MGL2+ Dermal Dendritic Cells Are Sufficient to Initiate Contact Hypersensitivity In Vivo

BACKGROUND:Dendritic cells (DCs) are the most potent antigen-presenting cells in the mammalian immune system. In the skin, epidermal Langerhans cells (LCs) and dermal dendritic cells (DDCs) survey for invasive pathogens and present antigens to T cells after migration to the cutaneous lymph nodes (LNs). So far, functional and phenotypic differences between these two DC subsets remain unclear due to lack of markers to identify DDCs. METHODOLOGY/PRINCIPAL FINDINGS:In the present report, we demonstrated that macrophage galactose-type C-type lectin (MGL) 2 was exclusively expressed in the DDC subset in the skin-to-LN immune system. In the skin, MGL2 was expressed on the majority (about 88%) of MHCII(+)CD11c(+) cells in the dermis. In the cutaneous LN, MGL2 expression was restricted to B220(-)CD8alpha(lo)CD11b(+)CD11c(+)MHCII(hi) tissue-derived DC. MGL2(+)DDC migrated from the dermis into the draining LNs within 24 h after skin sensitization with FITC. Distinct from LCs, MGL2(+)DDCs localized near the high endothelial venules in the outer T cell cortex. In FITC-induced contact hypersensitivity (CHS), adoptive transfer of FITC(+)MGL2(+)DDCs, but not FITC(+)MGL2(-)DCs into naive mice resulted in the induction of FITC-specific ear swelling, indicating that DDCs played a key role in initiation of immune responses in the skin. CONCLUSIONS/SIGNIFICANCE:These results demonstrated the availability of MGL2 as a novel marker for DDCs and suggested the contribution of MGL2(+) DDCs for initiating CHS

Clinical outcome of patients with recurrent or refractory localized Ewing's sarcoma family of tumors: A retrospective report from the Japan Ewing Sarcoma Study Group

[Background] Patients with Ewing's sarcoma family of tumors (ESFT) who experience relapse or progression have a poor prognosis. [Aim] This study aimed to identify the prognostic and therapeutic factors affecting overall survival (OS) of patients with recurrent or refractory localized ESFT. [Methods and results] Thirty-eight patients with localized ESFT who experienced first relapse or progression between 2000 and 2018 were retrospectively reviewed. The 5-year OS rate of the entire cohort was 48.3% (95% confidence interval, 29.9%-64.5%). Multivariate analysis of OS identified time to relapse or progression, but not stem cell transplantation (SCT), as the sole independent risk factor (hazard ratio, 35.8; P = .002). Among 31 patients who received salvage chemotherapy before local treatment, 21 received chemotherapy regimens that are not conventionally used for newly diagnosed ESFT. The objective response rate to first-line salvage chemotherapy was 55.2% in the 29 evaluable patients. Time to relapse or progression was significantly associated with response to first-line salvage chemotherapy (P = .006). [Conclusions] The present study fails to demonstrate significant clinical benefit of SCT for recurrent or refractory localized ESFT. Recently established chemotherapy regimens may increase the survival rate of patients with recurrent or refractory localized ESFT while attenuating the beneficial effect of SCT

Pre-post changes in psychosocial functioning among relatives of patients with depressive disorders after Brief Multifamily Psychoeducation: A pilot study

<p>Abstract</p> <p>Background</p> <p>Depressive disorder is often chronic and recurrent, and results in a heavy psychosocial burden on the families of patients with this disorder. This study aims to examine the effectiveness of brief multifamily psychoeducation designed to alleviate their psychosocial burden.</p> <p>Methods</p> <p>Thirty-two relatives of patients with major depressive disorder participated in an open study testing the effectiveness of brief multifamily psychoeducation. The intervention consisted of four sessions over the course of 6 weeks. Outcome measures focused on emotional distress, care burden and Expressed Emotion (EE).</p> <p>Results</p> <p>The emotional distress, care burden and EE of the family all showed statistically significant improvements from baseline to after the family intervention. The proportion of relatives scoring 9 or more on K6, which indicates possible depressive or anxiety disorder, decreased from sixteen relatives (50.0%) at baseline, to only 3 relatives (9.3%) after the intervention.</p> <p>Conclusions</p> <p>This study suggests that brief multifamily psychoeducation is a useful intervention to reduce the psychosocial burden of the relatives of patients with depressive disorder. Further evaluation of family psychoeducation for relatives of patients with depressive disorder is warranted.</p

Induction of FITC-specific CHS in mice adoptively transferred with FITC⁺ MGL2⁺ DDC.

<p>(A) The left panels show the gating to sort FITC⁺ MGL2⁺ cells from the MACS-purified CD11c⁺ cells. The purity of FITC⁺ MGL2⁺ DDCs was approximately 98%. Ear swellings in mice transferred with FITC⁺ MGL2⁺ DDCs (“MGL2⁺”, <i>p</i><0.005), transferred with glutaraldehyde-fixed FITC⁺MGL2⁺ DDC (“fixed MGL2⁺”, difference not statistically significant), sensitized directly with FITC painting (“FITC painted”, <i>p</i><0.005), or untreated (“naive”) are shown in the graph. Each bar indicates mean±SD obtained from three independent recipients respectively. (B) The left panels show the gating strategy to sort FITC⁺MGL2⁺DDCs from the day 1 LNs or FITC⁺MGL2⁻DCs from the day 4 LNs. The purity of sorted cells was >90% when re-analyzed by FACS. Ear swelling in mice transferred with 5×10⁴ cells of FITC⁺MGL2⁺DDCs from the day 1 LNs (d1 DDC), glutaraldehyde-fixed FITC⁺MGL2⁺DDCs from the day 1 LNs (fixed d1 DDC) or FITC⁺MGL2⁻DCs from the day 4 LNs (d4 non-DDC) is shown in the right panel. The mice sensitized by FITC painting (FITC painted) and left untreated (naive) were used as positive and negative controls respectively. Each bar indicates the mean value from two recipients and the data are representative of two independent experiments.</p

Distribution of MGL2⁺ cells in draining LNs during the sensitization with FITC revealed by immunohistochemical analysis.

<p>(A) Serial sections of LNs stained with anti-MGL1, anti-MGL2 or anti-MGL1/2 cross-reactiv mAbs. MGL1 was strongly stained in the sinuses in both naive and FITC sensitized (24 h after sensitization) LNs. Arrows indicate the increased signals in the cortex after the sensitization, though the anti-MGL1 staining in the cortex was weaker than in the sinuses. Scale bar, 250 µm. (B) Serial LN sections 4 h after sensitization stained with mAbs to MØ/DC markers. Restricted localization of MGL2⁺ cells to the outer T cell cortex was unique compared to other APCs (MHCII) including B cells (B220), MØs (CD11b and F4/80), DCs (CD11c) or MGL1⁺ cells. S: subcapsular sinus, M: medulla, T: T cell cortex, B: B cell follicle. Scale bar, 200 µm. (C) Confocal microscopic observations of the distributions of MGL1 (mAb LOM-8,7) or MGL2 (mAb URA-1) (blue in each panel) with MØ markers (red) in the LN 24 h after FITC sensitization. FITC used is shown in green. Scale bar, 25 µm (×400).</p

Immunohistochemical localization of MGL2⁺ DDCs, LCs and LN conduits.

<p>(A) Distribution of MGL2 (red) and Langerin (blue) in a draining (top) or a non-draining (bottom) LNs 24 h after FITC (green) sensitization. Arrows indicate venule-like structures in the CR. Arrowheads indicate the Langerin⁺ cells loaded with FITC. Almost all the MGL2⁺ DDCs were FITC⁺ and shown in yellow when merged. B: B cell follicles. Scale bar, 100 µm. (B) Distribution of MGL2 (red), PNAd (HEV), Langerin (LC) or ER-TR7 antigen (FRN) (blue in each panel) in the sensitization process. Areas indicated by the squares are magnified in the lower panels. FITC is shown in green. CR associated with HEV in a naive LN is adjacent to MGL2⁺ DDC only at its follicular side but not at the cortical side. Arrowheads indicate association of MGL2⁺ DDCs with ER-TR7⁺ FRN around HEV-associated CR. Scale bar, 100 µm.</p

Expression of MGL1 and MGL2 in a DDC subset in skins of naive mice.

<p>(A) Binding of anti-MGL1 and anti-MGL2 mAbs to cryosections of skin (blue in each panel). The dashed line in the control panel (Ctrl. IgG) indicates the dermal-epidermal junction. Nuclei are shown in red. Scale bar, 100 µm. E: epidermis, D: dermis, F: hair follicle. (B) Surface staining by anti-MGL1 and anti-MGL2 mAbs of epidermal and dermal cell suspensions. (C) Intracellular staining of CD11c and surface staining of MHCII and MGL2 in the dermal cell suspension. Virtually all MGL2⁺ cells expressed CD11c. The proportion of MGL2⁺ cells in MHCII^hi CD11c⁺ DDC (cells in the rectangular gate) was 88.5±6.8%.</p

Hapten incorporation and migration kinetics of MGL2⁺ DDC after FITC sensitization revealed by the flow cytometric analysis.

<p>(A) FITC fluorescence associated with MGL2⁺ DDCs in LNs. MGL1⁺ cDC (blue) or MGL2⁺ DDCs (red), but not MGL1⁺ pDCs (purple) or cells nonspecifically bound by mAb URA-1 (orange) incorporate FITC. (B) Kinetics of MGL2⁺ DDCs in LNs after FITC sensitization. Top panels indicate the FITC fluorescence and staining with anti-MGL2 mAb on DCs purified from naive LNs (left) or LNs 24 h (middle) or 96 h (right) after sensitization. The bottom panel shows the ratio of total FITC⁺ DCs (closed diamond), FITC⁺ MGL2⁺ DDCs (open square) and FITC⁺ MGL2⁻ DCs (closed triangle) calculated by each quadrant shown in top panels. The data are shown as Mean±SD.</p