b403341489 "3"^^ . _:b403341456 . _:b403341457 . _:b403341458 . _:b403341732 . _:b403341459 . _:b403341488 "3"^^ . _:b403341468 . _:b403341469 . _:b403341470 . _:b403341853 . _:b403341471 . _:b403341491 "3"^^ . _:b403341464 . _:b403341465 . _:b403341466 . _:b403341467 . _:b403341490 "3"^^ . _:b403341444 . _:b403341445 . . _:b403341446 . . _:b403341447 . _:b403341548 . _:b403341501 "3"^^ . _:b403341440 . _:b403341441 . _:b403341442 . . _:b403341443 . _:b403341500 "3"^^ . . _:b403341452 . _:b403341453 . . _:b403341747 . _:b403341454 . . _:b403341593 . _:b403341503 "3"^^ . _:b403341455 . _:b403341448 . _:b403341449 . _:b5177915 . _:b403341450 . _:b403341653 . _:b403341451 . _:b403341451 . _:b403341502 "3"^^ . _:b403341492 . _:b403341493 . . _:b403341494 . _:b5177919 . _:b403341495 . _:b403341497 "3"^^ . . _:b403341488 . _:b403341748 . . _:b403341417 . _:b403341489 . . _:b403341490 . _:b403341491 . _:b5177940 . _:b403341770 . _:b403341625 . _:b403341496 "3"^^ . _:b403341500 . _:b403341712 . _:b403341501 . . . _:b403341502 . _:b403341503 . _:b403341499 "3"^^ . _:b403341496 . _:b403341497 . _:b403341883 . _:b403341498 . _:b403341499 . _:b403341498 "3"^^ . _:b403341476 . . _:b403341477 . _:b403341478 . _:b403341479 . _:b403341509 "3"^^ . _:b403341472 . _:b403341683 . _:b403341473 . _:b403341609 . . _:b403341474 . _:b403341475 . _:b403341508 "3"^^ . _:b403341484 . _:b403341662 . _:b403341485 . _:b403341486 . _:b403341550 . _:b403341487 . _:b403341511 "3"^^ . _:b403341480 . . _:b403341481 . _:b403341482 . _:b403341483 . _:b403341510 "3"^^ . _:b403341524 . _:b403341711 . . _:b403341525 . _:b403341526 . . _:b403341527 . _:b403341787 . _:b403341713 . _:b403341505 "3"^^ . _:b403341520 . _:b403341521 . _:b403341850 . _:b403341522 . . _:b403341523 . _:b403341504 "3"^^ . _:b403341532 . _:b403341533 . _:b403341534 . _:b403341535 . _:b403341507 "3"^^ . . _:b403341528 . _:b403341529 . . _:b403341530 . . _:b403341531 . _:b403341506 "3"^^ . . _:b403341508 . . . _:b403341509 . _:b403341510 . . _:b403341511 . _:b403341517 "3"^^ . _:b403341504 . _:b403341505 . . _:b403341506 . _:b403341507 . _:b5177933 "Interestingly, we showed that the intracellular pool of PAR2 was increased in the more differentiated layers of the E19.5 KO epidermis and colocalized with TSLP and hyperactive KLK5 in the upper layers of the KO epidermis (28, >>43<<). Therefore, these early events observed in utero confirmed that deregulated KLK5 in the upper GR layers initiates the proinflammatory response through PAR2 cleavage and TSLP production in vivo." . _:b403341516 "3"^^ . _:b403341516 . . _:b403341517 . _:b403341518 . _:b403341519 . _:b403341820 . _:b403341519 "3"^^ . _:b403341512 . . . _:b403341513 . . _:b403341514 . _:b403341515 . _:b403341518 "3"^^ . _:b403341556 . . _:b403341557 . . _:b403341558 . _:b403341756 . . _:b403341559 . _:b403341789 . _:b403341513 "3"^^ . _:b403341376 . _:b403341552 . . . _:b403341553 . . _:b403341554 . _:b403341555 . _:b403341512 "3"^^ . _:b403341564 . _:b403341565 . _:b403341566 . _:b403341567 . _:b403341515 "3"^^ . _:b403341560 . _:b403341561 . _:b403341562 . _:b403341563 . _:b403341514 "3"^^ . . _:b403341540 . _:b403341405 . _:b403341541 . _:b403341427 . _:b403341663 . . _:b403341542 . _:b403341543 . _:b403341525 "3"^^ . _:b403341536 . _:b403341537 . . . . . _:b403341538 . . _:b403341524 "3"^^ . _:b403341539 . _:b403341548 . . _:b403341507 . _:b403341466 . _:b403341549 . . _:b403341550 . _:b403341551 . _:b403341527 "3"^^ . _:b403341544 . _:b403341545 . _:b403341546 . _:b403341547 . _:b403341526 "3"^^ . _:b403341588 . _:b403341589 . _:b403341590 . _:b403341591 . _:b403341768 . _:b403341521 "3"^^ . _:b403341437 . _:b403341584 . _:b5177952 . _:b403341764 . _:b403341585 . _:b403341586 . _:b403341587 . _:b403341520 "3"^^ . _:b403341501 . . . _:b403341596 . . . . _:b403341597 . . . _:b403341598 . _:b403341599 . _:b403341523 "3"^^ . _:b403341592 . . _:b403341593 . _:b403341594 . _:b403341595 . _:b403341522 "3"^^ . _:b403341572 . _:b403341573 . _:b5177952 . . _:b403341574 . _:b403341575 . _:b403341628 . _:b403341533 "3"^^ . _:b403341441 . _:b403341568 . _:b403341569 . _:b403341570 . . _:b403341571 . _:b403341532 "3"^^ . _:b403341580 . . _:b403341431 . _:b403341581 . _:b403341582 . _:b403341583 . _:b403341535 "3"^^ . _:b403341576 . _:b403341577 . _:b403341578 . _:b403341579 . _:b403341534 "3"^^ . _:b403341620 . _:b403341621 . _:b403341622 . _:b403341623 . _:b403341529 "3"^^ . _:b403341616 . _:b403341704 . _:b403341617 . _:b403341618 . . . _:b403341619 . _:b403341528 "3"^^ . _:b403341628 . . _:b403341629 . . _:b403341630 . _:b403341631 . _:b403341531 "3"^^ . _:b403341370 . _:b403341624 . _:b403341625 . _:b403341626 . _:b403341503 . _:b403341627 . _:b403341530 "3"^^ . _:b403341604 . _:b403341605 . _:b403341645 . _:b403341573 . _:b403341606 . . _:b403341607 . _:b403341541 "3"^^ . _:b403341600 . _:b403341601 . _:b403341572 . _:b403341602 . _:b403341603 . _:b403341540 "3"^^ . _:b403341383 . _:b403341612 . _:b403341582 . _:b403341613 . _:b403341575 . . _:b403341614 . _:b403341615 . _:b403341560 . _:b403341543 "3"^^ . . _:b403341608 . _:b403341812 . _:b403341609 . _:b403341574 . . _:b403341610 . _:b403341611 . _:b403341542 "3"^^ . _:b403341652 . _:b403341653 . _:b5177924 "6 A), in accordance with previously published data, on NHK treated with trypsin (>>47<<), suggesting PAR2 cleavage and activation by KLK5 in our in vitro model." . _:b403341569 . _:b403341654 . _:b403341655 . _:b403341537 "3"^^ . _:b403341648 . . _:b403341568 . _:b403341649 . . _:b403341650 . _:b403341651 . _:b403341536 "3"^^ . . _:b403341660 . . _:b403341571 . _:b403341661 . _:b403341662 . _:b403341744 . . _:b403341539 "3"^^ . _:b403341663 . _:b403341443 . _:b403341656 . _:b403341657 . _:b403341570 . . _:b403341658 . _:b403341659 . _:b5177932 "Interestingly, we showed that the intracellular pool of PAR2 was increased in the more differentiated layers of the E19.5 KO epidermis and colocalized with TSLP and hyperactive KLK5 in the upper layers of the KO epidermis (>>28<<, 43). Therefore, these early events observed in utero confirmed that deregulated KLK5 in the upper GR layers initiates the proinflammatory response through PAR2 cleavage and TSLP production in vivo." . _:b403341538 "3"^^ . _:b403341636 . . . _:b403341581 . _:b403341637 . _:b403341638 . . _:b403341549 "3"^^ . _:b403341639 . _:b403341632 . _:b403341772 . _:b403341633 . _:b403341580 . _:b403341634 . _:b403341635 . _:b403341548 "3"^^ . . _:b403341644 . . _:b403341645 . _:b403341583 . . _:b403341646 . _:b403341647 . _:b403341551 "3"^^ . _:b403341640 . _:b403341641 . _:b403341582 . _:b403341642 . _:b403341643 . _:b403341550 "3"^^ . _:b403341684 . . _:b403341685 . _:b403341577 . . _:b403341686 . _:b403341687 . _:b403341545 "3"^^ . _:b403341680 . _:b403341614 . _:b403341681 . _:b403341576 . _:b403341375 . _:b403341682 . _:b403341683 . _:b403341544 "3"^^ . _:b403341692 . _:b403341799 . _:b403341693 . _:b403341579 . _:b403341694 . _:b403341695 . _:b403341547 "3"^^ . _:b403341688 . . _:b403341689 . _:b403341578 . _:b403341690 . _:b403341691 . _:b403341546 "3"^^ . . _:b403341668 . _:b403341669 . _:b403341589 . . _:b403341557 "2"^^ . _:b403341670 . _:b403341671 . _:b403341664 . _:b403341665 . _:b403341588 . _:b403341666 . _:b403341556 "2"^^ . _:b403341667 . _:b403341676 . _:b403341677 . _:b403341591 . _:b403341678 . _:b403341559 "2"^^ . _:b403341679 . _:b403341672 . _:b403341673 . _:b403341590 . . _:b403341558 "2"^^ . _:b403341674 . _:b403341675 . _:b403341716 . _:b403341717 . _:b403341585 . . _:b403341718 . _:b403341553 "2"^^ . . _:b403341719 . _:b403341712 . _:b5177936 "IL-1\u03B2 can be proteolytically activated by KLK7 (57), which is hyperactivated in Spink5\u2212/\u2212 skin (>>32<<). Thus, this protease could participate in the inflammatory process in our model as observed in KLK7 transgenic mice (58)." . _:b403341800 . _:b403341713 . _:b403341584 . "10.1084%2Fjem.20082242" . _:b403341714 . _:b403341715 . _:b403341552 "3"^^ . _:b403341724 . _:b403341725 . _:b403341631 . _:b403341587 . _:b403341726 . _:b403341555 "2"^^ . _:b403341727 . _:b403341788 . _:b403341720 . _:b403341721 . _:b403341586 . _:b403341722 . _:b403341554 "2"^^ . . _:b403341406 . _:b403341723 . _:b403341700 . _:b403341701 . _:b403341597 . _:b403341702 . _:b403341565 "2"^^ . _:b403341703 . _:b403341696 . . . _:b403341596 . _:b403341697 . _:b403341698 . _:b403341564 "2"^^ . . _:b403341699 . _:b403341387 . _:b403341567 . _:b403341708 . _:b403341709 . _:b403341599 . _:b403341710 . _:b403341567 "2"^^ . _:b403341711 . _:b403341596 . _:b403341515 . _:b403341704 . _:b403341705 . _:b403341598 . _:b403341456 . _:b403341566 "2"^^ . _:b403341706 . . _:b403341707 . _:b403341748 . _:b403341749 . _:b403341593 . _:b403341750 . _:b403341561 "2"^^ . _:b403341751 . . _:b403341744 . _:b403341745 . _:b403341592 . _:b403341746 . _:b403341560 "2"^^ . _:b403341747 . _:b403341756 . _:b403341757 . _:b403341595 . _:b403341758 . _:b403341563 "2"^^ . . _:b403341759 . _:b403341827 . _:b403341497 . _:b403341752 . _:b403341403 . _:b403341753 . _:b403341594 . . _:b403341754 . _:b403341562 "2"^^ . _:b403341475 . . . _:b5177949 . _:b403341457 . _:b403341755 . _:b403341732 . _:b5177946 . _:b403341733 . _:b403341605 . _:b403341734 . _:b403341573 "2"^^ . _:b403341735 . _:b403341668 . . . _:b5177910 "In addition to the protease hyperactivities at 20 and 30 kD, which has been previously described in embryos (>>32<<), casein gel zymography experiments showed a KO graft-specific activity migrating at 34 kD." . _:b403341551 . _:b403341728 . _:b403341729 . _:b403341604 . . _:b403341572 "2"^^ . _:b403341730 . _:b403341731 . _:b403341740 . _:b403341741 . _:b403341607 . _:b403341742 . _:b403341575 "2"^^ . _:b403341743 . . _:b403341736 . _:b403341603 . _:b403341450 . _:b403341606 . _:b403341737 . _:b403341738 . _:b403341574 "2"^^ . . _:b403341774 . _:b403341739 . _:b403341780 . _:b403341781 . _:b403341601 . _:b403341782 . _:b403341569 "2"^^ . _:b403341783 . _:b403341776 . _:b403341777 . _:b403341600 . _:b403341778 . _:b403341568 "2"^^ . _:b403341779 . _:b403341788 . _:b403341584 . _:b403341388 . _:b403341789 . _:b403341603 . _:b403341790 . _:b403341571 "2"^^ . _:b403341791 . _:b403341784 . _:b403341785 . _:b403341602 . _:b403341468 . . _:b403341786 . _:b5177954 . _:b403341570 "2"^^ . _:b403341787 . _:b403341606 . _:b403341764 . _:b403341588 . _:b403341765 . _:b403341613 . _:b403341766 . _:b403341581 "2"^^ . _:b403341767 . _:b403341874 . _:b403341533 . _:b403341760 . . _:b403341612 . _:b403341761 . . . _:b403341740 . _:b403341580 "2"^^ . _:b403341762 . _:b403341763 . _:b403341772 . _:b403341773 . _:b403341615 . _:b403341774 . _:b403341583 "2"^^ . _:b403341775 . _:b403341768 . _:b403341769 . _:b403341614 . . _:b403341770 . _:b403341582 "2"^^ . _:b403341771 . _:b403341600 . _:b403341812 . _:b403341813 . _:b403341609 . . _:b403341577 "2"^^ . _:b403341814 . _:b403341815 . _:b403341808 . _:b403341809 . _:b403341608 . _:b403341810 . _:b403341576 "2"^^ . _:b403341811 . _:b403341820 . _:b403341389 . _:b403341611 . _:b403341821 . _:b403341822 . _:b403341579 "2"^^ . _:b403341823 . _:b403341816 . _:b403341783 . . _:b403341817 . _:b403341610 . . _:b403341818 . _:b5177928 "6 C), which is in accordance with already published data (>>47<<). Moreover, this treatment also strongly reduced KLK5 efficiency in inducing TSLP, IL8, and TNF-\u03B1 transcription (mean of 81, 59, and 40% decrease, respectively; Fig." . _:b403341578 "2"^^ . _:b403341819 . _:b403341378 . _:b403341796 . . _:b403341797 . _:b403341638 . _:b403341621 . _:b403341798 . _:b403341589 "2"^^ . _:b403341799 . _:b403341792 . _:b5177952 "Several studies have reported significant genetic association of SPINK5 polymorphisms with AD in different populations (>>34<<\u201338), whereas other studies did not confirm these results (39\u201341)." . _:b403341578 . _:b403341561 . _:b403341452 . _:b403341727 . _:b403341620 . _:b403341793 . _:b403341794 . _:b403341588 "2"^^ . _:b403341795 . _:b403341804 . _:b403341805 . _:b403341623 . _:b403341806 . _:b403341591 "2"^^ . _:b403341807 . _:b403341800 . _:b403341801 . _:b403341622 . . _:b403341802 . _:b403341590 "2"^^ . _:b403341803 . . _:b403341844 . _:b403341845 . _:b403341617 . . _:b403341384 . _:b403341585 "2"^^ . _:b403341486 . _:b403341846 . _:b403341847 . _:b403341840 . _:b403341835 . _:b403341769 . _:b403341841 . _:b403341616 . _:b403341842 . _:b403341584 "2"^^ . _:b403341843 . . . _:b403341852 . _:b403341853 . _:b403341619 . _:b403341854 . _:b5177941 . _:b403341587 "2"^^ . . . _:b403341855 . . _:b403341848 . _:b403341849 . _:b403341618 . _:b403341850 . _:b403341586 "2"^^ . _:b403341851 . _:b403341828 . _:b403341829 . _:b403341629 . . _:b403341371 . _:b403341597 "2"^^ . _:b403341830 . _:b403341831 . . _:b403341824 . _:b403341825 . _:b403341628 . _:b403341826 . _:b403341596 "2"^^ . . _:b403341827 . . _:b403341836 . _:b403341837 . _:b403341631 . _:b403341838 . _:b403341599 "2"^^ . . _:b403341839 . _:b403341832 . _:b403341636 . . _:b403341833 . _:b403341630 . _:b403341544 . . _:b403341834 . _:b403341598 "2"^^ . _:b403341835 . _:b403341876 . _:b403341877 . _:b403341625 . _:b403341878 . _:b403341593 "2"^^ . _:b403341879 . _:b403341872 . _:b403341869 . _:b403341873 . _:b403341669 . _:b403341624 . _:b403341874 . _:b403341592 "2"^^ . _:b403341875 . _:b403341884 . _:b403341885 . _:b403341627 . _:b403341886 . _:b403341802 . _:b403341595 "2"^^ . _:b403341880 . _:b403341881 . _:b403341626 . . . _:b403341594 "2"^^ . _:b403341882 . . _:b403341883 . _:b403341512 . . . _:b403341860 . _:b403341861 . _:b403341637 . _:b403341862 . _:b403341605 "2"^^ . _:b403341448 . _:b403341863 . _:b403341856 . _:b403341857 . _:b403341775 . _:b403341636 . _:b403341858 . _:b403341706 . _:b403341604 "2"^^ . _:b403341859 . _:b5177948 "In addition, in an immune-competent system, TSLP has the potential to activate dendritic cells and epidermal LCs in particular, triggering the generation of proallergic CD4+ Th2 cells (>>19<<, 20, 64)." . _:b403341868 . _:b403341869 . _:b403341639 . _:b403341870 . _:b403341607 "2"^^ . _:b403341871 . . _:b403341864 . _:b403341865 . _:b403341638 . . _:b403341606 "2"^^ . _:b403341379 . . _:b403341866 . _:b403341867 . . _:b403341633 . _:b403341601 "2"^^ . _:b403341632 . _:b403341865 . . _:b403341600 "2"^^ . . _:b403341635 . . _:b403341603 "2"^^ . _:b5177909 "In summary, the abnormal differentiation pattern observed in KO graft, together with the desmosomal cleavage, confirmed that the KO grafted skin reproduces all major histological and ultrastructural anomalies seen in NS patients skin (>>43<<), thus providing a suitable model to understand the long-term effect of LEKTI deficiency in the skin." . _:b403341808 . . _:b403341634 . _:b403341602 "2"^^ . _:b403341790 . _:b403341645 . _:b403341613 "2"^^ . _:b403341658 . _:b403341854 . _:b403341382 . _:b403341644 . _:b403341612 "2"^^ . . _:b5177947 "mast cells in Spink5\u2212/\u2212 grafted skin could also directly participate in the inflammatory response and favor a Th2-permissive microenvironment through PAR2 activation on mast cells (60), dendritic cells (61), and eosinophils (62, >>63<<). In addition, in an immune-competent system, TSLP has the potential to activate dendritic cells and epidermal LCs in particular, triggering the generation of proallergic CD4+ Th2 cells (19, 20, 64)." . _:b403341647 . . _:b403341615 "2"^^ . _:b403341613 . _:b403341758 . _:b403341646 . _:b403341614 "2"^^ . _:b5177951 "5) who display KLK5 and PAR2 deregulation (>>43<<). Thus, TSLP provides an important link between the absence of LEKTI protease inhibitor and the AD-like phenotype constantly seen in NS patients." . _:b403341641 . _:b403341609 "2"^^ . _:b403341686 . _:b403341640 . _:b403341680 . _:b403341608 "2"^^ . . _:b403341643 . _:b403341611 "2"^^ . _:b403341671 . _:b403341642 . _:b403341610 "2"^^ . _:b5177903 . _:b403341825 . . _:b5177948 . _:b403341653 . _:b403341621 "2"^^ . _:b403341521 . _:b403341652 . _:b403341620 "2"^^ . _:b403341655 . _:b403341623 "2"^^ . _:b5177953 . _:b403341632 . . _:b403341654 . _:b403341473 . _:b403341622 "2"^^ . _:b5177911 . _:b403341649 . _:b403341617 "2"^^ . _:b403341648 . . _:b403341616 "2"^^ . _:b403341651 . _:b403341517 . _:b403341619 "2"^^ . _:b5177939 . _:b403341650 . _:b403341618 "2"^^ . . _:b403341527 . _:b403341661 . _:b403341415 . _:b403341629 "2"^^ . _:b403341660 . _:b403341628 "2"^^ . _:b403341726 . _:b403341663 . _:b403341631 "2"^^ . _:b403341662 . _:b403341396 . _:b403341630 "2"^^ . _:b403341801 . _:b403341508 . _:b403341657 . _:b403341625 "2"^^ . . _:b403341656 . . _:b403341624 "2"^^ . . _:b403341784 . _:b403341659 . _:b403341762 . _:b403341627 "2"^^ . . _:b403341658 . _:b403341626 "2"^^ . _:b403341669 . . _:b403341637 "2"^^ . . . _:b5177912 . _:b403341668 . . _:b403341636 "2"^^ . _:b403341685 . . _:b403341671 . _:b403341639 "2"^^ . _:b403341670 . _:b5177957 "An emerging hypothesis regarding asthma and AD is that they are epithelial diseases initiated by the epithelial cells themselves through the production of TSLP (>>66<<). In the present study, we have identified a mechanism leading to the production of the major pro-Th2 cytokine TSLP by skin epithelial cells, despite the absence of microbe products or physical injury." . _:b403341828 . _:b403341638 "2"^^ . _:b5177937 "Thus, this protease could participate in the inflammatory process in our model as observed in KLK7 transgenic mice (>>58<<)." . . . . _:b403341665 . . _:b403341633 "2"^^ . . _:b403341810 . _:b403341664 . _:b403341632 "2"^^ . . _:b403341780 . _:b403341667 . _:b403341635 "2"^^ . _:b403341516 . . _:b403341666 . _:b403341377 . _:b403341805 . _:b403341634 "2"^^ . . _:b403341804 . . _:b403341677 . _:b403341645 "2"^^ . _:b403341676 . _:b403341644 "2"^^ . _:b403341679 . . _:b403341647 "2"^^ . . _:b403341678 . _:b403341571 . _:b403341646 "2"^^ . _:b403341705 . _:b403341666 . _:b5177925 "Previous studies have shown that up-regulation of ICAM1 by PAR2 signaling was mediated by NF-\u03BAB pathway (>>47<<). Moreover, a functional NF-\u03BAB binding site was previously identified in the TSLP promoter in airway epithelial cells (54, 55). We therefore assessed the involvement of NF-\u03BAB signaling in TSLP, IL8, and TNF-\u03B1 up-regulation by KLK5 using" . _:b403341868 . _:b403341673 . _:b403341641 "2"^^ . . . . _:b403341672 . _:b403341640 "2"^^ . _:b5177954 "The abundant and epidermally restricted TSLP secretion could induce an AD-like phenotype associated with elevated serum levels of IgE, as illustrated in Tslp skin-specific transgenic mice (>>21<<, 65). Thereby, the effect of LEKTI deficiency on triggering PAR2 signaling could be a primary event inducing atopic manifestations in NS before skin barrier defect. In parallel, as the result of protease hyperactivities, FILAGGRIN is also" . _:b403341461 . _:b403341373 . _:b403341675 . _:b403341643 "2"^^ . . _:b403341674 . _:b403341870 . _:b403341642 "2"^^ . . _:b403341685 . _:b403341653 "2"^^ . _:b403341866 . _:b403341684 . . _:b403341652 "2"^^ . _:b403341687 . . _:b5177917 . _:b403341655 "2"^^ . . _:b403341686 . _:b403341654 "2"^^ . _:b403341420 . _:b403341612 . _:b403341681 . _:b403341649 "2"^^ . _:b403341680 . _:b403341648 "2"^^ . _:b403341683 . _:b403341651 "2"^^ . . _:b403341682 . _:b403341650 "2"^^ . _:b403341693 . _:b403341661 "2"^^ . _:b403341692 . _:b403341660 "2"^^ . _:b403341695 . _:b403341397 . _:b403341663 "2"^^ . _:b403341385 . . _:b403341694 . _:b403341662 "2"^^ . _:b403341407 . _:b403341675 . _:b403341689 . . _:b403341657 "2"^^ . . _:b403341778 . _:b403341688 . _:b403341656 "2"^^ . _:b403341691 . _:b403341659 "2"^^ . _:b403341482 . _:b5177916 . _:b403341690 . _:b403341658 "2"^^ . _:b403341477 . _:b403341842 . _:b403341703 . _:b403341701 . _:b403341669 "2"^^ . _:b403341700 . _:b403341668 "2"^^ . _:b403341807 . _:b403341703 . _:b403341671 "2"^^ . . _:b403341702 . _:b403341670 "2"^^ . _:b403341843 . _:b403341697 . _:b403341665 "2"^^ . . . . _:b403341696 . _:b403341553 . _:b403341664 "2"^^ . _:b403341699 . . _:b403341667 "2"^^ . _:b403341698 . . _:b403341666 "2"^^ . . _:b403341581 . _:b403341709 . . _:b403341677 "2"^^ . _:b403341798 . _:b403341708 . . _:b403341676 "2"^^ . . _:b403341391 . . _:b403341711 . _:b403341679 "2"^^ . . . _:b403341710 . _:b403341678 "2"^^ . . _:b403341699 . _:b403341705 . . _:b403341673 "2"^^ . _:b403341704 . _:b403341672 "2"^^ . _:b403341707 . _:b403341675 "2"^^ . _:b403341706 . _:b403341674 "2"^^ . _:b403341814 . . . _:b403341717 . _:b403341416 . _:b403341873 . _:b403341685 "2"^^ . _:b403341436 . . _:b403341619 . _:b403341716 . _:b403341684 "2"^^ . _:b403341776 . _:b403341773 . _:b403341719 . _:b403341687 "2"^^ . _:b403341372 . _:b403341718 . _:b403341686 "2"^^ . . . _:b403341713 . . _:b403341681 "2"^^ . _:b403341523 . _:b403341767 . _:b403341483 . _:b403341712 . _:b403341680 "2"^^ . _:b403341855 . _:b403341707 . _:b403341715 . _:b403341683 "2"^^ . . _:b403341692 . _:b403341714 . _:b403341682 "2"^^ . _:b403341826 . _:b403341725 . _:b403341728 . _:b403341693 "2"^^ . _:b5177936 . _:b403341724 . . _:b403341742 . _:b403341692 "2"^^ . . . . . . _:b403341727 . _:b403341623 . _:b403341710 . _:b403341695 "2"^^ . _:b403341422 . _:b403341535 . . _:b403341495 . _:b403341536 . _:b403341726 . _:b403341694 "2"^^ . _:b403341524 . _:b403341518 . _:b403341881 . . _:b403341729 . _:b403341721 . _:b403341421 . _:b403341689 "2"^^ . . _:b403341720 . _:b403341688 "2"^^ . . _:b403341723 . _:b403341691 "2"^^ . . _:b403341722 . _:b403341690 "2"^^ . . _:b403341861 . . _:b403341733 . . _:b403341701 "2"^^ . _:b403341737 . . _:b403341849 . _:b403341732 . . _:b403341700 "2"^^ . . _:b403341735 . _:b403341703 "2"^^ . _:b403341734 . _:b403341702 "2"^^ . _:b403341447 . . _:b403341729 . _:b403341697 "2"^^ . . _:b403341611 . _:b403341728 . . _:b403341696 "2"^^ . _:b403341731 . _:b403341699 "2"^^ . . _:b403341730 . _:b403341698 "2"^^ . . . _:b403341741 . _:b403341709 "2"^^ . . . _:b403341740 . _:b403341694 . _:b403341708 "2"^^ . . _:b403341743 . _:b403341711 "2"^^ . _:b403341886 . . _:b403341742 . _:b403341710 "2"^^ . _:b403341737 . . _:b403341705 "2"^^ . _:b403341867 . _:b403341736 . _:b403341580 . _:b403341704 "2"^^ . . _:b5177914 . _:b403341585 . _:b403341739 . _:b403341707 "2"^^ . _:b403341738 . . _:b403341706 "2"^^ . _:b403341414 . _:b5177901 "Each NS patient was a homozygote or a compound heterozygote for SPINK5 mutations, leading to the absence of detectable LEKTI on immunohistochemical analysis of skin sections as previously described (>>27<<). Informed consent was obtained from the patients. This study was approved by the medical ethical committee of the Purpan Hospital in Toulouse." . _:b403341670 . _:b403341488 . _:b403341749 . . _:b403341717 "2"^^ . . _:b403341748 . _:b403341591 . _:b403341716 "2"^^ . _:b403341851 . . _:b403341751 . _:b403341719 "2"^^ . _:b403341750 . _:b403341718 "2"^^ . . _:b403341745 . _:b403341569 . _:b403341713 "2"^^ . . _:b403341744 . _:b403341712 "2"^^ . _:b403341747 . _:b403341715 "2"^^ . _:b403341746 . _:b403341714 "2"^^ . _:b403341478 . _:b403341757 . . _:b403341725 "2"^^ . . . _:b403341648 . _:b403341455 . _:b403341756 . . . _:b403341724 "2"^^ . _:b403341797 . _:b403341759 . _:b403341727 "2"^^ . _:b403341758 . _:b403341726 "2"^^ . _:b403341753 . _:b403341721 "2"^^ . _:b403341752 . _:b403341720 "2"^^ . . _:b403341755 . _:b403341723 "2"^^ . _:b403341754 . _:b403341722 "2"^^ . _:b403341765 . _:b403341733 "2"^^ . _:b403341528 . _:b403341795 . _:b403341764 . _:b403341732 "2"^^ . . _:b403341752 . _:b403341661 . _:b5177950 . _:b403341767 . _:b403341529 . _:b403341735 "2"^^ . . _:b403341766 . _:b5177904 _:b5177908 . _:b403341734 "2"^^ . _:b5177904 _:b5177909 . . _:b5177904 _:b5177910 . . _:b403341761 . _:b5177904 _:b5177911 . . _:b403341729 "2"^^ . . _:b5177904 _:b5177905 . _:b5177904 _:b5177906 . _:b5177904 _:b5177907 . _:b403341760 . _:b5177904 _:b5177916 . _:b403341728 "2"^^ . _:b5177904 _:b5177917 . _:b403341651 . . _:b5177904 _:b5177918 . _:b5177904 _:b5177919 . _:b403341763 . _:b5177904 _:b5177912 . _:b403341731 "2"^^ . . _:b5177904 _:b5177913 . _:b5177904 _:b5177914 . _:b5177904 _:b5177915 . _:b403341762 . _:b403341730 "2"^^ . _:b403341773 . _:b403341418 . _:b403341857 . _:b403341741 "2"^^ . . _:b403341772 . _:b403341740 "2"^^ . . _:b403341449 . _:b403341519 . _:b403341775 . . _:b403341743 "2"^^ . _:b403341474 . . _:b403341543 . _:b403341774 . _:b403341742 "2"^^ . _:b403341769 . _:b403341737 "2"^^ . . _:b403341768 . . _:b5177903 "NHKs were seeded on 6-well plates and grown in half EpiLife 0.06 mM CaCl2 (Invitrogen) and half Green medium (>>68<<) until confluency. NHK were treated with 400 nM KLK5 (1 U trypsin-like equivalent) in serum-free Green medium for 3 h." . _:b403341736 "2"^^ . . _:b403341771 . _:b403341739 "2"^^ . _:b403341770 . _:b403341738 "2"^^ . _:b403341781 . _:b403341749 "2"^^ . . . _:b403341780 . _:b403341748 "2"^^ . . _:b403341783 . _:b403341751 "2"^^ . . . _:b403341782 . _:b403341750 "2"^^ . . _:b403341681 . _:b403341777 . _:b403341745 "2"^^ . . _:b5177947 . . _:b403341374 . _:b403341776 . _:b403341885 . . _:b403341744 "2"^^ . _:b403341779 . _:b5177930 "discussion" . _:b403341747 "2"^^ . _:b403341380 . . _:b403341778 . _:b5177904 _:b5177924 . _:b403341746 "2"^^ . _:b5177904 _:b5177925 . . _:b5177904 _:b5177926 . _:b5177904 _:b5177927 . _:b403341789 . _:b5177899 _:b5177900 . _:b403341757 "2"^^ . _:b5177904 _:b5177920 . _:b5177904 _:b5177921 . _:b5177899 _:b5177901 . _:b5177904 _:b5177922 . _:b5177899 _:b5177902 . _:b5177904 _:b5177923 . _:b403341788 . _:b5177899 _:b5177903 . _:b403341756 "2"^^ . . _:b403341520 . _:b403341791 . _:b5177904 _:b5177928 . _:b403341759 "2"^^ . _:b5177904 _:b5177929 . . _:b403341809 . _:b403341542 . _:b403341790 . _:b403341758 "2"^^ . . _:b403341785 . _:b403341840 . . _:b5177945 "active \u03B21-tryptase by activated mast cells in Spink5\u2212/\u2212 grafted skin could also directly participate in the inflammatory response and favor a Th2-permissive microenvironment through PAR2 activation on mast cells (60), dendritic cells (>>61<<), and eosinophils (62, 63). In addition, in an immune-competent system, TSLP has the potential to activate dendritic cells and epidermal LCs in particular, triggering the generation of proallergic CD4+ Th2 cells (19, 20, 64)." . _:b403341753 "2"^^ . _:b403341657 . _:b403341784 . . _:b403341752 "2"^^ . _:b5177957 . . . _:b403341787 . _:b403341755 "2"^^ . . _:b403341786 . _:b403341754 "2"^^ . . _:b403341714 . _:b403341797 . _:b403341765 "2"^^ . . _:b403341630 . _:b403341796 . _:b403341764 "2"^^ . _:b403341799 . _:b403341767 "2"^^ . . _:b403341667 . _:b5177919 "We also analyzed the expression of two pro-Th2 chemokines, thymus and activation-regulated chemokine (Tarc; Ccl17), which was found to be up-regulated in Tslp-transgenic mice skin (>>21<<), and macrophage-derived chemokine (Mdc; Ccl22). These two chemokines are known to preferentially attract CCR4+ Th2 cell (51)." . _:b403341798 . _:b403341766 "2"^^ . _:b403341793 . _:b403341806 . . _:b403341761 "2"^^ . _:b5177939 "In addition, IL-1\u03B2 and TNF-\u03B1 overexpression in Spink5\u2212/\u2212 epidermis may induce TSLP expression in keratinocytes (>>19<<, 54) and attract innate immune cells." . _:b403341554 . _:b403341792 . _:b403341760 "2"^^ . _:b5177921 "Indeed, KLK5, but not KLK7, has previously been shown to cleave and activate PAR2 (>>52<<). Therefore, the unregulated KLK5 observed in NS and in Spink5\u2212/\u2212 mice could induce a proinflammatory cascade independently of external stimuli." . _:b403341557 . _:b403341795 . _:b403341763 "2"^^ . _:b403341872 . _:b403341794 . _:b403341762 "2"^^ . . _:b403341656 . _:b403341805 . _:b403341773 "2"^^ . . . _:b403341804 . _:b403341772 "2"^^ . _:b403341400 . . _:b403341807 . . _:b403341775 "2"^^ . . _:b403341399 . . _:b403341629 . _:b403341806 . _:b403341774 "2"^^ . _:b5177917 "proinflammatory cytokine TNF-\u03B1, which is also produced in response to cutaneous barrier disruption (17) and tightly associated with inflammatory deviation of the immune response, such as asthma and probably allergic inflammation (48, >>49<<). Despite unsuccessful immunostaining of TNF-\u03B1 on skin cross sections, we analyzed its transcriptional expression and showed that Tnf-\u03B1 mRNA was strongly overexpressed in the KO epidermis and also slightly induced in the dermis of KO" . _:b403341801 . _:b403341769 "2"^^ . _:b5177900 . _:b5177912 . _:b403341800 . . _:b403341768 "2"^^ . _:b403341615 . _:b403341803 . _:b403341771 "2"^^ . _:b5177956 "In parallel, as the result of protease hyperactivities, FILAGGRIN is also decreased (agranulosis) and the SC is often detached from the underlying cell layers in the epidermis of NS patients (>>43<<). Both anomalies impair skin barrier and favor allergen and pathogen penetration in NS epidermis, thus contributing to the aggravation of the atopic phenotype." . _:b403341802 . . _:b403341770 "2"^^ . _:b403341531 . _:b403341813 . _:b403341781 "2"^^ . _:b403341812 . _:b403341780 "2"^^ . . _:b403341815 . _:b403341783 "2"^^ . _:b403341814 . _:b403341782 "2"^^ . _:b403341813 . _:b403341719 . _:b403341809 . _:b403341777 "2"^^ . . . _:b403341446 . _:b403341808 . . _:b403341776 "2"^^ . _:b403341811 . _:b403341779 "2"^^ . _:b403341736 . _:b403341810 . _:b403341778 "2"^^ . _:b5177905 . _:b403341586 . _:b403341821 . _:b403341789 "2"^^ . _:b403341820 . _:b403341788 "2"^^ . _:b5177907 . . _:b403341823 . _:b403341537 . _:b5177906 . _:b403341791 "2"^^ . . _:b403341822 . _:b403341790 "2"^^ . _:b403341817 . . _:b403341785 "2"^^ . . . _:b403341816 . _:b403341784 "2"^^ . _:b403341819 . _:b403341787 "2"^^ . _:b5177910 . . _:b403341818 . _:b403341786 "2"^^ . _:b5177942 . _:b5177952 . _:b403341633 . _:b403341797 "2"^^ . _:b403341715 . _:b5177943 . _:b403341796 "2"^^ . . _:b403341479 . _:b5177907 "S5), which is in agreement with its degradation as previously described in newborn KO mice and NS skin (>>32<<, 43). In summary, the abnormal differentiation pattern observed in KO graft, together with the desmosomal cleavage, confirmed that the KO grafted skin reproduces all major histological and ultrastructural anomalies seen in NS patients" . . _:b403341799 "2"^^ . . . _:b403341798 "2"^^ . _:b403341522 . . _:b403341793 "2"^^ . . _:b403341792 "2"^^ . _:b403341795 "2"^^ . . _:b403341794 "2"^^ . _:b5177900 . . _:b403341805 "2"^^ . _:b5177901 . . _:b403341811 . _:b403341804 "2"^^ . _:b403341592 . _:b403341819 . _:b5177902 . . _:b403341807 "2"^^ . . _:b403341465 . _:b5177903 . . _:b403341806 "2"^^ . . . . _:b403341801 "2"^^ . . _:b403341579 . _:b403341514 . _:b403341800 "2"^^ . _:b5177944 . . _:b403341803 "2"^^ . _:b5177899 . _:b403341802 "2"^^ . . _:b5177908 . _:b403341695 . . _:b5177932 . . _:b403341813 "2"^^ . _:b5177909 . _:b403341739 . _:b403341812 "2"^^ . _:b403341573 . . _:b5177910 . . _:b403341815 "2"^^ . . _:b5177902 "WT and Spink5\u2212/\u2212 grafted skin samples were prepared as described previously (>>32<<)." . _:b5177953 "Several studies have reported significant genetic association of SPINK5 polymorphisms with AD in different populations (34\u201338), whereas other studies did not confirm these results (>>39<<\u201341). This suggests that SPINK5 genetic variations may account, to some extent, for predisposition to AD in some patients, depending on other genetic and/or environmental factors." . _:b5177911 . _:b403341605 . _:b403341622 . _:b403341814 "2"^^ . _:b5177940 "In addition, IL-1\u03B2 and TNF-\u03B1 overexpression in Spink5\u2212/\u2212 epidermis may induce TSLP expression in keratinocytes (19, >>54<<) and attract innate immune cells." . _:b5177953 . _:b5177904 . _:b403341809 "2"^^ . _:b403341500 . _:b403341564 . _:b403341693 . _:b5177905 . . _:b403341676 . _:b403341493 . _:b403341808 "2"^^ . _:b5177906 . . _:b5177902 . _:b403341811 "2"^^ . . _:b403341765 . _:b5177907 . _:b403341810 "2"^^ . _:b5177916 . . _:b403341821 "2"^^ . _:b403341731 . _:b5177917 . . _:b403341820 "2"^^ . _:b5177918 . _:b5177929 "IL8, and TNF-\u03B1 transcripts observed in NSK transfected with KLK5 siRNA could be the result of uncompleted knockdown of KLK5 mRNA but could also be the result of the activation of PAR2 by other hyperactivated proteases, such as KLK14 (>>52<<) and/or other signaling pathway induction." . _:b403341823 "2"^^ . _:b5177919 . _:b403341822 "2"^^ . _:b403341635 . _:b5177912 . _:b403341817 "2"^^ . _:b5177913 . _:b403341816 "2"^^ . _:b5177914 . . _:b403341819 "2"^^ . _:b5177915 . . _:b5177930 _:b5177932 . _:b403341818 "2"^^ . _:b5177930 _:b5177933 . _:b403341875 . _:b5177930 _:b5177934 . _:b5177924 . _:b5177930 _:b5177935 . _:b403341460 . _:b5177925 . _:b5177930 _:b5177931 . _:b403341823 . . _:b5177926 . . _:b5177927 . _:b403341506 . _:b5177930 _:b5177948 . _:b5177930 _:b5177949 . _:b403341558 . _:b5177930 _:b5177950 . _:b5177920 . _:b5177930 _:b5177951 . _:b5177930 _:b5177944 . _:b5177930 _:b5177945 . _:b5177930 _:b5177946 . _:b5177921 . _:b5177930 _:b5177947 . _:b5177930 _:b5177940 . _:b5177930 _:b5177941 . . _:b5177930 _:b5177942 . _:b5177922 . _:b5177930 _:b5177943 . _:b5177930 _:b5177936 . _:b5177930 _:b5177937 . _:b5177930 _:b5177938 . _:b5177923 . _:b5177930 _:b5177939 . _:b5177932 . _:b403341696 . . _:b5177933 . _:b5177930 _:b5177956 . _:b5177955 . _:b403341469 . . _:b5177930 _:b5177957 . _:b5177934 . . _:b5177930 _:b5177952 . _:b403341607 . _:b5177930 _:b5177953 . _:b5177930 _:b5177954 . _:b5177935 . _:b5177930 _:b5177955 . . _:b5177905 "As newborn Spink5\u2212/\u2212 (KO) mice died a few hours after birth (>>32<<), total dorsal skin of embryonic day (E) 19.5 embryos was grafted on nude mice to analyze the consequences of Spink5 inactivation in adult skin." . _:b403341834 . _:b5177928 . _:b5177929 . . _:b5177899 "materials and methods" . _:b5177930 . . _:b5177931 . . _:b5177940 . . _:b5177941 . _:b5177942 . . . _:b5177943 . . _:b403341572 . _:b403341845 . _:b5177936 . . . _:b5177937 . _:b5177953 . _:b403341394 . _:b5177938 . _:b403341425 . . _:b5177939 . . . _:b5177948 . _:b403341373 . . _:b403341716 . _:b5177949 . _:b403341372 . . _:b403341830 . _:b5177950 . _:b403341375 . _:b5177951 . _:b403341374 . _:b5177944 . _:b5177945 . . . _:b403341435 . _:b403341877 . _:b5177946 . _:b403341371 . _:b403341659 . _:b5177923 . _:b5177935 "IL-1\u03B2 can be proteolytically activated by KLK7 (>>57<<), which is hyperactivated in Spink5\u2212/\u2212 skin (32)." . _:b5177947 . _:b403341370 . . _:b5177955 "The abundant and epidermally restricted TSLP secretion could induce an AD-like phenotype associated with elevated serum levels of IgE, as illustrated in Tslp skin-specific transgenic mice (21, >>65<<). Thereby, the effect of LEKTI deficiency on triggering PAR2 signaling could be a primary event inducing atopic manifestations in NS before skin barrier defect. In parallel, as the result of protease hyperactivities, FILAGGRIN is also" . _:b5177956 . _:b403341381 . . _:b5177957 . _:b403341380 . . _:b403341708 . _:b403341383 . _:b403341577 . _:b403341846 . _:b403341382 . _:b403341698 . _:b403341413 . . _:b5177952 . _:b403341377 . _:b5177943 "TSLP, synergistically with IL-1\u03B2 and TNF-\u03B1, stimulates the production of pro-Th2 cytokines by activated mast cells (>>59<<). Massive production of active \u03B21-tryptase by activated mast cells in Spink5\u2212/\u2212 grafted skin could also directly participate in the inflammatory response and favor a Th2-permissive microenvironment through PAR2 activation on mast cells" . . _:b5177953 . _:b403341376 . . _:b403341429 . . _:b5177954 . _:b403341379 . _:b403341721 . _:b5177955 . _:b403341378 . _:b403341389 . . _:b403341388 . _:b403341391 . _:b403341390 . . _:b403341385 . _:b403341467 . _:b403341384 . . . _:b403341682 . _:b403341757 . _:b403341387 . _:b403341594 . _:b403341432 . _:b403341386 . . . _:b403341397 . . _:b403341396 . _:b403341602 . . _:b403341777 . _:b403341399 . . _:b403341766 . _:b403341654 . _:b403341398 . _:b403341393 . _:b403341803 . . _:b403341392 . . _:b403341487 . _:b403341395 . _:b403341689 . _:b403341394 . _:b403341491 . _:b403341405 . .