Thomas M. Kundig

Background: Intralymphatic immunotherapy (ILIT) has been investigated for two decades in small clinical trials, typically using native allergens adsorbed to aluminium hydroxide (alum). This randomised, double‐blind, placebo‐controlled study evaluated the safety and efficacy of ILIT with a grass pollen allergoid formulated with microcrystalline tyrosine (MCT). Methods: Sixty adults with grass pollen–induced allergic rhinoconjunctivitis (ARC) were randomised to receive ILIT with grass allergoid (equivalent to 60 ng Phl p 5; n = 31) or placebo ( n = 29). Three ultrasound‐guided injections (100 μL) were administered into inguinal lymph nodes at 4‐week intervals in early 2022. The primary endpoint was the combined symptom and medication score (cSMS), recorded daily during the 2022 and 2023 grass pollen seasons. Secondary endpoints included safety, Rhinitis Quality of Life Questionnaire (RQLQ), serology, spirometry, fractional exhaled nitric oxide (FeNO), and skin‐prick testing (SPT). An open‐label extension, in which the placebo participants received active ILIT, was conducted in 2024. Results: ILIT was well tolerated, with no severe adverse events (SAEs). Mild local reactions (wheal, erythema, swelling) occurred after 50 of 93 active injections (53.8%), and three moderate reactions (3.2%) were reported. In the placebo group, one mild and no moderate reactions occurred. No allergen‐specific systemic reactions were observed. Compared with placebo, ILIT reduced cSMS by 33% in 2022 ( p > 0.05) and 50% in 2023 ( p < 0.05). RQLQ improved with relative reductions of 22% and 13% ( p > 0.05), respectively. Conclusion: ILIT with a grass pollen allergoid formulated with MCT was well tolerated and associated with clinically relevant reductions in symptom and medication use in grass pollen–induced ARC. In this small study cohort, ILIT proved to be efficient and safe, but further optimisation of dosing regimens should follow.

Synthetic mRNA produced by in vitro transcription is the active pharmaceutical ingredient of approved vaccines and of many drugs under development. It typically contains a 3′ poly(A) tail required for optimal stabilization and translation of the mRNA. The average length of the poly(A) tail in the produced mRNA cannot be precisely predicted and consequently must be measured using complicated technologies such as reverse transcription and sequencing or cleavage of the poly(A) tail and analysis by capillary electrophoresis or mass spectrometry. We report an accelerated method to evaluate the average poly(A) tail length in synthetic mRNA, which benefits from the fact that thiazole orange is fluorescent only when in close proximity of nucleic acid. An oligo(dT)₁₂ oligonucleotide having thiazole orange at its extremities emits a fluorescence signal proportional to the amount of poly(A) sequence available. Using a titration curve made with known amounts of poly(A) RNA oligonucleotide, the thiazole orange oligo(dT)₁₂ oligonucleotide can instantly indicate the average length of the poly(A) tail in an mRNA sample. This fast and easy method can be used in any laboratory to determine the size of the poly(A) tail in in vitro-transcribed mRNA produced for research, pre-clinical studies and clinical trials.

Insect bite hypersensitivity (IBH) is the most common allergic skin disease in horses, but existing clinical scoring systems lack extensive validation, limiting standardized disease monitoring and therapeutic evaluation. Develop and validate a standardized, sensitive, and reproducible clinical scoring system for IBH in horses. Forty-four privately-owned horses with clinically diagnosed IBH examined under field conditions in Switzerland. In this prospective field validation study, the equine IBH severity score (EqIS) integrates a lesion severity score and an area score using a multiplicative algorithm. Six trained evaluators assessed horses independently. Primary outcomes were intra- and interobserver reliability of the lesion severity score and EqIS assessed using intraclass correlation coefficients (ICC). Secondary analyses included agreement of the area score (Pearson correlation), correlations between owner-reported visual analog scale (VAS) scores and EqIS, and comparison between EqIS and a simplified EqIS developed for field use. Intraobserver reliability was excellent (ICC = 0.995 for the lesion severity score and 0.989 for EqIS), and interobserver reliability was similarly high (ICC = 0.98 for both). The area score showed strong interobserver agreement (Pearson r = 0.86-0.94). Owner VAS scores correlated moderately with EqIS (ρ = 0.49-0.51) and strongly with each other (ρ = 0.88). The simplified EqIS demonstrated strong agreement with the validated version (R2 = 0.86). The EqIS is the first extensively validated IBH-specific scoring system and demonstrates excellent reproducibility across evaluators. It provides an objective tool for standardized disease assessment and therapeutic monitoring, whereas the simplified EqIS offers a practical alternative for routine field use.

Synthetic mRNA produced by in vitro transcription is the active pharmaceutical ingredient of approved vaccines and of many drugs under development. It typically contains a 3’ poly-A tail required for optimal stabilization and translation of the mRNA. The average length of the poly-A tail in the produced mRNA cannot be precisely predicted and consequently must be measured using complicated technologies such as reverse transcription and sequencing or cleavage of the poly-A tail and analysis by capillary electrophoresis or mass spectrometry. We report an accelerated method to evaluate the average poly-A tail length in synthetic mRNA, which benefits from the fact that thiazole orange is fluorescent only when in close proximity of nucleic acid. A poly-dT(12) oligonucleotide having thiazole orange at its extremities emits a fluorescence signal proportional to the amount of poly-A sequence available. Using a titration curve made with known amounts of poly-A RNA oligonucleotide, the thiazole orange oligo dT probe can instantly indicate the average length of the poly-A tail in an mRNA sample. This fast and easy method can be used in any laboratory to determine the size of the poly-A tail in in vitro transcribed mRNA produced for research, pre-clinical studies and clinical trials.

Background: Lichen planus (LP) is a chronic inflammatory dermatosis with multiple clinical variants involving the skin, mucous membranes, nails, and hair follicles. Methods: We conducted a retrospective, descriptive study of patients diagnosed with LP at a tertiary referral center from January 2011 to December 2024. Inclusion required concordance between clinical presentation and histopathologic findings. Demographic characteristics, LP subtypes, anatomical involvement, comorbidities, therapeutic approaches, and treatment outcomes were extracted from electronic health records. In addition, an exploratory sensitivity analysis restricted to patients with a single LP subtype was performed to allow for independent subgroup comparisons, and a modified Charlson Comorbidity Index (CCI) based on available comorbidity domains was calculated. Pairwise multivariable logistic regression models adjusted for age, sex, and outcome-specific modified CCI were performed for selected comorbidities. Results: A total of 754 patients were included (mean age 53.1 years), with cutaneous LP (cLP), oral LP (oLP), genital LP (gLP), and lichen planopilaris (LPP) being the most frequent subtypes; a total of 620 had a single major LP subtype and were included in the mutually exclusive analysis. In these groups, modified CCI, age-adjusted modified CCI, and overall comorbidity count differed significantly across subtypes (Kruskal–Wallis p < 0.001). After adjustment in pairwise models, cLP-only showed higher odds of malignancy compared with oLP-only, gLP-only, and LPP-only and higher odds of diabetes mellitus compared with all other pure subtypes. Most other comorbidity comparisons were non-significant or imprecise because of low event numbers. Topical glucocorticoids were the most frequently used treatment, and treatment responses varied by subtype, being more effective in cLP and gLP compared to LPP. Topical calcineurin inhibitors demonstrated the highest response rates in gLP. Acitretin was most effective in cLP, whereas isotretinoin showed favorable responses in oLP. Conclusions: This large, histopathologically confirmed cohort highlights distinct differences in comorbidity patterns, anatomical involvement, and therapeutic response across LP subtypes. Treatment outcomes vary substantially by subtype, underscoring the need for individualized management strategies. Prospective studies are warranted to further elucidate subtype-specific disease associations and optimize treatment approaches.

Background/Objectives: Neoadjuvant and perioperative treatment regimens for melanoma have demonstrated significantly longer event-free survival (EFS) compared with adjuvant therapy in the NADINA and SWOG S1801 trials. While these studies yielded promising results, real-world effectiveness and safety remain to be clarified. Methods: We performed a retrospective, real-world study of all patients with advanced, resectable cutaneous or mucosal melanoma stage III/IV who received neoadjuvant treatment at the Department of Dermatology, University Hospital Zurich, Switzerland between April 2023 and September 2025. Primary endpoints were pathologic and radiologic response, EFS, recurrence-free survival (RFS), and safety. Results: In total, 31 patients were analyzed (52% female; median age 65 years), including 5 patients without lymph node involvement. Eighteen patients (58%) with cutaneous melanoma received neoadjuvant immunotherapy according to the NADINA protocol, three patients (10%) with mucosal melanoma received ipilimumab (1 mg/kg) and nivolumab (3 mg/kg), and ten patients (32%) were treated according to the SWOG S1801 protocol. A major pathologic response (MPR) was achieved in 12 of 31 patients (38%) overall, including 5 of 18 (28%) in the NADINA cohort, 6 of 10 (60%) in the SWOG S1801 cohort, and 1 of 3 (33%) in the mucosal cohort. We observed a pathologic partial response (pPR) in 7 of 31 patients (23%) overall, including 6 of 18 (33%) in the NADINA cohort and 1 of 3 (33%) in the mucosal cohort. A pathologic non-response (pNR) was seen in 9 of 31 patients (29%) overall, including 5 of 18 (28%) in the NADINA cohort, 3 of 10 (30%) in the SWOG S1801 cohort, and 1 of 3 (33%) in the mucosal cohort. Among all patients without lymph node involvement, 1 of 5 achieved MPR (20%), 2 had pPR (40%), and 2 showed pNR (40%). At data cutoff (median follow-up, 9.2 months), the 9-month EFS was 77% in the NADINA cohort, 74% in the SWOG S1801 cohort, and 33% in the mucosal cohort. In the whole cohort, 6-month RFS for the subgroups of MPR, pPR and pNR was 72.9%, 85.7% and 72.9%. Radiologic response evaluation with FDG-PET/CT after neoadjuvant therapy correlated significantly with pathologic response (p = 0.02). No patient with complete metabolic response (CMR) or partial metabolic response (PMR) recurred until data cutoff. In total, 6 of 31 patients (19%) showed stable metabolic disease (SMD), and 8 of 31 patients (26%) showed progressive metabolic disease (PMD). The 6-month RFS in the subgroups of SMD and PMD was 62.5% in each case. Adverse events (AEs) of grade 3 or higher were reported in 13 of 31 patients (42%) in the total real-world cohort, 8 of 18 in the NADINA cohort (44%), 2 of 10 (20%) in the SWOG S1801 cohort (20%), and 3 of 3 (100%) in the mucosal cohort. The most frequent grade 3/4 toxicities were immune-related (ir) Colitis (n = 3, 10%), irHepatitis (n = 2, 6%) and irMyocarditis (n = 2, 6%). Conclusions: Neoadjuvant immunotherapy is effective in real-world practice with a similar safety profile as shown in the clinical studies. Nevertheless, MPR rates in the NADINA real-world cohort were lower compared to the phase III trial. Larger multicenter studies are needed to validate our findings and to better understand response patterns, even in patients without lymph node involvement and in rare melanoma subtypes.

Vitiligo is an autoimmune disorder marked by the progressive loss of skin melanocytes, increasingly linked to immune dysregulation as a key driver of disease onset and progression. Regulatory T (Treg) cells are essential for maintaining immune homeostasis by suppressing autoreactive immune responses. Mounting evidence implicates functional and numerical alterations in Treg cells in the pathogenesis of vitiligo. This study reviews findings on lesional and circulating Treg cells in vitiligo patients compared to healthy controls (HCs), examining Treg cell frequency, their ability to suppress CD4 ⁺ and CD8 ⁺ T cell activity, levels of the immunoregulatory cytokines interleukin‐10 (IL‐10) and transforming growth factor‐β (TGF‐β), expression of the key suppressive marker FOXP3, as well as levels of the pro‐inflammatory cytokines interleukin‐17 (IL‐17) and interleukin‐22 (IL‐22). A comprehensive systematic search was performed across Embase, MEDLINE/PubMed, and Scopus databases to identify eligible studies. A total of 21 studies comprising 1016 vitiligo patients and 846 HCs were included in the review. The analysis revealed a significant reduction in peripheral Treg cell counts ( p = 0.01), impaired overall suppressive capacity of CD4+ and CD8+ T cells ( p = 0.01), reduced levels of IL‐10 ( p = 0.02), increased levels of IL‐17 ( p ≤ 0.01) and IL‐22 ( p ≤ 0.01) in the blood of vitiligo patients compared to HCs. No statistically significant difference was observed in circulating TGF‐β levels ( p = 0.1). Most studies reported reduced FOXP3 expression in both skin and blood of vitiligo patients. Current evidence suggests vitiligo involves both reduced numbers and impaired function of Treg cells, supporting further study of Treg pathways as targets for immunomodulatory therapy.

The Interleukin-36 (IL-36) cytokine family have emerged as important players in mounting an inflammatory response at epithelial barriers and tailoring appropriate adaptive immune responses. As members of the Interleukin-1 superfamily, IL-36 cytokines lack a signal peptide for conventional secretion and require extracellular proteolysis to generate bioactive cytokines. Although the IL-36 family plays an important role in the pathogenesis of plaque and pustular psoriasis, little is known about the release mechanisms of these cytokines from keratinocytes and the physiological stimuli involved. Nucleic acid released from damaged or dying keratinocytes initiates early inflammatory signals that result in the breaking of tolerance associated with psoriasis pathogenesis onset. Cathelicidin peptide, LL37 binds to DNA or double-stranded RNA (dsRNA) and activates a type I Interferon responses in plasmacytoid dendritic cells and keratinocytes. Here, we demonstrate that LL37 binds to dsRNA and induces IL-36γ release from human primary keratinocytes. LL37/dsRNA complexes activate RIG-I-like Receptor signalling, resulting in Caspase-3 and Gasdermin E (GSDME) cleavage. Subsequent GSDME pore formation facilitates IL-36γ release. This response is magnified by priming with psoriasis-associated cytokines, IL-17A and IFNγ. IL-36γ release in this manner is largely independent of cell death in primary keratinocytes and lacked extracellular proteolysis of IL-36γ. Conversely, transfection of keratinocytes directly with dsRNA synthetic analogue, Poly(I:C) induces NLRP1 inflammasome activation, which facilitates IL-36γ expression and release in a GSDMD-dependent manner. Inflammasome-associated cell death also enables extracellular processing of IL-36γ by the release of keratinocyte-derived proteases. These data highlight the distinct responses triggered by dsRNA sensors in keratinocytes. Depending on the inflammatory context and magnitude of the exogenous threat, keratinocytes will release IL-36γ coupled with cell death and extracellular cleavage or release the inactive pro-form, which requires subsequent processing by neutrophil proteases to unleash full biological activity, as occurring in psoriatic skin.

Lichen planus (LP) is a highly prevalent inflammatory skin disease. While various clinical subtypes have been defined, detailed comparisons of these variants are lacking. This study aimed to elucidate differences in gene expression and cellular composition across LP subtypes. Lesional skin biopsies from 28 LP patients (classical, oral, genital, and lichen planopilaris) and seven non-diseased skin controls (NDC) were analyzed. Gene expression profiling of 730 inflammation-related genes was conducted using NanoString. Immune cell compositions were assessed by multiplex immunohistochemistry. Gene expression profiles revealed unique inflammatory signatures for each LP subtype. Lichen planopilaris exhibited the most divergence, with downregulated gene expression and upregulation of complement pathway genes (C5-7), along with elevated M2 macrophages. Oral and genital LP demonstrated similar profiles with strong upregulation of TNF-related and Toll-like receptor-associated genes. Oral LP showed the highest upregulation of cytotoxicity-associated genes, as well as high numbers of CD8+ IL-17A+ (Tc17) cells (8.02%). Interferon gene signatures were strongly upregulated in oral and classical LP. The study highlights distinct differences in inflammatory gene expression and cell composition across LP subtypes, emphasizing the need for tailored therapeutic approaches.