Recurrence-free survival of patients in the two separate trials.

Flowcytometric gating strategies for dendritic cells in the melanoma sentinel lymph node

Major leukocyte subset frequencies in melanoma SLN grouped by stage

Flowcytometric gating strategies for regulatory T cells in the melanoma sentinel lymph node and correlation between Treg frequencies in the melanoma SLN as measured by FoxP3 or by CTLA4 expression

Supplementary Figure S1 from Sunitinib-Induced Myeloid Lineage Redistribution in Renal Cell Cancer Patients: CD1c+ Dendritic Cell Frequency Predicts Progression-Free Survival

Gating strategies for analysis of A) plasmacytoid DC (pDC) and B) skin-migratory conventional DC (cDC) subsets: Dermal DC (DDC) and Langerhans Cells (LC) and C) LN-resident cDC subsets in melanoma Sentinel Lymph Nodes (SLN).

CD4+CD25hiFoxP3+ Treg rates in melanoma SLN, grouped by tumor stage

SLN characteristics of patients with 2 SLNs

Differences in SLN-resident DCs between patients with and without distant recurrence

Expression of the cross-priming associated markers BDCA3/CD141 and CLEC9A on the four identified cDC subsets in single-cell suspensions of untreated melanoma Sentinel Lymph Nodes SLN (mean {plus minus} SEM of at least three separate experiments are shown).

Differences in frequency and phenotype of skin-derived SLN DCs between patients with and without local recurrence

Placebo-administered vs off-trial patients

Background We previously reported CpG-B injection at the primary tumor excision site prior to re-excision and sentinel node biopsy to result in immune activation of the sentinel lymph node (SLN), increased melanoma-specific CD8(+) T cell rates in peripheral blood, and prolonged recurrence-free survival. Here, we assessed recruitment and activation of antigen-presenting cell (APC) subsets in the SLN and at the injection site in relation to T cell infiltration. Methods Re-excision skin specimens from patients with clinical stage I-II melanoma, collected 7 days after intradermal injection of either saline (n=10) or 8 mg CpG-B (CPG7909, n=12), were examined by immunohistochemistry, quantifying immune subsets in the epidermis, papillary, and reticular dermis. Counts were related to flow cytometric data from matched SLN samples. Additional in vitro cultures and transcriptional analyses on peripheral blood mononuclear cells (PBMCs) were performed to ascertain CpG-induced APC activation and chemokine profiles. Results Significant increases in CD83(+), CD14(+), CD68(+), and CD123(+) APC were observed in the reticular dermis of CpG-B-injected skin samples. Fluorescent double/triple staining revealed recruitment of both CD123(+)BDCA2(+) plasmacytoid dendritic cells (DCs) and BDCA3/CD141(+)CLEC9A(+) type-1 conventional DC (cDC1), of which only the cDC1 showed considerable levels of CD83 expression. Simultaneous CpG-B-induced increases in T cell infiltration were strongly correlated with both cDC1 and CD14 counts. Moreover, cDC1 and CD14(+) APC rates in the reticular dermis and matched SLN suspensions were positively correlated. Flow cytometric, transcriptional, and chemokine release analyses of PBMC, on in vitro or in vivo exposure to CpG-B, indicate a role for the activation and recruitment of both cDC1 and CD14(+) monocyte-derived APCs in the release of CXCL10 and subsequent T cell infiltration. Conclusion The CpG-B-induced concerted recruitment of cDC1 and CD14(+) APC to the injection site and its draining lymph nodes may allow for both the (cross-)priming of T cells and their subsequent homing to effector sites.

In autoimmune diseases like rheumatoid arthritis (RA), multidrug resistance (MDR) transporters of the ATP-binding cassette (ABC) transporter superfamily harbor dual functions by extruding pro-inflammatory mediators and exporting disease modifying anti-rheumatics drugs (DMARDs), hence contributing to diminished treatment response. Herein we determined the expression (mRNA/protein) and functional efflux activities of multiple selected ABC transporters in immune-effector cells of RA patients in relation to DMARD response. ABC transporter profiling included ABCB1 (P-glycoprotein), ABCC1-6/ABCC10-12 (multidrug resistance proteins 1-9) and ABCG2 (Breast Cancer Resistance Protein). Analyses were performed in peripheral blood lymphocytes (PBL) and monocyte-derived macrophages (MDM) obtained from 52 RA patients (DMARD-naive and DMARD (non)-responders) and HC (n = 19) using PCR, immunohistochemistry and flow cytometry. Notwithstanding the large inter-patient variabilities, PBLs from RA patients displayed significantly higher mRNA levels of ABCC1 (2.1-fold), ABCC4 (1.6-fold) and ABCC10 (1.9-fold) compared with HC. Expression levels of ABCB1, ABCC1, ABCC4 and ABCC10 were significantly and positively correlated with each other. Furthermore, significantly increased ABCG2 mRNA (2.8-fold) and protein levels (2.4-fold) were observed in MDM from RA patients compared to HC. Additional analyses revealed that a 1.8-fold increased functional activity of ABCB1 in CD3+ cells in RA patients receiving DMARD treatment versus DMARD-naive patients, was exclusively contributed by DMARD non-responders. Although up to 1.7-fold higher levels of MDR mRNA levels were noted in PBL of DMARD non-responders over DMARD responders, these differences were not statistically significant. Together, these results underscore the involvement of multiple ABC transporters in immune-competent cells in relation to RA and DMARD response.

Keloids constitute an abnormal fibroproliferative wound healing response in which raised scar tissue grows excessively and invasively beyond the original wound borders. This review provides a comprehensive overview of several important themes in keloid research: namely keloid histopathology, heterogeneity, pathogenesis, and model systems. Although keloidal collagen versus nodules and α-SMA-immunoreactivity have been considered pathognomonic for keloids versus hypertrophic scars, conflicting results have been reported which will be discussed together with other histopathological keloid characteristics. Importantly, histopathological keloid abnormalities are also present in the keloid epidermis. Heterogeneity between and within keloids exists which is often not considered when interpreting results and may explain discrepancies between studies. At least two distinct keloid phenotypes exist, the superficial-spreading/flat keloids and the bulging/raised keloids. Within keloids, the periphery is often seen as the actively growing margin compared to the more quiescent center, although the opposite has also been reported. Interestingly, the normal skin directly surrounding keloids also shows partial keloid characteristics. Keloids are most likely to occur after an inciting stimulus such as (minor and disproportionate) dermal injury or an inflammatory process (environmental factors) at a keloid-prone anatomical site (topological factors) in a genetically predisposed individual (patient-related factors). The specific cellular abnormalities these various patient, topological and environmental factors generate to ultimately result in keloid scar formation are discussed. Existing keloid models can largely be divided into in vivo and in vitro systems including a number of subdivisions: human/animal, explant/culture, homotypic/heterotypic culture, direct/indirect co-culture, and 3D/monolayer culture. As skin physiology, immunology and wound healing is markedly different in animals and since keloids are exclusive to humans, there is a need for relevant human in vitro models. Of these, the direct co-culture systems that generate full thickness keloid equivalents appear the most promising and will be key to further advance keloid research on its pathogenesis and thereby ultimately advance keloid treatment. Finally, the recent change in keloid nomenclature will be discussed, which has moved away from identifying keloids solely as abnormal scars with a purely cosmetic association toward understanding keloids for the fibroproliferative disorder that they are.

增生性瘢痕和瘢痕疙瘩是隆起性、异常性瘢痕的两种类型。增生性瘢痕生长在原创面边界内,最终生长变小,但瘢痕疙瘩生长超出原创面边界,不自行变小,治疗困难。瘢痕疙瘩通常发生在肤色较深的人。有时仍难以确定隆起的瘢痕是增生性瘢痕还是瘢痕疙瘩。 这项荷兰研究旨在确定这两种异常瘢痕是否都形成,因为它们保留了幼稚化、不成熟瘢痕的特征,而不是以正常的方式成熟。 作者研究了存在于皮肤外层(表皮)和内层(真皮)的细胞类型,发现增生性和瘢痕疙瘩与未成熟瘢痕而不是正常健康瘢痕非常相似,存在两个重要的方面:外层细胞没有适当成熟(外皮蛋白表达增加);正常皮肤的内层细胞表达一种称为 CD34 的蛋白,不具有收缩组织的细胞 (α-SMA),但在异常瘢痕中,这种情况发生逆转,CD34 缺失,存在收缩细胞 (α-SMA)。 与异常瘢痕不同,未成熟瘢痕内外层细胞表现为细胞生长增加 (Ki67)。异常瘢痕的外层要厚得多,内层有许多细胞(波形蛋白),但这些细胞已经停止生长(称为 p16)。 根据内层的外观很难将增生性瘢痕与瘢痕疙瘩分开,但增生性瘢痕有更多的细胞收缩,瘢痕疙瘩内层有很厚的结缔组织带。 总之,本研究结果提示,增生性瘢痕和瘢痕疙瘩保持了未成熟瘢痕的特征,不像正常瘢痕那样成熟。 本摘要涉及研究:增生性瘢痕和瘢痕疙瘩未能从 CD34 - α-平滑肌肌动蛋白 (SMA) + 未成熟瘢痕表型进展,并且表现出 α-SMA 和 p16 表达的梯度差异