Luiz Carlos Rodrigues Junior

Aging is accompanied by numerous interlinked cellular, genetic, and molecular changes, one of the most consequential being inflammaging: a chronic, low-grade systemic inflammatory state linked to metabolic dysregulation and pleiotropic effects on immune function. Although diverse intrinsic and extrinsic factors are known to drive inflammaging, the contribution of persistent viral infection remains incompletely understood. Here, we evaluate evidence that human herpesviruses, including cytomegalovirus (CMV), Epstein–Barr virus (EBV), herpes simplex viruses (HSV-1/2), and varicella zoster virus (VZV), may act as chronic antigenic and inflammatory stimuli capable of amplifying inflammaging and shaping the trajectory of aging. We review the molecular, cellular, and systems-level mechanisms by which herpesvirus latency and periodic reactivation sustain innate and adaptive immune activation, promote cellular senescence, and contribute to immune dysregulation and exhaustion. We further examine epidemiological evidence linking herpesvirus infection to dementia and other age-associated outcomes and highlight emerging data suggesting that antiviral treatment and vaccination, such as against VZV, may mitigate these effects. We conclude that herpesviruses are best understood as adjuvant contributors, rather than primary drivers, of inflammaging, and propose that persistent herpesvirus infection be considered an addressable risk factor in strategies aimed at promoting healthy aging.

Aging is accompanied by numerous interlinked cellular, genetic, and molecular changes, one of the most consequential being inflammaging: a chronic, low-grade systemic inflammatory state linked to metabolic dysregulation and pleiotropic effects on immune function. Although diverse intrinsic and extrinsic factors are known to drive inflammaging, the contribution of persistent viral infection remains incompletely understood. Here, we synthesize evidence that human herpesviruses, including cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex viruses (HSV-1/2), and varicella zoster virus (VZV), act as chronic antigenic and inflammatory stimuli capable of amplifying inflammaging and shaping the trajectory of aging. We review the molecular, cellular, and systems-level mechanisms by which herpesvirus latency and periodic reactivation sustain innate and adaptive immune activation, promote cellular senescence, and contribute to immune dysregulation and exhaustion. We further examine epidemiological evidence linking herpesvirus infection to dementia and other age-associated outcomes and highlight emerging data suggesting that antiviral treatment and vaccination, such as against VZV, may mitigate these effects. We conclude that herpesviruses are best understood as adjuvant contributors, rather than primary drivers, of inflammaging, and propose that persistent herpesvirus infection be considered an addressable risk factor in strategies aimed at promoting healthy aging.

BACKGROUND Mosquito saliva contains bioactive molecules that modulate host immunity and may influence arboviral infection. The contribution of Aedes aegypti salivary gland extract (SGE) to viral replication and immune regulation during Zika virus (ZIKV) infection remains poorly understood. OBJECTIVES To investigate the immunomodulatory effects of Ae. aegypti SGE during ZIKV infection. METHODS Peripheral blood mononuclear cells (PBMCs) and murine antigen-presenting cell lines were exposed to ZIKV with or without SGE. Viral replication was measured by quantitative polymerase chain reaction (qPCR), cell death and immune subsets by flow cytometry, oxidative stress markers by biochemical assays, and cytokine production by enzyme-linked immunosorbent assay (ELISA). FINDINGS SGE enhanced ZIKV replication, particularly in PBMCs, with increased RNA copies (median Δ = +1,779), reduced late apoptosis of CD4⁺ T cells (p = 0.0055), dendritic cell death (p < 0.01), and impaired T-cell proliferation. SGE attenuated ZIKV-induced oxidative damage by restoring glutathione levels, reducing lipid and protein oxidation (p < 0.001), and increasing nitric oxide (NO) production. Cytokine profiling revealed suppression of interferon-y (IFN-γ) (p < 0.001) and induction of interleukin-4 (IL-4) (p < 0.0001), indicating a Th2-skewed response. Murine cell lines confirmed SGE-driven cytokine modulation. MAIN CONCLUSIONS Ae. aegypti SGE alters host immune homeostasis, favouring ZIKV infection by weakening antiviral defences and redirecting immune and redox pathways, thereby facilitating viral expansion.

The induction of immune responses in tissues and mucosa has emerged as one of the most promising strategies for the development of more effective vaccines and immunotherapies. In this context, CD8 ⁺ resident memory T cells (CD8 ⁺ TRM) have arisen as key players in local immune surveillance, acting persistently within non-lymphoid tissues. These cells represent a new and promising frontier in local immune responses and as potential clinical tools. CD8 ⁺ TRM are being extensively investigated as therapeutic targets against viral infections and cancer, although their clinical applications have yet to be fully established. Understanding the molecular signals that regulate their generation, differentiation, maintenance, and activation is crucial for the precise targeting of their immune functions. This review explores the main mechanisms involved in the formation and maintenance of CD8 ⁺ TRM, from the strength of MHC: TCR interactions to the coordinated role of cytokines, chemokines, and transcription factors in tissue retention and the expression of markers such as CD69, CD103, and CD49a. By integrating this knowledge, we discuss strategies to manipulate these pathways with the goal of developing more effective vaccines and personalized therapies based on resident memory T cells. We also examine how these molecular signals and pathways, either independently or in combination, can be explored both in the fight against viral infections and cancer, and in identifying CD8 ⁺ TRM predictive biomarkers for response to anticancer immunotherapies across various tumor types.

This study aimed to investigate the therapeutic potential of a nanoparticle formulation containing the immunodominant peptide SSIEFARL from Herpes simplex virus 2 glycoprotein B A against genital herpes. A nanoparticle formulation (NanoSSIEFARL) was engineered and characterized for its physicochemical and immunomodulatory properties. NanoSSIEFARL displayed mean particle size of 212 ± 5 nm, polydispersity index of 0.12 ± 0.01 and zeta potential of -7.4 ± 2.5 mV, exhibiting spherical morphology. pH stability remained consistent over 30 days (day 0: 4.5 ± 0.5; day 30: 5.0 ± 0.5). A novel high-performance liquid chromatography method was validated for SSIEFARL quantification. Peptide association efficiency reached 98.3% ± 2.1, with 41.6% ± 4.4 peptide release from nanoparticles after 4 hours. Cytotoxicity assessment revealed cellular viability exceeding 90%, with macrophage uptake observed after 4 hours. Altogether, these results suggest that NanoSSIEFARL is a promising candidate for effective immunotherapy against genital herpes.

Purinergic signaling modulates immune function and is involved in the immunopathogenesis of several viral infections. This study aimed to investigate alterations in purinergic pathways in coronavirus disease 2019 (COVID‐19) patients. Mild and severe COVID‐19 patients had lower extracellular adenosine triphosphate and adenosine levels, and higher cytokines than healthy controls. Mild COVID‐19 patients presented lower frequencies of CD4⁺CD25⁺CD39⁺(activated/memory regulatory T cell [mTreg]) and increased frequencies of high‐differentiated (CD27⁻CD28⁻) CD8⁺T cells compared with healthy controls. Severe COVID‐19 patients also showed higher frequencies of CD4⁺CD39⁺, CD4⁺CD25⁻CD39⁺(memory T effector cell), and high‐differentiated CD8⁺T cells (CD27⁻CD28⁻), and diminished frequencies of CD4⁺CD73⁺, CD4⁺CD25⁺CD39⁺mTreg cell, CD8⁺CD73⁺, and low‐differentiated CD8⁺T cells (CD27⁺CD28⁺) in the blood in relation to mild COVID‐19 patients and controls. Moreover, severe COVID‐19 patients presented higher expression of PD‐1 on low‐differentiated CD8⁺T cells. Both severe and mild COVID‐19 patients presented higher frequencies of CD4⁺Annexin‐V⁺and CD8⁺Annexin‐V⁺T cells, indicating increased T‐cell apoptosis. Plasma samples collected from severe COVID‐19 patients were able to decrease the expression of CD73 on CD4⁺and CD8⁺T cells of a healthy donor. Interestingly, the in vitro incubation of peripheral blood mononuclear cell from severe COVID‐19 patients with adenosine reduced the nuclear factor‐κB activation in T cells and monocytes. Together, these data add new knowledge to the COVID‐19 immunopathology through purinergic regulation.

Herpes simplex virus (HSV) 1 and 2 are viruses that infect individuals worldwide and for which there is no cure or vaccine available. The protective response against herpes is mostly mediated by CD8 T lymphocytes that respond to the immunodominant SSIEFARL epitope. However, there are some obstacles concerning the use of free SSIEFARL for vaccine or immunotherapy. The aim of this study was to evaluate the feasibility of nanoencapsulation of SSIEFARL and its immunostimulatory properties. Nano/SSIEFARL was produced by interfacial polymerization in methylmetacrylate, and the physico‐chemical properties, morphology and immunobiological parameters were evaluated. To evaluate the ex vivo capacity of Nano/SSIEFARL, we used splenocytes from HSV‐1‐infected mice to enhance the frequency of SSIEFARL‐specific CD8 T lymphocytes. The results indicate that Nano/SSIEFARL has a spherical shape, an average diameter of 352 ± 22 nm, the PDI was 0.361 ± 0.009 and is negatively charged (−26.30 ± 35). The stability at 4°C was 28 days. Also, Nano/SSIEFARL is not toxic for cells at low concentrations in vitro and it is taken up by JAWS II dendritic cells. No histopathological changes were observed in kidneys, liver and lymph nodes of animals treated with Nano/SSIEFARL. Nan/SSIEFARL increased the production of IL‐1β, TNF‐α and IL‐12 by the dendritic cells. Finally, Nano/SSIEFARL expanded the frequency of SSIEFARL‐specific CD8+T lymphocytes at the same rate as free SSIEFARL. In conclusion all data together indicate that SSIEFARL is suitable for nanoencapsulation, and the system produced presents some immunoadjuvant properties that can be used to improve the immune response against herpes.

Sepsis is characterized by a dysregulated immune response to infection characterized by an early hyperinflammatory and oxidative response followed by a subsequent immunosuppression phase. Although there have been some advances in the treatment of sepsis, mortality rates remain high, urging for the search of new therapies. β-Lapachone (β-Lap) is a natural compound obtained from Tabebuia avellanedae Lorentz ex Griseb. with several pharmacological properties including bactericidal, anti-inflammatory, and antioxidant activity. Thus, the aim of this study was to evaluate the effects of β-Lap in a mouse sepsis model. To this, we tested two therapeutic protocols in mice submitted to cecal ligation and puncture- (CLP-) induced sepsis. First, we found that in pretreated animals, β-Lap reduced the systemic inflammatory response and improved bacterial clearance and mouse survival. Moreover, β-Lap also decreased lipid peroxidation and increased the total antioxidant capacity in the serum and peritoneal cavity of septic animals. In the model of severe sepsis, the posttreatment with β-Lap was able to increase the survival of animals and maintain the antioxidant defense function. In conclusion, the β-Lap was able to increase the survival of septic animals by a mechanism involving immunomodulatory and antioxidant protective effects.

Background: Leishmaniasis reaches millions of people around the world. The control of the disease is difficult due to the restricted access to the diagnosis and medication, and low adherence to the treatment. Thus, more efficient drugs are needed and natural products are good alternatives. Iridoids, natural products with reported leishmanicidal activity, can be exploited for the development of anti- Leishmania drugs. The aim of this study was to isolate and to investigate the in vitro activity of iridoids against Leishmania amazonensis and to compare the activity in silico of these compounds with those reported as active against this parasite.Methods: Iridoids were isolated by chromatographic methods. The in vitro activity of asperuloside (1) and geniposide (2) from Escalonia bifida, galiridoside (3) from Angelonia integerrima and theveridoside (4) and ipolamiide (5) from Amphilophium crucigerum was investigated against promastigote forms of Leishmania amazonensis. Molecular modeling studies of 1-5 and iridoids cited as active against Leishmania spp. were performed.Results: Compounds 1-5 (5-100 µM) did not inhibit the parasite survival. Physicochemical parameters predicted for 1-5 did not show differences compared to those described in literature. The SAR and the pharmacophoric model confirmed the importance of maintaining the cyclopentane[C]pyran ring of the iridoid, of oxygen-linked substituents at the C1 and C6 positions and of bulky substituents attached to the iridoid ring to present leishmanicidal activity.Conclusion: The results obtained in this study indicate that iridoids are a promising group of secondary metabolites and should be further investigated in the search for new anti-Leishmania drugs.