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Fish & Shellfish Immunology Nov 2023NK-lysins are one of the most abundant antimicrobial peptides produced by cytotoxic T lymphocytes (CTLs) and natural killer cells (NKs), and identified as a new class of...
NK-lysins are one of the most abundant antimicrobial peptides produced by cytotoxic T lymphocytes (CTLs) and natural killer cells (NKs), and identified as a new class of intrinsically disordered proteins, playing critical roles in the cell-mediated cytotoxicity response, as well as immunomodulatory and antimicrobial activities upon a significant range of pathogens. In the present study, an NK-lysin was identified from Obscure puffer Takifugu obscurus (ToNK-lysin). The open reading frame of ToNK-lysin sequence spans 423 bp, encoding a peptide with 140 amino acids which shares a moderate residue identity (18%-60%) with NK-lysin of mammals and other teleost species. Phylogenetic analysis revealed that ToNK-lysin was most closely related to NK-lysins from the Pleuronectiformes (Bastard halibut Paralichthys olivaceus and Pacific halibut Hippoglossus stenolepis). Comprehensive computational analysis revealed that ToNK-lysin have substantial level of intrinsic disorder, which might be contribute to its multifunction. The transcripts of the ToNK-lysin were detected in multiple examined tissues and most abundant in gills. After bacterial and Poly I:C challenge, the transcriptional levels of ToNK-lysin were significantly up-regulated in the head kidney, liver and spleen at different time points. The recombinant ToNK-lysin showed significant antibacterial activity against Vibrio harveyi and Escherichia coli, and the ToNK-lysin treatment not only reduced the bacterial loads in liver and head kidney, but also alleviated the pathogen-mediated upregulation of immune-related genes. In addition, the co-incubation with rToNK-lysin protein remarkably degraded bacterial genomic DNA, suggesting the potential mechanism of ToNK-lysin against microbes. These results suggest that ToNK-lysin possess antibacterial and immunoregulatory function both in vivo and in vitro, which may allow it a potential applicability to the aquaculture industry.
Topics: Animals; Amino Acid Sequence; Phylogeny; Anti-Bacterial Agents; Adjuvants, Immunologic; Immunologic Factors; Tetraodontiformes; Proteolipids; Mammals
PubMed: 37748586
DOI: 10.1016/j.fsi.2023.109080 -
Food Chemistry Feb 2024Naringenin (NG) belongs to the class of flavanones having impressive pharmacological properties. Unfortunately, the in vivo bioavailability of NG is very low due to its...
Naringenin (NG) belongs to the class of flavanones having impressive pharmacological properties. Unfortunately, the in vivo bioavailability of NG is very low due to its higher hydrophobicity, which limits its practical use. Thus, in this study, we tried to develop NG-loaded macrophage membrane-coated liposome-based biomimetic nanoparticles with distinct physicochemical compositions and biological attributes for improving their bioavailability at the target site. The developed biomimetic nanoparticle (BNP) has shown good biocompatibility, stability, satisfactory particle size, pH-responsive drug (NG) release kinetics, and higher cellular uptake in vitro. The anti-metastatic efficacy of NGBNP has confirmed in syngeneic athymic BALB/c nude experimental models. By western blot analysis, semi-quantitative PCR, real-time PCR, and IHC, we conclude that NGBNP gets localized on the metastatic niche via its surface receptor α4, β1 integrin, and VCAM1 of metastatic cells and reduces the number of metastatic colonies in the lungs via regulating the apoptotic signaling axis.
PubMed: 37741236
DOI: 10.1016/j.foodchem.2023.137445 -
Fish & Shellfish Immunology Nov 2023The NK-lysin antimicrobial peptide, first identified in mammals, possesses both antibacterial and cytotoxic activity against cancer cell lines. Homologue peptides...
The NK-lysin antimicrobial peptide, first identified in mammals, possesses both antibacterial and cytotoxic activity against cancer cell lines. Homologue peptides isolated from different fish species have been examined for their functional characteristics in the last few years. In this study, a NK-lysin transcript was identified in silico from the head kidney transcriptome of the Antarctic teleost Trematomus bernacchii. The corresponding amino acid sequence, slightly longer than NK-lysins of other fish species, contains six cysteine residues that in mammalian counterparts form three disulphide bridges. Real time-PCR analysis indicated its predominant expression in T. bernacchii immune-related organs and tissues, with greatest mRNA abundance detected in gills and spleen. Instead of focusing on the full T. bernacchii derived NK-lysin mature molecule, we selected a 27 amino acid residue peptide (named NKL-WT), corresponding to the potent antibiotic NK-2 sequence found in human NK-lysin. Moreover, we designed a mutant peptide (named NKL-MUT) in which two alanine residues substitute the two cysteines found in the NKL-WT. The two peptides were obtained by solid phase organic synthesis to investigate their functional features. NKL-WT and NKL-MUT displayed antibacterial activity against the human pathogenic bacterium Enterococcus faecalis and the ESKAPE pathogen Acinetobacter baumannii, respectively. Moreover, at the determined Minimum Inhibitory Concentration and Minimum Bactericidal Concentration values against these pathogens, both peptides showed high selectivity as they did not exhibit any haemolytic activity on erythrocytes or cytotoxic activity against mammalian primary cell lines. Finally, the NKL-MUT selectively triggers the killing of the melanoma cell line B16F10 by means of a pro-apoptotic pathway at a concentration range in which no effects were found in normal mammalian cell lines. In conclusion, the two peptides could be considered as promising candidates in the fight against antibiotic resistance and tumour proliferative action, and also be used as innovative adjuvants, either to decrease chemotherapy side effects or to enhance anticancer drug activity.
Topics: Humans; Animals; Antarctic Regions; Fish Proteins; Peptides; Anti-Bacterial Agents; Perciformes; Proteolipids; Fishes; Mammals
PubMed: 37734650
DOI: 10.1016/j.fsi.2023.109099 -
Journal of Autoimmunity Nov 2023Sensitization to self-peptides induces various immunological responses, from autoimmunity to tumor immunity, depending on the peptide sequence; however, the underlying...
Harnessing autoimmunity with dominant self-peptide: Modulating the sustainability of tissue-preferential antigen-specific Tregs by governing the binding stability via peptide flanking residues.
Sensitization to self-peptides induces various immunological responses, from autoimmunity to tumor immunity, depending on the peptide sequence; however, the underlying mechanisms remain unclear, and thus, curative therapeutic options considering immunity balance are limited. Herein, two overlapping dominant peptides of myelin proteolipid protein, PLP136-150 and PLP139-151, which induce different forms of experimental autoimmune encephalomyelitis (EAE), monophasic and relapsing EAE, respectively, were investigated. Mice with monophasic EAE exhibited highly resistant to EAE re-induction with any encephalitogenic peptides, whereas mice with relapsing EAE were susceptible, and progressed, to EAE re-induction. This resistance to relapse and re-induction in monophasic EAE mice was associated with the maintenance of potent CD69CD103CD4CD25 regulatory T-cells (Tregs) enriched with antigen specificity, which expanded preferentially in the central nervous system with sustained suppressive activity. This tissue-preferential sustainability of potent antigen-specific Tregs was correlated with the antigenicity of PLP136-150, depending on its flanking residues. That is, the flanking residues of PLP136-150 enable to form pivotally arranged strong hydrogen bonds that secured its binding stability to MHC-class II. These potent Tregs acting tissue-preferentially were induced only by sensitization of PLP136-150, not by its tolerance induction, independent of EAE development. These findings suggest that, for optimal therapy, "benign autoimmunity" can be critically achieved through inverse vaccination with self-peptides by manipulating their flanking residues.
Topics: Animals; T-Lymphocytes, Regulatory; Encephalomyelitis, Autoimmune, Experimental; Mice; Myelin Proteolipid Protein; Autoimmunity; Autoantigens; Female; Peptide Fragments; Protein Binding; Peptides; Disease Models, Animal; Mice, Inbred C57BL
PubMed: 37716077
DOI: 10.1016/j.jaut.2023.103094 -
Cellular and Molecular Biology... Jul 2023The roles of apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3C (A3C) in various human malignancies are not consistent. A3C expression is correlated with...
The roles of apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3C (A3C) in various human malignancies are not consistent. A3C expression is correlated with early-stage breast cancer and is presented as a good prognostic factor; however, it induces fewer therapeutic effects of cytotoxic drugs in low-grade gliomas. To explore the impact of A3C on gliomas, a statistical analysis of several public databases was conducted. The results showed that enhanced A3C expression was associated with advanced tumor grades and poor expression of prognostic factors. Similarly, our in vitro study revealed that glioblastoma (GBM) cell lines had higher A3C mRNA and protein expression than that of normal brain tissue cDNA and lysates. We first performed an immunohistochemical stain (IHC) to prove that gliomas with high A3C expression presented the wild type-Isocitrate dehydrogenase 1 (IDH1), and they had an unfavorable prognosis in human glioma tissues. In addition, the oncological factors associated with A3C expression suggested that DNA repair pathways are important mechanisms for inducing tumorigenesis and chemoresistance in gliomas. Moreover, a significant correlation was observed between A3C expression and proteolipid protein 2 (PLP2). Reactive oxygen species (ROS) -activated PLP2 prevents DNA damage-induced cell apoptosis. Compared to high immunostaining scores for A3C and/or PLP2 expression, combined low immunostaining scores for A3C and PLP2 correlated with improved survival in gliomas; however, the detailed mechanism is to be elucidated. In conclusion, our results not only confirmed A3C played an important role in glioma development, but the A3C IHC test could successfully predict the therapeutic effects and disease prognosis.
Topics: Female; Humans; Apoptosis; Brain; Glioblastoma; MARVEL Domain-Containing Proteins; Proteolipids; Prognosis
PubMed: 37715423
DOI: 10.14715/cmb/2023.69.7.12 -
Frontiers in Bioscience (Landmark... Aug 2023Plasmolipin (PLLP) is a membrane protein located in lipid rafts that participates in the formation of myelin. It is also implicated in many pathologies, such as...
BACKGROUND
Plasmolipin (PLLP) is a membrane protein located in lipid rafts that participates in the formation of myelin. It is also implicated in many pathologies, such as neurological disorders, type 2 diabetes, and cancer metastasis. To better understand how PLLP interacts with raft components (gangliosides and cholesterol), we undertook a global study combining simulations and physicochemical measurements of molecular interactions in various PLLP-ganglioside systems.
METHODS
studies consisted of molecular dynamics simulations in reconstructed membrane environments. PLLP-ganglioside interaction measurements were performed by microtensiometry at the water-air interface on ganglioside monolayers.
RESULTS
We have elucidated the mode of interaction of PLLP with ganglioside GM1 and characterized this interaction at the molecular level. We showed that GM1 induces the structuring of the extracellular loops of PLLP and that this interaction propagates a conformational signal through the plasma membrane, involving a cholesterol molecule located between transmembrane domains. This conformational wave is finally transmitted to the intracellular domain of the protein, consistent with the role of PLLP in signal transduction.
CONCLUSIONS
This study is a typical example of the epigenetic dimension of protein structure, a concept developed by our team to describe the chaperone effect of gangliosides on disordered protein motifs which associate with lipid rafts. From a physiological point of view, these data shed light on the role of gangliosides in myelin formation. From a pathological point of view, this study will help to design innovative therapeutic strategies focused on ganglioside-PLLP interactions in various PLLP-associated diseases.
Topics: Humans; G(M1) Ganglioside; Gangliosides; Membrane Microdomains; Myelin Sheath; Proteolipids; Myelin and Lymphocyte-Associated Proteolipid Proteins
PubMed: 37664934
DOI: 10.31083/j.fbl2808157 -
PeerJ 2023Multiple sclerosis (MS) is a chronic inflammatory neurologic disease characterized by the demyelinating injury of the central nervous system (CNS). It was reported that...
Multiple sclerosis (MS) is a chronic inflammatory neurologic disease characterized by the demyelinating injury of the central nervous system (CNS). It was reported that the mutant peptide came from myelin proteolipid protein (PLP) and myelin basic protein (MBP) might play a critical role in immunotherapy function of MS. However, endogenous peptides in demyelinating brain tissue of MS and their role in the pathologic process of MS have not been revealed. Here, we performed peptidomic analysis of freshly isolated callosum (CC) from the brains of CPZ-treated mice and normal diet controls of male C57BL/6 mice by LC-MS/MS. Identified a total of 217 peptides were expressed at different levels in MS mice model compared with controls. By performed GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis, we found that the precursor protein of these differently expressed peptides (DEPs) were associated with myelin sheath and oxidative phosphorylation. Our study is the first brain peptidomic of MS mice model, revealing the distinct features of DEPs in demyelination brain tissue. These DPEs may provide further insight into the pathogenesis and complexity of MS, which would facilitate the discovery of the potential novel and effective strategy for the treatment of MS.
Topics: Male; Animals; Mice; Mice, Inbred C57BL; Chromatography, Liquid; Tandem Mass Spectrometry; Central Nervous System; Multiple Sclerosis; Disease Models, Animal; Peptides
PubMed: 37637167
DOI: 10.7717/peerj.15846 -
Clinical Case Reports Sep 2023This study aimed to characterize the clinical features, developmental milestones, and the natural history of Pelizaeus-Merzbacher disease (PMD) associated with gene...
This study aimed to characterize the clinical features, developmental milestones, and the natural history of Pelizaeus-Merzbacher disease (PMD) associated with gene duplications. The study examined 16 PMD Patients ranging in age from 7 to 48 years, who had a documented gene duplication. The study examined and analyzed the medical and developmental histories of the subjects utilizing a combination of resources that included medical history questionnaires, medical record reviews, and a 31-point functional disability scale that had been previously validated. The data extracted from the medical records and questionnaires for analysis included information related to medical and developmental histories, level of ambulation and cognition, and degree of functional disability. The summation of findings among the study population demonstrated that the presenting symptoms, developmental milestones achieved, and progression of symptoms reported are consistent with many previous studies of patients with duplications. All patients exhibited onset within the first year of life, with nystagmus predominating as the first symptom noticed. All patients exhibited delays in both motor and language development; however, many individuals were able to meet several developmental milestones. They exhibited some degree of continued motor impairment with none having the ability to walk independently. All patients were able to complete at least some of the cognition achievements and although not all were verbal, a number were able to use communication devices to complete these tasks. A critical tool of the study was the functional disability scale which provided a major advantage in helping quantify the clinical course of PMD, and for several, we were able to gather this information at more than one point in time. These reported findings in our cohort contribute important insight into the clinical heterogeneity and potential underlying mechanisms that define the molecular pathogenesis of the disease. This is one of only a small number of natural history studies examining the clinical course of a cohort of patients with duplications within the context of a validated functional disability scoring system. This study is unique in that it is limited to subjects with gene duplications. This study demonstrated many commonalities to other studies that have characterized the features of PMD and other PLP1-related disorders but also provide significant new insights into the evolving story that marks the natural history.
PubMed: 37636890
DOI: 10.1002/ccr3.7814 -
Nutrients Aug 2023Human genetic studies have associated Neuronatin gene variants with anorexia nervosa (AN) and obesity. Studies on the expression of the Neuronatin gene product, a...
Human genetic studies have associated Neuronatin gene variants with anorexia nervosa (AN) and obesity. Studies on the expression of the Neuronatin gene product, a proteolipid, are lacking. We investigated the relationship between circulating Neuronatin, body mass index (BMI), body composition (BC), physical activity (PA), and psychometric outcomes in patients with AN, normal weight, and obesity. Plasma Neuronatin was measured by ELISA in (1) 79 subjects of five BMI categories (AN/BMI < 17.5 kg/m; normal weight/BMI 18.5-25 kg/m; obesity/BMI 30-40 kg/m; obesity/BMI 40-50 kg/m; obesity/BMI > 50 kg/m) with assessment of BC (bioimpedance analysis; BIA); (2) 49 women with AN (BMI 14.5 ± 1.8 kg/m) with measurements of BC (BIA) and PA (accelerometry); (3) 79 women with obesity (BMI 48.8 ± 7.8 kg/m) with measurements of anxiety (GAD-7), stress (PSQ-20), depression (PHQ-9) and eating behavior (EDI-2). Overall, a positive correlation was found between Neuronatin and BMI ( = 0.006) as well as total fat mass (FM; = 0.036). In AN, Neuronatin did not correlate with BMI, FM, or PA ( > 0.05); no correlations were found between Neuronatin and psychometric outcomes in obesity ( > 0.05). The findings suggest an FM-dependent peripheral Neuronatin expression. The decreased Neuronatin expression in AN provides evidence that Neuronatin is implicated in the pathogenesis of eating disorders.
Topics: Humans; Female; Body Mass Index; Accelerometry; Anorexia Nervosa; Obesity; Adipose Tissue
PubMed: 37630847
DOI: 10.3390/nu15163657 -
International Journal of Molecular... Aug 2023Anti-glycolipid antibodies have been reported to play pathogenic roles in peripheral inflammatory neuropathies, such as Guillain-Barré syndrome. On the other hand, the...
Anti-Glycolipid Antibody Examination in Five EAE Models and Theiler's Virus Model of Multiple Sclerosis: Detection of Anti-GM1, GM3, GM4, and Sulfatide Antibodies in Relapsing-Remitting EAE.
Anti-glycolipid antibodies have been reported to play pathogenic roles in peripheral inflammatory neuropathies, such as Guillain-Barré syndrome. On the other hand, the role in multiple sclerosis (MS), inflammatory demyelinating disease in the central nervous system (CNS), is largely unknown, although the presence of anti-glycolipid antibodies was reported to differ among MS patients with relapsing-remitting (RR), primary progressive (PP), and secondary progressive (SP) disease courses. We investigated whether the induction of anti-glycolipid antibodies could differ among experimental MS models with distinct clinical courses, depending on induction methods. Using three mouse strains, SJL/J, C57BL/6, and A.SW mice, we induced five distinct experimental autoimmune encephalomyelitis (EAE) models with myelin oligodendrocyte glycoprotein (MOG), MOG, or myelin proteolipid protein (PLP), with or without an additional adjuvant curdlan injection. We also induced a viral model of MS, using Theiler's murine encephalomyelitis virus (TMEV). Each MS model had an RR, SP, PP, hyperacute, or chronic clinical course. Using the sera from the MS models, we quantified antibodies against 11 glycolipids: GM1, GM2, GM3, GM4, GD3, galactocerebroside, GD1a, GD1b, GT1b, GQ1b, and sulfatide. Among the MS models, we detected significant increases in four anti-glycolipid antibodies, GM1, GM3, GM4, and sulfatide, in PLP-induced EAE with an RR disease course. We also tested cellular immune responses to the glycolipids and found CD1d-independent lymphoproliferative responses only to sulfatide with decreased interleukin (IL)-10 production. Although these results implied that anti-glycolipid antibodies might play a role in remissions or relapses in RR-EAE, their functional roles need to be determined by mechanistic experiments, such as injections of monoclonal anti-glycolipid antibodies.
Topics: Animals; Mice; Mice, Inbred C57BL; Multiple Sclerosis; Encephalomyelitis, Autoimmune, Experimental; Theilovirus; Sulfoglycosphingolipids; Neoplasm Recurrence, Local; Antibodies; Myelin-Oligodendrocyte Glycoprotein; Glycolipids
PubMed: 37629117
DOI: 10.3390/ijms241612937