Poor Sleep Quality Associated With Enlarged Perivascular Spaces in Patients With Lacunar Stroke.

BACKGROUND AND OBJECTIVE: Enlarged perivascular spaces (EPVSs) are considered as an MRI marker of cerebral small vessel diseases and were reported to be associated with brain waste clearance dysfunction. A previous study found that interstitial fluid clearance in the mouse brain occurred mainly during sleep. However, the relationship between sleep quality and EPVS in humans has not been well-understood. Thus, we aimed to investigate the relationship between sleep and EPVS in humans. METHODS: This retrospective study was conducted in patients with lacunar stroke in the Neurology Department of Beijing Chaoyang Hospital. Patients with EPVS >10 on one side of the basal ganglia (BG) and white matter slice containing the maximum amount were defined as the BG-EPVS group and the white matter (WM)-EPVS group, respectively. Patients with EPVS <10 in the slice containing the maximum amount were defined as the control group. Sleep quality was evaluated by the Pittsburgh Sleep Quality Index (PSQI) including seven components, where a score of 6 or higher indicated poor sleep quality. Spearman's correlation analysis and the binary logistic regression analysis were performed to analyze the relationship between poor sleep quality and BG-EPVS and WM-EPVS, respectively. RESULTS: A total of 398 patients were enrolled in this study, including 114 patients in the BG-EPVS group and 85 patients in the WM-EPVS group. The proportion of poor sleep quality in the BG-EPVS group was higher than that in the control group (58.8 vs. 32.5%, p < 0.001). The score of PSQI, subjective sleep quality, sleep latency, sleep duration, and sleep efficiency were higher in the BG-EPVS group than that in the control group (p < 0.05). The proportion of poor sleep quality was also higher in the WM-EPVS group than that in the control group (50.6 vs. 35.3%, p = 0.031). The score of sleep duration and sleep disturbances was higher in the WM-EPVS group than that in the control group. Spearman's correlation analysis showed that poor sleep quality was positively associated with BG-EPVS (ρ = 0.264, p < 0.001) and WM-EPVS (ρ = 0.154, p = 0.044). The binary logistic regression analysis showed that poor sleep quality, longer sleep latency, and less sleep duration were independently related to BG-EPVS and poor sleep quality, less sleep duration, and more serious sleep disturbances were independently related to WM-EPVS after adjusting for confounders (P < 0.05). CONCLUSION: Poor sleep quality was independently associated with EPVS in BG and WM.

The Relationship Between Chronic Obstructive Pulmonary Disease and Cerebral Small Vessel Disease Assessed by Magnetic Resonance Imaging: A Case-Control Study from a Single Center in Beijing, China.

BACKGROUND Chronic obstructive pulmonary disease (COPD) and cerebral small vessel disease (CSVD) reportedly share similar risk factors and pathogenesis. However, the relationship between these 2 diseases is not clear. This study aimed to investigate the association between COPD and CSVD. MATERIAL AND METHODS Patients with stable COPD and matched healthy control participants were recruited for this study. Clinical characteristics were collected based on medical history, serological tests, brain magnetic resonance imaging, and pulmonary function tests. Individual CSVD imaging markers (white matter hyperintensities [WMH], enlarged perivascular space [EPVS], and brain atrophy) were assessed to determine their severity. Logistic analysis was used to test the relationship between CSVD markers and COPD. RESULTS Significant differences in WMH, basal ganglia EPVS (BG-EPVS), and centrum semiovale EPVS (CSO-EPVS) were found between COPD and control groups (P0.05). CONCLUSIONS A significant correlation exists between COPD and imaging markers of CSVD, including WMH, BG-EPVS, and CSO-EPVS. In addition, the severity of WMH and BG-EPVS is positively related to the duration of COPD, suggesting that COPD may be a risk factor for CSVD.

Dual-functional NiCo2S4 polyhedral architecture with superior electrochemical performance for supercapacitors and lithium-ion batteries.

Dual-functional NiCo2S4 polyhedral architectures with outstanding electrochemical performance for supercapacitors and lithium-ion batteries (LIBs) have been rationally designed and successfully synthesized by a hydrothermal method. The as-synthesized NiCo2S4 electrode for supercapacitor exhibits an outstanding specific capacitance of 1298Fg-1 at 1Ag-1 and an excellent rate capability of ~80.4% at 20Ag-1. Besides, capacitance retention of 90.44% is realized after 8000 cycles. In addition, the NiCo2S4 as anode in LIBs delivers high initial charge/discharge capacities of 807.6 and 972.8mAhg-1 at 0.5C as well as good rate capability. In view of these points, this work provides a feasible pathway for assembling electrodes and devices with excellent electrochemical properties in the next generation energy storage applications.

Constructing highly dispersed 0D Co3S4 quantum dots/2D g-C3N4 nanosheets nanocomposites for excellent photocatalytic performance.

The development of noble-metal-free catalysts with high efficiency photocatalytic properties is critical to the heterogeneous catalysis. Herein, zero-dimensional (0D) metal sulfide quantum dots/two-dimensional (2D) g-C3N4 nanosheets (Co3S4/CNNS) nanocomposites are synthesized by a two-step method, including the ways of in-situ deposition and water bath. The highly dispersed Co3S4 quantum dots (particle size is 2-4 nm) are evenly and tightly fixed on CNNS, which can be used as co-catalyst to effectively replace noble metals to improve the photocatalytic properties of CNNS. Co3S4/CNNS-900 has the apparent quantum efficiency, which is up to 7.85% at 400 nm. At the same time, the H2 evolution rate of Co3S4/CNNS-900 is 20,536.4 µmol g-1 h-1, which is 555 times than CNNS. The excellent photocatalytic performance is due to the highly dispersed Co3S4 quantum dots on 2D CNNS, which facilitate the formation of more active sites, Co3S4/CNNS promotes the separation and migration of photogenerated carriers, shortens the migration distance of photogenerated carriers, and eventually leads to an increase of the photocatalytic performance.

Hedgehog-inspired nanostructures for hydrogel-based all-solid-state hybrid supercapacitors with excellent flexibility and electrochemical performance.

High-security deformable energy-storage devices that are mechanically robust, with considerable energy and power densities are becoming desirable for smart wearable electronics. Here, a highly flexible hydrogel-based all-solid-state hybrid supercapacitor was rationally designed and assembled, with unique NiCo2O4@NixCoyMoO4 (x : y = 3 : 1) nanostructures as the electrode, which was bio-inspired by the curling up and relaxation of hedgehogs. The hybrid supercapacitor shows no obvious decay in capacitance during bending to different states, indicating its outstanding flexibility and mechanical stability. The capacitance was still maintained at 92.0% of the initial value, even after continuous bending for 3000 cycles. The highly monodisperse NiCo2O4@NixCoyMoO4 nanostructures releasing stress during bending is responsible for the favorable stability and flexibility. Furthermore, the hybrid supercapacitor displayed outstanding electrochemical performance, with a high specific capacitance of 207 F g-1 at 1 A g-1, a high energy density of 64.7 W h kg-1 at 749.6 W kg-1, and favorable cycling stability (nearly 100% after 10 000 cycles). The flexible hybrid supercapacitor could be charged with a solar cell and served as the power source to light up LEDs. This simple and reliable hybrid supercapacitor, with extraordinary mechanical stability and electrochemical performance, is a promising power source for smart wearable electronics.

A stable layered P3/P2 and spinel intergrowth nanocomposite as a long-life and high-rate cathode for sodium-ion batteries.

Layered sodium transition-metal oxides, NaxMeO2, with large theoretical capacity are regarded as an important class of cathode materials for sodium-ion batteries (SIBs). However, they usually exhibit inferior thermodynamic stability and sluggish Na+ kinetics due to the unwanted phase transitions and large Na+-ionic radius. In this work, considering the beneficial synergistic effects of layered P2/P3 and Fd3[combining macron]m spinel phases, a stable layered/spinel intergrowth nanocomposite Na0.5[Ni0.2Co0.15Mn0.65]O2 is rationally designed and successfully prepared via a co-precipitation route and a subsequent solid-state reaction, and the triphase synergy in this layered/spinel nanocomposite is demonstrated. In Na/Na0.5[Ni0.2Co0.15Mn0.65]O2 half-cells, it delivers a high specific capacity of ∼180 mA h g-1 and a good cycling stability, with a capacity retention of 87.6% after 100 cycles, at a rate of 0.1C between 1.5 and 4.0 V (vs. Na/Na+). The large reversible capacity of 105 mA h g-1 is also achieved even at a high rate of 10C, indicating high-rate capability. Besides, the full-cells using this nanocomposite as the cathode and hard carbon as the anode exhibit long-term cycle-life and high-power properties, indicating the expected merits of layered/spinel mixed phases. The superior sodium storage performance of this layered P3/P2 and spinel intergrowth nanocomposite makes it a promising candidate as a long-life and high-rate cathode for SIBs.

Assessment of Homocysteine as a Diagnostic and Early Prognostic Biomarker for Patients with Acute Lacunar Infarction.

BACKGROUND: Although increasing evidence has demonstrated that elevated homocysteine (Hcy) levels may be an important contributor for the development of cerebral infarction, rare studies focused on its diagnostic and early prognostic roles in acute lacunar infarction. METHODS: A total of 197 patients with acute lacunar infarction and 192 to form the control group were prospectively recruited between January 2013 and February 2017. Early neurological deterioration was defined as an increase of ≥2 points in National Institutes of Health Stroke Scale or the decrease in Barthel index (BI) score at discharge. RESULTS: Univariate and multivariate logistic regression analyses revealed that higher levels of fibrinogen and Hcy were independently clinical predictors associated with lacunar infarction. Receiver operating characteristic curves analysis demonstrated that the diagnosis value of Hcy was superior to fibrinogen, with the area under the curve of 0.881 and 0.688 respectively. Using the optimal cutoff value of 15.5 µmol/L of Hcy, a sensitivity of 65% and a specificity of 100% were achieved for predicting lacunar infarction. Hcy was only significantly related with BI reduction in the males (30.5 [15.5-65.5] vs. 18 [15-24], p = 0.034) in the univariate analysis but not in the females and the multivariate analysis. CONCLUSIONS: Serum Hcy may be an independent diagnostic and not an early prognostic biomarker for patients with acute lacunar infarction.

Serum Uric Acid is Independently Associated with Enlarged Perivascular Spaces.

Enlarged perivascular spaces (EPVS) are reported to be associated with impaired cognitive function and sleep disorders. It is of clinical importance to understand the risk factors for EPVS. Hyperuricemia increases the risk of hypertension and endothelial dysfunction, which are well recognized to be associated with EPVS. Therefore, we postulated that serum uric acid (SUA) might be associated with EPVS. A total of 665 lacunar stroke patients were enrolled in this study. The SUA concentrations of patients with severe EPVS were much higher than those of patients with mild EPVS (for basal ganglia: 5.25 ± 1.40 mg/dl vs. 4.75 ± 1.40 mg/dl, p < 0.001; for white matter: 5.31 ± 1.41 mg/dl vs. 4.88 ± 1.37 mg/dl, p = 0.009). The percentage of subjects with severe EPVS tended to be higher in the highest quartile of SUA (chi-square test: P = 0.002 for basal ganglia and 0.006 for white matter). Spearman correlation analysis indicated that the SUA concentrations were positively correlated with the severity of EPVS (rho > 0, p < 0.05). Multivariate logistic regression analysis showed that high normal SUA was independently associated with a higher severity of EPVS. This finding suggests that high SUA levels might be an independent risk factor for EPVS in lacunar stroke patients.

Surface/Interfacial Structure and Chemistry of High-Energy Nickel-Rich Layered Oxide Cathodes: Advances and Perspectives.

The urgent prerequisites of high energy-density and superior electrochemical properties have been the main inspiration for the advancement of cathode materials in lithium-ion batteries (LIBs) in the last two decades. Nickel-rich layered transition-metal oxides with large reversible capacity as well as high operating voltage are considered as the most promising candidate for next-generation LIBs. Nonetheless, the poor long-term cycle-life and inferior thermal stability have limited their broadly practical applications. In the research of LIBs, it is observed that surface/interfacial structure and chemistry play significant roles in the performance of cathode cycling. This is due to the fact that they are basically responsible for the reversibility of Li+ intercalation/deintercalation chemistries while dictating the kinetics of the general cell reactions. In this Review, the surface/interfacial structure and chemistry of nickel-rich layered cathodes involving structural defects, redox mechanisms, structural evolutions, side-reactions among others are initially demonstrated. Recent advancements in stabilizing the surface/interfacial structure and chemistry of nickel-rich cathodes by surface modification, core-shell/concentration-gradient structure, foreign-ion substitution, hybrid surface, and electrolyte additive are presented. Then lastly, the remaining challenges such as the fundamental studies and commercialized applications, as well as the future research directions are discussed.

The relationship between ambulatory blood pressure variability and enlarged perivascular spaces: a cross-sectional study.

OBJECTIVES: Recent studies reported that 24-hour ambulatory blood pressure variability (ABPV) was associated with lacunar infarction and white matter hyperintensities (WMH). However, the relationship between ABPV and enlarged perivascular spaces (EPVS) has not been investigated. Thus, our study aimed to investigate whether ABPV is associated with EPVS by 24-hour ambulatory blood pressure monitoring (ABPM). DESIGN: We conducted this study as a cross-sectional study. SETTINGS: The study was based on patients who presented for physical examinations in our hospital from May 2013 to June 2016. PARTICIPANTS: Patients with both brain MRI scans and 24-hour ABPM were included and patients with acute stroke, a history of severe stroke and some other severe diseases were excluded. A total of 573 Chinese patients were prospectively enrolled in this study. PRIMARY AND SECONDARY OUTCOME MEASURES: EPVS in basal ganglia (BG) and white matter (WM) were identified on MRI and classified into three categories by the severity. WMH were scored by the Fazekas scale. Coefficient of variation (CV) and SD were considered as metrics of ABPV. Spearman correlation analysis and ordinal logistic regression analysis were used to assess the relationship between ABPV and EPVS. RESULTS: There were statistical differences among the subgroups stratified by the severity of EPVS in BG in the following ABPV metrics: SD and CV of systolic blood pressure (SBP), CV of diastolic blood pressure (DBP) in 24 hours, daytime and nighttime and SD of DBP in nighttime. The above ABPV metrics were positively associated with the degree of EPVS. The association was unchanged after adjusting for confounders. Spearman correlation analysis showed ABPV was not related to the degree of EPVS in the WM. CONCLUSION: ABPV was independently associated with EPVS in BG after controlling for blood pressure, but not in the WM. Pathogenesis of EPVS in BG and WM might be different.

Higher ambulatory systolic blood pressure independently associated with enlarged perivascular spaces in basal ganglia.

BACKGROUNDS: Enlarged perivascular spaces (EPVS) have been identified as a marker of cerebral small vessel diseases (CSVD). Ambulatory blood pressure (ABP) is the strongest predictor of hypertension-related brain damage. However, the relationship between ABP levels and EPVS is unclear. OBJECTIVES: This study aimed to investigate the association between ABP levels and EPVS by 24-hour ambulatory blood pressure monitoring (ABPM). METHODS: We prospectively recruited inpatients for physical examinations in our hospital from May 2013 to Jun 2016. 24-hour ABPM data and cranial magnetic resonance imaging information were collected. EPVS in basal ganglia (BG) and centrum semiovale (CSO) were identified and classified into three categories by the severity. White matter hyperintensities were scored by Fazekas scale. Spearman correlation analysis and multiple logistic regression analysis were used to determine the relationship between ABP levels and EPVS. RESULTS: A total of 573 subjects were enrolled in this study. 24-hour, day and night systolic blood pressure (SBP) levels were positively related to higher numbers of EPVS in BG (24-hour SBP: r = 0.23, p < 0.01; day SBP: r = 0.25, p < 0.01; night SBP: r = 0.30, p < 0.01). The association was unchanged after controlling for confounders by multiple logistic regression analysis. 24-hour and day diastolic blood pressure (DBP) levels increased with an increasing degree of EPVS in CSO (p = 0.04 and 0.049, respectively). But the association disappeared after adjusting for confounders. Spearman correlation analysis indicated that ABP levels were not associated with higher numbers of EPVS in CSO (p > 0.05). DBP levels were not independently associated with the severity of EPVS in BG and CSO. CONCLUSION: Higher SBP levels were independently associated with EPVS in BG, but not in CSO, which supported EPVS in BG to be a marker of CSVD. Pathogenesis of EPVS in BG and CSO might be different.

IL-4 and IFN-γ induced by human immunodeficiency virus vaccine in a schistosome infection model.

The co-infection of HIV and helminth parasites, such as Schistosoma spp, has increased in sub-Saharan Africa. Many HIV vaccine candidate studies have been completed or are in ongoing clinical trials, but it is not clear how HIV vaccines might affect the course of schistosome infections. In this study, we immunized S. mansoni-infected mice with an efficient DNA vaccine that included HIV gag. Using this model, we found that Th2 cytokines, such as IL-4 and IL-13, were highly induced after schistosome infection. Treatment of infected mice with the HIV DNA vaccine resulted in a significant attenuation of this rise in IL-13 expression and an increase in expression of the Th1 cytokine, TNF-α. However, vaccine administration did not significantly influence the expression of IL-4, or IFN-γ, and did not affect T cell proliferative capacity. Interestingly, the IL-4 (+) IFN-γ (+) phenotype appears in schistosome-infected mice that received HIV vaccination, and is associated with the expression of transcription factors GATA3 (+) T-bet (+) in these mice. These studies indicate that DNA vaccination can have an impact on ongoing chronic infection.

Plasmodium inui infection reduces the efficacy of a simian immunodeficiency virus DNA vaccine in a rhesus macaque model through alteration of the vaccine-induced immune response.

Human immunodeficiency virus type 1 and malaria are co-endemic in many areas. We evaluated the effects of Plasmodium inui infection on the performance of a simian immunodeficiency virus (SIV) DNA vaccine. Rhesus macaques were infected with P. inui by transfusion of whole blood from a persistently infected animal. Animals with and animals without P. inui infection were then vaccinated 4 times with an SIV DNA vaccine encoding SIVgag, SIVpol, and SIVenv. Animals were subsequently challenged with thirty 50% rhesus monkey infectious doses of SIVmac251 6 weeks after the last vaccination. P. inui-infected immunized animals showed a significantly higher viral load than animals without P. inui infection (P = .010, by the Wilcoxon rank sum test). The higher viral loads in the P. inui-infected animals were durable and were observed at all sampling time points across the study (P = .00245, by the Wilcoxon rank test). The P. inui-infected animals also had correspondingly lower CD4(+) cell counts. There were fewer vaccine-specific CD4(+) and CD8(+) cells in the P. inui-infected animals, compared with uninfected animals. Of importance, P. inui infection seemed to decrease the number of CD8(+) cells that could proliferate or secrete interferon γ, although the number of CD8(+) cells capable of secreting tumor necrosis factor α following in vitro stimulation was increased. This study demonstrated that P. inui infection had an influence on the immune response to an SIV DNA vaccine and decreased the vaccine's efficacy.

Long-term programming of antigen-specific immunity from gene expression signatures in the PBMC of rhesus macaques immunized with an SIV DNA vaccine.

While HIV-1-specific cellular immunity is thought to be critical for the suppression of viral replication, the correlates of protection have not yet been determined. Rhesus macaques (RM) are an important animal model for the study and development of vaccines against HIV/AIDS. Our laboratory has helped to develop and study DNA-based vaccines in which recent technological advances, including genetic optimization and in vivo electroporation (EP), have helped to dramatically boost their immunogenicity. In this study, RMs were immunized with a DNA vaccine including individual plasmids encoding SIV gag, env, and pol alone, or in combination with a molecular adjuvant, plasmid DNA expressing the chemokine ligand 5 (RANTES), followed by EP. Along with standard immunological assays, flow-based activation analysis without ex vivo restimulation and high-throughput gene expression analysis was performed. Strong cellular immunity was induced by vaccination which was supported by all assays including PBMC microarray analysis that identified the up-regulation of 563 gene sequences including those involved in interferon signaling. Furthermore, 699 gene sequences were differentially regulated in these groups at peak viremia following SIVmac251 challenge. We observed that the RANTES-adjuvanted animals were significantly better at suppressing viral replication during chronic infection and exhibited a distinct pattern of gene expression which included immune cell-trafficking and cell cycle genes. Furthermore, a greater percentage of vaccine-induced central memory CD8+ T-cells capable of an activated phenotype were detected in these animals as measured by activation analysis. Thus, co-immunization with the RANTES molecular adjuvant followed by EP led to the generation of cellular immunity that was transcriptionally distinct and had a greater protective efficacy than its DNA alone counterpart. Furthermore, activation analysis and high-throughput gene expression data may provide better insight into mechanisms of viral control than may be observed using standard immunological assays.

High antibody and cellular responses induced to HIV-1 clade C envelope following DNA vaccines delivered by electroporation.

BACKGROUND: Clade C is the predominant HIV-1 strain infecting people in sub-Saharan Africa, India, and China and there is a critical need for a vaccine targeted to these areas. In this study we tested a DNA based vaccine that encodes the SIVgag, SIVpol and HIV-1 envelope clade C. METHODS: Rhesus macaques were immunized by electroporation with the DNA plasmid encoding optimized SIVgag, SIVpol and an HIV-1 env clade C with or without the adjuvant RANTES. Animals were monitored for immune responses and challenged following the final immunization with 25 animal infectious doses (AID) of SHIV-1157ipd3N4. RESULTS: We found that the vaccine induced high levels of antigen specific IFN-γ producing effector cells and the capacity for CD4+ and CD8+ to proliferate upon antigen stimulation. Importantly, we found that the vaccine induced antibody titers as high as 1/4000. These antibodies were capable of neutralizing tier 1 HIV-1 viruses. Finally, when macaques were challenged with SHIV, viral loads were controlled in vaccinated groups. CONCLUSION: We conclude that immunization with a simian/human immunodeficiency virus DNA-based vaccine delivered by electroporation can induce cellular and humoral immune responses that are able to control viral replication.

Viral reservoir is suppressed but not eliminated by CD8 vaccine specific lymphocytes.

It has long been postulated that while CD8 lymphocytes are capable of suppressing human immunodeficiency virus (HIV)-1 replication it is unlikely that the viral reservoirs once formed can be cleared. Our previous studies demonstrate that co-immunizing cynomologous macaques with a simian/human immunodeficiency virus (SHIV) DNA-based vaccines induces a strong cellular immune response that is able to suppress viral replication. We further demonstrated that interleukin (IL)-12 could significantly enhance the vaccine specific CD8 lymphocyte response. In this manuscript cynomologous macaques were vaccinated with a SHIV DNA-based vaccine co-delivered with IL-12. The macaques were then challenged with SHIV89.6p. Two years post-immunization and viral challenge we transiently depleted CD8(+) T cells. Plasma viral load increased, demonstrating the central role of CD8(+) T cells in viral suppression yet an inability to clear the viral reservoirs. Furthermore, in the data presented here, we found a higher number of IFN-gamma producing vaccine specific cells did not enhance suppression of viral replication.

High dose of plasmid IL-15 inhibits immune responses in an influenza non-human primates immunogenicity model.

Interleukin (IL)-15, is a cytokine that is important for the maintenance of long-lasting, high-avidity T cell response to invading pathogens and has, therefore, been used in vaccine and therapeutic platforms as an adjuvant. In addition to pure protein delivery, plasmids encoding the IL-15 gene have been utilized. However, it is critical to determine the appropriate dose to maximize the adjuvanting effects. We immunized rhesus macaques with different doses of IL-15 expressing plasmid in an influenza non-human primate immunogenicity model. We found that co-immunization of rhesus macaques with a Flu DNA-based vaccine and low doses of plasmid encoding macaque IL-15 enhanced the production of IFN-gamma (0.5 mg) and the proliferation of CD4(+) and CD8(+) T cells, as well as T(CM) levels in proliferating CD8(+) T cells (0.25 mg). Whereas, high doses of IL-15 (4 mg) decrease the production of IFN-gamma and the proliferation of CD4(+) and CD8(+) T cells and T(CM) levels in the proliferating CD4(+) and CD8(+) T cells. In addition, the data of hemagglutination inhibition (HI) antibody titer suggest that although not significantly different, there appears to be a slight increase in antibodies at lower doses of IL-15. Importantly, however, the higher doses of IL-15 decrease the antibody levels significantly. This study demonstrates the importance of optimizing DNA-based cytokine adjuvants.

Sustained suppression of SHIV89.6P replication in macaques by vaccine-induced CD8+ memory T cells.

OBJECTIVE: We previously demonstrated that a strategy of co-immunizing cynomologous macaques with a simian/human immunodeficiency virus DNA-based vaccine and a plasmid encoding macaque interleukin (IL)-15 induces a strong CD8 and CD4 effector T-cell response that, upon subsequent challenge with SHIV89.6P, controls viral replication and protects immunized animals against ongoing infection. In this follow-up study, we measured viral replication 2 years after vaccination challenge and determined the mechanism by which antigen-specific CD8 T cells suppress viral replication. METHOD: From the original group of 18, we assessed the immune response in the 13 surviving animals. In addition, using cM-T807, we depleted CD8 lymphocytes to assess the role CD8 cells play in suppression of viral replication. RESULT: We found that peripheral blood mononuclear cells from vaccinated animals had a robust simian immunodeficiency virus Gag-specific IFN-gamma response. In addition, in the DNA and IL-15 group, we observed higher levels of simian immunodeficiency virus Gag-specific, proliferating CD8 T cells. The profile of these cells revealed more central memory than effector cells. When we transiently depleted animals of CD8 T cells, plasma viral load increased, and peak viral load was lower in the DNA and IL-15 group compared with the DNA alone and control groups. As CD8 T cells recovered, viral replication was controlled and we observed an increase in the number of antigen-specific effector CD8 T cells. CONCLUSION: We conclude that co-immunization with a simian/human immunodeficiency virus DNA-based vaccine and IL-15 achieves sustained viral suppression and that vaccine-induced CD8 memory T cells, which differentiate into effector cells, are central to that suppression.

Synergistic effects of adjuvants interferon-gamma and levamisole on DNA vaccination against infection with Newcastle disease virus.

Both humoral and cell-mediated immune responses are important to protect animals from initial acute viral infection and establishment of chronic infection. Adjuvants for DNA vaccines can influence the balance between humoral and cell-mediated immunities. In this study, a DNA vaccine encoding the hemagglutinin-neuraminidase and fusion genes of Newcastle disease virus (NDV) incorporated with chicken interferon(provax-chIFN-gamma) cDNA as a molecular adjuvant and levamisole (LMS) as a chemical adjuvant was tested for its efficacy in protection against NDV lethal challenge. Compared with DNA vaccine alone, the DNA vaccine with provax-chIFN-gamma plus LMS induced significantly higher humoral and cell-mediated responses, as shown by higher levels of hemagglutination inhibition (HI) titers and T cell proliferation. In addition, the DNA vaccine with provax-chIFN-gamma plus LMS formulation increased the expression of IFN-gamma, interleukin (IL)-2, IL-4, IL-12, and IL-13, suggesting that the effectiveness of the IFN-gamma and LMS formulation is partly due to the enhancement of balanced cytokine production. Furthermore, the two adjuvants yielded 80% protection in chickens against challenge with a lethal dose of the virulent NDV strain. This study demonstrates that the synergistic effects of provax-chIFN-gamma plus LMS as the adjuvants in NDV DNA vaccination could be used to improve protective efficacy in chickens.

An automated process to extract plasmid DNA by alkaline lysis.

With advances in the development of DNA vaccines and gene therapy, there is a growing need for plasmid DNA with high quality for fundamental research and clinical trials. In this report, a scalable automated process for large-scale preparation of plasmid is described. This process is based on alkaline lysis and can be easily scaled up to meet demands for larger quantities. In the process, harvested bacteria are passed through two mixing chambers at controlled speeds to affect lysis and control alkalinity. The resulting solution is passed through a series of filters to remove contaminants, and ethanol precipitated. System parameters are examined to maximize the quantity and quality of the prepared plasmid. Using this procedure, plasmid can be extracted and purified from 1 l of Escherichia coli cultures at an OD600 nm of 50 in less than 45 min. The plasmid yields are approximately 90 mg/l culture.