Naiura Vieira Pereira

Epidermodysplasia verruciformis (EV) is characterized by susceptibility to cutaneous β -human papillomaviruses, persistent viral lesions, and an increased risk of cutaneous squamous cell carcinoma (cSCC). We investigated whether expression of Toll-like receptors 3, 4, 5, and 9 (TLR3, TLR4, TLR5, and TLR9) differs across histological states within EV. Immunoreactivity was quantified by digital H -score using QuPath in 97 specimens from 28 patients, comprising 13 lesion-free skin samples, 15 flat warts, and 69 cSCCs. Prespecified comparisons evaluated flat wart versus lesion-free skin, cSCC versus non-neoplastic tissue, and invasive versus in situ cSCC, with invasive tumor differentiation assessed secondarily. Patient-clustered generalized estimating equations and false-discovery-rate (FDR) correction were used for inference. After correction for multiple testing, TLR9 H -scores were higher in flat warts than in lesion-free skin ( β = 60.58; q = 0.0332). Compared with non-neoplastic tissue, cSCC showed lower TLR3 ( β = −81.40; q = 0.00745), higher TLR4 ( β = 55.59; q = 0.04793), and lower TLR9 expression ( β = −34.80; q = 0.03412). TLR5 showed a nominal increase in cSCC that did not meet the FDR threshold. None of the four receptors differed significantly between invasive and in situ cSCC, and no differentiation trend remained significant after FDR correction. These findings show receptor-specific differences in TLR expression across histological states of EV, particularly involving TLR3, TLR4, and TLR9, although the diagnostic and prognostic significance of these tissue patterns remains to be established.

Epidermodysplasia verruciformis (EV) is characterized by susceptibility to cutaneous β-human papillomaviruses, persistent viral lesions and an increased risk of cutaneous squamous cell carcinoma (cSCC). We investigated whether expression of Toll-like receptors 3, 4, 5 and 9 (TLR3, TLR4, TLR5 and TLR9) differs across histological states within EV. Immunoreactivity was quantified by digital H-score using QuPath in 97 specimens from 28 patients, comprising 13 lesion-free skin samples, 15 flat warts and 69 cSCCs. Prespecified comparisons evaluated flat wart versus lesion-free skin, cSCC versus non-neoplastic tissue and invasive versus in situ cSCC, with invasive tumour differentiation assessed secondarily. Patient-clustered generalized estimating equations and false-discovery-rate (FDR) correction were used for inference. After correction for multiple testing, TLR9 H-scores were higher in flat warts than in lesion-free skin (β=60.58; q=0.0332). Compared with non-neoplastic tissue, cSCC showed lower TLR3 (β=−81.40; q=0.00745), higher TLR4 (β=55.59; q=0.04793) and lower TLR9 expression (β=−34.80; q=0.03412). TLR5 showed a nominal increase in cSCC that did not meet the FDR threshold. None of the four receptors differed significantly between invasive and in situ cSCC, and no differentiation trend remained significant after FDR correction. These findings show receptor-specific differences in TLR expression across histological states of EV, particularly involving TLR3, TLR4 and TLR9, although the diagnostic and prognostic significance of these tissue patterns remains to be established.

Epidermodysplasia verruciformis (EV) is a rare genodermatosis characterized by persistent cutaneous β-human papillomavirus infection and increased risk of cutaneous squamous cell carcinoma (cSCC). The cyclic GMP-AMP synthase (cGAS)–stimulator of interferon genes (STING) pathway links cytosolic DNA sensing, genomic instability and innate immune activation, but its expression in EV-associated cSCC remains poorly defined. We quantified STING, p53 and BCL2 immunohistochemical expression in tissue microarrays containing 47 EV-associated and 83 non-EV cSCCs. Tumor H-scores were medians across valid cores; primary comparisons used patient-clustered generalized estimating equations (GEE). STING expression was higher in EV-associated tumors (median H-score 192.4 vs. 149.5); in grade-adjusted GEE, EV status was associated with a + 37.2-unit difference (95% confidence interval [CI] 23.4–51.0; p < 0.001). The EV–non-EV STING difference varied by histological grade (interaction p = 0.0067), with the largest estimated difference in grade 1 tumors. p53 expression was also higher in EV tumors (97.7 vs. 36.3; adjusted difference + 43.8, 95% CI 18.8–68.8; p < 0.001), and EV tumors had higher odds of belonging to a higher p53 tertile (odds ratio [OR] 3.54, 95% CI 1.98–6.32; p < 0.001). BCL2 remained low and was not significantly associated with EV (adjusted difference − 1.93, 95% CI − 4.84 to 0.98; p = 0.193). STING and p53 remained positively associated after adjustment for histological grade, EV status and patient clustering. These findings define an altered STING protein-expression phenotype in EV-associated cSCC, accompanied by increased p53 expression but no independent BCL2 association, and support functional investigation of cGAS–STING signaling.

Zika virus (ZIKV) infection during pregnancy is a critical driver of Congenital Zika Syndrome (CZS), yet the mechanisms of pathogenesis at the placental barrier remain incompletely understood. This article is a translational, observational, and experimental study combining clinical placental analyses, placental explants cultures and in vivo murine model to investigate the mechanisms involved in ZIKV infection. We evaluate the histopathological analyses to verify presence of inflammation in ZIKV-infected human placentas from newborns with CZS and without CZS (N-CZS), identifying more intense Hofbauer cell hyperplasia, villitis and decidual inflammation in CZS group. Moreover, placental immunohistochemistry analyses identified decreased TLR4 expression in the villi and reduced TNF and IL-10 levels across placental layers of CZS group. Next, we investigated the effects of inflammation on viral replication and explored whether the flavonoid Naringenin (NGN) could modulate this inflammation. Using a placental villous explant model, we verified that inflammation induced by LPS exacerbates viral replication and pathological markers. Notably, treatment with the NGN rescued the inflammatory and virological outcomes. These findings were further validated in a murine model of congenital infection, where NGN administration alleviated microcephaly-related structural alterations in ZIKV-exposed neonates. Our results indicate that placental inflammation is a key provocateur of ZIKV replication and subsequent fetal brain malformation. Furthermore, we identify NGN as a promising bifunctional antiviral and anti-inflammatory candidate for mitigating the developmental impacts of ZIKV infection.

Epidermodysplasia verruciformis (EV) is a rare genodermatosis characterized by lifelong β‐human papillomavirus (β‐HPV) persistence, extensive flat warts, and increased risk of cutaneous squamous cell carcinoma. While TMC6, TMC8, and CIB1 mutations are recognized as genetic drivers, innate immune mechanisms contributing to HPV persistence remain incompletely defined. This study quantified the expression of Toll‐like receptors (TLRs) 3, 4, 5, and 9 in normal skin and flat warts from patients with EV and immunocompetent individuals without EV (NEV). We performed immunohistochemical analysis on 135 formalin‐fixed, paraffin‐embedded specimens using standardized digital morphometry of epidermal keratinocytes. EV patients exhibited significantly reduced TLR3 and TLR9 expression in both normal skin and flat warts relative to controls, whereas TLR4 and TLR5 levels were comparable. Notably, flat warts from NEV individuals showed marked TLR3 upregulation relative to matched normal skin, whereas this response was absent in EV patients. These findings are consistent with an EV‐associated epithelial innate‐sensing phenotype. Our data suggest innate immune deficiencies may interact with previously described keratinocyte abnormalities, amplifying local immune dysfunction. These findings provide a framework for investigating TLR‐based therapeutic approaches in EV.

SARS-CoV-2, a β-coronavirus, primarily affects the lungs, with non-specific lesions and no cytopathic viral effect in the skin. Cutaneous antiviral mechanisms include activation of TLR/IRF pathways and production of type I IFN. We evaluated the antiviral mechanisms involved in the skin of COVID-19 patients, including skin samples from 35 deceased patients who had contracted COVID-19 before the launch of the vaccine. Detection of SARS-CoV-2 in the skin was performed using transmission electron microscopy and RT-qPCR. Microscopic and molecular effects of the virus in skin were evaluated by histopathology, RT-qPCR, and immunohistochemistry (IHC). The results revealed the presence of SARS-CoV-2 and microscopic changes, including microvascular hyaline thrombi, perivascular dermatitis, and eccrine gland necrosis. There was increased transcription of TBK1 and a reduction in transcription of TNFα by RT-qPCR in the COVID-19 group. IHC revealed reduced expression of ACE2, TLR7, and IL-6, and elevated expression of IFN-β by epidermal cells. In the dermis, there was decreased expression of STING, IFN-β, and TNF-α and increased expression of IL-6 in sweat glands. Our results highlight the role of type I IFN in the skin of COVID-19 patients, which may modulate the cutaneous response to SARS-CoV-2.

Chronic urticaria (CU) is a skin disease characterized by recurrent episodes of urticaria and/or angioedema, persisting for more than six weeks. Chronic urticaria is classified as chronic spontaneous urticaria (CSU) and chronic inducible urticaria (CIndU). Histamine, released by mast cells and basophils, is the central mediator in the development of signs and symptoms, especially pruritus. IL-31 activates pruritus via IL-31RA. Th2 cytokines also contributes to the inflammatory environment, releasing further IL-31. We aimed to investigate the quantification of mast cells, expression of pro-inflammatory cytokines in skin samples from individuals with CU and cell activation by the basophil activation test. Thirteen patients with CSU, 11 patients with CIndU and 10 healthy controls (HC) were enrolled in the study. We performed histologic quantification of the mast cells, metachromatic stained with toluidine blue, in CSU and CIndU lesional samples; IL-31, IL-31RA, IL-4 and IFN-γ expression by immunohistochemistry; and basophil activation test (BAT) by flow cytometry. The main findings were increased mast cell quantification in CSU; increased epidermal and dermal expression of IL-31 in CSU lesional samples and increased dermal expression in CIndU samples; augmented IL-31RA expression at epidermis and dermis of CSU group; augmented expression IL-4 at epidermis of CSU patients. We also found enhanced BAT positivity in CSU, reinforcing the autoimmune profile of our CSU patients. The analysis of the proinflammatory cytokines related to pruritus, showed increased expression of the evaluated components. These remarkable findings in CSU emphasize their relevance as potential disease biomarkers and targets for immunomodulatory interventions.

Atopic dermatitis (AD) is an inflammatory skin disease with intense pruritus, and chronic skin colonization by Staphylococcus aureus. To understand the inflammatory status in AD, we investigated the inflammasome complex, that activates ASC (Apoptosis-associated speck-like protein containing a CARD), caspase-1 and GSDMD (gasdermin-D), and production of IL-1β and IL-18. We aimed to evaluate the expression of the inflammasome pathway in the skin of adults with AD. Thirty patients with moderate to severe AD and 20 healthy controls were enrolled in the study. We performed the analysis of the inflammasome components NLRP1, NLRP3, AIM-2, IL-1β, IL-18, Caspase-1, ASC, GSDMD, and CD68 expression (macrophage marker) by immunohistochemistry and immunofluorescence. The main findings included increased expression of NLRP3, NLRP1 and AIM-2 at dermal level of severe AD; augmented IL-18 and IL-1β expression at epidermis of moderate and severe patients, and in the dermis of severe AD; augmented expression of ASC, caspase-1 and GSDMD in both epidermis and dermis of moderate and severe AD. We detected positive correlation between caspase-1, GSDMD and IL-1β (epidermis) and caspase-1 (dermis) and AD severity; NLRP3, AIM-2 and IL-1β, and NLRP3 with IL-18 in the epidermis; ASC, GSDMD and IL-1β, and NLRP3, AIM-2, caspase-1, and IL-18 in the dermis. We also evidenced the presence of CD68⁺ macrophages secreting GSDMD, ASC and IL-1β in moderate and severe AD. Cutaneous macrophages, early detected in moderate AD, have its role in the disease inflammatory mechanisms. Our study indicates a canonical activation pathway of inflammasomes, reinforced by the chronic status of inflammation in AD. The analysis of the inflammasome complex evidenced an imbalance in its regulation, with increased expression of the evaluated components, which is remarkably in severe AD, emphasizing its relevance as potential disease biomarkers and targets for immunomodulatory interventions.