Blanket antimicrobial resistance gene database with structural information, BOARDS, provides insights on historical landscape of resistance prevalence and effects of mutations in enzyme structure.

Antimicrobial resistance (AMR) in pathogenic bacteria poses a significant threat to public health, yet there is still a need for development in the tools to deeply understand AMR genes based on genetic or structural information. In this study, we present an interactive web database named Blanket Overarching Antimicrobial-Resistance gene Database with Structural information (BOARDS, sbml.unist.ac.kr), a database that comprehensively includes 3,943 reported AMR gene information for 1,997 extended spectrum beta-lactamase (ESBL) and 1,946 other genes as well as a total of 27,395 predicted protein structures. These structures, which include both wild-type AMR genes and their mutants, were derived from 80,094 publicly available whole-genome sequences. In addition, we developed the rapid analysis and detection tool of antimicrobial-resistance (RADAR), a one-stop analysis pipeline to detect AMR genes across whole-genome sequencing (WGSs). By integrating BOARDS and RADAR, the AMR prevalence landscape for eight multi-drug resistant pathogens was reconstructed, leading to unexpected findings such as the pre-existence of the MCR genes before their official reports. Enzymatic structure prediction-based analysis revealed that the occurrence of mutations found in some ESBL genes was found to be closely related to the binding affinities with their antibiotic substrates. Overall, BOARDS can play a significant role in performing in-depth analysis on AMR.IMPORTANCEWhile the increasing antibiotic resistance (AMR) in pathogen has been a burden on public health, effective tools for deep understanding of AMR based on genetic or structural information remain limited. In this study, a blanket overarching antimicrobial-resistance gene database with structure information (BOARDS)-a web-based database that comprehensively collected AMR gene data with predictive protein structural information was constructed. Additionally, we report the development of a RADAR pipeline that can analyze whole-genome sequences as well. BOARDS, which includes sequence and structural information, has shown the historical landscape and prevalence of the AMR genes and can provide insight into single-nucleotide polymorphism effects on antibiotic degrading enzymes within protein structures.

Assessment of the association between dust storms and COVID-19 infection rate in southwest Iran.

This study assesses a plausible correlation between a dust intrusion episode and a daily increase in COVID-19 cases. A surge in COVID-19 cases was observed a few days after a Middle East Dust (MED) event that peaked on 25th April 2020 in southwest Iran. To investigate potential causal factors for the spike in number of cases, cross-correlations between daily combined aerosol optical depths (AODs) and confirmed cases were computed for Khuzestan, Iran. Additionally, atmospheric stability data time series were assessed by covering before, during, and after dust intrusion, producing four statistically clustered distinct city groups. Groups 1 and 2 had different peak lag times of 10 and 4-5 days, respectively. Since there were statistically significant associations between AOD levels and confirmed cases in both groups, dust incursion may have increased population susceptibility to COVID-19 disease. Group 3 was utilized as a control group with neither a significant level of dust incursion during the episodic period nor any significant associations. Group 4 cities, which experienced high dust incursion levels, showed no significant correlation with confirmed case count increases. Random Forest Analysis assessed the influence of wind speed and AOD, showing relative importance of 0.31 and 0.23 on the daily increase percent of confirmed cases, respectively. This study may serve as a reference for better understanding and predicting factors affecting COVID-19 transmission and diffusion routes, focusing on the role of MED intrusions.

Laboratory Evaluation of Mechanical Properties of Modified Asphalt and Mixture Using Graphene Platelets (GnPs).

Recently, nanomaterials have attracted attention in the field of pavement construction as modifiers to endure heavy loads and climate changes. In this study, conventional asphalt (bitumen) of penetration grade AC (60/70) was modified with graphene platelets (GnPs) at three different contents: 0.5%, 1.0%, and 1.5% by weight of asphalt content. Kinematic viscosity, softening point, penetration, and dynamic shear rheology tests were performed to evaluate the mechanical properties of modified binder. The results showed that adding GnPs improves the mechanical properties of asphalt binder; the kinematic viscosities, softening points, and rutting parameters increased but penetrations decreased with the contents of GnPs. Hot mix asphalt specimens with GnPs-modified asphalt were prepared and characterized with Marshall tests, thermal stress restrained specimen tests (TSRST), wheel tracking tests, and indirect tensile tests. Similar to the results of asphalt binder, the mechanical properties of asphalt mixture were improved by GnPs. Marshall stability increased by 21% and flow decreased by 24% with accepted value of 2.8 mm in penetration when the mixture was modified with 1.0 wt% of GnPs. At the same GnPs content, modified asphalt mixture led to lower failure temperature by 2 °C in comparison with unmodified asphalt mixture and the cryogenic failure stress was improved by 12%. The wheel tracking tests showed that GnPs-modified asphalt mixture has outstanding deformation resistance in comparison with unmodified asphalt mixtures: after 5000 cycles, 1.0 wt% of GnPs reduced the rut depth of asphalt mixture by 60%-the rut depth of unmodified asphalt mixture was 6.9 mm compared to 2.75 mm for modified asphalt mixture. After 10,000 cycles, the modified asphalt mixture showed rut depth of 3.24 mm in comparison with 8.12 mm in case of unmodified asphalt mixture. Addition of GnPs into asphalt mixture significantly improved the indirect tensile strength: 1.0 wt% of GnPs increased the indirect tensile strength of unmodified asphalt mixture from 0.79 to 1.1 MPa recording ~40% increment. The results of this study can confirm that graphene platelets enhance the mechanical properties of asphalt mixture and its performance.

Statistical and Reliability Study on Shear Strength of Recycled Coarse Aggregate Reinforced Concrete Beams.

The consumption of structural concrete in the construction industry is rapidly growing, and concrete will remain the main construction material for increasing urbanization all over the world in the near future. Meanwhile, construction and demolition waste from concrete structures is also leading to a significant environmental problem. Therefore, a proper sustainable solution is needed to address this environmental concern. One of the solutions can be using recycled coarse aggregates (RCA) in reinforced concrete (RC) structures. Extensive research has been conducted in this area in recent years. However, the usage of RCA concrete in the industry is still limited due to the absence of structural regulations appropriate to the RCA concrete. This study addresses a safety margin of RCA concrete beams in terms of shear capacity which is comparable to natural coarse aggregates (NCA) concrete beams. To this end, a database for reinforced concrete beams made of recycled coarse aggregates with and without shear reinforcement was established, collecting the shear specimens available from various works in the existing literature. The database was used to statistically identify the strength margin between RCA and NCA concrete beams and to calculate its safety margin based on reliability analysis. Moreover, a comparability study of RCA beams was conducted with its control specimens and with a database for conventional RC beams.

Investigation of the Optic Nerve Head Morphology Influence to the Optic Nerve Head Biomechanics - Patient Specific Model.

Glaucoma is associated with damage and death of optic nerve fibers within the Lamina Cribrosa (LC) region of the Optic Nerve Head. The pathogenesis of the disease is unclear, and the anterior LC surface morphology of different individuals can be one of the possible contributor of glaucoma development and progression. The current study evaluates the relationship between the LC surface curvature and distribution of shear stresses on the LC surface. The patient-specific reconstructed ocular model was developed and analyzed in a finite element analysis software. In addition, the effect of elastic modulus of both sclera and LC on the shear stress was examined. Results showed that there is a correlation between the shear stress distribution and the curvature of the anterior LC surface. This finding highlights the potential significance of the LC morphology on the distribution of LC shear stress and require further investigation.

Investigation of the Optic Nerve Head Morphology Influence to the Optic Nerve Head Biomechanics - Population Specific Model.

Glaucoma is an eye disorder potentially leading to permanent blindness through the damage of optic nerves at the optic nerve head (ONH). As a critical region of optic nerve damage, the porous Lamina Cribrosa (LC) in the ONH plays a critical role in determining whether optic nerves passing through will experience apoptosis in response to shear stress. The primary cause of shear stress to the LC is the increase in intraocular pressure (IOP). Since morphology governs how mechanical stresses distributes, LC morphology could be an important factor in determining the risks of glaucoma development and progression. The current project aims at investigating how anterior LC surface morphology impacts its response to shear stress caused by IOP. Results of the current study show that steeper LC morphologies could be associated with increased average shear stress on the anterior LC surface. The effect of LC morphology was comparable to that of IOP. This highlights the potential significance of LC morphology on the distribution of IOP-induced shear stress and calls for further investigation in this area.

Strategies to enhance the stability of nanoscale zero-valent iron (NZVI) in continuous BrO3- reduction.

The reduction of bromate to bromide was successfully achieved by bimetallic catalysts with NZVI support in continuous-flow reactors. The stability of NZVI-supported bimetallic catalysts was enhanced by decelerating the iron corrosion and sequential rapid passivation of the iron-Cu-Pd ensembles under optimized reaction conditions. Thus >99% bromate removal can be continuously achieved for 11 h. The lifetime of the bimetallic catalyst was further enhanced and tested under different hydraulic retention time, catalyst loading, and initial bromate concentrations. At the optimized operation conditions, the catalyst showed a complete bromate reduction by 24 h and then the reactivity slowly decreased to 20% over the next 100 h. X-ray diffraction and X-ray photoelectron spectroscopy showed that the reactive NZVI support was oxidized to Fe(II) and Fe(III) along with Cu(0) oxidation to CuO, while the oxidation state of Pd did not change. Therefore, bromate reduction occurred on the surface of reactive NZVI support and Cu(0) particle, while Pd played a role as a hydrogenation catalyst that prolonged the lifetime of the bimetallic catalyst.

Sudden Event Monitoring of Civil Infrastructure Using Demand-Based Wireless Smart Sensors.

Wireless smart sensors (WSS) have been proposed as an effective means to reduce the high cost of wired structural health monitoring systems. However, many damage scenarios for civil infrastructure involve sudden events, such as strong earthquakes, which can result in damage or even failure in a matter of seconds. Wireless monitoring systems typically employ duty cycling to reduce power consumption; hence, they will miss such events if they are in power-saving sleep mode when the events occur. This paper develops a demand-based WSS to meet the requirements of sudden event monitoring with minimal power budget and low response latency, without sacrificing high-fidelity measurements or risking a loss of critical information. In the proposed WSS, a programmable event-based switch is implemented utilizing a low-power trigger accelerometer; the switch is integrated in a high-fidelity sensor platform. Particularly, the approach can rapidly turn on the WSS upon the occurrence of a sudden event and seamlessly transition from low-power acceleration measurement to high-fidelity data acquisition. The capabilities of the proposed WSS are validated through laboratory and field experiments. The results show that the proposed approach is able to capture the occurrence of sudden events and provide high-fidelity data for structural condition assessment in an efficient manner.

Protein nanoparticle vaccine based on flagellin carrier fused to influenza conserved epitopes confers full protection against influenza A virus challenge.

Currently marketed influenza vaccines only confer protection against matching influenza virus strains. The influenza A composition of these vaccines needs to be annually updated. Vaccines that target conserved epitopes of influenza viruses would in principle offer broad cross-protection against influenza A viruses. In our study, we investigated the specific immune responses and protective efficacy of protein nanoparticles based on fusion proteins of flagellin carrier linked to conserved influenza epitopes. We designed fusion proteins by replacing the hyperimmunogenic region of flagellin (FliC) with four tandem copies of the ectodomain of matrix protein 2 (f4M2e), H1 HA2 domain (fHApr8) or H3 HA2 domain (fHAaichi). Protein nanoparticles fabricated from these fusion proteins by using DTSSP crosslinking retained Toll-like receptor 5 agonist activity of FliC. Intranasal immunization with f4M2e, f4M2e/fHApr8 or f4M2e/fHAaichi nanoparticles induced vaccine antigen-specific humoral immune responses. It was also found that the incorporation of the H1 HA2 domain into f4M2e/fHApr8 nanoparticles boosted M2e specific antibody responses. Immunized mice were fully protected against lethal doses of virus challenge.

Coated protein nanoclusters from influenza H7N9 HA are highly immunogenic and induce robust protective immunity.

Recurring influenza viruses pose an annual threat to public health. A time-saving, cost-effective and egg-independent influenza vaccine approach is important particularly when responding to an emerging pandemic. We fabricated coated, two-layer protein nanoclusters from recombinant trimeric hemagglutinin from an avian-origin H7N9 influenza A virus as an approach for vaccine development in response to an emerging pandemic. Assessment of the virus-specific immune responses and protective efficacy in mice immunized with the nanoclusters demonstrated that the vaccine candidates were highly immunogenic, able to induce protective immunity and long-lasting humoral antibody responses to this virus without the use of adjuvants. Because the advantages of the highly immunogenic coated nanoclusters also include rapid productions in an egg-independent system, this approach has great potential for influenza vaccine production not only in response to an emerging pandemic, but also as a replacement for conventional seasonal influenza vaccines.

A Novel R848-Conjugated Inactivated Influenza Virus Vaccine Is Efficacious and Safe in a Neonate Nonhuman Primate Model.

Influenza virus infection of neonates poses a major health concern, often resulting in severe disease and hospitalization. At present, vaccines for this at-risk population are lacking. Thus, development of an effective vaccine is an urgent need. In this study, we have used an innovative nonhuman primate neonate challenge model to test the efficacy of a novel TLR 7/8 agonist R848-conjugated influenza virus vaccine. The use of the intact virus represents a step forward in conjugate vaccine design because it provides multiple antigenic targets allowing for elicitation of a broad immune response. Our results show that this vaccine induces high-level virus-specific Ab- and cell-mediated responses in neonates that result in increased virus clearance and reduced lung pathology postchallenge compared with the nonadjuvanted virus vaccine. Surprisingly, the addition of a second TLR agonist (flagellin) did not enhance vaccine protection, suggesting that combinations of TLR that provide increased efficacy must be determined empirically. These data support further exploration of this new conjugate influenza vaccine approach as a platform for use in the at-risk neonate population.

Blimp-1/PRDM1 regulates the transcription of human CS1 (SLAMF7) gene in NK and B cells.

CS1 (CRACC/CD319/SLAMF7) is a member of SLAM (Signaling Lymphocyte Activation Molecule) family receptors and is expressed on NK cells, a subset of CD8(+) T lymphocytes, activated monocytes, mature dendritic cells and activated B cells. In NK cells, CS1 signaling induces cytolytic function of NK cells against targets whereas in B cells CS1 induces proliferation and autocrine cytokine production. CS1 is upregulated in multiple myeloma cells and contributes to clonogenic growth and tumorigenicity. However, the mechanism of CS1 upregulation is unknown. In this study, we analyzed the transcriptional regulation of human CS1 gene in NK and B cells. The promoter region of CS1 contains a Blimp-1/PRDM1 binding site and relative luciferase activities of successive deletion mutants of CS1 promoter were different between Blimp-1/PRDM1-positive and Blimp-1/PRDM1-negative cells. Proximal region of CS1 promoter contains a CAAT box and atypical TATA-box that might result in common transcription initiation at -29 nucleotides upstream of the ATG translation start codon. Electrophoretic Mobility Shift Assay (EMSA) and Chromatin Immunoprecipitation (ChIP) assays revealed Blimp-1/PRDM1 binds to the CS1 promoter region. Mutating the Blimp-1/PRDM1 site at -750 to -746 decreased the transcriptional activity of CS1 promoter implicating a trans-activating function of Blimp-1/PRDM1 in human CS1 gene regulation. The finding that Blimp-1/PRDM1 enhances transcription of CS1 gene in multiple myeloma cells may help in developing novel strategies for therapeutic intervention in multiple myeloma.

Inclusion of Flagellin during Vaccination against Influenza Enhances Recall Responses in Nonhuman Primate Neonates.

UNLABELLED: Influenza virus can cause life-threatening infections in neonates and young infants. Although vaccination is a major countermeasure against influenza, current vaccines are not approved for use in infants less than 6 months of age, in part due to the weak immune response following vaccination. Thus, there is a strong need to develop new vaccines with improved efficacy for this vulnerable population. To address this issue, we established a neonatal African green monkey (AGM) nonhuman primate model that could be used to identify effective influenza vaccine approaches for use in young infants. We assessed the ability of flagellin, a Toll-like receptor 5 (TLR5) agonist, to serve as an effective adjuvant in this at-risk population. Four- to 6-day-old AGMs were primed and boosted with inactivated PR8 influenza virus (IPR8) adjuvanted with either wild-type flagellin or inactive flagellin with a mutation at position 229 (m229), the latter of which is incapable of signaling through TLR5. Increased IgG responses were observed following a boost, as well as at early times after challenge, in infants vaccinated with flagellin-adjuvanted IPR8. Inclusion of flagellin during vaccination also resulted in a significantly increased number of influenza virus-specific T cells following challenge compared to the number in infants vaccinated with the m229 adjuvant. Finally, following challenge infants vaccinated with IPR8 plus flagellin exhibited a reduced pathology in the lungs compared to that in infants that received IPR8 plus m229. This study provides the first evidence of flagellin-mediated enhancement of vaccine responses in nonhuman primate neonates. IMPORTANCE: Young infants are particularly susceptible to severe disease as a result of influenza virus infection. Compounding this is the lack of effective vaccines for use in this vulnerable population. Here we describe a vaccine approach that results in improved immune responses and protection in young infants. Incorporation of flagellin during vaccination resulted in increased antibody and T cell responses together with reduced disease following virus infection. These results suggest that flagellin may serve as an effective adjuvant for vaccines targeted to this vulnerable population.

CS1 (SLAMF7) inhibits production of proinflammatory cytokines by activated monocytes.

OBJECTIVE AND DESIGN: CS1 (CRACC, CD319, SLAMF7) is a member of the Signaling Lymphocyte Activation Molecule family expressed on immune cells mediating host defense. CS1 is a self-ligand and has both activating and inhibitory functions in Natural Killer cells. However, the function of CS1 in human monocytes is currently unknown. The objective of this study was to evaluate the control of CS1 surface expression in activated monocytes and to assess the effect of CS1 triggering on proinflammatory cytokine production by monocytes. MATERIAL, METHODS AND TREATMENT: Human monocytes were isolated from PBMC of healthy volunteers by magnetic depletion method or FACS sorting. The monocytes were cultured with or without LPS (1 µg/ml) in the presence or absence of various pharmacological inhibitors to inhibit NF-кB and PI3K signaling pathways. The cells were stimulated with anti-CS1 antibody or isotype control. Total RNA was extracted and RT-PCR was performed using specific primers for CS1 and EAT-2. Cell supernatants were collected and cytokine levels (TNF-α and IL-12p70) were determined by sandwich ELISA. RESULTS: Our study revealed that adherent or LPS-activated monocytes express CS1, and CS1 induction is via NF-кB and PI3K pathways. Importantly, cross-linking CS1 resulted in reduced production of proinflammatory cytokines TNF-α and IL-12p70 by LPS-activated monocytes. CONCLUSIONS: Our study demonstrated that CS1 plays an inhibitory role in human monocytes to control proinflammatory immune responses.

Mineralisation of reconstituted collagen using polyvinylphosphonic acid/polyacrylic acid templating matrix protein analogues in the presence of calcium, phosphate and hydroxyl ions.

The complex morphologies of mineralised collagen fibrils are regulated through interactions between the collagen matrix and non-collagenous extracellular proteins. In the present study, polyvinylphosphonic acid, a biomimetic analogue of matrix phosphoproteins, was synthesised and confirmed with FTIR and NMR. Biomimetic mineralisation of reconstituted collagen fibrils devoid of natural non-collagenous proteins was demonstrated with TEM using a Portland cement-containing resin composite and a phosphate-containing fluid in the presence of polyacrylic acid as sequestration, and polyvinylphosphonic acid as templating matrix protein analogues. In the presence of these dual biomimetic analogues in the mineralisation medium, intrafibrillar and extrafibrillar mineralisation via bottom-up nanoparticle assembly based on the non-classical crystallisation pathway could be identified. Conversely, only large mineral spheres with no preferred association with collagen fibrils were observed in the absence of biomimetic analogues in the medium. Mineral phases were evident within the collagen fibrils as early as 4 h after the initially-formed amorphous calcium phosphate nanoprecursors were transformed into apatite nanocrystals. Selected area electron diffraction patterns of highly mineralised collagen fibrils were nearly identical to those of natural bone, with apatite crystallites preferentially aligned along the collagen fibril axes.

Functional role of human NK cell receptor 2B4 (CD244) isoforms.

2B4 (CD244), a member of the signaling lymphocyte-activation molecule (SLAM/CD150), is expressed on all NK cells, a subpopulation of T cells, monocytes and basophils. Human NK cells express two isoforms of 2B4, h2B4-A and h2B4-B that differ in a small portion of the extracellular domain. In the present investigation, we have studied the functions of h2B4-A and h2B4-B. Our study demonstrated that these two isoforms differ in their binding affinity for CD48, which results in differential cytotoxic activity as well as intracellular calcium release by NK cells upon target cell recognition. Analysis of the predicted 3-D structure of the two isoforms showed conformational differences that could account for their differences in binding affinity to CD48. h2B4-A was able to mediate natural cytotoxicity against CD48-expressing K562 target cells and induce intracellular calcium release, whereas h2B4-B showed no effects. NK-92MI, U937, THP-1, KU812, primary monocytes, basophils and NK cells showed expression of both h2B4-A and h2B4-B whereas YT and IL-2-activated NK cells did not show any h2B4-B expression. Stimulation of NK cells through 2B4 resulted in decreased mRNA levels of both h2B4-A and h2B4-B indicating that down-regulation of 2B4 isoforms may be an important factor in controlling NK cell activation during immune responses.

Human natural killer cell receptor 2B4 (CD244) down-regulates its own expression by reduced promoter activity at an Ets element.

2B4 (CD244), a member of the CD2 subset of the immunoglobulin superfamily, is important for stimulating human natural killer (NK) cell cytotoxicity and cytokine production. It is expressed on all NK cells, a subpopulation of T cells, monocytes, basophils and eosinophils. 2B4 interaction with its ligand CD48 regulates NK, T and B lymphocyte functions and thus plays a central role in various immune responses. Previous study indicated a role for AP-1 and Ets in the transcription of the 2B4 gene. In this study we report that stimulation of NK cells through surface 2B4 down-regulates its own expression due to a reduction in the promoter activity at the Ets element. The down-regulation of 2B4 could be a mechanism to attenuate the co-stimulatory signal from 2B4--CD48 interactions.

Human CD57+ germinal center-T cells are the major helpers for GC-B cells and induce class switch recombination.

BACKGROUND: The function of CD57+ CD4+ T cells, constituting a major subset of germinal center T (GC-Th) cells in human lymphoid tissues, has been unclear. There have been contradictory reports regarding the B cell helping function of CD57+ GC-Th cells in production of immunoglobulin (Ig). Furthermore, the cytokine and co-stimulation requirement for their helper activity remains largely unknown. To clarify and gain more insight into their function in helping B cells, we systematically investigated the capacity of human tonsil CD57+ GC-Th cells in inducing B cell Ig synthesis. RESULTS: We demonstrated that CD57+ GC-Th cells are highly efficient in helping B cell production of all four subsets of Ig (IgM, IgG, IgA and IgE) compared to other T-helper cells located in germinal centers or interfollicular areas. CD57+ GC-Th cells were particularly more efficient than other T cells in helping GC-B cells but not naive B cells. CD57+ GC-Th cells induced the expression of activation-induced cytosine deaminase (AID) and class switch recombination in developing B cells. IgG1-3 and IgA1 were the major Ig isotypes induced by CD57+ GC-Th cells. CD40L, but not IL-4, IL-10 and IFN-gamma, was critical in CD57+ GC-Th cell-driven B cell production of Ig. However, IL-10, when added exogenously, significantly enhanced the helper activity of CD57+ GC-Th cells, while TGF-beta1 completely and IFN-gamma partially suppressed the CD57+ GC-Th cell-driven Ig production. CONCLUSIONS: CD57+CD4+ T cells in the germinal centers of human lymphoid tissues are the major T helper cell subset for GC-B cells in Ig synthesis. Their helper activity is consistent with their capacity to induce AID and class switch recombination, and can be regulated by CD40L, IL-4, IL-10 and TGF-beta.

Unique gene expression program of human germinal center T helper cells.

Gene expression profiling was used to compare the gene expression patterns of human germinal center (GC) T helper (Th) cells with other CD4+ T-cell subsets (naive, central, and effector memory T cells). GC-Th cells, specifically localized in germinal centers to help B cells, are distantly related to central and effector memory T cells in global gene expression profiles. GC-Th cells displayed substantial differences in mRNA for adhesion molecules, chemoattractant receptors, and cytokines compared with other populations. Distinct expression of transcriptional factors by GC-Th cells is consistent with the hypothesis that they may be different from other T cells in cell lineage. Interestingly, CXCL13, a critical chemokine for B-cell entry to lymphoid follicles, is one of the most highly up-regulated genes in GC-Th cells. GC-Th cells (but not other T cells) produce and secrete large amounts of functional CXCL13 upon T-cell receptor activation, a process that is dependent on costimulation, requires translation and transcription, and is dramatically enhanced by activation in the presence of GC-B cells. This study revealed for the first time the unique gene expression program of GC-Th cells.