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The use of immune checkpoint inhibitors (ICI) is expanding with the approval for advanced/metastatic keratinocyte carcinoma; however, most tumors are non-aggressive. Local administration could broaden ICI, but adequate immune response might require an immune-attractive adjuvant such as ablative fractional laser (AFL). Accordingly, this study aimed to explore intratumoral injection of anti-PD1 with and without AFL in basal cell carcinoma (BCC), exploring anti-PD1 concentration, immune cell infiltration, tumor response, and safety. This open-label, proof-of-concept trial investigated intratumoral anti-PD1 + AFL combination therapy versus anti-PD1 or AFL monotherapy in 28 BCC patients. The primary endpoints were immune cell infiltration evaluated immunohistochemically and clinical tumor response after 3 months. The secondary outcomes were tumoral drug concentration and safety. The most robust response was obtained following intervention with combined anti-PD1+AFL, leading to a ~2.5-fold increase in CD3+ cells (p = 0.027), and tumor reduction ≥25% in 73%, including two tumors with complete remission. Upon anti-PD1 monotherapy, a slight decrease in CD3+ cells was observed while a non-significant increase following AFL was seen. Tumor reduction ≥25% was seen in 45% and 50%, respectively, after anti-PD1 and AFL monotherapy. The CD8/CD3 ratio remained unchanged after anti-PD1+AFL and anti-PD1 monotherapy, while AFL led to a decreased ratio. A non-significant decline in the Foxp3/CD3 ratio was observed for all groups. Side-effects were mild with no systemic drug concentration detected. Intratumoral anti-PD1 injection is feasible, and a single exposure to locally injected anti-PD1 with adjuvant AFL increased immune cell infiltration and reduction in BCC with limited side-effects.

Presentation with severe acute kidney injury due to cast nephropathy (CN) is a medical emergency in multiple myeloma (MM), with high risk of dialysis-dependent renal failure and death. Accrual of patients with CN into interventional studies is difficult, while phase III trials exclude patients with severe renal insufficiency. Real-world data are warranted. We assessed 2252 patients from the population-based Danish Multiple Myeloma Registry (DMMR) who were diagnosed between 2013 and 2017. We identified 204 patients with clinically-suspected CN, defined as serum creatinine concentration >177 μmol/L and serum free light chain (sFLC) concentration >1000 mg/L at the time of diagnosis. The median age was 72 years. Thirty-one percent of patients presented with dialysis-dependent renal failure. Kidney biopsies were performed in 19% of patients and showed CN in 74% of cases. Despite prompt initiation of bortezomib-based therapy in 94% of patients, 33% of patients died in the first year after diagnosis. Compared with the rest of the patients in the DMMR with symptomatic MM, patients with clinically-suspected CN had worse overall survival (OS) irrespective of transplant eligibility. Achievement of renal recovery was associated with deep reductions of involved sFLC. Achievement of very good partial response or better in the first line of therapy and/or deep reduction of involved sFLC at 3 months after initiation of therapy were associated with superior OS. In conclusion, MM patients presenting with clinically-suspected CN have an alarmingly high one-year mortality when treated with current standards of care. Early and deep hematologic response is crucial for survival.

Background and Aims Accelerated vascular calcification in different vascular beds is common in patients with chronic kidney disease (CKD). A severe form of vascular calcifications is calcific uremic arteriolopathy (CUA) presenting with painful ischemic skin lesions and high mortality. The prognostic value of skin biopsies in relation to CUA is unclear and the prevalence of skin vascular calcifications in different stages of CKD is sparsely described. The aim of the study was to describe the occurence of small vesssel vascular calcifications in unaffected skin biopsies in relation to calcifications in other vascular beds across the spectrum of CKD including CUA. Method A cross-sectional cohort (total, n=39) comprising dialysis patients with current or previous CUA (CKD5D+CUA, n=9), dialysis patients without CUA (CKD5D-CUA, n=12), patients with CKD stage 3-4 (CKD3-4, n=12), and healthy kidney controls (control, n=6). The presence of vascular calcifications in the dermis and subcutis were assessed in 4 mm punch biopsies of unaffected skin from the lateral thigh. The presence of vascular calcification was evaluated by H&E, von Kossa and Alizarin staining. The presence of breast arterial calcifications (BAC) was evaluated by mammography, the abdominal aortic calcification score (AAC) by lateral lumbar X-ray and calcification propensity was measured by T50 reflecting the calcification propensity in blood. Results None of the included patients showed vascular calcifications in skin biopsies. Declining kidney function was associated with presence of BAC, increased AAC and reduced T50 (Table). No significant difference was found between CKD5D+CUA and CKD5D-CUA regarding BAC (p=1.000), AAC (p=0.815) or T50 (p=0.165). CKD5D+CUA compared to all other groups had no difference in BAC (p=0.109) and AAC (p=0.141) but reduced T50 (p=0.004). Dialysis patients (CKD5D±CUA) had significant more BAC (p=0.003), higher AAC (p Conclusion No vascular calcifications were found in punch biopsies from unaffected skin in patients with different stages of CKD including CUA. Despite imaging verified vessel calcification by BAC and AAC and increased calcification propensity by T50. This suggest that conventional punch biopsies cannot be used to identify skin vascular calcification and thereby patients at risk for developing CUA.

Tissue injury can lead to fibrotic disease and chronic inflammation, for example, in the lungs of patients with idiopathic pulmonary fibrosis. Jürgensen et al. report that fibroblasts regulate immune responses during tissue injury by clearing collectins with pro- or antiinflammatory activity via uPARAP, an endocytic collectin receptor identified in this work., Collectins such as mannose-binding lectin (MBL) and surfactant protein D (SP-D) become temporarily deposited in extravascular compartments after tissue injury and perform immune-stimulatory or inflammation-limiting functions. However, their turnover mechanisms, necessary to prevent excessive tissue damage, are virtually unknown. In this study, we show that fibroblasts in injured tissues undertake the clearance of collectins by using the endocytic collagen receptor uPARAP. In cellular assays, several types of collectins were endocytosed in a highly specific uPARAP-dependent process, not shared by the closely related receptor MR/CD206. When introduced into dermis or bleomycin-injured lungs of mice, collectins MBL and SP-D were endocytosed and routed for lysosomal degradation by uPARAP-positive fibroblasts. Fibroblast-specific expression of uPARAP governed endogenous SP-D levels and overall survival after lung injury. In lung tissue from idiopathic pulmonary fibrosis patients, a strong up-regulation of uPARAP was observed in fibroblasts adjacent to regions with SP-D secretion. This study demonstrates a novel immune-regulatory function of fibroblasts and identifies uPARAP as an endocytic receptor in immunity.

Background The transcription factor Forkhead Box P3 (FoxP3) is a marker of regulatory T cells (Tregs) — a subset of T cells known to suppress a wide range of immune responses. These cells are considered to be pivotal for the induction of tolerance to donor antigens in human allografts. We aimed to correlate the number of lymphocytes expressing FoxP3 in transbronchial biopsies from lung allografts with the FoxP3 expression in bronchoalveolar lavage fluid (BALF). In addition, we aimed to correlate the number of FoxP3 + cells in transbronchial biopsies with the degree of acute cellular rejection in lung allografts. Materials and methods The expression of FoxP3 was evaluated using immunohistochemical staining in 40 lung allograft biopsies obtained from 23 patients. The number of Tregs was related to the FoxP3 mRNA levels as determined using qRT-PCR in corresponding BALF samples from the same patients. Furthermore, the number of Tregs was related to the degree of acute allograft rejection (according to ISHLT criteria, A0–A4). Results Regression analysis showed a significant concordance between the number of Tregs in lung tissue and the level of FoxP3 mRNA relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA levels in BALF (n = 40, p = 0.0001). In addition, we found a significant increase in the number of Tregs during acute allograft rejections of grades A2 and higher (median: 32.6 Tregs/mm 2 ) when compared to those of grades A1 and A0 (median: 4.9 Tregs/mm 2 ) (p = 0.0002). Discussion and conclusion The association between the distribution of Tregs in transbronchial biopsies and the level of FoxP3 mRNA in BALF indicates that assessment of FoxP3 mRNA in BALF is a reliable non-invasive method for evaluating the number of Tregs in lung tissue. Furthermore, the association between the number of Tregs in lung tissue and the degree of acute cellular rejection shows that Tregs are recruited to the site of inflammation and may be involved in the regulation of acute rejection. Thus, Tregs may play a role in the cellular processes that affect lung allograft outcome.

Rheumatic disease is the dominant cause of amyloid A (AA) amyloidosis, but other chronic inflammatory diseases may have similar consequences. Hidradenitis suppurativa (HS) is a relatively common, but little known skin disease characterized by chronic inflammation. Here we present a case of chronic HS leading to biopsy-verified severe renal AA amyloidosis and dialysis dependency.

Background Regulatory T lymphocytes (Tregs) play an important role in acute rejection after lung transplantation. However, the importance of the time elapsed after transplantation on the Treg response requires further investigation. We aim to evaluate the change over time in the frequency of Tregs in lung allograft biopsies and to assess how Tregs relate to simultaneous and subsequent acute cellular rejection. Materials and methods A total of 258 biopsy samples obtained 0.5, 1, 3, 12 and 24 months after transplantation from 58 consecutive lung transplant patients were included. The biopsies were scored for acute rejection according to the ISHLT criteria (A0–A4) and immunohistochemically stained with antibodies against FoxP3. Results There was a tendency for a decrease in the number of Tregs/mm 2 with time. However, the previous levels of Tregs/mm 2 did not have any significant effect on future levels of Tregs/mm 2 . For biopsies taken 0.5 and 1 month after transplantation, a significant correlation between Tregs/mm 2 and the degree of acute rejection was found, and logistic regression analysis using updated values for Tregs/mm 2 showed a significant relationship between Tregs/mm 2 at 2 weeks and an A-score ≥ 2 after 1 and 3 months. At later time points, this correlation disappeared. Discussion and conclusion Our data indicate that the time elapsed after transplantation is an important parameter influencing the Treg response after lung transplantation. This observation is in accordance with studies indicating a narrow therapeutic window for induction of tolerance by specifically targeting T-cells. The results also indirectly indicate that Tregs early after transplantation could have an impact on the long-term outcome.

A high prevalence of cervical cancer associated high-risk types of human papillomavirus (hrHPV) has been demonstrated in premalignant and invasive squamous cell lesions of the penis, but large studies correlating histological characteristics with HPV status are few in number. Tumour tissues from 145 patients with invasive (n = 116) or in situ (n = 29) penile squamous cell carcinoma were subjected to systematic histological evaluation and were PCR-tested for 14 hrHPV types and 23 low-risk HPV types. Around half (52%) of invasive and nine-tenths (90%) of in situ lesions were positive for an hrHPV type, of which HPV 16 was by far the predominant type (91% of hrHPV-positive lesions). In relation to histological characteristics, hrHPV positivity was statistically significantly more common in high-grade tumours, lesions dominated by small tumour cells, lesions with a high number of multinucleated cells and mitoses, and lesions with a small amount of parakeratosis. In conclusion, about half of invasive penile squamous carcinomas in this study were hrHPV-positive, most notably to HPV 16, and probably arose through in situ lesions whereas the other half of invasive penile lesions appeared to be unrelated to hrHPV. A number of histological characteristics differed significantly between hrHPV-positive and -negative invasive penile carcinomas.

Background:Elastin is a signature protein of the arteries and lungs, thus it was hypothesized that elastin is subject to enzymatic degradation during cardiovascular and pulmonary diseases. The aim was to investigate if different fragments of the same protein entail different information associated to two different diseases and if these fragments have the potential of being diagnostic biomarkers.Methods:Monoclonal antibodies were raised against an identified fragment (the ELM-2 neoepitope) generated at the amino acid position '552 in elastin by matrix metalloproteinase (MMP) -9/-12. A newly identified ELM neoepitope was generated by the same proteases but at amino acid position '441. The distribution of ELM-2 and ELM, in human arterial plaques and fibrotic lung tissues were investigated by immunohistochemistry. A competitive ELISA for ELM-2 was developed. The clinical relevance of the ELM and ELM-2 ELISAs was evaluated in patients with acute myocardial infarction (AMI), no AMI, high coronary calcium, or low coronary calcium. The serological release of ELM-2 in patients with chronic obstructive pulmonary disease (COPD) or idiopathic pulmonary fibrosis (IPF) was compared to controls.Results:ELM and ELM-2 neoepitopes were both localized in diseased carotid arteries and fibrotic lungs. In the cardiovascular cohort, ELM-2 levels were 66% higher in serum from AMI patients compared to patients with no AMI (p

Purpose Regulatory T lymphocytes (Tregs) play an important role in acute rejection after lung transplantation - pressumably inhibiting the immunologic response. However, the importance of time after transplantation on the Treg response needs further investigation. The purpose of this study is to assess whether the correlation between regulatory T cells and the degree of acute rejection changes over time. Methods and Materials A total of 227 biopsy samples from 58 lung transplanted patients in the surveillance follow-up programme were included. These comprised available biopsies obtained 0.5, 1, 3, 12 and 24 months after transplantation. The biopsies were scored for acute rejection according to the ISHLT criteria (A0-A4). The transcription factor FoxP3 was used as a specific immunohistochemical marker of Tregs on formalin-fixed, paraffin-embedded tissue. Results were expressed as the number of positive cells per square millimeter. Results For the 5 different time points studied, Spearman’s rank correlation coefficient is shown in table 1 below. For the biopsies taken 0.5, 1 and 3 months after transplantation a significant correlation between Treg frequency and the degree of acute rejection was found for each time point (p=0.05, p Conclusions Our data indicate that time after transplantation is an important parameter influencing the Treg response after lung transplantation. This observation is in keeping with other studies indicating a narrow therapeutic window for inducing tolerance by specifically targeting T-cells. The results also indirectly indicate that T-regs have an impact on chronic rejection and long term outcome. Table 1 Time after transplantation (months) 0.5 1 3 12 24 Number of samples 46 49 50 46 36 Median rejection grade 1 1 0 0 0 Median number of Tregs/mm2 11.99 14.25 9.56 10.24 Spearman’s rank corr. coeff. 0.29 0.59 0.34 −0.04 0.10