ABSTRACT
A ruthenium-catalyzed C-H activation and a concomitant [4+2] annulation of sulfoxonium ylide with maleimide have been demonstrated. This tandem reaction results in the formation of fused benzo[e]isoindole-1,3,5-trione. The method employs mild conditions and is free of metal oxidants. The reaction pathway predominantly involves protodemetalation over ß-hydride elimination due to the lack of syn ß-hydrogens.
ABSTRACT
Bruton's tyrosine kinase (BTK) regulates diverse cellular signaling of the innate and adaptive immune system in response to microbial pathogens. Downregulation or constitutive activation of BTK is reported in patients with autoimmune diseases or various B-cell leukemias. BTK is a multidomain protein tyrosine kinase that adopts an Src-like autoinhibited conformation maintained by the interaction between the kinase and PH-TH domains. The PH-TH domain plays a central role in regulating BTK function. BTK is activated by binding to PIP3 at the plasma membrane upon stimulation by the B-cell receptor (BCR). The PIP3 binding allows dimerization of the PH-TH domain and subsequent transphosphorylation of the activation loop. Alternatively, a recent study shows that the multivalent T-cell-independent (TI) antigen induces BCR response by activating BTK independent of PIP3 binding. It was proposed that a transiently stable IP6-dependent PH-TH dimer may activate BTK during BCR activation by the TI antigens. However, no IP6-dependent PH-TH dimer has been identified yet. Here, we investigated a constitutively active PH-TH mutant (E41K) to determine if the elusive IP6-dependent PH-TH dimer exists. We showed that the constitutively active E41K mutation activates BTK by stabilizing the IP6-dependent PH-TH dimer. We observed that a downregulating mutation in the PH-TH domain (R28H) linked to X-linked agammaglobulinemia impairs BTK activation at the membrane and in the cytosol by preventing PH-TH dimerization. We conclude that the IP6 dynamically remodels the BTK active fraction between the membrane and the cytoplasm. Stimulating with IP6 increases the cytosolic fraction of the activated BTK.
Subject(s)
Agammaglobulinaemia Tyrosine Kinase , Phytic Acid , Agammaglobulinaemia Tyrosine Kinase/metabolism , Agammaglobulinaemia Tyrosine Kinase/genetics , Agammaglobulinaemia Tyrosine Kinase/chemistry , Humans , Phytic Acid/metabolism , Animals , Protein-Tyrosine Kinases/metabolism , Protein-Tyrosine Kinases/genetics , Protein-Tyrosine Kinases/chemistry , Protein Multimerization , Receptors, Antigen, B-Cell/metabolism , Receptors, Antigen, B-Cell/genetics , Enzyme Activation , Mutation, Missense , Mice , Agammaglobulinemia/genetics , Agammaglobulinemia/metabolism , PhosphorylationABSTRACT
Corn silk, often considered as a waste material in sweet corn processing, is typically discarded by most food manufacturing industries. This study aims to maximize the utilization of corn silk by evaluating its phytochemical, physicochemical and sensory characteristics. The development of food products with enhanced nutritional value is a pressing concern for both scientists and food producers in the industry. Therefore, this research focuses on the creation of highly nutritious muffins incorporating underutilized corn silk powder (CSP). In the muffin preparation, CSP was used to partially replace refined wheat flour at levels of 10%, 20%, 30% and 40%. As the proportion of CSP increased, the protein and crude fiber content of the muffins gradually increased. Additionally, the total phenolic content and antioxidant activity of the muffins significantly increased (p ≤ 0.05) with the inclusion of CSP, reaching their maximum values when CSP was used to replace 40% of the refined wheat flour. The incorporation of CSP led to a decrease in the L* (lightness) value, resulting in lower a* (redness) and b* (yellowness) values in the muffins. Texture analysis revealed that the cohesiveness, chewiness and gumminess of the muffins increased as the amount of CSP in the recipe was raised. A sensory evaluation was conducted to assess the acceptability of the corn silk muffins. The addition of CSP in muffins improved the sensory characteristics including colour, aroma, mouthfeel, texture and overall acceptability. These findings indicate that CSP has the potential to be used in the development of bakery food products, instant mixes, infant food formulas and value-added items.
ABSTRACT
Photocatalysis is recognized to be one of the most promising ways to address energy and environmental issues by utilizing visible light. Graphitic carbon nitride (g-C3N4), with a moderate band gap (â¼2.7 eV) has been the flashpoint in environmental photocatalysis as it can work better under visible light, can be synthesized by a facile synthesis process using low-cost materials, thermally and chemically stable. Still the photocatalytic performance of g-C3N4 is not satisfactory because of certain limitations such as insufficient visible light absorption capacity, low electron-hole separation efficiency, high recombination rate, poor surface area. Introduction of doping, band structure engineering, defecting and designing of heterojunction, composites etc. were investigated to amplify its applications. Among all these modifications, elemental doping is a suitable and successful alternative for the enhancement of the photocatalytic activity by changing the optical and electronic properties. This review emphasizes on advancement and trends of elemental doping and its application on photocatalytic organic pollutant remediation in aqueous medium. The fundamental photocatalytic activity of heterogeneous photocatalysis and specifically g-C3N4-based photocatalysis have been discussed. The benfits of non-metal doping, enhanced photocatalytic performance by doping element, mechanism invloved in doping, advantages of co-doping has been explained. Mono, bi, and tri non-metal doped g-C3N4 and their application for the removal of organic pollutants from water medium by visible light photocatalysis has been summerized. Life cycle assessment (LCA) of photocatalytic system has been highlighted. Future research should focus on the large-scale application of the photocatalysis process considering the economic aspects. A rigorous life cycle assessment for deploying the non-metal doped g-C3N4-based photocatalysis technology for successful commercial application is recommended.
Subject(s)
Graphite , Catalysis , Graphite/chemistry , Environmental Restoration and Remediation/methods , Nitrogen Compounds/chemistry , Water Pollutants, Chemical/chemistry , Water Pollutants, Chemical/analysis , Photochemical Processes , Light , Nitriles/chemistryABSTRACT
Nuclear factor kappa beta (NF-κB) plays a pivotal role in breast cancer, particularly triple-negative breast cancer, by promoting inflammation, proliferation, epithelial-mesenchymal transition, metastasis, and drug resistance. Upregulation of NF-κB boosts vascular endothelial growth factor (VEGF) expression, assisting angiogenesis. The Ru(II) complexes of methyl- and dimethylpyrazolyl-benzimidazole N,N donors inhibit phosphorylation of ser536 in p65 and translocation of the NF-κB heterodimer (p50/p65) to the nucleus, disabling transcription to upregulate inflammatory signaling. The methyl- and dimethylpyrazolyl-benzimidazole inhibit VEGFR2 phosphorylation at Y1175, disrupting downstream signaling through PLC-γ and ERK1/2, ultimately suppressing Ca(II)-signaling. Partial release of the antiangiogenic ligand in a reactive oxygen species-rich environment is possible as per our observation to inhibit both NF-κB and VEGFR2 by the complexes. The complexes are nontoxic to zebrafish embryos up to 50 µM, but the ligands show strong in vivo antiangiogenic activity at 3 µM during embryonic growth in Tg(fli1:GFP) zebrafish but no visible effect on the adult phase.
Subject(s)
NF-kappa B , Triple Negative Breast Neoplasms , Humans , Animals , NF-kappa B/metabolism , Zebrafish/metabolism , Transcription Factor RelA/metabolism , Triple Negative Breast Neoplasms/drug therapy , Vascular Endothelial Growth Factor A , Ligands , Benzimidazoles/pharmacologyABSTRACT
Ligand-independent activation of VEGFRs is a hallmark of diabetes and several cancers. Like EGFR, VEGFR2 is activated spontaneously at high receptor concentrations. VEGFR1, on the other hand, remains constitutively inactive in the unligated state, making it an exception among VEGFRs. Ligand stimulation transiently phosphorylates VEGFR1 and induces weak kinase activation in endothelial cells. Recent studies, however, suggest that VEGFR1 signaling is indispensable in regulating various physiological or pathological events. The reason why VEGFR1 is regulated differently from other VEGFRs remains unknown. Here, we elucidate a mechanism of juxtamembrane inhibition that shifts the equilibrium of VEGFR1 towards the inactive state, rendering it an inefficient kinase. The juxtamembrane inhibition of VEGFR1 suppresses its basal phosphorylation even at high receptor concentrations and transiently stabilizes tyrosine phosphorylation after ligand stimulation. We conclude that a subtle imbalance in phosphatase activation or removing juxtamembrane inhibition is sufficient to induce ligand-independent activation of VEGFR1 and sustain tyrosine phosphorylation.
Subject(s)
Endothelial Cells , Vascular Endothelial Growth Factor Receptor-1 , Endothelial Cells/metabolism , Ligands , Vascular Endothelial Growth Factor Receptor-1/genetics , Vascular Endothelial Growth Factor Receptor-1/metabolism , Vascular Endothelial Growth Factor Receptor-2/genetics , Vascular Endothelial Growth Factor Receptor-2/metabolism , Signal Transduction/physiology , Cell Membrane/metabolism , Tyrosine/metabolismABSTRACT
Malaria parasite lacks canonical pathways for amino acid biosynthesis and depends primarily on hemoglobin degradation and extracellular resources for amino acids. Interestingly, a putative gene for glutamine synthetase (GS) is retained despite glutamine being an abundant amino acid in human and mosquito hosts. Here we show Plasmodium GS has evolved as a unique type I enzyme with distinct structural and regulatory properties to adapt to the asexual niche. Methionine sulfoximine (MSO) and phosphinothricin (PPT) inhibit parasite GS activity. GS is localized to the parasite cytosol and abundantly expressed in all the life cycle stages. Parasite GS displays species-specific requirement in Plasmodium falciparum (Pf) having asparagine-rich proteome. Targeting PfGS affects asparagine levels and inhibits protein synthesis through eIF2α phosphorylation leading to parasite death. Exposure of artemisinin-resistant Pf parasites to MSO and PPT inhibits the emergence of viable parasites upon artemisinin treatment.
Subject(s)
Artemisinins , Parasites , Animals , Humans , Glutamate-Ammonia Ligase/genetics , Glutamate-Ammonia Ligase/metabolism , Asparagine/genetics , Amino Acids , Glutamine/metabolism , Plasmodium falciparum/genetics , Plasmodium falciparum/metabolism , Artemisinins/pharmacology , Parasites/genetics , Parasites/metabolismABSTRACT
Age-related immune dysfunctions, such as decreased T-cell output, are closely related to pathologies like cancers and lack of vaccine efficacy among the elderly. Engineered fusokine, GIFT-7, a fusion of interleukin 7 (IL-7) and GM-CSF, can reverse aging-related lymphoid organ atrophy. We generated a GIFT-7 fusokine tumor vaccine and employed it in aged syngeneic mouse models of glioblastoma and found that peripheral vaccination with GIFT-7TVax resulted in thymic regeneration and generated durable long-term antitumor immunity specifically in aged mice. Global cytokine analysis showed increased pro-inflammatory cytokines including IL-1ß in the vaccinated group that resulted in hyperactivation of dendritic cells. In addition, GIFT-7 vaccination resulted in increased T-cell trafficking to the brain and robust Th-17 long-term effector memory T-cell formation. TCR-seq analysis showed increased productive frequency among detected rearrangements within the vaccinated group. Overall, our data demonstrate that aging immune system can be therapeutically augmented to generate lasting antitumor immunity.
Subject(s)
Cancer Vaccines , Glioblastoma , Mice , Animals , Cytokines , Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology , Interleukin-7/pharmacology , Glioblastoma/therapyABSTRACT
The food vacuole plays a central role in the blood stage of parasite development by digesting host hemoglobin acquired from red blood cells and detoxifying the host heme released during hemoglobin digestion into hemozoin. Blood-stage parasites undergo periodic schizont bursts, releasing food vacuoles containing hemozoin. Clinical studies in malaria-infected patients and in vivo animal studies have shown the association of hemozoin with disease pathogenesis and abnormal host immune responses in malaria. Here, we perform a detailed in vivo characterization of putative Plasmodium berghei amino acid transporter 1 localized in the food vacuole to understand its significance in the malaria parasite. We show that the targeted deletion of amino acid transporter 1 in Plasmodium berghei leads to a swollen food vacuole phenotype with the accumulation of host hemoglobin-derived peptides. Plasmodium berghei amino acid transporter 1-knockout parasites produce less hemozoin, and the hemozoin crystals display a thin morphology compared with wild-type parasites. The knockout parasites show reduced sensitivity to chloroquine and amodiaquine by showing recrudescence. More importantly, mice infected with the knockout parasites are protected from cerebral malaria and display reduced neuronal inflammation and cerebral complications. Genetic complementation of the knockout parasites restores the food vacuole morphology with hemozoin levels similar to that of wild-type parasites, causing cerebral malaria in the infected mice. The knockout parasites also show a significant delay in male gametocyte exflagellation. Our findings highlight the significance of amino acid transporter 1 in food vacuole functionality and its association with malaria pathogenesis and gametocyte development. IMPORTANCE Food vacuoles of the malaria parasite are involved in the degradation of red blood cell hemoglobin. The amino acids derived from hemoglobin degradation support parasite growth, and the heme released is detoxified into hemozoin. Antimalarials such as quinolines target hemozoin formation in the food vacuole. Food vacuole transporters transport hemoglobin-derived amino acids and peptides from the food vacuole to the parasite cytosol. Such transporters are also associated with drug resistance. Here, we show that the deletion of amino acid transporter 1 in Plasmodium berghei leads to swollen food vacuoles with the accumulation of hemoglobin-derived peptides. The transporter-deleted parasites generate less hemozoin with thin crystal morphology and show reduced sensitivity to quinolines. Mice infected with transporter-deleted parasites are protected from cerebral malaria. There is also a delay in male gametocyte exflagellation, affecting transmission. Our findings uncover the functional significance of amino acid transporter 1 in the life cycle of the malaria parasite.
ABSTRACT
The Chenopodium genus includes >250 species, among which only quinoa, pigweed, djulis, and kaniwa have been explored for starches. Chenopodium is a non-conventional and rich source of starch, which has been found effective in producing different classes of food. Chenopodium starches are characterized by their smaller granule size (0.4-3.5 µm), higher swelling index, shorter/lower gelatinization regions/temperature, good emulsifying properties, and high digestibility, making them suitable for food applications. However, most of the investigations into Chenopodium starches are in the primary stages (isolation, modification, and characterization), except for quinoa. This review comprehensively explores the major developments in Chenopodium starch research, emphasizing isolation, structural composition, functionality, hydrolysis, modification, and application. A critical analysis of the trends, limitations, and scope of these starches for novel food applications has also been provided to promote further scientific advancement in the field.
Subject(s)
Amaranthus , Chenopodium quinoa , Chenopodium , Starch/chemistry , Chenopodium quinoa/chemistry , Temperature , Amylose/chemistryABSTRACT
T cell signaling starts with assembling several tyrosine kinases and adapter proteins to the T cell receptor (TCR), following the antigen binding to the TCR. The stability of the TCR-antigen complex and the delay between the recruitment and activation of each kinase determines the T cell response. Integration of such delays constitutes a kinetic proofreading mechanism to regulate T cell response to the antigen binding. However, the mechanism of these delays is not fully understood. Combining biochemical experiments and kinetic modeling, here we report a thermodynamic brake in the regulatory module of the tyrosine kinase ZAP-70, which determines the ligand selectivity, and may delay the ZAP-70 activation upon antigen binding to TCR. The regulatory module of ZAP-70 comprises of a tandem SH2 domain that binds to its ligand, doubly-phosphorylated ITAM peptide (ITAM-Y2P), in two kinetic steps: a fast step and a slow step. We show the initial encounter complex formation between the ITAM-Y2P and tandem SH2 domain follows a fast-kinetic step, whereas the conformational transition to the holo-state follows a slow-kinetic step. We further observed a thermodynamic penalty imposed during the second phosphate-binding event reduces the rate of structural transition to the holo-state. Phylogenetic analysis revealed the evolution of the thermodynamic brake coincides with the divergence of the adaptive immune system to the cell-mediated and humoral responses. In addition, the paralogous kinase Syk expressed in B cells does not possess such a functional thermodynamic brake, which may explain the higher basal activation and lack of ligand selectivity in Syk.
Subject(s)
Evolution, Molecular , Receptors, Antigen, T-Cell , T-Lymphocytes , ZAP-70 Protein-Tyrosine Kinase , Ligands , Phosphorylation , Phylogeny , Receptors, Antigen, T-Cell/genetics , Receptors, Antigen, T-Cell/metabolism , T-Lymphocytes/enzymology , Thermodynamics , Animals , ZAP-70 Protein-Tyrosine Kinase/chemistry , src Homology DomainsABSTRACT
Adipogenic differentiation of visceral adipose tissue-resident multipotent mesenchymal stromal cells (VA-MSC) into adipocytes is metabolically protective. Under chronic inflammatory stress, this neoadipogenesis process is suppressed by various pro-inflammatory cytokines and growth factors. However, the underlying mechanism(s) regulating VA-MSC plasticity remains largely unexplored. Using an adipogenic differentiation screen, we identified IFNγ and TGFß as key inhibitors of primary human VA-MSC differentiation. Further studies using human and mouse VA-MSCs and a chronic high-fat diet-fed murine model revealed that IFNγ/JAK2-activated STAT5 transcription factor is a central regulator of VA-MSC differentiation under chronic inflammatory conditions. Furthermore, our results indicate that under such conditions, IFNγ-activated STAT5 and TGFß-activated Smad3 physically interact via Smad4. This STAT5-Smad4-Smad3 complex plays a crucial role in preventing the early adipogenic commitment of VA-MSCs by suppressing key pro-adipogenic transcription factors, including CEBPδ, CEBPα, and PPARγ. Genetic or pharmacological disruption of IFNγ-TGFß synergy by inhibiting either STAT5 or Smad3 rescued adipogenesis under chronic inflammatory stress. Overall, our study delineates a central mechanism of MSC plasticity regulation by the convergence of multiple inflammatory signaling pathways.
ABSTRACT
Heme-biosynthetic pathway of malaria parasite is dispensable for asexual stages, but essential for mosquito and liver stages. Despite having backup mechanisms to acquire hemoglobin-heme, pathway intermediates and/or enzymes from the host, asexual parasites express heme pathway enzymes and synthesize heme. Here we show heme synthesized in asexual stages promotes cerebral pathogenesis by enhancing hemozoin formation. Hemozoin is a parasite molecule associated with inflammation, aberrant host-immune responses, disease severity and cerebral pathogenesis. The heme pathway knockout parasites synthesize less hemozoin, and mice infected with knockout parasites are protected from cerebral malaria and death due to anemia is delayed. Biosynthetic heme regulates food vacuole integrity and the food vacuoles from knockout parasites are compromised in pH, lipid unsaturation and proteins, essential for hemozoin formation. Targeting parasite heme synthesis by griseofulvin-a FDA-approved antifungal drug, prevents cerebral malaria in mice and provides an adjunct therapeutic option for cerebral and severe malaria.
Subject(s)
Malaria, Cerebral , Parasites , Animals , Griseofulvin/pharmacology , Heme/metabolism , Hemoglobins , Malaria, Cerebral/drug therapy , Malaria, Cerebral/prevention & control , Mice , Parasites/metabolismABSTRACT
Lung squamous-cell carcinoma originates as a consequence of oncogenic molecular variants arising from diverse mutagenic processes such as tobacco, defective homologous recombination, aging, and cytidine deamination by APOBEC proteins. Only some of the many variants generated by these processes actually contribute to tumorigenesis. Therefore, molecular investigation of mutagenic processes such as cytidine deamination by APOBEC should also determine whether the mutations produced by these processes contribute substantially to the growth and survival of cancer. Here, we determine the processes that gave rise to mutations of 681 lung squamous-cell carcinomas, and quantify the probability that each mutation was the product of each process. We then calculate the contribution of each mutation to increases in cellular proliferation and survival. We performed in vitro experiments to determine cytidine deamination activity of APOBEC3B against oligonucleotides corresponding with genomic sequences that give rise to variants of high cancer effect size. The largest APOBEC-related cancer effects are attributable to mutations in PIK3CA and NFE2L2. We demonstrate that APOBEC effectively deaminates NFE2L2 at the locations that confer high cancer effect. Overall, we demonstrate that APOBEC activity can lead to mutations in NFE2L2 that have large contributions to cancer cell growth and survival, and that NFE2L2 is an attractive potential target for therapeutic intervention.
Subject(s)
Carcinoma, Squamous Cell , Lung Neoplasms , Carcinogenesis , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Squamous Cell/genetics , Cytidine/metabolism , Cytidine Deaminase/genetics , Cytidine Deaminase/metabolism , Humans , Lung/metabolism , Lung Neoplasms/genetics , Minor Histocompatibility Antigens/genetics , Mutagenesis , Mutation/genetics , NF-E2-Related Factor 2/geneticsABSTRACT
Allogeneic islet transplantation is a promising experimental therapy for poorly controlled diabetes. Despite pharmacological immunosuppression, long-term islet engraftment remains elusive. Here, we designed a synthetic fusion transgene coupling PD-L1 and indoleamine dioxygenase [hereafter PIDO] whose constitutive expression prevents immune destruction of genetically engineered islet allograft transplanted in immunocompetent mice. PIDO expressing murine islets maintain robust dynamic insulin secretion in vitro and when transplanted in allogeneic hyperglycemic murine recipients reverse pre-existing streptozotocin-induced and autoimmune diabetes in the absence of pharmacological immunosuppression for more than 50 and 8 weeks, respectively, and is dependent on host CD4 competence. Additionally, PIDO expression in allografts preserves endocrine functional viability of islets and promotes a localized tolerogenic milieu characterized by the suppression of host CD8 T cell and phagocyte recruitment and accumulation of FOXP3+ Tregs. Furthermore, in the canine model of xenogeneic islet transplantation, muscle implanted PIDO-expressing porcine islets displayed physiological glucose-responsive insulin secretion competency in euglycemic recipient for up to 20 weeks. In conclusion, the PIDO transgenic technology enables host CD4+ T cell-modulated immune evasiveness and long-term functional viability of islet allo- and xenografts in immune-competent recipients without the need for pharmacological immune suppression and would allow for improved outcomes for tissue transplantation.
Subject(s)
Diabetes Mellitus, Type 1 , Islets of Langerhans Transplantation , Islets of Langerhans , Animals , Dogs , Humans , Mice , Allografts , B7-H1 Antigen/metabolism , Graft Rejection/prevention & control , Graft Survival , Immunosuppression Therapy , Islets of Langerhans/metabolism , Mice, Inbred C57BL , Swine , Indoleamine-Pyrrole 2,3,-DioxygenaseABSTRACT
Preserving islet health and function is critical during pretransplant culture to improve islet transplantation outcome and for ex vivo modeling of diabetes for pharmaceutical drug discovery. The limited islet engraftment potential is primarily attributable to loss of extracellular matrix (ECM) support and interaction. Multipotent cells with ECM depositing competency improve islet survival during short coculture period. However, role of pancreatic stellate cells (PSCs) and their ECM support in preserving ex vivo islet physiology remains largely unknown. Here, we report novel cytoprotective effects of culture-adapted porcine PSCs and role of their ECM-mediated intercellular communication on pig, mouse and human islets ex vivo. Using direct-contact coculture system, we demonstrate that porcine PSCs preserve and significantly prolong islet viability and function from 7 ± 3 days to more than 28 ± 5 (P < .001) days in vitro. These beneficial effects of PSCs on islet health are not species-specific. Using NSC47924 to specifically inhibit 37/67 kDa laminin receptor (LR), we identified that LR-mediated intercellular communication is essential for PSCs to protect functional viability of islets in vitro. Finally, our results demonstrate that PSC co-transplantation improved function and enhanced capacity of syngeneic islets to reverse hyperglycemia in mice with preexisiting diabetes. Cumulatively, our findings unveil novel effects of culture-adapted PSCs on islet health likely mirroring in vivo niche interaction. Furthermore, islet and PSC coculture may aid in development of ex vivo diabetes modeling and also suggests that a combined islet-PSC tissue engineered implant may significantly improve islet transplantation outcome.
Subject(s)
Islets of Langerhans Transplantation , Islets of Langerhans , Animals , Coculture Techniques , Extracellular Matrix , Islets of Langerhans Transplantation/methods , Mice , Pancreatic Stellate Cells , SwineABSTRACT
BACKGROUND: For any effective vaccination strategy, the willingness of the beneficiaries and its contributing factors are important. This study was conducted among the health-care workers (HCWs) and community members to find the perceptions regarding the COVID-19 vaccine and understand the influencers and the barriers of vaccine acceptance. MATERIALS AND METHODS: A qualitative study was conducted from October 2020 to December 2020 in two primary care settings in an urban area. Eighteen in-depth interviews (IDIs) after taking consent were conducted with the help of IDI guide developed and validated beforehand by the experts. IDIs were done among the ten community members and eight HCWs selected conveniently. Data collection were continued till data saturation when no new information yielded from the interviews. Thematic analysis was performed. RESULTS: All the participants were hopeful about availability of the vaccine. The key influencers identified for promoting willingness to accept the vaccine among both the groups were opinion of the health-care providers, colleagues' and other people's acceptance of the vaccine, effectiveness of vaccine on other people, and perceived risk of the disease. Fear of adverse reactions was the most important barrier among all the respondents. The prevalent perception was that other preventive practices and vaccine together can only be the best solution to prevent COVID-19 illness. The HCWs perceived that acceptance of vaccine among the community members would be good overall but apprehended some initial difficulties. Mass campaign to promote COVID-19 vaccination and sensitization events are the need of the hour. CONCLUSIONS: Since opinion of health-care personnel emerged as an important influencer of vaccine acceptance, mass campaign and sensitization programs spearheaded by the health-care providers can bring about change by increasing the vaccine acceptance among the beneficiaries at large. Re-enforcement regarding practice of preventive measures should be made among the population irrespective of the vaccination status.
ABSTRACT
The cell-mediated immune response constitutes a robust host defense mechanism to eliminate pathogens and oncogenic cells. T cells play a central role in such a defense mechanism and creating memories to prevent any potential infection. T cell recognizes foreign antigen by its surface receptors when presented through antigen-presenting cells (APCs) and calibrates its cellular response by a network of intracellular signaling events. Activation of T-cell receptor (TCR) leads to changes in gene expression and metabolic networks regulating cell development, proliferation, and migration. TCR does not possess any catalytic activity, and the signaling initiates with the colocalization of several enzymes and scaffold proteins. Deregulation of T cell signaling is often linked to autoimmune disorders like severe combined immunodeficiency (SCID), rheumatoid arthritis, and multiple sclerosis. The TCR remarkably distinguishes the minor difference between self and non-self antigen through a kinetic proofreading mechanism. The output of TCR signaling is determined by the half-life of the receptor antigen complex and the time taken to recruit and activate the downstream enzymes. A longer half-life of a non-self antigen receptor complex could initiate downstream signaling by activating associated enzymes. Whereas, the short-lived, self-peptide receptor complex disassembles before the downstream enzymes are activated. Activation of TCR rewires the cellular metabolic response to aerobic glycolysis from oxidative phosphorylation. How does the early event in the TCR signaling cross-talk with the cellular metabolism is an open question. In this review, we have discussed the recent developments in understanding the regulation of TCR signaling, and then we reviewed the emerging role of metabolism in regulating T cell function.