Inhibitory Phenotype of HBV-Specific CD4⁺ T-Cells Is Characterized by High PD-1 Expression but Absent Coregulation of Multiple Inhibitory Molecules

Background: T-cell exhaustion seems to play a critical role in CD8(+) T-cell dysfunction during chronic viral infections. However, up to now little is known about the mechanisms underlying CD4(+) T-cell dysfunction during chronic hepatitis B virus (CHB) infection and the role of inhibitory molecules such as programmed death 1 (PD-1) for CD4(+) T-cell failure. Methods: The expression of multiple inhibitory molecules such as PD-1, CTLA-4, TIM-3, CD244, KLRG1 and markers defining the grade of T-cell differentiation as CCR7, CD45RA, CD57 and CD127 were analyzed on virus-specific CD4(+) T-cells from peripheral blood using a newly established DRB1*01-restricted MHC class II Tetramer. Effects of in vitro PD-L1/2 blockade were defined by investigating changes in CD4(+) T-cell proliferation and cytokine production. Results: CD4(+) T-cell responses during chronic HBV infection was characterized by reduced Tetramer(+)CD4(+) T-cell frequencies, effector memory phenotype, sustained PD-1 but low levels of CTLA-4, TIM-3, KLRG1 and CD244 expression. PD-1 blockade revealed individualized patterns of in vitro responsiveness with partly increased IFN-gamma, IL-2 and TNF-alpha secretion as well as enhanced CD4(+) T-cell expansion almost in treated patients with viral control. Conclusion: HBV-specific CD4(+) T-cells are reliably detectable during different courses of HBV infection by MHC class II Tetramer technology. CD4(+) T-cell dysfunction during chronic HBV is basically linked to strong PD-1 upregulation but absent coregulation of multiple inhibitory receptors. PD-L1/2 neutralization partly leads to enhanced CD4(+) T-cell functionality with heterogeneous patterns of CD4(+) T-cell rejunivation

Review on retrospective procedures to correct retinal motion artefacts in OCT imaging

Motion artefacts from involuntary changes in eye fixation remain a major imaging issue in optical coherence tomography (OCT). This paper reviews the state-of-the-art of retrospective procedures to correct retinal motion and axial eye motion artefacts in OCT imaging. Following an overview of motion induced artefacts and correction strategies, a chronological survey of retrospective approaches since the introduction of OCT until the current days is presented. Pre-processing, registration, and validation techniques are described. The review finishes by discussing the limitations of the current techniques and the challenges to be tackled in future developments

Tolerability of inhaled N-chlorotaurine in an acute pig streptococcal lower airway inflammation model

<p>Abstract</p> <p>Background</p> <p>Inhalation of N-chlorotaurine (NCT), an endogenous new broad spectrum non-antibiotic anti-infective, has been shown to be very well tolerated in the pig model recently. In the present study, inhaled NCT was tested for tolerability and efficacy in the infected bronchopulmonary system using the same model.</p> <p>Methods</p> <p>Anesthetized pigs were inoculated with 20 ml of a solution containing approximately 10⁸CFU/ml <it>Streptococcus pyogenes </it>strain d68 via a duodenal tube placed through the tracheal tube down to the carina. Two hours later, 5 ml of 1% NCT aqueous solution (test group, n = 15) or 5 ml of 0.9% NaCl (control group, n = 16) was inhaled via the tracheal tube connected to a nebulizer. Inhalation was repeated every hour, four times in total. Lung function and haemodynamics were monitored. Bronchoalveolar lavage samples were removed for determination of colony forming units (CFU), and lung samples for histology.</p> <p>Results</p> <p>Arterial pressure of oxygen (PaO₂) decreased rapidly after instillation of the bacteria in all animals and showed only a slight further decrease at the end of the experiment without a difference between both groups. Pulmonary artery pressure increased to a peak 1-1.5 h after application of the bacteria, decreased in the following hour and remained constant during treatment, again similarly in both groups. Histology demonstrated granulocytic infiltration in the central parts of the lung, while this was absent in the periphery. Expression of TNF-alpha, IL-8, and haemoxygenase-1 in lung biopsies was similar in both groups. CFU counts in bronchoalveolar lavage came to 170 (10; 1388) CFU/ml (median and 25 and 75 percentiles) for the NCT treated pigs, and to 250 (10; 5.5 × 10⁵) CFU/ml for NaCl treated pigs (p = 0.4159).</p> <p>Conclusions</p> <p>Inhaled NCT at a concentration of 1% proved to be very well tolerated also in the infected bronchopulmonary system. This study confirms the tolerability in this delicate body region, which has been proven in healthy pigs previously. Regarding efficacy, no conclusions can be drawn, mainly because of the limited test period of the model.</p

Maturation-Dependent Licensing of Naive T Cells for Rapid TNF Production

The peripheral naïve T cell pool is comprised of a heterogeneous population of cells at various stages of development, which is a process that begins in the thymus and is completed after a post-thymic maturation phase in the periphery. One hallmark of naïve T cells in secondary lymphoid organs is their unique ability to produce TNF rapidly after activation and prior to acquiring other effector functions. To determine how maturation influences the licensing of naïve T cells to produce TNF, we compared cytokine profiles of CD4+ and CD8+ single positive (SP) thymocytes, recent thymic emigrants (RTEs) and mature-naïve (MN) T cells during TCR activation. SP thymocytes exhibited a poor ability to produce TNF when compared to splenic T cells despite expressing similar TCR levels and possessing comparable activation kinetics (upregulation of CD25 and CD69). Provision of optimal antigen presenting cells from the spleen did not fully enable SP thymocytes to produce TNF, suggesting an intrinsic defect in their ability to produce TNF efficiently. Using a thymocyte adoptive transfer model, we demonstrate that the ability of T cells to produce TNF increases progressively with time in the periphery as a function of their maturation state. RTEs that were identified in NG-BAC transgenic mice by the expression of GFP showed a significantly enhanced ability to express TNF relative to SP thymocytes but not to the extent of fully MN T cells. Together, these findings suggest that TNF expression by naïve T cells is regulated via a gradual licensing process that requires functional maturation in peripheral lymphoid organs

OCTA multilayer and multisector peripapillary microvascular modeling for diagnosing and staging of glaucoma

Purpose: To develop and assess an automatic procedure for classifying and staging glaucomatous vascular damage based on optical coherence tomography angiography (OCTA) imaging. Methods: OCTA scans (Zeiss Cirrus 5000 HD-OCT) from a random eye of 39 healthy subjects and 82 glaucoma patients were used to develop a new classification algorithm based on multilayer and multisector information. The averaged circumpapillary retinal nerve fiber layer (RNFL) thickness was also collected. Three models, support vector machine (SVM), random forest (RF), and gradient boosting (xGB), were developed and optimized for classifying between healthy and glaucoma patients, primary open-angle glaucoma (POAG) and normal-tension glaucoma (NTG), and glaucoma severity groups. Results: All the models, the SVM (area under the receiver operating characteristic [AUROC] 0.89 ± 0.06), the RF (AUROC 0.86 ± 0.06), and the xGB (AUROC 0.85 ± 0.07), with 26, 22, and 29 vascular features obtained after feature selection, respectively, presented a similar performance to the RNFL thickness (AUROC 0.85± 0.06) in classifying healthy and glaucoma patients. The superficial vascular plexus was the most informative layer with the infero temporal sector as the most discriminative region of interest. No significant differentiation was obtained in discriminating the POAG from the NTG group. The xGB model, after feature selection, presented the best performance in classifying the severity groups (AUROC 0.76± 0.06), outperforming the RNFL (AUROC 0.67± 0.06). Conclusions: OCTA multilayer and multisector information has similar performance to RNFL for glaucoma diagnosis, but it has an added value for glaucoma severity classification, showing promising results for staging glaucoma progression. Translational Relevance: OCTA, in its current stage, has the potential to be used in clinical practice as a complementary imaging technique in glaucoma management

Genetic Detection and Characterization of Lujo Virus, a New Hemorrhagic Fever–Associated Arenavirus from Southern Africa

Lujo virus (LUJV), a new member of the family Arenaviridae and the first hemorrhagic fever–associated arenavirus from the Old World discovered in three decades, was isolated in South Africa during an outbreak of human disease characterized by nosocomial transmission and an unprecedented high case fatality rate of 80% (4/5 cases). Unbiased pyrosequencing of RNA extracts from serum and tissues of outbreak victims enabled identification and detailed phylogenetic characterization within 72 hours of sample receipt. Full genome analyses of LUJV showed it to be unique and branching off the ancestral node of the Old World arenaviruses. The virus G1 glycoprotein sequence was highly diverse and almost equidistant from that of other Old World and New World arenaviruses, consistent with a potential distinctive receptor tropism. LUJV is a novel, genetically distinct, highly pathogenic arenavirus

Phase retardation and corneal sublayer thickness repeatability using ultrahigh resolution polarization-sensitive OCT

PURPOSE: To assess phase retardation and corneal sublayer thickness repeatability using ultrahigh resolution polarization-sensitive OCT (PS-OCT).SETTING: Narayana Nethralaya eye hospital, Bangalore.DESIGN: ObservationalMethods: In this study, all eyes were imaged using our custom-built ultrahigh resolution PS-OCT and high-resolution hybrid OCT (MS-39, CSO, Italy). We evaluated the repeatability of phase retardation en face maps and corneal sublayer thickness profiles. The reflectivity and phase retardation were calculated from the two orthogonal polarization channels to generate en face maps of phase retardation and corneal sublayer thicknesses. Three consecutive measurements of all subjects were acquired for each eye. For each measurement, the subject was asked to sit back and was re-aligned again. The repeatability was assessed using intraclass correlation coefficient (ICC).RESULTS: The phase retardation en face maps showed preferential arrangement of collagen fibrils with least retardation in apex and maximum retardation in the periphery. The phase retardation showed excellent repeatability (ICC&gt;0.95) in all zones. The Bowman's layer and stromal layer thicknesses were measured with excellent repeatability (ICC&gt;0.93 and &gt;0.99, respectively). Significant differences (p&lt;0.05) in stromal layer thickness were observed between MS-39 and PS-OCT. The repeatability of epithelium thickness measurements was better with PS-OCT than MS-39.CONCLUSION: The combinational assessment of corneal birefringence and sublayer thicknesses shows the advanced potential of ultrahigh resolution PS-OCT in routine clinical practice over current OCT devices.</p