Total body CD4+ T cell dynamics in treated and untreated SIV infection revealed by in vivo imaging

The peripheral blood represents only a small fraction of the total number of lymphocytes in the body. To develop a more thorough understanding of T cell dynamics, including the effects of SIV/SHIV/HIV infection on immune cell depletion and immune reconstitution following combination antiretroviral therapy (cART), one needs to utilize approaches that allow direct visualization of lymphoid tissues. In the present study, noninvasive in vivo imaging of the CD4+ T cell pool has revealed that the timing of the CD4+ T cell pool reconstitution following initiation of ART in SIV-infected nonhuman primates (NHPs) appears seemingly stochastic among clusters of lymph nodes within the same host. At 4 weeks following initiation or interruption of cART, the changes observed in peripheral blood (PB) are primarily related to changes in the whole-body CD4 pool rather than changes in lymphocyte trafficking. Lymph node CD4 pools in long-term antiretroviral-treated and plasma viral load-suppressed hosts appear suboptimally reconstituted compared with healthy controls, while splenic CD4 pools appear similar between the 2 groups

Viral dissemination and immune activation modulate antiretroviral drug levels in lymph nodes of SIV-infected rhesus macaques

Introduction and methodsTo understand the relationship between immunovirological factors and antiretroviral (ARV) drug levels in lymph nodes (LN) in HIV therapy, we analyzed drug levels in twenty-one SIV-infected rhesus macaques subcutaneously treated with daily tenofovir (TFV) and emtricitabine (FTC) for three months.ResultsThe intracellular active drug-metabolite (IADM) levels (TFV-dp and FTC-tp) in lymph node mononuclear cells (LNMC) were significantly lower than in peripheral blood mononuclear cells (PBMC) (P≤0.005). Between Month 1 and Month 3, IADM levels increased in both LNMC (P≤0.001) and PBMC (P≤0.01), with a steeper increase in LNMC (P≤0.01). The viral dissemination in plasma, LN, and rectal tissue at ART initiation correlated negatively with IADM levels at Month 1. Physiologically-based pharmacokinetic model simulations suggest that, following subcutaneous ARV administration, ART-induced reduction of immune activation improves the formation of active drug-metabolites through modulation of kinase activity and/or through improved parent drug accessibility to LN cellular compartments.ConclusionThese observations have broad implications for drugs that need to phosphorylate to exert their pharmacological activity, especially in the settings of the pre-/post-exposure prophylaxis and efficacy of antiviral therapies targeting pathogenic viruses such as HIV or SARS-CoV-2 replicating in highly inflammatory anatomic compartments

Microchannel cooling for the LHCb VELO Upgrade I

The LHCb VELO Upgrade I, currently being installed for the 2022 start of LHC Run 3, uses silicon microchannel coolers with internally circulating bi-phase \cotwo for thermal control of hybrid pixel modules operating in vacuum. This is the largest scale application of this technology to date. Production of the microchannel coolers was completed in July 2019 and the assembly into cooling structures was completed in September 2021. This paper describes the R\&D path supporting the microchannel production and assembly and the motivation for the design choices. The microchannel coolers have excellent thermal peformance, low and uniform mass, no thermal expansion mismatch with the ASICs and are radiation hard. The fluidic and thermal performance is presented.Comment: 31 pages, 27 figure

Skull Anatomy of the Bizarre Crocodylian Mourasuchus nativus (Alligatoridae, Caimaninae)

Mourasuchus is a Miocene alligatorid endemic to South America, and is represented by four species. Together with the closely related Purussaurus, it is a peculiar crocodylian taxon of neogene Caimaninae and one of the most bizarre forms among eusuchian crocodiles. The phylogenetic relationships between Mourasuchus species have not been explored, and detailed skull descriptions are scarce. The goal of this study is to provide new data on skull morphology and cranial recesses in Mourasuchus nativus, including a new tomography analysis (3D modeling). We observed that several diagnostic characters of Purussaurus, such as lack of contact between the nasal and lacrimal, separation of the nasal and frontal by the prefrontals, and the posterior dorsal margin of the skull table, are shared with Mourasuchus. M. nativus is characterized by the presence of solid transverse squamosal eminences, large posttemporal fenestrae, and a quadrate laterocaudal bridge separating V2-V3 trigeminal openings. Compared with other crocodylians, the endocast of M. nativus is similar in shape but quite sigmoid in lateral view, the canal of the supraorbital ramus of V2 is more vertically oriented, the thick tympanic branch canal opens in a large foramen aligned with trigeminal foramen, and the canal of the vagal (X) tympanic ramus is also very wide. Contrary to extant alligatorids, the median pharyngeal recess remains paired throughout its course and only connects its opposite fellow near the external ventral opening. The knowledge of the internal skull anatomy of Mourasuchus contributes to the understanding of the general morphology of alligatorids, Caimaninae, and their variation.Fil: Bona, Paula. Universidad Nacional de la Plata. Facultad de Ciencias Naturales y Museo. División Paleontología Vertebrados; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Degrange, Federico Javier. Universidad Nacional de la Plata. Facultad de Ciencias Naturales y Museo. División Paleontología Vertebrados; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Ciencias de la Tierra. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Centro de Investigaciones en Ciencias de la Tierra; ArgentinaFil: Fernández, Marta Susana. Universidad Nacional de la Plata. Facultad de Ciencias Naturales y Museo. División Paleontología Vertebrados; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

CD4+ levels control the odds of induction of humoral immune responses to tracer doses of therapeutic antibodies.

Rapidly increasing number of therapeutic antibodies are being repurposed to imaging probes for noninvasive diagnosis, as well as monitoring during treatment or disease recurrence. Though antibody-based imaging involves tracer doses (~3 log lower than therapeutic doses), and immune responses are severely reduced in patients with impaired immunity, formation of anti-tracer antibodies (ATA) has been observed hampering further diagnostic monitoring. Here, we explored the potential to develop humoral responses to intravenously administered tracer dose of a monoclonal antibody F(ab΄)2 fragment, and associated with host related immune measures in 49 rhesus macaques categorized into healthy (uninfected controls), SIV-progressors, SIV non-progressors, or total body irradiated (TBI). Antibody fragment administered in tracer amount (~100μg) induced immune responses with significantly lower odds in SIV-progressors or TBI macaques (P<0.005) as compared to healthy animals. Peripheral blood (PB) CD4+ cell counts, but not CD20+ cell levels, were associated with significantly higher risk of developing a humoral response (P<0.001). Doubling the PB CD4+ counts is associated with an odds ratio of developing an immune response of 1.73. Among SIV-infected animals, CD4+ cell count was a stronger predictor of immune response than plasma SIV-RNA levels. Both SIV-progressors and TBI macaques showed higher odds of responses with increasing CD4+ counts, however when compared to healthy or SIV non-progressors with similar CD4+ count, they were still functionally incompetent in generating a response (P<0.01). Moreover, presence of ATA in systemic circulation altered the in vivo biodistribution by increasing hepatic uptake and decreasing plasma radiotracer clearance, with minimal to no binding detected in targeted tissues

Maximum intensity projection SPECT images.

<p>Rhesus macaques that developed antibody response to the radiotracer after baseline exposure were subsequently imaged while anti-tracer antibodies were present in the plasma in (A) healthy macaque imaged at 48 hours post-radiotracer injection with intact-huOKT4A labelled with ¹¹¹In and scanned using Triad88 (Trionix) camera, (B) SIV-TK infected non-progressor imaged at 4 hours post-radiotracer injection with F(ab΄)₂-huOKT4A labelled with ^99mTc and scanned using Triad88 (Trionix) camera, and (C) healthy macaque that underwent total body irradiation prior to 2^nd exposure and imaged at 4 hours post-radiotracer injection with F(ab΄)₂-CD4R1 labelled with ^99mTc and scanned using Symbia T2 (Siemens) camera. Images show increased hepatic uptake and altered biodistribution with minimal to no binding observed in secondary lymphoid organs when imaged in the presence of anti-tracer antibodies. Within each panel, images were adjusted (standardized) for injected dose and body weight. Tissue uptakes were converted to RAINBOW color scale as shown in color bar.</p

Immune response outcome of total 118 exposures for all 49 macaques receiving F(ab΄)₂ radiotracers.

<p>Immune response outcome of total 118 exposures for all 49 macaques receiving F(ab΄)₂ radiotracers.</p

Correlation between radio-HPLC and plasma binding assay.

<p>Strong correlation was observed between plasma binding assay and HPLC assay. Samples were assayed with either F(ab΄)₂-CD4R1 antibody (brown dots) or F(ab΄)₂-huOKT4A antibody (red dots). For samples assayed with F(ab΄)₂-CD4R1, the bootstrap estimate and 95% confidence interval of the Spearman rank correlation between PBA-ratio and HPLC-HM-ratio was -0.88 (-0.91, -0.84), and between PBA-ratio and HPLC-Fab’2-ratio was 0.90 (0.85, 0.93). Similar results were observed for samples assayed with F(ab΄)₂-huOKT4A.</p