A third (booster) dose of the inactivated SARS-CoV-2 vaccine elicits immunogenicity and T follicular helper cell responses in people living with HIV

IntroductionThis study sought to explore the immunogenicity of a booster dose of an inactivated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine in people living with human immunodeficiency virus (HIV) and identify the factors affecting the magnitude of anti-SARS-CoV-2 antibody levels.Materials and methodsA total of 34 people living with HIV (PLWH) and 34 healthy donors (HD) were administered a booster dose of the same SARS-CoV-2 vaccine. Anti-SARS-CoV-2 antibody and immunoglobulin G (IgG) levels were measured using the SARS-CoV-2 S protein neutralizing antibody Enzyme-Linked Immunosorbent Assay (ELISA) and 2019-nCov IgG Chemiluminescent Immunoassay Microparticles, respectively. Spearman correlation analysis was used to measure the correlation between laboratory markers and neutralizing antibody and IgG levels. Peripheral blood mononuclear cells (PBMCs) were extracted from each subject using density gradient centrifugation and the numbers of memory T and T follicular helper (Tfh) cells were determined using flow cytometry.ResultsPLWH had a marked reduction in CD4 and B cell levels that was accompanied by a lower CD4/CD8 T cell ratio. However, those who received a supplementary dose of inactivated SARS-CoV-2 vaccines exhibited antibody positivity rates that were analogous to levels previously observed. The booster vaccine led to a reduction in IgG and neutralizing antibody levels and the amplitude of this decline was substantially higher in the PLWH than HD group. Correlation analyses revealed a strong correlation between neutralizing antibody levels and the count and proportion of CD4 cells. Anti-SARS-CoV-2 IgG antibody levels followed a similar trend. The expression of memory T and Tfh cells was considerably lower in the PLWH than in the HD group.DiscussionPLWH had an attenuated immune response to a third (booster) administration of an inactivated SARS-CoV-2 vaccine, as shown by lower neutralizing antibody and IgG levels. This could be attributed to the reduced responsiveness of CD4 cells, particularly memory T and cTfh subsets. CD4 and cTfh cells may serve as pivotal markers of enduring and protective antibody levels. Vaccination dose recalibration may be critical for HIV-positive individuals, particularly those with a lower proportion of CD4 and Tfh cells

Elucidating Electrocatalytic Oxygen Reduction Kinetics via Intermediates by Time-Dependent Electrochemiluminescence

Facile evaluation of oxygen reduction reaction (ORR) kinetics for massive electrocatalysts is critical for sustainable fuel cells development and industrial H2O2 production. Despite great success in ORR studies by mainstream strategies, such as membrane electrode assembly, rotation electrode technique and advanced surface-sensitive spectroscopy, the time/spatial distribution of reactive oxygen species (ROS) intermediates in the diffusion layer is still unknown. By time-dependent electrochemiluminescence (Td-ECL), here we report an intermediate-oriented methodology for ORR kinetics analysis. Thanks to multiple ultra-sensitive stoichiometric reactions between ROS and the ECL emitter, except for electron transfer numbers and rate constants, the potential-dependent time/spatial distribution of ROS was successfully obtained for the first time. Such uncovered exclusive information would guide fuel cells and H2O2 production with maximized activity and durability, for instance, a larger overpotential would be beneficial to electrocatalysts of 2e reduction for H2O2 production, because of the high yield of H2O2 and low concentration of attackable O2•. This work would pave the exploration of not only the fundamentals of unambiguous ORR mechanism but also the durability of electrocatalysts for practical applications

Ultrafast Humidity Sensing by Anisotropic Deformation of Carbon Nitride Nanoribbons for Real-Time Respiratory Monitoring

Developing humidity sensing materials with fast response and high sensitivity was of great interest for many applications from industrial field to human healthcare. Here, we report a carbon nitride nanoribbons (CNNRs)-based humidity sensor. Thanks to the delicate humidity-responsive anisotropic deformation and well-balanced hydrophilic surfaces/hydrophobic framework with rapid adsorption/desorption of water molecules by CNNRs, this humidity sensor possessed an ultrafast response of ca. 50 ms, high reproducibility and selectivity, and linearity in an almost full humidity range. As an example, this sensor was successfully applied to real-time breathing detection, and the as-obtained breathing graphic waveforms exhibited a higher sensitivity than that by the traditional clinic measurements. This work would pave a new way for ultrafast and sensitive humidity sensing by using anisotropic deformation of CNNRs and introduce a new application scheme of humidity sensors in more user-friendly respiratory monitoring with higher resolution.<br /

Highly Sensitive and Quality Self-Testable Electrochemiluminescence Assay of DNA Methyltransferase Activity Using Multifunctional Sandwich-Assembled Carbon Nitride Nanosheets

DNA methylation catalyzed by methylase plays a key role in many biological activities. However, developing a highly sensitive, simple, and reliable way for evaluation of DNA methyltransferase (MTase) activity is still a challenge. Here, we report a sandwich-assembled electrochemiluminescence (ECL) biosensor using multifunctional carbon nitride nanosheets (CNNS) to evaluate the Dam MTase activity. The CNNS could not only be used as an excellent substrate to conjugate a large amount of hairpin probe DNA to improve the sensitivity but also be utilized as an internal reliability checker and an analyte reporter in the bottom and top layers of the biosensor, respectively. Such a unique sandwich configuration of CNNS well coupled the advantages of ECL luminophor that were generally assembled in the bottom or top layer in a conventional manner. As a result, the biosensor exhibited an ultralow detection limit down to 0.043 U/mL and a linear range between 0.05 and 80 U/mL, superior to the MTase activity assay in most previous reports. We highlighted the great potential of emerging CNNS luminophor in developing highly sensitive and smart quality self-testable ECL sensing systems using a sandwiched configuration for early disease diagnosis, treatment, and management

Label-Free Detection of Telomerase Activity in Urine Using Telomerase-Responsive Porous Anodic Alumina Nanochannels

Telomerase is closely related to cancers, which makes it one of the most widely known tumor marker. Recently, many methods have been reported for telomerase activity measurement in which complex label procedures were commonly used. In this paper, a label-free method for detection of telomerase activity in urine based on steric hindrance changes induced by confinement geometry in the porous anodic alumina (PAA) nanochannels was proposed. Telomerase substrate (TS) primer was first assembled on the inside wall of PAA nanochannels by Schiff reaction under mild conditions. Then, under the action of telomerase, TS primer was amplified and extended to repeating G-rich sequences (TTAGGG)_<i>x</i>, which formed multiplex G-quadruplex in the presence of potassium ions (K⁺). This configurational change led to the increment of steric hindrance in the nanochannels, resulting in the decrement of anodic current of potassium ferricyanide (K₃[Fe­(CN)₆]). Compared with previously reported methods based on PAA nanochannels (usually one G-quadruplex formed), multiplex repeating G-quadruplex formed on one TS primer in this work. As a result, large current drop (∼3.6 μA, 36%) was obtained, which gave facility to improve the detection sensitivity. The decreased ratio of anodic current has a linear correlation with the logarithm of HeLa cell number in the range of 10–5000 cells, with the detection limit of seven cells. The method is simple, reliable, and has been successfully applied in the detection of telomerase in urine with good accuracy, selectivity and reproducibility. In addition, the method is nondestructive test compared to blood analysis and pathology tests, which is significant for cancer discovery, development, and prognosis

Competitive Multiple-Mechanism-Driven Electrochemiluminescent Detection of 8‑Hydroxy-2′-deoxyguanosine

Natural selection over billions of years has developed highly effective in vivo signal transduction that is often governed by a series of competitive multiple mechanisms. Several artificial signal transduction pathways have inspired numerous biosensing systems, but most of these are driven by a single mechanism. Herein we describe a multiple-mechanism-driven electrochemiluminescent (ECL) biosensor that utilizes competitive catalytic and steric hindrance effects by assembling hemin/G-quadruplex on carbon nitride nanosheets. Taking the detection of 8-hydroxy-2′-deoxyguanosine (8-OHdG) as example, the dynamic ranges of the detectable concentrations from the different mechanisms were integrated into a single sensor interface. Moreover, the detection sensitivity was more precisely controlled by the competition between the two mechanisms and inherently boosted compared with that of single-mechanism-driven detection. Going beyond the conventional single-mechanism-driven biosensing, the elaborate biomimetic coupling of multiple mechanisms in a single interface may open a new approach for future multiplexed biosensing

Coupled Fluorometer-Potentiostat System and Metal-Free Monochromatic Luminophores for High-Resolution Wavelength-Resolved Electrochemiluminescent Multiplex Bioassay

The sensitive simultaneous detection of multiple biomarkers is critical for the early diagnosis of diseases. Electrochemiluminescence (ECL) offers outstanding advantages, e.g., low background, over other optical sensing techniques. However, multiplexed ECL bioassay is hindered not only by the lack of generally available ECL spectrometers but also by the limited number of biocompatible monochromatic ECL luminophores for decades. Herein, we report addressing these issues by re-examination of the recent tabletop spectrofluorometer coupled potentiostat as a high-resolution ECL spectrum acquisition system and using carbon nitrides as monochromatic luminophores. A wavelength-resolved multiplexing ECL biosensor is demonstrated to simultaneously detect CA19-9 and mesothelin, two pancreatic cancer biomarkers, at a single-electrode interface. This work could initiate new opportunities for more general multiplex ECL biosensors with competitive performances

Simultaneous Noncovalent Modification and Exfoliation of 2D Carbon Nitride for Enhanced Electrochemiluminescent Biosensing

As an emerging nitrogen-rich 2D carbon material, graphitic carbon nitride (CN) has drawn much attention for applications ranging from photo-/electro­catalysts to biosensors. Interfacial modification of CN is fundamentally vital but is still in its infancy and remains challenging due to the low reactivity of CN. Here we report that, in conjunction with a π-π stacking interaction, bulk CN could be simultaneously exfoliated via facile mechanical grinding. The obtained CN nanosheets (m-CNNS) not only retained the pristine opto­electronic properties of bulk CN but also enriched a friendly interface for further coupling biomolecules with advanced properties, overcoming the deficiencies of CN in surface science. The m-CNNS were further covalently linked to a DNA probe, and the resultant electro­chemiluminescent biosensor for the target DNA exhibited much enhanced sensitivity with respect to that obtained by direct physical absorption of the DNA probe on unmodified CNNS. This noncovalent exfoliation and interfacial modification should greatly expand the scope of potential applications of CN in areas such as biosensing and should also be applicable to other 2D materials in interface modulation

Metal-Free All-Carbon Nanohybrid for Ultrasensitive Photoelectrochemical Immunosensing of alpha-Fetoprotein

C₆₀ can accept up to six electrons reversibly and show exceptional light absorption over the entire UV–vis spectrum, making it a potential photoactive probe for photoelectrochemical (PEC) bioassay. However, few successful works have been reported to apply fullerenes in PEC biosensing, partially because of the low electronic conductivity and poor interfacial interactions with targeted biomolecules. Herein, we report the addressing of these two obstacles by coupling high conductive graphite flake (Gr), graphene oxide (GO) with sufficient oxygen-containing functional groups, and an alkylated C₆₀ (AC₆₀) into a metal-free all-carbon nanohybrid (AC₆₀-Gr-GO) via harnessing delicate noncovalent interactions among them through a facile mechanical grinding. It was revealed that the as-obtained AC₆₀-Gr-GO nanohybrid not only showed conspicuous enhancement of photocurrent up to 35 times but also offered rich anchors for bioconjugation. With detection of alpha-fetoprotein as an example, the AC₆₀-Gr-GO based PEC immunosensor demonstrated a broad linear detection range (1 pg·mL^–1 to 100 ng·mL^–1) and a detection limit as low as 0.54 pg·mL^–1, superior/competitive to PEC immunosensors for AFP in previous reports. By a proper reinforcement in conductivity and biointerface engineering, this work may provide a new way to use fullerenes as photoactive materials in more general PEC biosensing

Visual, Label-Free Telomerase Activity Monitor via Enzymatic Etching of Gold Nanorods

Early diagnosis and life-long surveillance are clinically important to improve the long-term survival of cancer patients. Telomerase activity is a valuable biomarker for cancer diagnosis, but its measurement often used complex label procedures. Herein, we designed a novel, simple, visual and label-free method for telomerase detection by using enzymatic etching of gold nanorods (GNRs). First, repeating (TTAGGG)_<i>x</i> sequences were extented on telomerase substrate (TS) primer. It formed G-quadruplex under the help of Hemin and K⁺. Second, the obtained horseradish peroxidase mimicking hemin/G-quadruplex catalyzed the H₂O₂-mediated etching of GNRs to the short GNRs, even to gold nanoparticles (GNPs), generating a series of distinct color changes due to their plasmon-related optical response. Thus, this enzymatic reaction can be easily coupled to telomerase activity, allowing for the detection of telomerase activity based on vivid colors. This can be differentiated sensitively by naked eyes because human eyes are more sensitive to color variations rather than the optical density variations. As a result, telomerase activity can be quantitatively detected ranging from 200 to 15000 HeLa cells mL^–1. The detection limit was 90 HeLa cells mL^–1 (<i>S</i>/<i>N</i> = 3). Importantly, the application of this method in bladder cancer samples was in agreement with the clinical results. Thus, this method was considerably suitable for point-of-care diagnostics in resource-constrained regions because of the easy readout of results without the use of sophisticated apparatus