Simian varicella virus infection and reactivation in rhesus macaques trigger cytokine and Aß40/42 alterations in serum and cerebrospinal fluid.

Simian varicella virus (SVV) produces peripheral inflammatory responses during varicella (primary infection) and zoster (reactivation) in rhesus macaques (RM). However, it is unclear if peripheral measures are accurate proxies for central nervous system (CNS) responses. Thus, we analyzed cytokine and Aß42/Aß40 changes in paired serum and cerebrospinal fluid (CSF) during the course of infection. During varicella and zoster, every RM had variable changes in serum and CSF cytokine and Aß42/Aß40 levels compared to pre-inoculation levels. Overall, peripheral infection appears to affect CNS cytokine and Aß42/Aß40 levels independent of serum responses, suggesting that peripheral disease may contribute to CNS disease.

The association of dietary antioxidants and the inflammatory potential of the diet with poor physical function and disability in older Australian men: the Concord Health and Ageing in Men Project.

Our objective was to evaluate the association of antioxidant intake and the inflammatory potential of the diet with functional decline in older men. A diet history questionnaire was used to collect dietary intake data from men aged ≥ 75 years (n 794) participating in the Concord Health and Aging in Men Project cohort study. Intake of vitamins A, C, E and Zn were compared with the Australian Nutrient Reference Values to determine adequacy. The Energy-adjusted Dietary Inflammatory Index (E-DIITM) was used to assess the inflammatory potential of the diet. Physical performance data were collected via handgrip strength and walking speed tests, and activities of daily living (ADL) and instrumental activities of daily living (IADL) questionnaires, at baseline and 3-year follow-up (n 616). Logistic regression analysis was used to identify associations between diet and incident poor physical function and disability. Both poor antioxidant intake and high E-DII scores at baseline were significantly associated with poor grip strength and ADL disability at 3-year follow-up. No significant associations with walking speed or IADL disability were observed. Individual micronutrient analysis revealed a significant association between the lowest two quartiles of vitamin C intake and poor grip strength. The lowest quartiles of intake for vitamins A, C, E and Zn were significantly associated with incident ADL disability. The study observed that poor antioxidant and anti-inflammatory food intake were associated with odds of developing disability and declining muscle strength in older men. Further interventional research is necessary to clarify the causality of these associations.

Reproductive futures within a context of uterus transplants in India.

Reproduction is considered a core event in a cisgender heterosexual woman's life. Several hierarchies exist within the field of reproduction, with reproduction being privileged among people from dominant races, classes, castes, ages, and abilities. In this paper, I explore uterine transplantion as an emerging mode of reproduction, which privileges the experience of pregnancy in addition to genetic relatedness, specifically within the Indian context. The focus is on the socio-cultural and economic discourses surrounding the marketisation of reproductive technologies and how these recalibrate social and familial dynamics concerning reproduction. I argue that the mobilisation of the language of reproduction as a right could potentially transform into reproduction as a duty. Reproductive biopolitics is used as a lens to think through the value of wombs in relation to the bodies they inhabit, and the pressures the marketisation of wombs puts on both recipients and donors.

Effects of anti-inflammatory dietary patterns on non-alcoholic fatty liver disease: a systematic literature review.

PURPOSE: Non-alcoholic fatty liver disease (NAFLD) is the leading chronic hepatic condition. Low-grade chronic inflammation contributes to disease progression. Diet has protective effects on hepatic health and inflammatory pathways. The purpose of this review is to systematically review and describe the effects of anti-inflammatory dietary patterns on NAFLD. METHODS: The Cochrane CENTRAL Library, Cumulative Index of Nursing and Allied Health Literature, Embase, MEDLINE and Web of Science databases were searched. A total of 252 records were identified, 7 of which were included in this review. The revised Cochrane risk-of-bias tool was used to conduct a quality assessment for randomised trials. Certainty of evidence was assessed using the grading of recommendations, assessment, development, and evaluation tool. RESULTS: Of the 7 included studies, 6 were classified as low risk of bias and studies ranged from high to very low certainty of evidence. In the randomised-controlled studies systematically reviewed, either adherence to the Mediterranean, DASH, or FLiO diet was studied, against usual care or energy matched controls, with a total of 255 participants. Anti-inflammatory dietary pattern adherence significantly reduced the severity of most hepatic and inflammatory markers, and secondary outcomes. A minority of outcomes were improved significantly more than controls. CONCLUSION: Anti-inflammatory dietary patterns showed benefits to NAFLD risk factors, severity markers and inflammatory markers compared to the control diet. It is unclear whether reductions in the evaluated parameters are related solely to the anti-inflammatory diet or weight loss resulting from caloric restriction, as improvements in control groups were also evidenced. Current limited body of evidence indicates need for further research including isocaloric dietary patterns, longer interventions, measures of inflammatory markers, and studies including normal-weight subjects to confirm findings at higher certainty. PROSPERO REGISTRATION: CRD42021269382.

Genome­wide identification of CAMTA gene family members in rice (Oryza sativa L.) and in silico study on their versatility in respect to gene expression and promoter structure.

The calmodulin-binding transcription activator (CAMTA) is a family of transcriptional factors containing a cluster of calmodulin-binding proteins that can activate gene regulation in response to stresses. The presence of this family of genes has been reported earlier, though, the comprehensive analyses of rice CAMTA (OsCAMTA) genes, their promoter regions, and the proteins were not deliberated till date. The present report revealed the existence of seven CAMTA genes along with their alternate transcripts in five chromosomes of rice (Oryza sativa) genome. Phylogenetic trees classified seven CAMTA genes into three clades indicating the evolutionary conservation in gene structure and their association with other plant species. The in silico study was carried out considering 2 kilobases (kb) promoter regions of seven OsCAMTA genes regarding the distribution of transcription factor binding sites (TFbs) of major and plant-specific transcription factors whereas OsCAMTA7a was identified with highest number of TFbs, while OsCAMTA4 had the lowest. Comparative modelling, i.e., homology modelling, and molecular docking of the CAMTA proteins contributed the thoughtful comprehension of protein 3D structures and protein-protein interaction with probable partners. Gene ontology annotation identified the involvement of the proteins in biological processes, molecular functions, and localization in cellular components. Differential gene expression study gave an insight on functional multiplicity to showcase OsCAMTA3b as most upregulated stress-responsive gene. Summarization of the present findings can be interpreted that OsCAMTA gene duplication, variation in TFbs available in the promoters, and interactions of OsCAMTA proteins with their binding partners might be linked to tolerance against multiple biotic and abiotic cues.

An enzyme-linked immunosorbent assay (ELISA) to determine Simian Varicella Virus antibody titers in infected rhesus monkeys (Macaca mulatta).

BACKGROUND: Simian varicella virus (SVV) is a primate herpesvirus that causes a natural varicella-like disease in Old World monkeys and may cause epizootics in facilities housing nonhuman primates. SVV infection of nonhuman primates is used as an experimental model to investigate varicella pathogenesis and to develop antiviral strategies. METHODS: An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect SVV antibodies in infected rhesus macaque monkeys. RESULTS: An ELISA determined SVV antibody titers following experimental infection. SVV IgG was detected by day 14 post-infection and remained elevated for at least 84 days. CONCLUSIONS: The SVV ELISA is a safe and rapid approach to confirm SVV seropositivity and to determine SVV antibody titers in naturally and experimentally SVV-infected monkeys. In addition to being a useful diagnostic assay to rapidly confirm acute disease or past SVV infection, the SVV ELISA is a valuable epidemiological tool to determine the incidence of SVV in non-human primate facilities.

Associations between dietary intake of total protein and sources of protein (plant vs. animal) and risk of all-cause and cause-specific mortality in older Australian men: The Concord Health and Ageing in Men Project.

BACKGROUND: The association between dietary protein intake and the risk of mortality is still controversial. The present study aimed to examine the associations between dietary total, animal and plant protein intake and all-cause and cause-specific mortality. METHODS: Community-dwelling men aged ≥ 70 years were recruited from local government areas surrounding Concord Hospital in Sydney, New South Wales for the Concord Health and Ageing in Men Project (CHAMP). The research dietitian administered a standardised validated diet history questionnaire to capture baseline dietary intake. In total, 794 men participated in a detailed diet history interview at the third wave. Adequacy of protein intake was assessed by comparing participant intake with the Nutrient Reference Values. Total protein intake was categorised into quintiles. Sources of protein were also captured. Mortality was ascertained through the New South Wales death registry. Cox proportional hazard models were used to assess the association between dietary total, animal and plant protein intake and risk of mortality. RESULTS: The mean age of the CHAMP men was 81 years. In total, 162 men died during a median follow-up of 3.7 years. Of these, 54 (33.3%) and 49 (30.2%) men died due to cancer and cardiovascular disease, respectively. There were U-shaped associations between protein intake and all-cause and cancer mortality. In multiple adjusted analysis, the second (hazard ratio [HR] = 0.38; 95% confidence interval [CI] = 0.18-0.82) and third (HR = 0.36; 95% CI = 0.16-0.82) quintiles of protein intakes were significantly associated with reduced risk of all-cause and only second quintile (HR = 0.47; 95% CI = 0.10-0.93) of protein intake was significantly associated with cancer mortality. Each serve increase in animal protein was significantly associated with 12% (HR = 1.12; 95% CI = 1.00-1.26) and 23% (HR = 1.23; 95% CI = 1.02-1.49) increased risk of all-cause mortality and cancer mortality respectively. Conversely, each serve increase in plant protein intake was significantly associated with 25% (HR = 0.75; 95% CI 0.61-0.92) and 28% (HR = 0.72; 95% CI = 0.53-0.97) reduced risk of all-cause and cancer mortality, respectively. No such associations were observed for cardiovascular disease mortality. CONCLUSIONS: Both second and third quintiles of total protein intake were associated with reduced all-cause and cancer mortality. Plant protein was inversely associated with all-cause and cancer mortality, whereas animal protein intake was positively associated with mortality.

Vaccine for Diabetes-Where Do We Stand?

Diabetes is an endocrinological disorder with a rapidly increasing number of patients globally. Over the last few years, the alarming status of diabetes has become a pivotal factor pertaining to morbidity and mortality among the youth as well as middle-aged people. Current developments in our understanding related to autoimmune responses leading to diabetes have developed a cause for concern in the prospective usage of immunomodulatory agents to prevent diabetes. The mechanism of action of vaccines varies greatly, such as removing autoreactive T cells and inhibiting the interactions between immune cells. Currently, most developed diabetes vaccines have been tested in animal models, while only a few human trials have been completed with positive outcomes. In this review, we investigate the undergoing clinical trial studies for the development of a prototype diabetes vaccine.

Low-glycemic foods with wheat, barley and herbs (Terminalia chebula, Terminalia bellerica and Emblica officinalis) inhibit α-amylase, α-glucosidase and DPP-IV activity in high fat and low dose streptozotocin-induced diabetic rat.

Wheat, barley or wheat + barley and herbs (Terminalia chebula, Terminalia bellerica and Emblica officinalis) based low-glycemic-index (low-GI) foods were developed and studied α-amylase, α-glucosidase and DPP-IV inhibition property in vitro and in the streptozotocin-induced diabetic rats. The GI of products ranged from 47 to 53 than control white bread (GI = 95). Total phenolic (20.1 ± 1 mg gallic acid/g dry wt.) and flavonoids (15.2 ± 1 mg quercetin/g dry wt.) were higher in wheat + barley than barley (17.2 ± 1; 13.6 ± 2) and wheat (16.9 ± 1; 14.9 ± 2) products. The in vitro α-amylase (4-10%), α-glucosidase (5-17%) and DPP-IV (3-26%) inhibition (IC50) of methanol extracts were higher than the aqueous extracts. The fasting blood glucose (50.85, 33.22 and 24.52%) and oral glucose tolerance (AUC = 32.1, 36.04, and 27.73%) was lower in barley, wheat, and wheat + barley fed diabetic groups than diabetic control group (1571.5 ± 13.5 mg/dL/120 min). Feeding wheat, barley, and W + B foods for 60 days inhibited the intestinal α-amylase (1.2, 1.1 and 1.5-folds), α-glucosidase (1.3, 1.2 and 1.7-folds) and DPP-IV (1.6, 1.5 and 2.1-folds) activity compared to diabetic control. Low-GI foods lower the systemic glucose level, inhibit the glycolytic enzymes and DPP-IV activity and hence desirable for diabetes management. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05231-0.

The association between antioxidant intake, dietary pattern and depressive symptoms in older Australian men: the Concord Health and Ageing in Men Project.

PURPOSE: The objectives of the study were to evaluate the associations between antioxidant intake, dietary patterns and depressive symptoms among older men. METHOD: 794 men participated in a detailed diet history interview at the Concord Health and Ageing in Men Project 3rd wave (considered baseline nutrition) and 781 men participated at the 4th wave (considered 3-year follow-up). Depressive symptoms were measured using the Geriatric Depression Scale (GDS ≥ 5). Dietary adequacy of antioxidant intake was assessed by comparing participants' median intake of vitamin A, E, C and zinc to the Nutrient Reference Values for Australia. Attainment of NRVs of antioxidant was categorised into a dichotomised variable 'poor' (meeting ≤ 2) or 'good' (meeting ≥ 3). Individual antioxidant nutrient was categorised into quartiles. The Australian and Mediterranean diet scores were assessed as predictor variables. RESULTS: The prevalence of GDS ≥ 5 was 12.8% at baseline nutrition and 13.2% of men developed GDS ≥ 5 at a 3-year follow-up. There was a significant cross-sectional association between poor antioxidant intake and GDS ≥ 5 in adjusted analyses [OR: 1.95 (95% CI 1.03, 3.70)]. Poor antioxidant intake at baseline nutrition remained prospectively associated with incident GDS ≥ 5 [OR: 2.46 (95% CI 1.24, 4.88)] in adjusted analyses. This association was also found for the lowest quartile of zinc [OR 2.72 (95% CI 1.37, 5.42)] and vitamin E intake [OR 2.18 (95% CI 1.05, 4.51)]. None of the other antioxidants and dietary patterns had a significant association with incident depressive symptoms. CONCLUSION: Inadequacy of antioxidant intake, particularly zinc and vitamin E, is associated with increased risk of clinically significant depressive symptoms in older men.

Dietary and supplemental antioxidant intake and risk of major adverse cardiovascular events in older men: The concord health and ageing in men project.

BACKGROUND AND AIMS: The role of antioxidant intake in cardiovascular disease remains inconclusive. This study evaluates the association between antioxidant intake and the risk of major adverse cardiovascular events (MACE) among older Australian men. METHODS AND RESULTS: 794 men aged ≥75 years participated in the 3rd wave of the Concord Health and Ageing in Men Project. Dietary adequacy of antioxidant intake was assessed by comparing participants' intake of vitamins A, E, C and zinc to the Nutrient Reference Values (NRV) for Australia. Attainment of NRVs of antioxidants was categorised into a dichotomised variable 'inadequate' (meeting≤2 of 4 antioxidants) or 'adequate' (meeting≥3 of 4 antioxidants). The usage of antioxidant supplements was assessed. The outcome measure was MACE. The composite MACE endpoint was defined as having one of the following: death, myocardial infarction, ischemic stroke, congestive cardiac failure (CCF), and revascularization during the period of observation. There was no significant association between dietary (HR: 1.03, 95% CI: 0.71, 1.48) or supplemental antioxidant intake (HR: 1.10, 95% CI: 0.75, 1.63) and overall MACE. However, a significant association was observed between inadequate antioxidant intake and CCF (HR: 1.32; 95% CI: 1.16, 1.50). The lowest quartile of zinc intake (<11.00 mg/d) was significantly associated with CCF (HR 2.36; 95% CI: 1.04, 5.34). None of the other antioxidants were significantly associated with CCF or other MACE components. CONCLUSION: Inadequate dietary antioxidant intake, particularly zinc, is associated with increased risk of CCF in older Australian men but not associated with overall MACE.

Associations between nutrient intakes and dietary patterns with different sarcopenia definitions in older Australian men: the concord health and ageing in men project.

OBJECTIVE: To assess the associations between nutrient intake and dietary patterns with different sarcopenia definitions in older men. DESIGN: Cross-sectional study. SETTING: Sarcopenia was defined using the Foundation for the National Institutes of Health (FNIH), the European Working Group on Sarcopenia in Older People (EWGSOP) and the European Working Group on Sarcopenia in Older People 2 (EWGSOP2). Dietary adequacy of fourteen nutrients was assessed by comparing participants' intakes with the Nutrient Reference Values (NRV). Attainment of NRV for nutrients was incorporated into a variable 'poor' (meeting ≤ 9) v. 'good' (meeting ≥ 10) using the cut-point method. Also, two different dietary patterns, monounsaturated:saturated fat and n-6:n-3 fatty acids ratio and individual nutrients were used as predictor variables. PARTICIPANTS: A total of 794 men aged ≥75 years participated in this study. RESULTS: The prevalence of sarcopenia by the FNIH, EWGSOP and EWGSOP2 definitions was 12·9 %, 12·9 % and 19·6 %, respectively. With the adjustment, poor nutrient intake was significantly associated with FNIH-defined sarcopenia (OR: 2·07 (95 % CI 1·16, 3·67)), but not with EWGSOP and EWGSPOP2 definitions. The lowest and second-lowest quartiles of protein, Mg and Ca and the lowest quartiles of n-6 PUFA and n-3 PUFA intakes were significantly associated with FNIH-defined sarcopenia. Each unit decrease in n-6:n-3 ratio was significantly associated with a 9 % increased risk of FNIH-defined sarcopenia (OR: 1·09 (95 % CI 1·04, 1·16)). CONCLUSIONS: Inadequate intakes of nutrients are associated with FNIH-defined sarcopenia in older men, but not with the other two sarcopenia definitions. Further studies are required to understand these relationships.

Changes in micronutrient intake and factors associated with this change among older Australian men: the Concord Health and Ageing in Men Project.

OBJECTIVES: To examine changes in micronutrient intake over 3 years and identify any associations between socio-economic, health, lifestyle and meal-related factors and these changes in micronutrient intakes among older men. DESIGN: Prospective study. SETTING: Dietary adequacy of individual micronutrient was compared to the estimated average requirement of the nutrient reference values (NRV). Attainment of the NRV for twelve micronutrients was incorporated into a dichotomised variable 'not meeting' (meeting ≤ 6) or 'meeting' (meeting ≥ 7) and categorised into four categories to assess change in micronutrient intake over 3 years. The multinomial logistic regression analyses were conducted to model predictors of changes in micronutrient intake. PARTICIPANTS: Seven hundred and ninety-four men participated in a detailed diet history interview at the third wave (baseline nutrition) and 718 men participated at the fourth wave (3-year follow-up). RESULTS: The mean age was 81 years (range 75-99 years). Median intakes of the majority of micronutrients decreased significantly over a 3-year follow-up. Inadequacy of the NRV for thiamine, dietary folate, Zn, Mg, Ca and I were significantly increased at a 3-year follow-up than baseline nutrition. The incidence of inadequate micronutrient intake was 21 % and remained inadequate micronutrient intake was 16·4 % at 3-year follow-up. Changes in micronutrient intakes were significantly associated with participants born in the UK and Italy, low levels of physical activity, having ≥2 medical conditions and used meal services. CONCLUSIONS: Micronutrient intake decreases with age in older men. Our results suggest that strategies to improve some of the suboptimal micronutrient intakes might need to be developed and implemented for older men.

Elevated serum substance P during simian varicella virus infection in rhesus macaques: implications for chronic inflammation and adverse cerebrovascular events.

Varicella and zoster, produced by varicella-zoster virus (VZV), are associated with an increased risk of stroke that may be due to persistent inflammation and hypercoagulability. Because substance P is associated with inflammation, hypercoagulability, and atherosclerotic plaque rupture that may contribute to increased stroke risk after VZV infection, we measured serum substance P in simian varicella virus-infected rhesus macaques. We found significantly increased and persistent serum substance P concentrations during varicella and zoster compared with pre-inoculation, supporting the hypothesis that VZV-induced increases in serum substance P may contribute to increased stroke risk associated with VZV infection.

'Aching to be a boy': A preliminary analysis of gender assignment of intersex persons in India in a culture of son preference.

Intersexuality, particularly in the global South, remains an under-researched field of study. In my in-progress doctoral research project, I explore the cultural, social, and medical discourses that influence how key stakeholders such as healthcare providers make decisions about the sex and gender assignment of the intersex child in India. In this paper I interrogate some of these ideas around gender assignment of intersex people in India, paying particular attention to the context of son preference. I am interested in exploring how decisions of gender assignment by medical professionals are guided by ideas of son preference. Focusing on four qualitative, semi-structured, in-depth interviews across two cities with medical doctors from different specializations, this paper is a preliminary attempt to examine some of the factors that guide medical professionals in making decisions about gender assignment of intersex children and explore the dynamics of the decision-making process. Specifically, I explore the factors that inform doctors' decision-making and locate these decision-making processes within the broader socio-cultural context of India.

Metagenomic Insights into Rhizospheric Microbiome Profiling in Lentil Cultivars Unveils Differential Microbial Nitrogen and Phosphorus Metabolism under Rice-Fallow Ecology.

The plant rhizosphere interfaces an array of microbiomes related to plant growth and development. Cultivar-specific soil microbial communities with respect to their taxonomic structure and specific function have not been investigated explicitly in improving the adaptation of lentil cultivars under rice-fallow ecology. The present study was carried out to decipher the rhizosphere microbiome assembly of two lentil cultivars under rice-fallow ecology for discerning the diversity of microbial communities and for predicting the function of microbiome genes related to nitrogen (N) and phosphorus (P) cycling processes deploying high-throughput whole (meta) genome sequencing. The metagenome profile of two cultivars detected variable microbiome composition with discrete metabolic activity. Cyanobacteria, Bacteroidetes, Proteobacteria, Gemmatimonadetes, and Thaumarchaeota were abundant phyla in the "Farmer-2" rhizosphere, whereas Actinobacteria, Acidobacteria, Firmicutes, Planctomycetes, Chloroflexi, and some incompletely described procaryotes of the "Candidatus" category were found to be robustly enriched the rhizosphere of "Moitree". Functional prediction profiles of the microbial metagenomes between two cultivars revealed mostly house keeping genes with general metabolism. Additionally, the rhizosphere of "Moitree" had a high abundance of genes related to denitrification processes. Significant difference was observed regarding P cycling genes between the cultivars. "Moitree" with a profuse root system exhibited better N fixation and translocation ability due to a good "foraging strategy" for improving acquisition of native P under the nutrient depleted rice-fallow ecology. However, "Farmer-2" revealed a better "mining strategy" for enhancing P solubilization and further transportation to sinks. This study warrants comprehensive research for explaining the role of microbiome diversity and cultivar-microbe interactions towards stimulating microbiome-derived soil reactions regarding nutrient availability under rice-fallow ecology.

Lack of T-cell-mediated IL-2 and TNFα production is linked to decreased CD58 expression in intestinal tissue during acute simian immunodeficiency virus infection.

For an effective T-cell activation and response, co-stimulation is required in addition to the antigen-specific signal from their antigen receptors. The CD2/CD58 interaction is considered as one of the most important T-cell co-stimulatory pathways for T-cell activation and proliferation, and its role in regulating intestinal T-cell function in acute and chronic SIV -infected macaques is poorly documented. Here, we demonstrated a significant reduction of CD58 expression in both T- and B-cell populations during acute SIV infection along with high plasma viral load and a loss of intestinal CD4+ T cells compared to SIV-uninfected control macaques. The reduction of CD58 expression in T cells was correlated with the reduced expression of T-cell-mediated IL-2 and TNFα production. Together, these results indicate that reduction in the CD2/CD58 interaction pathway in mucosal lymphocytes might play a crucial role in mucosal T-cell dysfunction during acute SIV/HIV infection.

Maintenance of AP-2-Dependent Functional Activities of Nef Restricts Pathways of Immune Escape from CD8 T Lymphocyte Responses.

Nef-specific CD8+ T lymphocytes (CD8TL) are linked to extraordinary control of primate lentiviral replication, but the mechanisms underlying their efficacy remain largely unknown. The immunodominant, Mamu-B*017:01+-restricted Nef195-203MW9 epitope in SIVmac239 partially overlaps a sorting motif important for interactions with host AP-2 proteins and, hence, downmodulation of several host proteins, including Tetherin (CD317/BST-2), CD28, CD4, SERINC3, and SERINC5. We reasoned that CD8TL-driven evolution in this epitope might compromise Nef's ability to modulate these important molecules. Here, we used deep sequencing of SIV from nine B*017:01+ macaques throughout infection with SIVmac239 to characterize the patterns of viral escape in this epitope and then assayed the impacts of these variants on Nef-mediated modulation of multiple host molecules. Acute variation in multiple Nef195-203MW9 residues significantly compromised Nef's ability to downregulate surface Tetherin, CD4, and CD28 and reduced its ability to prevent SERINC5-mediated reduction in viral infectivity but did not impact downregulation of CD3 or major histocompatibility complex class I, suggesting the selective disruption of immunomodulatory pathways involving Nef AP-2 interactions. Together, our data illuminate a pattern of viral escape dictated by a selective balance to maintain AP-2-mediated downregulation while evading epitope-specific CD8TL responses. These data could shed light on mechanisms of both CD8TL-driven viral control generally and on Mamu-B*017:01-mediated viral control specifically.IMPORTANCE A rare subset of humans infected with HIV-1 and macaques infected with SIV can control the virus without aid of antiviral medications. A common feature of these individuals is the ability to mount unusually effective CD8 T lymphocyte responses against the virus. One of the most formidable aspects of HIV is its ability to evolve to evade immune responses, particularly CD8 T lymphocytes. We show that macaques that target a specific peptide in the SIV Nef protein are capable of better control of the virus and that, as the virus evolves to escape this response, it does so at a cost to specific functions performed by the Nef protein. Our results help show how the virus can be controlled by an immune response, which could help in designing effective vaccines.

Attenuation of Simian Varicella Virus Infection by Enhanced Green Fluorescent Protein in Rhesus Macaques.

Simian varicella virus (SVV), the primate counterpart of varicella-zoster virus, causes varicella (chickenpox), establishes latency in ganglia, and reactivates to produce zoster. We previously demonstrated that a recombinant SVV expressing enhanced green fluorescent protein (rSVV.eGFP) is slightly attenuated both in culture and in infected monkeys. Here, we generated two additional recombinant SVVs to visualize infected cells in vitro and in vivo One harbors eGFP fused to the N terminus of open reading frame 9 (ORF9) (rSVV.eGFP-2a-ORF9), and another harbors eGFP fused to the C terminus of ORF66 (rSVV.eGFP-ORF66). Both recombinant viruses efficiently expressed eGFP in cultured cells. Both recombinant SVV infections in culture were comparable to that of wild-type SVV (SVV.wt). Unlike SVV.wt, eGFP-tagged SVV did not replicate in rhesus cells in culture. Intratracheal (i.t.) or i.t. plus intravenous (i.v.) inoculation of rhesus macaques with these new eGFP-tagged viruses resulted in low viremia without varicella rash, although SVV DNA was abundant in bronchoalveolar lavage (BAL) fluid at 10 days postinoculation (dpi). SVV DNA was also found in trigeminal ganglia of one monkey inoculated with rSVV.eGFP-ORF66. Intriguingly, a humoral response to both SVV and eGFP was observed. In addition, monkeys inoculated with the eGFP-expressing viruses were protected from superinfection with SVV.wt, suggesting that the monkeys had mounted an efficient immune response. Together, our results show that eGFP expression could be responsible for their reduced pathogenesis.IMPORTANCE SVV infection in nonhuman primates has served as an extremely useful animal model to study varicella-zoster virus (VZV) pathogenesis. eGFP-tagged viruses are a great tool to investigate their pathogenesis. We constructed and tested two new recombinant SVVs with eGFP inserted into two different locations in the SVV genome. Both recombinant SVVs showed robust replication in culture but reduced viremia compared to that with SVV.wt during primary infection in rhesus macaques. Our results indicate that conclusions on eGFP-tagged viruses based on in vitro results should be handled with care, since eGFP expression could result in attenuation of the virus.

Acute Viral Escape Selectively Impairs Nef-Mediated Major Histocompatibility Complex Class I Downmodulation and Increases Susceptibility to Antiviral T Cells.

Nef-specific CD8(+) T lymphocytes (CD8TL) are associated with control of simian immunodeficiency virus (SIV) despite extensive nef variation between and within animals. Deep viral sequencing of the immunodominant Mamu-B*017:01-restricted Nef165-173IW9 epitope revealed highly restricted evolution. A common acute escape variant, T170I, unexpectedly and uniquely degraded Nef's major histocompatibility complex class I (MHC-I) downregulatory capacity, rendering the virus more vulnerable to CD8TL targeting other epitopes. These data aid in a mechanistic understanding of Nef functions and suggest means of immunity-mediated control of lentivirus replication.