BLT1-dependent alveolar recruitment of CD4(+)CD25(+) Foxp3(+) regulatory T cells is important for resolution of acute lung injury.

RATIONALE: Recent study has demonstrated that CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs) present in bronchoalveolar lavage fluid (BALF) contribute to the resolution of an experimental acute lung injury (ALI). However, the molecular mechanism underlying the alveolar recruitment of Treg remains unclear. OBJECTIVES: To determine the role of BLT1, a chemotactic receptor for leukotriene B4 (LTB4), in Treg recruitment to BALF of LPS-induced ALI. METHODS: We examined BLT1 expression in mouse and human Tregs and evaluated its role in mediating Treg migration in vitro and in vivo. MEASUREMENTS AND MAIN RESULTS: We found that BLT1 expression was strongly up-regulated in Tregs on activation, and that BLT1 mediated the migration of activated, but not resting, Tregs toward LTB4 in vitro. LTB4 levels were persistently elevated in BALF of LPS-induced ALI. Blockade of LTB4-BLT1 pathway by administrating antagonists 1 day after LPS exposure significantly decreased BALF Treg numbers and impaired resolution of ALI characterized by persistent BALF protein, neutrophilic infiltrates, and elevated proinflammatory cytokines. Furthermore, there were significantly less BLT1(-/-) Tregs than wild-type Tregs migrating to BALF of LPS-exposed recipient Rag-1(-/-) mice after adoptive transfer (point estimate 299.73; 95% confidence interval, 255.77-343.69; P < 0.00001), and the impaired alveolar recruitment of BLT1(-/-) Tregs caused the inability to restore the resolution of ALI. CONCLUSIONS: Our findings reveal a novel antiinflammatory role of BLT1 in the resolution of ALI by mediating the alveolar recruitment of Tregs, and indicate that therapies aimed at interrupting the LTB4-BLT1 pathway after ALI onset could be harmful to the resolution of ALI.

Generation of Dhx9-deficient clones in T-cell development with a mitotic recombination technique.

Mitotic recombination is an effective tool for generating mutant clones in somatic tissues. Because of difficulties associated with detecting and quantifying mutant clones in mice, this technique is limited to analysis of growth-related phenotypes induced by loss function of tumor suppressor genes. Here, we used the polymorphic CD45.1/CD45.2 alleles on chromosome 1 as pan-hematopoietic markers to track mosaic clones generated through mitotic recombination in developing T cells. We show that lineage-specific mitotic recombination can be induced and reliably detected as CD45.1 or CD45.2 homozygous clones from the CD45.1/CD45.2 heterozygous background. We have applied this system in the analysis of a lethal mutation in the Dhx9 gene. Mosaic analysis revealed a stage-specific role for Dhx9 during T-cell maturation. Thus, the experimental system described in this study offers a practical means for mosaic analysis of germline mutations in the hematopoietic system.

Ultraviolet B irradiation induces skin accumulation of plasmacytoid dendritic cells: a possible role for chemerin.

Photosensitivity represents a common feature for most forms of lupus erythematosus (LE) including cutaneous LE. Skin inflammatory infiltrates in response to ultraviolet (UV) exposure are closely involved in the development of skin lesions of LE patients. Skin-infiltrating plasmacytoid dendritic cells (pDCs), considered as a hallmark of cutaneous LE, contribute to its pathogenesis via the production of type I interferons (IFNs). Chemerin, a recently identified chemoattractant for pDCs through its functional receptor chemR23, was found to be elevated in skin lesions of LE patients. The aim of this study was to investigate the effect of UVB irradiation on skin pDC recruitment and chemerin expression. We found that UVB irradiation induced a rapid but transient influx of pDCs as well as a persistent infiltration of neutrophils and macrophages in the mouse skin. The mRNA expression levels of IRF-7, IFN-α and chemR23 were increased in UVB-irradiated skin. Furthermore, UVB irradiation up-regulated skin chemerin production and pDC accumulation in parallel, both of which reached their peaks simultaneously (24 h post-irradiation). Dermal fibroblasts seemed to be the major source of chemerin as evidenced by significantly increased chemerin secretion by UVB-irradiated dermal fibroblasts. More importantly, LE-prone MRL/lpr mouse exhibited greatly increased skin pDC accumulation and chemerin production in response to UVB irradiation, indicating their contributions to increased susceptibility of photosensitivity in the MRL/lpr mouse. Thus, our findings demonstrated that elevated chemerin expression positively correlates with pDC accumulation in UVB-irradiated skin, suggesting a role of chemerin in mediating skin recruitment of pDCs in response to UVB exposure.