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Leopoldo Spadea, Emanuele Tonti, Rita Napolitano Eye Clinic, Department Sensory Organs, Policlinico “Umberto I”, “Sapienza” University of Rome, Rome, Italy Objective: To describe a very unique case of two Descemet stripping automated endothelial keratoplasty (DSAEK) surgeries performed in both eyes of the same patient with an extremely different graft thickness and overall corneal thickness but with the same corrected distance visual acuity (CDVA) 2 years after surgery.Case presentation: A 75-year-old woman with bilateral bullous keratopathy (BK) was submitted to DSAEK surgeries in both eyes, first in right and after 6 months in left eye. CDVA was 20/160 in the right eye and 20/63 in the left eye. Corneal thickness evaluated by anterior segment optical coherence tomography was 569 µm in the right eye and 560 µm in the left eye. The root mean square (RMS) was 2.1 in the right and left eyes. Endothelial cell densities were not detectable in both eyes. The estimated precut donor graft thickness from eye bank was 250 and 40 µm in the right and in the left graft, respectively. Two years after surgery CDVA was 20/25 in both eyes. Corneal thickness was 633 µm with a lenticule thickness of 206 µm in the right eye and 439 µm with a lenticule thickness of 48 µm in the left eye. The RMS was 1.7 in the right eye and 1.4 in the left eye. Endothelial cell density was 2.272 cells/mm2 in the right and 2.154 cells/mm2 in the left eye.Conclusion: DSAEK was safe and effective in the treatment of BK. In our report, the visual outcome resulted to be poorly related either to donor graft thickness or to postoperative corneal thickness. Keywords: anterior segment OCT, bullous keratopathy, Descemet stripping endothelial keratoplasty, graft thickness, visual acuity

Leopoldo Spadea, Emanuele Tonti, Enzo Maria Vingolo Department of Medico-Surgical Sciences and Biotechnologies, Sapienza University of Rome, Latina, Italy Abstract: Collagen cross-linking (CXL) is a relatively new conservative approach for progressive corneal ectasia, which is able to strengthen corneal tissue reforming new covalent bonds. Subjective and objective results following this method seem to be promising. In recent years, newer CXL protocols have been developed to perform more effective and less invasive procedures. The increasing diffusion of CXL in the corneal ectatic disease has increased the need to have actual indices regarding the efficacy of the treatment. Evaluation of demarcation line (DL), a transition zone between the cross-linked anterior corneal stroma and the untreated posterior corneal stroma, is considered a measurement of the depth of CXL treatment into the stroma. Some evidence in the literature emphasize that DL could be a measure of effectiveness of the CXL. On the contrary, some authors believe that the “the deeper, the better” principle is rather a simplistic approach for interpreting the clinical importance of the corneal stromal DL. Keywords: corneal cross-linking, demarcation line, keratoconus, ultraviolet A

Purpose. To evaluate safety and efficacy of customized central corneal regularization (CCR), together with simultaneous accelerated corneal collagen cross-linking (A-CXL) - CCR-CXL protocol, to treat keratoconus-related corneal ectasia. Design. Retrospective, comparative observational case series. Methods. Patients that had undergone combined CCR-CXL protocol. Main inclusion criteria were keratoconus visual acuity deterioration and contact lens intolerance. All patients underwent complete ophthalmological evaluation, corrected distance visual acuity (CDVA) and Scheimpflug-corneal tomography. Central corneal regularization was performed by ablation using flying spot laser. Subsequently, the stroma was saturated with 0.17% riboflavin-5-phosphate added every 2 minutes, followed by A-CXL 9 mW/cm2 for 10 minutes. CDVA, medium keratometry value (Kmed), and total corneal morphological irregularity index (CMI) of patients were analyzed before surgery and after 1, 3 and 12 months. A P value of .05 or less was considered statistically significant. Results. 46 eyes of 39 keratoconus patients were treated. At 1 month, the mean CDVA (LogMar) increased from 0.19 ± 0.02 to 0.12 ± 0.02 (P < .05), and the difference remained stable at month 12. Kmax decrease was statistically significant from 57.02 ± 5.65 to 50.21 ± 4.48 (P < .05). CMI decreased significantly from 47.8 ± 2.84 to 30.1 ± 2.4 (P < .01). Conclusions. CCR-CXL protocol is safe and effective in arresting keratectasia progression and increasing corneal optic regularity in keratoconus. These findings showed a significant improvement in CDVA, keratometry values and corneal optical aberrations after being treated with the CCR-CXL protocol. [ABSTRACT FROM AUTHOR]

The paper deals with a method, based on the finite formulation approach for the solution of a quasi-static eddy-current problem (ECP). As an example, the method has been tested against a two-dimensional axisymmetric ECP and the obtained results have been compared with those from well-established methods. The finite formulation is a new formulation, complementary to the traditional differential formulation, consisting of a discrete, i.e., algebraic, formulation obtained directly from experimental facts, without applying a discretization procedure to a differential equation. It adopts the global variables together with the primal-dual cellulization of the space and time domain as starting point. Index Terms--Eddy currents, quasi-static magnetic field.

Background: Glaucoma is a leading cause of irreversible blindness worldwide, necessitating precise management strategies tailored to individual patient characteristics. Artificial intelligence (AI) holds promise in revolutionizing the approach to glaucoma care by providing personalized interventions., Aim: This review explores the current landscape of AI applications in the personalized management of glaucoma patients, highlighting advancements, challenges, and future directions., Methods: A systematic search of electronic databases, including PubMed, Scopus, and Web of Science, was conducted to identify relevant studies published up to 2024. Studies exploring the use of AI techniques in personalized management strategies for glaucoma patients were included., Results: The review identified diverse AI applications in glaucoma management, ranging from early detection and diagnosis to treatment optimization and prognosis prediction. Machine learning algorithms, particularly deep learning models, demonstrated high accuracy in diagnosing glaucoma from various imaging modalities such as optical coherence tomography (OCT) and visual field tests. AI-driven risk stratification tools facilitated personalized treatment decisions by integrating patient-specific data with predictive analytics, enhancing therapeutic outcomes while minimizing adverse effects. Moreover, AI-based teleophthalmology platforms enabled remote monitoring and timely intervention, improving patient access to specialized care., Conclusions: Integrating AI technologies in the personalized management of glaucoma patients holds immense potential for optimizing clinical decision-making, enhancing treatment efficacy, and mitigating disease progression. However, challenges such as data heterogeneity, model interpretability, and regulatory concerns warrant further investigation. Future research should focus on refining AI algorithms, validating their clinical utility through large-scale prospective studies, and ensuring seamless integration into routine clinical practice to realize the full benefits of personalized glaucoma care.

The terminology "vitreous cortex remnants" (VCR) indicates the outermost lamellae of vitreous cortex that remains attached to the retinal surface as a consequence of vitreoschisis. The relevance of VCR removal in eyes with rhegmatogenous retinal detachment (RRD) is unknown. We conducted a review from January 1, 2000, to July 30, 2023, examining 1493 eyes. Outcome measures included: prevalence of VCR, relationship between VCR and detachment recurrence due to proliferative vitreoretinopathy (PVR), and relationship between VCR and epiretinal membrane (ERM) formation. A meta-analysis was performed with data reported as odds ratios (OR) or mean difference and 95 % confidence intervals. Prevalence of macular and peripheral VCR was 53.4 and 46.8, respectively, with an overall VCR prevalence of 50.8 (95 % CI 42.6, 59.1) Given the scarcity of available data, meta-analysis regarding the relationship between peripheral VCR and redetachment due to PVR was not feasible. The odds of developing ERM were not statistically different between eyes that had had macular VCR removal vs. eyes that had not (log OR -0.08 [95 % CI -1.06, 0.89 p= 0.89]. Additional prospective studies are required to verify whether removal of VCR may reduce the odds of recurrence of RRD due to PVR and the development of ERM., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Regulatory approval of immune checkpoint inhibitors (ICIs) was based on results of large, randomized clinical trials, resulting in limited outcomes data in patient cohorts typically underrepresented in such trials. The objective of this study was to evaluate the efficacy and safety of ICIs in these unique patient cohorts. This is a multicenter, retrospective analysis of real-world data at six academic and community clinics in the United States from 1 January 2011 to 1 April 2018. Patients were included if they had received at least one cycle of ICI treatment. Unique patient cohorts included age > 75 years, non-White race, positive smoking history, ECOG performance status (PS) ≥ 2, BMI ≥ 30 kg/m 2 , autoimmune diseases (AIDs), chronic viral infections (CVI), extensive prior lines of therapy (LOTs), or >three metastatic sites. Immune-related adverse events (irAEs), overall survival (OS), and time to treatment failure were evaluated in the entire cohort and in NSCLC patients treated with PD-(L)1 monotherapy. Outcomes and their association with unique patient cohorts were compared on univariate analysis and multivariate analysis to those without a particular characteristic in the entire NSCLC PD-(L)1 monotherapy cohorts. In total, 1453 patients were included: 56.5%-smokers, 30.4%-non-White, 22.8%-elderly, 20.8%-ECOG PS ≥ 2, 15.7%-history of AIDs, and 4.7%-history of CVI. The common ICIs were nivolumab (37.1%) and pembrolizumab (22.2%). Black patients, compared to White patients, experienced fewer irAEs (OR 0.54, p < 0.001). An ECOG PS of ≥2 (HR = 2.01, p < 0.001) and an increased number of previous LOTs were associated with poor OS (the median OS of 26.2 vs. 16.2 vs. 9.6 months for one vs. two vs. three prior LOTs, p < 0.001). The above results were confirmed in anti-PD-(L)1 monotherapy non-small cell lung cancer patients (n = 384). Overall, ICIs were safe and efficacious in these typically underrepresented patient cohorts. We noted ECOG PS ≥ 2 and an increased prior LOTs were associated with poor ICI efficacy, and Black patients, compared to White patients, experienced fewer irAEs.

Glaucoma, a complex and multifactorial neurodegenerative disorder, is a leading cause of irreversible blindness worldwide. Despite significant advancements in our understanding of its pathogenesis and management, early diagnosis and effective treatment of glaucoma remain major clinical challenges. Epigenetic modifications, encompassing deoxyribonucleic acid (DNA) methylation, histone modifications, and non-coding RNAs, have emerged as critical regulators of gene expression and cellular processes. The aim of this comprehensive review focuses on the emerging field of epigenetics and its role in understanding the complex genetic and molecular mechanisms underlying glaucoma. The review will provide an overview of the pathophysiology of glaucoma, emphasizing the intricacies of intraocular pressure regulation, retinal ganglion cell dysfunction, and optic nerve damage. It explores how epigenetic modifications, such as DNA methylation and histone modifications, can influence gene expression, and how these mechanisms are implicated in glaucomatous neurodegeneration and contribute to glaucoma pathogenesis. The manuscript discusses evidence from both animal models and human studies, providing insights into the epigenetic alterations associated with glaucoma onset and progression. Additionally, it discusses the potential of using epigenetic modifications as diagnostic biomarkers and therapeutic targets for more personalized and targeted glaucoma treatment.

Objective: Evidence for pharmacogenomic (PGx) guided treatment in child and adolescent psychiatry is growing. This study evaluated the impact of PGx testing on psychotropic medication prescribing in an ambulatory child and adolescent psychiatry and a developmental pediatrics clinic. Methods: This was a single-center, retrospective, descriptive analysis of patients who underwent PGx testing between January 2015 and October 2022 at a child and adolescent psychiatry clinic or developmental pediatrics clinic. The primary outcome was the proportion of patients with at least one psychotropic medication modification made 6-month posttesting that could be attributed to CYP2C19, CYP2D6, HLA-B*15:02, or HLA-A*31:01. Secondary outcomes included reason for testing, types of therapeutic modifications made, and whether the therapeutic modifications concorded with PGx guidelines. Results: A total of 193 patients were analyzed. The average age was 10 ± 4 years old, 60% were male, 78% were Caucasian. Sixty-eight percent had a primary diagnosis of a neurodevelopmental disorder, namely autism spectrum disorder (51%), and attention-deficit/hyperactivity disorder (14%). The reasons for PGx testing included medication inefficacy (34%), medication intolerance (20%), and family request (19%). At the time of PGx testing, 37% of patients were taking ≥1 psychotropic medication with PGx annotation. Overall, 35 PGx-related therapeutic modifications were made in 32 (17%) patients. These included continuing current PGx medication (6.2%) and starting PGx medication (5.2%). These modifications mainly involved antidepressants. Out of these 35 PGx-related therapeutic modifications, 94% were concordant with PGx guidelines. Among 29 patients who were prescribed at least one CYP2D6 inhibitor, 25 (86%) underwent CYP2D6 phenoconversion. Conclusions: It is critical to apply pediatric age-specific considerations when utilizing PGx testing in child and adolescent psychiatry. PGx testing stewardship could provide a framework to guide the clinical utility of PGx in a pediatric population with mental health conditions, including neurodevelopmental disorders.

Limbal stem cell deficiency (LSCD) causes severe vision impairment and can lead to blindness, representing one of the most challenging ocular surface disorders. Stem cell deficiency can be congenital or, more often, acquired. The categorization of ocular surface transplantation techniques is crucial to achieving treatment homogeneity and quality of care, according to the anatomic source of the tissue being transplanted, genetic source, autologous or allogenic transplantation (to reflect histocompatibility in the latter group), and cell culture and tissue engi neering techniques. The aim of this minireview is to provide a summary of the management of LSCD, from clinical characteristics and therapeutic outcomes to the development of novel therapeutic approaches. The manuscript also briefly summarizes recent findings in the current literature and outlines the future challenges to overcome in the management of the major types of ocular surface failure., Competing Interests: Conflict-of-interest statement: There is no conflict of interest associated with any of the senior authors or other coauthors who contributed their efforts to this manuscript., (©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.)

Brain plasticity is the capacity of cerebral neurons to change, structurally and functionally, in response to experiences. This is an essential property underlying the maturation of sensory functions, learning and memory processes, and brain repair in response to the occurrence of diseases and trauma. In this field, the visual system emerges as a paradigmatic research model, both for basic research studies and for translational investigations. The auditory system remains capable of reorganizing itself in response to different auditory stimulations or sensory organ modification. Acoustic biofeedback training can be an effective way to train patients with the central scotoma, who have poor fixation stability and poor visual acuity, in order to bring fixation on an eccentrical and healthy area of the retina: a pseudofovea. This review article is focused on the cellular and molecular mechanisms underlying retinal sensitivity changes and visual and auditory system plasticity.

Background: Sensitization to house dust mite (HDM) is a leading cause of allergic rhinitis and asthma. Despite more than 30 HDM-derived allergens having been identified to date, specific therapeutic approaches do not yet take into account the local sensitization profiles of patients. This study aimed to identify patterns of HDM sensitization in HDM-allergic adults living in distinct geographic areas, to inform the development of targeted diagnostic and therapeutic tools., Methods: Serum samples from 685 HDM-allergic subjects from Canada, Europe, South Africa, and the USA were tested for levels of IgE specific for 17 micro-arrayed HDM allergens by ImmunoCAP Immuno Solid-phase Allergen Chip (ISAC) technology., Results: The results confirmed significant geographical variability in sensitization patterns and levels of IgE. In all areas, the major sensitizers were the group 1 and group 2 allergens and Der p 23. Der p 23 was a frequent sensitizer: 64% of the subjects had IgE specific for Der p 23, and 2.3% were monosensitized to it. In South Africa, Der p 23 was the dominant HDM allergen (86% prevalence) and Der p 7 achieved major allergen status (56%). IgE sensitization to HDM was influenced by asthmatic status, levels of allergen exposure, age, race-ethnicity and smoking status, but not by BMI., Conclusion: Sensitization profiles to HDM allergens differ considerably among distinct geographic areas, with Der p 7 and Der p 23 being major sensitizers in South Africa. Such heterogeneity should be taken into account in the diagnosis and treatment of HDM-allergic patients., (© 2021 EAACI and John Wiley and Sons A/S. Published by John Wiley and Sons Ltd.)

Background: Allergen immunotherapy using synthetic peptide T-cell epitopes (Cat-PAD) from the major cat allergen Fel d 1 has been shown, in allergen exposure studies, to significantly reduce symptoms of allergic rhinoconjunctivitis in cat-allergic subjects. However, the immunological mechanisms underlying clinical benefit remain only partially understood. Since previous studies of whole allergen immunotherapy demonstrated a reduction in the frequency of allergen-specific (MHC II tetramer + ) CD4 + T cells expressing the chemokine receptor CRTh2, we assessed the impact of Cat-PAD on the frequency and functional phenotype of Fel d 1-specific CD4 + T cells., Methods: Using before and after treatment samples from subjects enrolled in a randomized, double-blind, placebo-controlled trial of Cat-PAD, we employed Fel d 1 MHC II tetramers and flow cytometry to analyze the expression of chemokine receptors CCR3, CCR4, CCR5, CXCR3, and CRTh2, together with markers of memory phenotype (CD27 and CCR7) on Fel d 1-specific CD4 + T cells., Results: No statistically significant change in the frequency of Fel d 1-specific CD4 + T cells, nor in their expression of chemokine receptors or memory phenotype, was observed. However, a significant reduction in cell surface expression of CRTh2 was observed between the placebo and active groups (P = 0.047)., Conclusions: Peptide immunotherapy with Cat-PAD does not significantly alter the frequency or phenotype of Fel d 1-CD4 + T cells, but may decrease their expression of CRTh2., (© 2019 The Authors. Allergy Published by John Wiley & Sons Ltd.)

Purpose: To measure foveal morphology using optical coherence tomography (OCT)., Methods: Three examiners classified normal subjects and patients with macular edema and epiretinal membranes from a normative database (299 OCT scans) and a study sample (80 OCT scans) on the basis of the foveal depression appearance (reference classification). The following parameters were measured for each OCT scan: Central Foveal Thickness (CFT), Central Subfield Thickness (CST), Central Subfield Volume (CSV), Centrofoveal Index (CI), Parafoveal Index (PI), and Foveal Indices (FI1, FI2, FI3). Foveal depression of OCT scans in the study sample were classified based on the values of OCT parameters using Bayesian statistics (test classifications)., Results: Agreement between reference and test classifications was of 70% for CFT, 58.7% for CST, 60% for CSV, 80% for CI, 72.5% for PI, 88.7% for FI1, 96.2% for FI2 and 97.5% for FI3., Conclusions: IF3 seems to provide a reliable measure of the foveal morphology.

Purpose: To use anterior segment optical coherence tomography (AS-OCT) to compare corneal stromal demarcation line depth after 4 treatment protocols of corneal crosslinking (CXL)., Setting: Eye Clinic, Sapienza University of Rome, Terracina (Latina), Italy., Design: Prospective case series., Methods: Patients with progressive keratoconus were delegated to one of the following CXL treatments: (1) conventional epithelium (epi)-off 3 mW/cm 2 according to the standard Dresden protocol (C-CXL group), (2) accelerated epi-off 10 mW/cm 2 (A-CXL group), (3) transepithelial epi-on 3 mW/cm 2 (TE-CXL group), or (4) transepithelial epi-on by iontophoresis 10 mW/cm 2 (I-CXL group). Two independent observers measured the corneal stromal demarcation line using AS-OCT., Results: The study comprised 70 patients (120 eyes, 30 eyes in each group). The corneal stromal demarcation line was identified on AS-OCT scans in 109 eyes (90.8%). One month after the treatment, the mean stromal demarcation line depth was 275.05 μm ± 41.83 (SD) in the C-CXL group, 279.35 ± 33.07 μm in the A-CXL group, 132.60 ± 22.14 μm in the TE-CXL group, and 235.40 ± 37.08 μm in the I-CXL group. The difference in stromal demarcation line depth was not statistically significant between the C-CXL and A-CXL group, but it was statistically significant (P < .05) between the epi-off and epi-on CXL groups and between the 2 epi-on groups, where the demarcation line was significantly deeper in the I-CXL group than in the TE-CXL group., Conclusion: The corneal stromal demarcation line was significantly deeper after epi-off 30-minute standard CXL treatment and after epi-off 9-minute accelerated CXL with high-intensity ultraviolet-A irradiation., (Copyright © 2018 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.)

Augmentation of immunogenicity can be achieved by particulate delivery of an antigen and by its co-administration with an adjuvant. However, many adjuvants initiate strong systemic inflammatory reactions in vivo, leading to potential adverse events and safety concerns. We have developed a synthetic vaccine particle (SVP) technology that enables co-encapsulation of antigen with potent adjuvants. We demonstrate that co-delivery of an antigen with a TLR7/8 or TLR9 agonist in synthetic polymer nanoparticles results in a strong augmentation of humoral and cellular immune responses with minimal systemic production of inflammatory cytokines. In contrast, antigen encapsulated into nanoparticles and admixed with free TLR7/8 agonist leads to lower immunogenicity and rapid induction of high levels of inflammatory cytokines in the serum (e.g., TNF-a and IL-6 levels are 50- to 200-fold higher upon injection of free resiquimod (R848) than of nanoparticle-encapsulated R848). Conversely, local immune stimulation as evidenced by cellular infiltration of draining lymph nodes and by intranodal cytokine production was more pronounced and persisted longer when SVP-encapsulated TLR agonists were used. The strong local immune activation achieved using a modular self-assembling nanoparticle platform markedly enhanced immunogenicity and was equally effective whether antigen and adjuvant were co-encapsulated in a single nanoparticle formulation or co-delivered in two separate nanoparticles. Moreover, particle encapsulation enabled the utilization of CpG oligonucleotides with the natural phosphodiester backbone, which are otherwise rapidly hydrolyzed by nucleases in vivo. The use of SVP may enable clinical use of potent TLR agonists as vaccine adjuvants for indications where cellular immunity or robust humoral responses are required., (Copyright © 2014. Published by Elsevier Ltd.)

Bisphosphonates are a class of drugs that are widely used to inhibit loss of bone mass in patients. We show here that the administration of clinically relevant doses of bisphosphonates in mice increases antibody responses to live and inactive viruses, proteins, haptens, and existing commercial vaccine formulations. Bisphosphonates exert this adjuvant-like activity in the absence of CD4(+) and γδ T cells, neutrophils, or dendritic cells, and their effect does not rely on local macrophage depletion, Toll-like receptor signaling, or the inflammasome. Rather, bisphosphonates target directly B cells and enhance B cell expansion and antibody production upon antigen encounter. These data establish bisphosphonates as an additional class of adjuvants that boost humoral immune responses., (Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.)

B cells require MHC class II (MHC II)-restricted cognate help and CD40 engagement by CD4(+) T follicular helper (T(FH)) cells to form germinal centers and long-lasting Ab responses. Invariant NKT (iNKT) cells are innate-like lymphocytes that jumpstart the adaptive immune response when activated by the CD1d-restricted lipid α-galactosylceramide (αGalCer). We previously observed that immunization of mice lacking CD4(+) T cells (MHC II(-/-)) elicits specific IgG responses only when protein Ags are mixed with αGalCer. In this study, we investigated the mechanisms underpinning this observation. We find that induction of Ag-specific Ab responses in MHC II(-/-) mice upon immunization with protein Ags mixed with αGalCer requires CD1d expression and CD40 engagement on B cells, suggesting that iNKT cells provide CD1d-restricted cognate help for B cells. Remarkably, splenic iNKT cells from immunized MHC II(-/-) mice display a typical CXCR5(hi)programmed death-1(hi)ICOS(hi)Bcl-6(hi) T(FH) phenotype and induce germinal centers. The specific IgG response induced in MHC II(-/-) mice has shorter duration than that developing in CD4-competent animals, suggesting that iNKT(FH) cells preferentially induce transient rather than long-lived Ab responses. Together, these results suggest that iNKT cells can be co-opted into the follicular helper function, yet iNKT(FH) and CD4(+) T(FH) cells display distinct helper features, consistent with the notion that these two cell subsets play nonredundant functions throughout immune responses.

Neutralizing antibodies have been thought to be required for protection against acutely cytopathic viruses, such as the neurotropic vesicular stomatitis virus (VSV). Utilizing mice that possess B cells but lack antibodies, we show here that survival upon subcutaneous (s.c.) VSV challenge was independent of neutralizing antibody production or cell-mediated adaptive immunity. However, B cells were absolutely required to provide lymphotoxin (LT) α1β2, which maintained a protective subcapsular sinus (SCS) macrophage phenotype within virus draining lymph nodes (LNs). Macrophages within the SCS of B cell-deficient LNs, or of mice that lack LTα1β2 selectively in B cells, displayed an aberrant phenotype, failed to replicate VSV, and therefore did not produce type I interferons, which were required to prevent fatal VSV invasion of intranodal nerves. Thus, although B cells are essential for survival during VSV infection, their contribution involves the provision of innate differentiation and maintenance signals to macrophages, rather than adaptive immune mechanisms., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Deciphering the signaling networks that underlie normal and disease processes remains a major challenge. Here, we report the discovery of signaling components involved in the Toll-like receptor (TLR) response of immune dendritic cells (DCs), including a previously unkown pathway shared across mammalian antiviral responses. By combining transcriptional profiling, genetic and small-molecule perturbations, and phosphoproteomics, we uncover 35 signaling regulators, including 16 known regulators, involved in TLR signaling. In particular, we find that Polo-like kinases (Plk) 2 and 4 are essential components of antiviral pathways in vitro and in vivo and activate a signaling branch involving a dozen proteins, among which is Tnfaip2, a gene associated with autoimmune diseases but whose role was unknown. Our study illustrates the power of combining systematic measurements and perturbations to elucidate complex signaling circuits and discover potential therapeutic targets., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Lymph nodes (LNs) capture microorganisms that breach the body's external barriers and enter draining lymphatics, limiting the systemic spread of pathogens. Recent work has shown that CD11b(+)CD169(+) macrophages, which populate the subcapsular sinus (SCS) of LNs, are critical for the clearance of viruses from the lymph and for initiating antiviral humoral immune responses. Here we show, using vesicular stomatitis virus (VSV), a relative of rabies virus transmitted by insect bites, that SCS macrophages perform a third vital function: they prevent lymph-borne neurotropic viruses from infecting the central nervous system (CNS). On local depletion of LN macrophages, about 60% of mice developed ascending paralysis and died 7-10 days after subcutaneous infection with a small dose of VSV, whereas macrophage-sufficient animals remained asymptomatic and cleared the virus. VSV gained access to the nervous system through peripheral nerves in macrophage-depleted LNs. In contrast, within macrophage-sufficient LNs VSV replicated preferentially in SCS macrophages but not in adjacent nerves. Removal of SCS macrophages did not compromise adaptive immune responses against VSV, but decreased type I interferon (IFN-I) production within infected LNs. VSV-infected macrophages recruited IFN-I-producing plasmacytoid dendritic cells to the SCS and in addition were a major source of IFN-I themselves. Experiments in bone marrow chimaeric mice revealed that IFN-I must act on both haematopoietic and stromal compartments, including the intranodal nerves, to prevent lethal infection with VSV. These results identify SCS macrophages as crucial gatekeepers to the CNS that prevent fatal viral invasion of the nervous system on peripheral infection.

CD4(+) T (Th)-cell help to B lymphocytes requires cognate interaction and CD40 engagement. Invariant natural killer T (iNKT) cells are innate-like T lymphocytes that recognize alphagalactosylceramide (alphaGalCer) presented by CD1d, and can help B-cell responses. We asked whether alphaGalCer-activated iNKT cells help B lymphocytes through cognate interaction, or indirectly, via enhancement of Th-B-cell interaction. After immunization with protein Ags and alphaGalCer, antibody titers were assessed in wild-type or splenectomized mice, and in bone marrow radiation chimeras lacking CD1d or CD40 expression on B lymphocytes, or expressing CD1d or MHC II disjointly on antigen-presenting cells (APCs). We find that alphaGalCer-dependent enhancement of B-cell response (1) can occur when B cells do not express CD1d but express CD40; (2) requires that iNKT and Th cells interact with the same APCs that coexpress both CD1d and MHC-II; and (3) takes place without spleen. These findings demonstrate alphaGalCer-induced help for antibody responses can occur without cognate iNKT/B-cell interaction, and suggest this help entails activation of APCs by iNKT cells, which in turn activate Th cells and their helper functions for B cells. Thus, the alphaGalCer-induced help recapitulates the function of classical adjuvants that stimulate the innate immune system to support adaptive immune responses.