RESUMEN
BACKGROUND: Metabolic reprogramming plays an essential role on lymphoma progression. Dysregulation of glutamine metabolism is implicated in natural-killer T-cell lymphoma (NKTCL) and tumor cell response to asparaginase-based anti-metabolic treatment. METHODS: To understand the metabolomic alterations and determine the potential therapeutic target of asparaginase, we assessed metabolomic profile using liquid chromatography-mass spectrometry in serum samples of 36 NKTCL patients, and integrated targeted metabolic analysis and RNA sequencing in tumor samples of 102 NKTCL patients. The biological function of solute carrier family 1 member 1 (SLC1A1) on metabolic flux, lymphoma cell growth, and drug sensitivity was further examined in vitro in NK-lymphoma cell line NK-92 and SNK-6, and in vivo in zebrafish xenograft models. FINDINGS: In NKTCL patients, serum metabolomic profile was characterized by aberrant glutamine metabolism and SLC1A1 was identified as a central regulator of altered glutaminolysis. Both in vitro and in vivo, ectopic expression of SLC1A1 increased cellular glutamine uptake, enhanced glutathione metabolic flux, and induced glutamine addiction, leading to acceleration of cell proliferation and tumor growth. Of note, SLC1A1 overexpression was significantly associated with PD-L1 downregulation and reduced cytotoxic CD3+/CD8+ T cell activity when co-cultured with peripheral blood mononuclear cells. Asparaginase treatment counteracted SLC1A1-mediated glutamine addiction, restored SLC1A1-induced impaired T-cell immunity. Clinically, high EAAT3 (SLC1A1-encoded protein) expression independently predicted superior progression-free and overall survival in 90 NKTCL patients treated with asparaginase-based regimens. INTERPRETATION: SLC1A1 functioned as an extracellular glutamine transporter, promoted tumor growth through reprogramming glutamine metabolism of NKTCL, while rendered tumor cells sensitive to asparaginase treatment. Moreover, SLC1A1-mediated modulation of PD-L1 expression might provide clinical rationale of co-targeting metabolic vulnerability and immunosuppressive microenvironment in NKTCL. FUNDING: This study was supported, in part, by research funding from the National Natural Science Foundation of China (82130004, 81830007 and 81900192), Chang Jiang Scholars Program, Shanghai Municipal Education Commission Gaofeng Clinical Medicine Grant Support (20152206 and 20152208), Clinical Research Plan of SHDC (2020CR1032B), Multicenter Clinical Research Project by Shanghai Jiao Tong University School of Medicine (DLY201601), Shanghai Chenguang Program (19CG15), Shanghai Sailing Program (19YF1430800), Medical-Engineering Cross Foundation of Shanghai Jiao Tong University (ZH2018QNA46), and Shanghai Yi Yuan Xin Xing Program.
Asunto(s)
Transportador 3 de Aminoácidos Excitadores/metabolismo , Glutamina/inmunología , Linfoma Extranodal de Células NK-T/metabolismo , Células T Asesinas Naturales/metabolismo , Animales , Asparaginasa/inmunología , Asparaginasa/metabolismo , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Línea Celular , Proliferación Celular/fisiología , Regulación hacia Abajo/inmunología , Transportador 3 de Aminoácidos Excitadores/inmunología , Femenino , Humanos , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Linfoma Extranodal de Células NK-T/inmunología , Linfoma Extranodal de Células NK-T/terapia , Masculino , Persona de Mediana Edad , Células T Asesinas Naturales/inmunología , Pez CebraRESUMEN
γδ T cell is one of the most important pathogenic immune cells in autoimmunity, especially in mucosal and epithelial diseases. Metabolism is essential for the maintenance of immune homeostasis. However, unlike αß T cells, the metabolic regulation of γδ T cell activation still remain unclear. Here, we identified glutamine metabolism as a critical regulator for the generation of IL-17-producing γδ T cells. Metabolic screening uncovered that amino acids related to glutamine metabolism increased most obviously during γδ T cell activation. Pharmaceutical blocking of glutamine impaired IL-17 production in γδ T cells both in vitro and in vivo. Mechanism studies further revealed that genes downregulated upon glutamine deprivation enriched in IL-17 and IL-23/STAT3 signaling pathways. Consistent with this, the activation of STAT3 was suppressed after glutamine blocking. More importantly, application of glutamine antagonist in vivo alleviated the progression of IL-23 induced psoriatic mice model. In addition, both the glutamine level and the expression of glutamine related enzymes were found higher in psoriasis patients when compared with healthy controls. Therefore, our work identified an important metabolic regulatory pathway in γδ T cell activation and suggested that glutamine metabolism could be used as a target for the treatment of γδ T cell related diseases.
Asunto(s)
Glutamina , Interleucina-17 , Receptores de Antígenos de Linfocitos T gamma-delta , Piel , Linfocitos T , Animales , Femenino , Glutamina/inmunología , Glutamina/metabolismo , Humanos , Inflamación/inmunología , Inflamación/metabolismo , Interleucina-17/inmunología , Interleucina-17/metabolismo , Activación de Linfocitos , Masculino , Ratones , Ratones Endogámicos BALB C , Receptores de Antígenos de Linfocitos T gamma-delta/inmunología , Receptores de Antígenos de Linfocitos T gamma-delta/metabolismo , Transducción de Señal/inmunología , Piel/inmunología , Piel/metabolismo , Linfocitos T/inmunología , Linfocitos T/metabolismoRESUMEN
Macrophages represent the first line of defence in innate immune responses and additionally serve important functions for the regulation of host inflammation and tissue homeostasis. The M1/M2 model describes the two extremes of macrophage polarization states, which can be induced by multiple stimuli, most notably by LPS/IFN-γ and IL-4/IL-13. Historically, the expression of two genes encoding for enzymes, which use the same amino acid as their substrate, iNOS and ARG1, has been used to define classically activated M1 (iNOS) and alternatively activated M2 (ARG1) macrophages. This 'arginine dichotomy' has recently become a matter of debate; however, in parallel with the emerging field of immunometabolism there is accumulating evidence that these two enzymes and their related metabolites are fundamentally involved in the intrinsic regulation of macrophage polarization and function. The aim of this review is to highlight recent advances in macrophage biology and immunometabolism with a specific focus on amino acid metabolism and their related metabolic pathways: iNOS/ARG1 (arginine), TCA cycle and OXPHOS (glutamine) as well as the one-carbon metabolism (serine, glycine).
Asunto(s)
Arginasa/metabolismo , Arginina/metabolismo , Glutamina/metabolismo , Glicina/inmunología , Macrófagos/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Serina/metabolismo , Arginasa/genética , Arginasa/inmunología , Arginina/inmunología , Diferenciación Celular/efectos de los fármacos , Ciclo del Ácido Cítrico/genética , Ciclo del Ácido Cítrico/inmunología , Regulación de la Expresión Génica , Glutamina/inmunología , Glicina/metabolismo , Humanos , Inmunidad Innata , Interferón gamma/farmacología , Lipopolisacáridos/farmacología , Activación de Macrófagos/efectos de los fármacos , Macrófagos/clasificación , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Mitocondrias/efectos de los fármacos , Mitocondrias/inmunología , Mitocondrias/metabolismo , Óxido Nítrico/inmunología , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/inmunología , Fosforilación Oxidativa , Serina/inmunologíaRESUMEN
Alteration in the Th17/Treg cell balance is implicated in various autoimmune diseases and these disease-associated pathologies. Increasing investigations have shown that glutamine metabolism regulates the differentiation of Th17 and Treg cells. Here we summarize the mechanisms by which glutamine metabolism regulates Th17/Treg cell fate. Some examples of a glutamine metabolism-dependent modulation of the development and progression of several Th17 Treg cell-associated diseases are provided afterward. This review will provide a comprehensive understanding of the importance of glutamine metabolism in the fate of Th17 Treg cell differentiation.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Diferenciación Celular/inmunología , Glutamina/inmunología , Linfocitos T Reguladores/inmunología , Células Th17/inmunología , Sistemas de Transporte de Aminoácidos/inmunología , Sistemas de Transporte de Aminoácidos/metabolismo , Animales , Enfermedades Autoinmunes/metabolismo , Autofagia/inmunología , Glutamina/metabolismo , Humanos , Transducción de Señal/inmunología , Linfocitos T Reguladores/citología , Linfocitos T Reguladores/metabolismo , Células Th17/citología , Células Th17/metabolismoRESUMEN
Rapidly proliferating tumor and immune cells need metabolic programs that support energy and biomass production. The amino acid glutamine is consumed by effector T cells and glutamine-addicted triple-negative breast cancer (TNBC) cells, suggesting that a metabolic competition for glutamine may exist within the tumor microenvironment, potentially serving as a therapeutic intervention strategy. Here, we report that there is an inverse correlation between glutamine metabolic genes and markers of T cell-mediated cytotoxicity in human basal-like breast cancer (BLBC) patient data sets, with increased glutamine metabolism and decreased T cell cytotoxicity associated with poor survival. We found that tumor cell-specific loss of glutaminase (GLS), a key enzyme for glutamine metabolism, improved antitumor T cell activation in both a spontaneous mouse TNBC model and orthotopic grafts. The glutamine transporter inhibitor V-9302 selectively blocked glutamine uptake by TNBC cells but not CD8+ T cells, driving synthesis of glutathione, a major cellular antioxidant, to improve CD8+ T cell effector function. We propose a "glutamine steal" scenario, in which cancer cells deprive tumor-infiltrating lymphocytes of needed glutamine, thus impairing antitumor immune responses. Therefore, tumor-selective targeting of glutamine metabolism may be a promising therapeutic strategy in TNBC.
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Antineoplásicos/farmacología , Linfocitos T CD8-positivos/inmunología , Proteínas Portadoras/antagonistas & inhibidores , Glutamina/inmunología , Inmunidad Celular , Linfocitos Infiltrantes de Tumor/inmunología , Neoplasias de la Mama Triple Negativas/inmunología , Animales , Linfocitos T CD8-positivos/metabolismo , Linfocitos T CD8-positivos/patología , Proteínas Portadoras/inmunología , Proteínas Portadoras/metabolismo , Línea Celular Tumoral , Femenino , Glutamina/metabolismo , Xenoinjertos , Humanos , Linfocitos Infiltrantes de Tumor/metabolismo , Linfocitos Infiltrantes de Tumor/patología , Ratones , Ratones Transgénicos , Trasplante de Neoplasias , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Neoplasias de la Mama Triple Negativas/metabolismo , Neoplasias de la Mama Triple Negativas/patologíaRESUMEN
Myeloid cells comprise a major component of the tumor microenvironment (TME) that promotes tumor growth and immune evasion. By employing a small-molecule inhibitor of glutamine metabolism, not only were we able to inhibit tumor growth, but we markedly inhibited the generation and recruitment of myeloid-derived suppressor cells (MDSCs). Targeting tumor glutamine metabolism led to a decrease in CSF3 and hence recruitment of MDSCs as well as immunogenic cell death, leading to an increase in inflammatory tumor-associated macrophages (TAMs). Alternatively, inhibiting glutamine metabolism of the MDSCs themselves led to activation-induced cell death and conversion of MDSCs to inflammatory macrophages. Surprisingly, blocking glutamine metabolism also inhibited IDO expression of both the tumor and myeloid-derived cells, leading to a marked decrease in kynurenine levels. This in turn inhibited the development of metastasis and further enhanced antitumor immunity. Indeed, targeting glutamine metabolism rendered checkpoint blockade-resistant tumors susceptible to immunotherapy. Overall, our studies define an intimate interplay between the unique metabolism of tumors and the metabolism of suppressive immune cells.
Asunto(s)
Inmunidad Celular , Macrófagos/inmunología , Células Supresoras de Origen Mieloide/inmunología , Neoplasias Experimentales/inmunología , Microambiente Tumoral/inmunología , Animales , Femenino , Glutamina/inmunología , Inmunoterapia , Macrófagos/patología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Células Supresoras de Origen Mieloide/patología , Neoplasias Experimentales/patología , Neoplasias Experimentales/terapiaRESUMEN
The only generally accepted treatment of coeliac disease (CD) is a lifelong gluten-free diet. Wheat gluten proteins include gliadins, low and high molecular weight glutenins. However, we have found significant structural variations within these protein families among different cultivars. To determine which structural motifs might be less toxic than others, we assessed five variants of α-gliadin immunodominant CD-toxic peptides synthesised as 16mers in CD T cell stimulation assays with gluten-sensitive T cell lines generated from duodenal biopsies from CD-affected individuals. The peptides harboured the overlapping T cell epitopes DQ 2.5-glia-α-2 and naturally occurring variants that differed in certain amino acids (AA). The results revealed that introduction of two selected AA substitutions in α-gliadin peptides reduced immunogenicity. A peptide with three AA substitutions involving two glutamic acids (E) and one glutamine residue (G) revealed the peptide was negative in 5:5 samples. We used CD small-intestinal organ culture to assess CD toxicity that revealed two peptides with selected substitution of both glutamic acid (E) and proline (P) residues abrogated evidence of CD toxicity.
Asunto(s)
Enfermedad Celíaca/inmunología , Gliadina/inmunología , Glútenes/inmunología , Péptidos/inmunología , Triticum/química , Aminoácidos , Duodeno/inmunología , Ácido Glutámico/inmunología , Glutamina/inmunología , Humanos , Fenómenos Inmunogenéticos , Prolina/inmunología , Linfocitos T/inmunologíaRESUMEN
PURPOSE: Diabetes is a chronic inflammatory disorder resulting in endothelial dysfunction which contributes to peripheral arterial disease and limb ischemia. Leukocytes play critical roles in vascular and tissue remodelling after ischemia. This study investigated the effects of dietary glutamine (GLN) supplementation on immune cell polarization in diabetic mice subjected to limb ischemia. METHODS: Diabetes was induced by an intraperitoneal injection of streptozotocin for 5 consecutive days in C57BL/6J mice. Diabetic mice were fed the AIN-93 diet or an AIN-93 diet in which a part of the casein was replaced by GLN. After 3 weeks of the dietary intervention, mice were subjected to unilateral femoral artery ligation to induce limb ischemia. RESULTS: GLN supplementation enhanced the proportion of anti-inflammatory monocytes and regulatory T cells in the blood. Expression of C-C motif chemokine receptor 5 by activated CD4+ T cells was promoted and prolonged in the GLN-supplemented group. GLN downregulated the percentage of M1 macrophages in muscle tissues which was correlated with lower levels of C-C motif chemokine ligand 2 in plasma. The muscle M1/M2 ratio was also reduced in the GLN group. Gene expression of interleukin-6 was suppressed by GLN supplementation, while expression levels of the peroxisome proliferator-activated receptor γ and myogenic differentiation 1 genes were elevated in post-ischemic muscles. Histological findings also indicated that muscle regeneration was accelerated in the GLN group. CONCLUSIONS: GLN supplementation in diabetic mice may exert more-balanced polarization of CD4+ T cells, monocytes, and macrophages, thus attenuating inflammatory responses and contributing to muscle regeneration after limb ischemia.
Asunto(s)
Polaridad Celular/efectos de los fármacos , Diabetes Mellitus Experimental/complicaciones , Suplementos Dietéticos , Glutamina/farmacología , Isquemia/dietoterapia , Músculo Esquelético/fisiología , Animales , Diabetes Mellitus Experimental/inmunología , Dieta/métodos , Modelos Animales de Enfermedad , Glutamina/administración & dosificación , Glutamina/inmunología , Miembro Posterior , Inmunidad/efectos de los fármacos , Inmunidad/inmunología , Isquemia/complicaciones , Isquemia/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Músculo Esquelético/inmunología , Regeneración/efectos de los fármacos , Regeneración/inmunologíaRESUMEN
Tumor-associated myeloid cells are one of the prominent components of solid tumors, serving as major immune regulators for the tumor microenvironment (TME) and an obstacle for immune-checkpoint blocking (ICB) therapy. However, it remains unclear how metabolic processes regulate the generation of suppressive myeloid cells in the TME. Here, we found that hematopoietic precursor cells are enriched in the tissues of several types of human cancer and can differentiate into immature myeloid cells (IMC). Tumor-infiltrating IMCs are highly immunosuppressive, glycolytic, and proliferative, as indicated by high levels of M-CSFR, Glut1, and Ki67. To elucidate the role of metabolism in regulating the generation of IMCs, we induced suppressive IMCs from hematopoietic precursor cells with GM-CSF and G-CSF in vitro We found that the generation of suppressive IMCs was accompanied by increased glycolysis, but not affected by glucose deprivation due to alternative catabolism. Generation of IMCs relied on glutaminolysis, regardless of glucose availability. Glutamine metabolism not only supported the expansion of IMCs with glutamine-derived α-ketoglutarate but also regulated the suppressive capacity through the glutamate-NMDA receptor axis. Moreover, inhibition of glutaminase GLS1 enhanced the therapeutic efficacy of anti-PD-L1 treatment, with reduced arginase 1+ myeloid cells, increased CD8+, IFNγ+ and granzyme B+ T cells, and delayed tumor growth in an ICB-resistant mouse model. Our work identified a novel regulatory mechanism of glutamine metabolism in controlling the generation of suppressive IMCs in the TME.
Asunto(s)
Glutamina/metabolismo , Glucólisis , Inmunosupresores/inmunología , Células Mieloides/inmunología , Neoplasias/inmunología , Neoplasias/metabolismo , Microambiente Tumoral/inmunología , Animales , Presentación de Antígeno/inmunología , Antígeno B7-H1/antagonistas & inhibidores , Línea Celular Tumoral , Proliferación Celular , Femenino , Glutamina/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Humanos , Ratones , Ratones Endogámicos BALB C , Células Mieloides/metabolismo , Neoplasias/patología , Transducción de Señal , Linfocitos T/inmunologíaRESUMEN
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease localized to China, Japan, and Korea that is characterized by severe hemorrhage and a high fatality rate. Currently, no specific vaccine or treatment has been approved for this disease. To develop a therapeutic agent for SFTS, we isolated antibodies from a phage-displayed antibody library that was constructed from a patient who recovered from SFTS virus (SFTSV) infection. One antibody, designated as Ab10, was reactive to the Gn envelope glycoprotein of SFTSV and protected host cells and A129 mice from infection in both in vitro and in vivo experiments. Notably, Ab10 protected 80% of mice, even when injected 5 days after inoculation with a lethal dose of SFTSV. Using cross-linker assisted mass spectrometry and alanine scanning, we located the non-linear epitope of Ab10 on the Gn glycoprotein domain II and an unstructured stem region, suggesting that Ab10 may inhibit a conformational alteration that is critical for cell membrane fusion between the virus and host cell. Ab10 reacted to recombinant Gn glycoprotein in Gangwon/Korea/2012, HB28, and SD4 strains. Additionally, based on its epitope, we predict that Ab10 binds the Gn glycoprotein in 247 of 272 SFTSV isolates previously reported. Together, these data suggest that Ab10 has potential to be developed into a therapeutic agent that could protect against more than 90% of reported SFTSV isolates.
Asunto(s)
Anticuerpos Neutralizantes/metabolismo , Phlebovirus/inmunología , Adulto , Animales , Anticuerpos Neutralizantes/fisiología , Anticuerpos Antivirales/metabolismo , Infecciones por Bunyaviridae/terapia , Epítopos/inmunología , Femenino , Fiebre , Glutamina/inmunología , Glutamina/metabolismo , Glicoproteínas/inmunología , Células HEK293 , Humanos , Leucopenia , Masculino , Ratones , Ratones Noqueados , Pruebas de Neutralización , Phlebovirus/patogenicidad , República de Corea , Trombocitopenia/inmunología , Proteínas del Envoltorio Viral/inmunologíaRESUMEN
High altitudes are a challenge for human physiology and for sports enthusiasts. Several reasons lead to deterioration in performance at high altitudes. Hypoxia owing to high altitude causes a breakdown of homeostasis with imbalance in several physiological systems, including the immune system. The reduction in mucosal immunity and inflammation and the predominance of the humoral immune response causes a condition of immunosuppression and an increased likelihood of infection. In addition, it is known that worsening of the immune response is associated with reduced performance. On the other hand, immunonutrition plays an important role in modulating the effects of physical exercise on the immune system. However, to our knowledge, few studies have evaluated the effect of nutrition on the immune system after exercise in hypoxia. Although the association between exercise and hypoxia has been shown to be more severe for the body owing to the sum of stressful agents, supplementation with carbohydrates and glutamine seems to play a relevant role in mitigating immunosuppressive effects. These findings, although limited by the fact that they are the result of very few studies, shed light on a relevant theme for sports physiology and nutrition and suggest that both supplements may be useful for athletes, visitors, and workers in high-altitude regions. The aim of this review was to discuss the effects of high-altitude hypoxia on the human body from the point of view of exercise immunology because it is known that transient immunosuppression after strenuous exercise and competition should be followed by reduction in training overload and worse performance.
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Mal de Altura/inmunología , Suplementos Dietéticos , Terapia Nutricional/métodos , Deportes/fisiología , Altitud , Mal de Altura/terapia , Rendimiento Atlético/fisiología , Carbohidratos de la Dieta/inmunología , Glutamina/inmunología , HumanosRESUMEN
Interleukin-13 (IL-13) is associated with allergic airway inflammation and airway remodeling. Our group found a variant with a single nucleotide polymorphism in the IL13 gene at position +2044G>A (rs20541) that was expected to result in the non-conservative replacement of a positively charged arginine (R) with a neutral glutamine (Q) at position 144. IL-13Q144 was associated with augmented allergic airway inflammation and bronchial asthma remodeling. There is some indication that anti-IL-13 monoclonal antibodies can demonstrate a positive effect on the clinical course of refractory asthmatic patients. To date, the binding stability of these agents for IL-13Q144 is unknown. The objective of this study was to investigate the prediction efficacy of the anti-IL-13 monoclonal antibodies tralokinumab and lebrikizumab in asthmatic patients with IL-13R144 and IL-13Q144. The three-dimensional (3-D) structure of tralokinumab was obtained from the Protein Data Bank (PDB ID: 5L6Y), and the complete 3-D structure of lebrikizumab was built through homology modeling. For the binding stability analysis, we performed and analyzed docking simulations of IL-13 with tralokinumab or lebrikizumab. The tralokinumab and lebrikizumab structures changed after binding to IL-13 to facilitate binding with IL-13Q144. The stability analysis with tralokinumab and lebrikizumab demonstrated that IL-13Q144 was more stable than IL-13R144 for both the Rosetta energy score and for the free energy of binding. IL-13Q144 might be a promising predictor of responsiveness to tralokinumab and lebrikizumab treatment for bronchial asthma.
Asunto(s)
Antiasmáticos/química , Anticuerpos Monoclonales/química , Arginina/química , Glutamina/química , Interleucina-13/antagonistas & inhibidores , Sustitución de Aminoácidos , Antiasmáticos/farmacología , Anticuerpos Monoclonales/farmacología , Arginina/inmunología , Asma/tratamiento farmacológico , Asma/genética , Asma/inmunología , Asma/patología , Sitios de Unión , Expresión Génica , Glutamina/inmunología , Humanos , Interleucina-13/química , Interleucina-13/genética , Interleucina-13/inmunología , Cinética , Pulmón/inmunología , Pulmón/patología , Modelos Moleculares , Simulación del Acoplamiento Molecular , Mutación , Unión Proteica , Conformación Proteica en Hélice alfa , Conformación Proteica en Lámina beta , Dominios y Motivos de Interacción de Proteínas , Homología Estructural de Proteína , TermodinámicaRESUMEN
KIR2DL2 and KIR2DL3 segregate as alleles of a single locus in the centromeric motif of the killer cell immunoglobulin-like receptor (KIR) gene family. Although KIR2DL2/L3 polymorphism is known to be associated with many human diseases and is an important factor for donor selection in allogeneic hematopoietic stem cell transplantation, the molecular determinant of functional diversity among various alleles is unclear. In this study we found that KIR2DL2/L3 with glutamic acid at position 35 (E(35)) are functionally stronger than those with glutamine at the same position (Q(35)). Cytotoxicity assay showed that NK cells from HLA-C1 positive donors with KIR2DL2/L3-E(35) could kill more target cells lacking their ligands than NK cells with the weaker -Q(35) alleles, indicating better licensing of KIR2DL2/L3(+) NK cells with the stronger alleles. Molecular modeling analysis reveals that the glutamic acid, which is negatively charged, interacts with positively charged histidine located at position 55, thereby stabilizing KIR2DL2/L3 dimer and reducing entropy loss when KIR2DL2/3 binds to HLA-C ligand. The results of this study will be important for future studies of KIR2DL2/L3-associated diseases as well as for donor selection in allogeneic stem cell transplantation.
Asunto(s)
Ácido Glutámico/genética , Glutamina/genética , Antígenos HLA-C/genética , Células Asesinas Naturales/inmunología , Polimorfismo de Nucleótido Simple , Receptores KIR2DL2/genética , Receptores KIR2DL3/genética , Alelos , Animales , Línea Celular , Citotoxicidad Inmunológica , Regulación de la Expresión Génica , Genotipo , Ácido Glutámico/química , Ácido Glutámico/inmunología , Glutamina/química , Glutamina/inmunología , Antígenos HLA-C/química , Antígenos HLA-C/inmunología , Humanos , Células Asesinas Naturales/citología , Ratones , Ratones Endogámicos NOD , Ratones SCID , Modelos Moleculares , Cultivo Primario de Células , Unión Proteica , Dominios Proteicos , Estructura Secundaria de Proteína , Receptores KIR2DL2/química , Receptores KIR2DL2/inmunología , Receptores KIR2DL3/química , Receptores KIR2DL3/inmunología , Transducción de SeñalRESUMEN
Osteoarthritis (OA) is a common form of arthritis in humans. It has long been regarded as a non-inflammatory disease, but a degree of inflammation is now recognized as being a vital inducer of subpopulation of OA. Besides inflammation, the establishment and development of OA are associated with alterations in metabolism and profiles of amino acids (AA), including glutamate- and arginine-family AA as well as their related metabolites (e.g., creatinine, hydroxyproline, γ-aminobutyrate, dimethylarginines and homoarginine). Functional AA (e.g., glutamine, arginine, glutamate, glycine, proline, and tryptophan) have various benefits (i.e., anti-inflammation and anti-oxidation) in treatment of inflammation-associated diseases, including OA. Thus, these AA have potential as immunomodulatory nutrients for patients with inflammation-induced OA.
Asunto(s)
Necesidades Nutricionales/inmunología , Estado Nutricional/inmunología , Osteoartritis/metabolismo , Arginina/análogos & derivados , Arginina/inmunología , Arginina/metabolismo , Creatinina/inmunología , Creatinina/metabolismo , Ácido Glutámico/inmunología , Ácido Glutámico/metabolismo , Glutamina/inmunología , Glutamina/metabolismo , Homoarginina/inmunología , Homoarginina/metabolismo , Humanos , Hidroxiprolina/inmunología , Hidroxiprolina/metabolismo , Factores Inmunológicos/metabolismo , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/patología , Osteoartritis/inmunología , Osteoartritis/patología , Prolina/inmunología , Prolina/metabolismo , Triptófano/inmunología , Triptófano/metabolismo , Ácido gamma-Aminobutírico/inmunología , Ácido gamma-Aminobutírico/metabolismoRESUMEN
T cell activation requires that the cell meet increased energetic and biosynthetic demands. We showed that exogenous nutrient availability regulated the differentiation of naïve CD4(+) T cells into distinct subsets. Activation of naïve CD4(+) T cells under conditions of glutamine deprivation resulted in their differentiation into Foxp3(+) (forkhead box P3-positive) regulatory T (Treg) cells, which had suppressor function in vivo. Moreover, glutamine-deprived CD4(+) T cells that were activated in the presence of cytokines that normally induce the generation of T helper 1 (TH1) cells instead differentiated into Foxp3(+) Treg cells. We found that α-ketoglutarate (αKG), the glutamine-derived metabolite that enters into the mitochondrial citric acid cycle, acted as a metabolic regulator of CD4(+) T cell differentiation. Activation of glutamine-deprived naïve CD4(+) T cells in the presence of a cell-permeable αKG analog increased the expression of the gene encoding the TH1 cell-associated transcription factor Tbet and resulted in their differentiation into TH1 cells, concomitant with stimulation of mammalian target of rapamycin complex 1 (mTORC1) signaling. Together, these data suggest that a decrease in the intracellular amount of αKG, caused by the limited availability of extracellular glutamine, shifts the balance between the generation of TH1 and Treg cells toward that of a Treg phenotype.
Asunto(s)
Diferenciación Celular/inmunología , Glutamina/inmunología , Ácidos Cetoglutáricos/inmunología , Linfocitos T Reguladores/inmunología , Células TH1/inmunología , Animales , Glutamina/metabolismo , Ácidos Cetoglutáricos/metabolismo , Diana Mecanicista del Complejo 1 de la Rapamicina , Ratones , Complejos Multiproteicos/inmunología , Complejos Multiproteicos/metabolismo , Linfocitos T Reguladores/metabolismo , Serina-Treonina Quinasas TOR/inmunología , Serina-Treonina Quinasas TOR/metabolismo , Células TH1/metabolismoRESUMEN
A hallmark of the gluten-driven enteropathy celiac disease is autoantibody production towards the enzyme transglutaminase 2 (TG2) that catalyzes the formation of covalent protein-protein cross-links. Activation of TG2-specific B cells likely involves gluten-specific CD4 T cells as production of the antibodies is dependent on disease-associated HLA-DQ allotypes and dietary intake of gluten. IgA plasma cells producing TG2 antibodies with few mutations are abundant in the celiac gut lesion. These plasma cells and serum antibodies to TG2 drop rapidly after initiation of a gluten-free diet, suggestive of extrafollicular responses or germinal center reactions of short duration. High antigen avidity is known to promote such responses, and is also important for breakage of self-tolerance. We here inquired whether TG2 avidity could be a feature relevant to celiac disease. Using recombinant enzyme we show by dynamic light scattering and gel electrophoresis that TG2 efficiently utilizes itself as a substrate due to conformation-dependent homotypic association, which involves the C-terminal domains of the enzyme. This leads to the formation of covalently linked TG2 multimers. The presence of exogenous substrate such as gluten peptide does not inhibit TG2 self-cross-linking, but rather results in formation of TG2-TG2-gluten complexes. The celiac disease autoantibody epitopes, clustered in the N-terminal part of TG2, are conserved in the TG2-multimers as determined by mass spectrometry and immunoprecipitation analysis. TG2 multimers are superior to TG2 monomer in activating A20 B cells transduced with TG2-specific B-cell receptor, and uptake of TG2-TG2-gluten multimers leads to efficient activation of gluten-specific T cells. Efficient catalytic self-multimerization of TG2 and generation of multivalent TG2 antigen decorated with gluten peptides suggest a mechanism by which self-reactive B cells are activated to give abundant numbers of plasma cells in celiac disease. Importantly, high avidity of the antigen could explain why TG2-specific plasma cells show signs of an extrafollicular generation pathway.
Asunto(s)
Autoantígenos/inmunología , Proteínas de Unión al GTP/inmunología , Receptores de Antígenos de Linfocitos B/inmunología , Proteínas Recombinantes/inmunología , Transglutaminasas/inmunología , Animales , Autoantígenos/metabolismo , Linfocitos B/inmunología , Linfocitos B/metabolismo , Sitios de Unión/genética , Sitios de Unión/inmunología , Catálisis , Enfermedad Celíaca/inmunología , Línea Celular Tumoral , Electroforesis en Gel de Poliacrilamida , Epítopos de Linfocito T/inmunología , Proteínas de Unión al GTP/genética , Proteínas de Unión al GTP/metabolismo , Glutamina/genética , Glutamina/inmunología , Glutamina/metabolismo , Glútenes/inmunología , Humanos , Lisina/genética , Lisina/inmunología , Lisina/metabolismo , Modelos Moleculares , Mutación , Conformación Proteica , Proteína Glutamina Gamma Glutamiltransferasa 2 , Multimerización de Proteína , Receptores de Antígenos de Linfocitos B/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Células Sf9 , Linfocitos T/inmunología , Linfocitos T/metabolismo , Transglutaminasas/genética , Transglutaminasas/metabolismoRESUMEN
As part of the innate immune response, neutrophils are at the forefront of defence against infection, resolution of inflammation and wound healing. They are the most abundant leucocytes in the peripheral blood, have a short lifespan and an estimated turnover of 10(10) to 10(11) cells per day. Neutrophils efficiently clear microbial infections by phagocytosis and by oxygen-dependent and oxygen-independent mechanisms. In 2004, a new neutrophil anti-microbial mechanism was described, the release of neutrophil extracellular traps (NETs) composed of DNA, histones and anti-microbial peptides. Several microorganisms, bacterial products, as well as pharmacological stimuli such as PMA, were shown to induce NETs. Neutrophils contain relatively few mitochondria, and derive most of their energy from glycolysis. In this scenario we aimed to analyse some of the metabolic requirements for NET formation. Here it is shown that NETs formation is strictly dependent on glucose and to a lesser extent on glutamine, that Glut-1, glucose uptake, and glycolysis rate increase upon PMA stimulation, and that NET formation is inhibited by the glycolysis inhibitor, 2-deoxy-glucose, and to a lesser extent by the ATP synthase inhibitor oligomycin. Moreover, when neutrophils were exposed to PMA in glucose-free medium for 3 hr, they lost their characteristic polymorphic nuclei but did not release NETs. However, if glucose (but not pyruvate) was added at this time, NET release took place within minutes, suggesting that NET formation could be metabolically divided into two phases; the first, independent from exogenous glucose (chromatin decondensation) and, the second (NET release), strictly dependent on exogenous glucose and glycolysis.
Asunto(s)
Trampas Extracelulares/metabolismo , Glucosa/metabolismo , Neutrófilos/metabolismo , Carcinógenos/farmacología , Desoxiglucosa/farmacología , Inhibidores Enzimáticos/farmacología , Trampas Extracelulares/inmunología , Glucosa/inmunología , Transportador de Glucosa de Tipo 1/inmunología , Transportador de Glucosa de Tipo 1/metabolismo , Glutamina/inmunología , Glutamina/metabolismo , Glucólisis/efectos de los fármacos , Humanos , Neutrófilos/inmunología , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
Antibody B4e8 exhibits modest cross-neutralizing activity, with preference for HIV subtype B. This preference might be explained by B4e8׳s extensive interaction with Arg315, which occurs at the center of most subtype B V3 sequences but is replaced by Gln in subtype C. The extent to which B4e8׳s ability to neutralize subtype C strains is hindered by Gln315 and/or other factors, e.g. epitope masking, is unclear. We confirmed here that an Arg315-to-Gln substitution in a subtype B virus abrogates B4e8 neutralizing activity. Conversely, B4e8-resistant subtype C viruses were rendered sensitive upon Gln 315-to-Arg substitution. V2 region swapping between B4e8-sensitive and- resistant subtype C strains revealed a role for V2 in limiting B4e8 access, but this was less significant than the absence of Arg315. Our findings, while illustrating the importance of Arg315 for B4e8, suggest that some subtype C strains may be vulnerable to B4e8 derivatives capable of binding stronger to Gln315-containing sequences.
Asunto(s)
Anticuerpos Neutralizantes/inmunología , Epítopos/inmunología , Glutamina/inmunología , Anticuerpos Anti-VIH/inmunología , VIH-1/inmunología , Productos del Gen env del Virus de la Inmunodeficiencia Humana/inmunología , Sustitución de Aminoácidos , Arginina/inmunología , VIH-1/genética , Productos del Gen env del Virus de la Inmunodeficiencia Humana/genéticaRESUMEN
The gastrointestinal tract has a major role in digestion and absorption of nutrients while playing a leading role in defense of the body. It forms a shield against the invasion of various microorganisms or their products (e.g. antigens, toxins) and therefore it is important to establish its integrity and functionality. That depends on the route of administration and the composition of the artificial nutrition. This study concentrates on the influences of different kinds of artificial nutrition in the functionality of the intestinal mucosal barriers. It seems that full macromolecular solutions of enteral nutrition ensure an adequate mucous immune response, while a lack of nutritional stimulus in the lumen leads rapidly to a dysfunction of gastric-associated lymphatic tissue and mucosal immune system. This dysfunction renders the patients susceptible to infections in distant organs, hospital pneumonia, and multiorgan failure of non-infectious etiology. In patients with indication of total parenteral nutrition administration, addition of bombesin or glutamine preserves mucosal immune response and may limit the adverse effects.