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1.
Am J Hum Genet ; 107(6): 1029-1043, 2020 12 03.
Artigo em Inglês | MEDLINE | ID: mdl-33202260

RESUMO

Genetic testing has increased the number of variants identified in disease genes, but the diagnostic utility is limited by lack of understanding variant function. CARD11 encodes an adaptor protein that expresses dominant-negative and gain-of-function variants associated with distinct immunodeficiencies. Here, we used a "cloning-free" saturation genome editing approach in a diploid cell line to simultaneously score 2,542 variants for decreased or increased function in the region of CARD11 associated with immunodeficiency. We also described an exon-skipping mechanism for CARD11 dominant-negative activity. The classification of reported clinical variants was sensitive (94.6%) and specific (88.9%), which rendered the data immediately useful for interpretation of seven coding and splicing variants implicated in immunodeficiency found in our clinic. This approach is generalizable for variant interpretation in many other clinically actionable genes, in any relevant cell type.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/genética , Variação Genética , Guanilato Ciclase/genética , Síndromes de Imunodeficiência/genética , Adenina/análogos & derivados , Adenina/farmacologia , Proteína 10 de Linfoma CCL de Células B/genética , Linfócitos B/citologia , Linhagem Celular , Diploide , Éxons , Genes Dominantes , Humanos , Células Jurkat , Linfoma/genética , Subunidade p50 de NF-kappa B/genética , Piperidinas/farmacologia , Polimorfismo de Nucleotídeo Único , Doenças da Imunodeficiência Primária/genética , Sensibilidade e Especificidade
2.
Sci Rep ; 10(1): 16298, 2020 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-33004909

RESUMO

To identify new candidate genes in osteoporosis, mainly involved in epigenetic mechanisms, we compared whole gene-expression in osteoblasts (OBs) obtained from women undergoing hip replacement surgery due to fragility fracture and severe osteoarthritis. Then, we analyzed the association of several SNPs with BMD in 1028 women. Microarray analysis yielded 2542 differentially expressed transcripts belonging to 1798 annotated genes, of which 45.6% (819) were overexpressed, and 54.4% (979) underexpressed (fold-change between - 7.45 and 4.0). Among the most represented pathways indicated by transcriptome analysis were chondrocyte development, positive regulation of bone mineralization, BMP signaling pathway, skeletal system development and Wnt signaling pathway. In the translational stage we genotyped 4 SNPs in DOT1L, HEY2, CARM1 and DNMT3A genes. Raw data analyzed against inheritance patterns showed a statistically significant association between a SNP of DNMT3A and femoral neck-(FN) sBMD and primarily a SNP of CARM1 was correlated with both FN and lumbar spine-(LS) sBMD. Most of these associations remained statistically significant after adjusting for confounders. In analysis with anthropometric and clinical variables, the SNP of CARM1 unexpectedly revealed a close association with BMI (p = 0.000082), insulin (p = 0.000085), and HOMA-IR (p = 0.000078). In conclusion, SNPs of the DNMT3A and CARM1 genes are associated with BMD, in the latter case probably owing to a strong correlation with obesity and fasting insulin levels.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/genética , DNA (Citosina-5-)-Metiltransferases/genética , Predisposição Genética para Doença/genética , Guanilato Ciclase/genética , Osteoporose/genética , Densidade Óssea/genética , Estudos de Casos e Controles , Perfilação da Expressão Gênica/métodos , Humanos , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos , Fraturas por Osteoporose/genética , Polimorfismo de Nucleotídeo Único/genética , Reação em Cadeia da Polimerase em Tempo Real , Transcriptoma
3.
PLoS Biol ; 18(9): e3000813, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32991574

RESUMO

Short-chain fatty acids (SCFAs) produced by gastrointestinal microbiota regulate immune responses, but host molecular mechanisms remain unknown. Unbiased screening using SCFA-conjugated affinity nanobeads identified apoptosis-associated speck-like protein (ASC), an adaptor protein of inflammasome complex, as a noncanonical SCFA receptor besides GPRs. SCFAs promoted inflammasome activation in macrophages by binding to its ASC PYRIN domain. Activated inflammasome suppressed survival of Salmonella enterica serovar Typhimurium (S. Typhimurium) in macrophages by pyroptosis and facilitated neutrophil recruitment to promote bacterial elimination and thus inhibit systemic dissemination in the host. Administration of SCFAs or dietary fibers, which are fermented to SCFAs by gut bacteria, significantly prolonged the survival of S. Typhimurium-infected mice through ASC-mediated inflammasome activation. SCFAs penetrated into the inflammatory region of the infected gut mucosa to protect against infection. This study provided evidence that SCFAs suppress Salmonella infection via inflammasome activation, shedding new light on the therapeutic activity of dietary fiber.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/metabolismo , Ácidos Graxos Voláteis/metabolismo , Inflamassomos/imunologia , Inflamassomos/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , Infecções por Salmonella/prevenção & controle , Animais , Proteínas Adaptadoras de Sinalização CARD/genética , Feminino , Microbioma Gastrointestinal/imunologia , Células HEK293 , Humanos , Imunidade Inata/fisiologia , Ativação de Macrófagos/genética , Ativação de Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ligação Proteica , Receptores Acoplados a Proteínas-G/genética , Infecções por Salmonella/genética , Infecções por Salmonella/imunologia , Infecções por Salmonella/metabolismo , Salmonella typhimurium/imunologia , Células U937
4.
Cardiovasc Pathol ; 49: 107261, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32771878

RESUMO

Cardiac inflammation in Coxsackievirus B3 (CVB3)-induced myocarditis is a consequence of viral-related cardiac injury and immune response. Caspase-associated recruitment domain 9 (CARD9) is a critical adaptor protein involved in transduction of signals from various innate pattern recognition receptors. In this study, the role of CARD9 in acute viral myocarditis was evaluated. CARD9-/- and C57BL/6 mice were infected with CVB3. On day 7 postinfection, myocardial tissue and blood samples were collected and examined. After CARD9 knockout, mRNA and protein levels of transforming growth factor-ß(TGF-ß), interleukin-17A(IL-17A), and CARD domain of B-cell CLL/lymphoma 10(BCL-10) in the myocardium were markedly lower in CARD9-/- mice than in C57BL/6 mice with CVB3-induced viral myocarditis. This trend was similar for the pathological scores for inflammation and serum levels of cytokines interleukin-6(IL-6), interleukin-10(IL-10), interferon -γ(IFN-γ), TGF-ß, and IL-17A. These results suggest that the CARD9-mediated secretion of pro-inflammatory cytokines plays an important role in the immune response to acute viral myocarditis.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/metabolismo , Infecções por Coxsackievirus/metabolismo , Enterovirus Humano B/imunologia , Miocardite/metabolismo , Miocárdio/metabolismo , Linfócitos T/metabolismo , Animais , Proteínas Adaptadoras de Sinalização CARD/deficiência , Proteínas Adaptadoras de Sinalização CARD/genética , Infecções por Coxsackievirus/genética , Infecções por Coxsackievirus/imunologia , Infecções por Coxsackievirus/virologia , Citocinas/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Interações Hospedeiro-Patógeno , Mediadores da Inflamação/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Knockout , Miocardite/genética , Miocardite/imunologia , Miocardite/virologia , Miocárdio/imunologia , Miocárdio/patologia , Transdução de Sinais , Linfócitos T/imunologia , Linfócitos T/virologia
5.
Life Sci ; 257: 118069, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32659370

RESUMO

AIM: Up-regulation of inflammasome proteins was reported in dystrophin-deficient muscles. However, it remains to be determined whether inflammasome activation plays a role in the pathogenesis of Duchenne muscular dystrophy. This study was therefore set out to investigate whether genetic disruption of the inflammasome pathway impacts the disease progression in mdx mice. MAIN METHODS: Mice deficient in both dystrophin and ASC (encoded by Pycard [PYD And CARD Domain Containing]) were generated. The impact of ASC deficiency on muscular dystrophy of mdx mice were assessed by measurements of serum cytokines, Western blot, real-time PCR and histopathological staining. KEY FINDINGS: The pro-inflammatory cytokines such as TNF-α, IL-6, KC/GRO and IL-10 were markedly increased in the sera of 8-week-old mdx mice compared to WT. Western blotting showed that P2X7, caspase-1, ASC and IL-18 were upregulated. Disruption of ASC and dystrophin expression in the mdx/ASC-/- mice was verified by Western blot analysis. Histopathological analysis did not find significant alterations in the muscular dystrophy phenotype in mdx/ASC-/- mice as compared to mdx mice. SIGNIFICANCE: Taken together, our results show that disruption of the central adaptor ASC of the inflammasome is insufficient to alleviate muscular dystrophy phenotype in mdx mice.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/genética , Distrofina/genética , Inflamassomos/metabolismo , Distrofia Muscular de Duchenne/fisiopatologia , Animais , Citocinas/sangue , Modelos Animais de Doenças , Progressão da Doença , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos mdx , Camundongos Knockout , Distrofia Muscular de Duchenne/genética , Regulação para Cima
7.
J Stroke Cerebrovasc Dis ; 29(8): 104874, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32689633

RESUMO

INTRODUCTION: Previous studies have reported the involvement of nucleotide-binding domain (NOD)-like receptor protein 3 (NLRP3) inflammasome in the inflammatory activation and pathophysiology of Ischemic Stroke (IS). Variations in genes encoding the constituent proteins of NLRP3 inflammasome can alter the risk of IS. OBJECTIVE: We investigated the role of the NLRP3 inflammasome in the pathogenesis of IS by establishing associations between combined polymorphisms of caspase recruitment domain-containing protein 8 (CARD8) rs2043211 and NLRP3 rs10754558 and the susceptibility to IS in a Chinese population. METHODS: Single nucleotide polymorphisms (SNPs) in CARD8 rs2043211 and NLRP3 rs10754558 were analyzed using TaqMan SNP genotyping assays in patients with IS (n=234) and healthy controls (n=115). Logistic regression analysis was carried out to evaluate potential interactions between CARD8 and NLRP3. RESULTS: Compared with healthy controls, there were no significant differences in the minor allele frequency (MAF) and the genotype frequency of NLRP3 rs10754558 or CARD8 rs2043211 in patients with IS(P>0.05). After stratification by gender, there was an increased risk for IS in men carrying heterozygous CARD8 rs2043211 when a co-dominant genetic model was applied (P=0.021, OR=3.83[1.22-12.03]). Logistic regression analysis indicated that men carrying both CARD8 rs2043211 AT and NLRP3 rs10754558 CG had a significantly higher risk of IS (P=0.046, OR=7.116[1.033-49.044]). CONCLUSIONS: Nucleotide variations in the genes encoding NLRP3 inflammasome proteins may be important to IS, and men carrying CARD8 rs2043211 and NLRP3 rs10754558, both heterozygous, confer a higher risk of IS.


Assuntos
Isquemia Encefálica/genética , Proteínas Adaptadoras de Sinalização CARD/genética , Inflamassomos/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteínas de Neoplasias/genética , Polimorfismo de Nucleotídeo Único , Acidente Vascular Cerebral/genética , Grupo com Ancestrais do Continente Asiático/genética , Isquemia Encefálica/diagnóstico , Isquemia Encefálica/etnologia , Estudos de Casos e Controles , China/epidemiologia , Feminino , Frequência do Gene , Interação Gene-Ambiente , Predisposição Genética para Doença , Heterozigoto , Humanos , Masculino , Fenótipo , Medição de Risco , Fatores de Risco , Fatores Sexuais , Acidente Vascular Cerebral/diagnóstico , Acidente Vascular Cerebral/etnologia
8.
Cell Immunol ; 353: 104129, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32473470

RESUMO

BENTA (B cell Expansion with NF-κB and T cell Anergy) is a novel lymphoproliferative disorder caused by germline, gain-of-function (GOF) mutations in the lymphocyte-restricted scaffolding protein CARD11. Similar somatic CARD11 mutations are found in lymphoid malignancies such as diffuse large B cell lymphoma (DLBCL). Normally, antigen receptor (AgR) engagement converts CARD11 into an active conformation that nucleates a signalosome required for IκB kinase (IKK) activation and NF-κB nuclear translocation. However, GOF CARD11 mutants drive constitutive NF-κB activity without AgR stimulation. Here we show that unlike wild-type CARD11, GOF CARD11 mutants can form large, peculiar cytosolic protein aggregates we term mCADS (mutant CARD11 dependent shells). MALT1 and phospho-IKK are reliably colocalized with mCADS, indicative of active signaling. Moreover, endogenous mCADS are detectable in ABC-DLBCL lines harboring similar GOF CARD11 mutations. The unique aggregation potential of GOF CARD11 mutants may represent a novel therapeutic target for treating BENTA or DLBCL.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/genética , Agregação Celular/fisiologia , Mutação com Ganho de Função , Guanilato Ciclase/genética , Proteínas Reguladoras de Apoptose/metabolismo , Proteínas Adaptadoras de Sinalização CARD/metabolismo , Guanilato Ciclase/metabolismo , Humanos , Quinase I-kappa B/metabolismo , Células Jurkat , Ativação Linfocitária , Linfócitos/metabolismo , Linfócitos/fisiologia , Linfoma Difuso de Grandes Células B/genética , Linfoma Difuso de Grandes Células B/metabolismo , Linfoma Difuso de Grandes Células B/patologia , Proteína de Translocação 1 do Linfoma de Tecido Linfoide Associado à Mucosa/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais
9.
Curr Top Microbiol Immunol ; 429: 177-203, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32415389

RESUMO

Caspase recruitment domain protein 9 (CARD9) forms essential signaling complexes in the innate immune system that integrate cues from C-type lectin receptors and specific intracellular pattern recognition receptors. These CARD9-mediated signals are pivotal for host defense against fungi, and they mediate immunity against certain bacteria, viruses and parasites. Furthermore, CARD9-regulated pathways are involved in sterile inflammatory responses critical for immune homeostasis and can control pro- and antitumor immunity in cancer microenvironments. Consequently, multiple genetic alterations of human CARD9 are connected to primary immunodeficiencies or prevalent inflammatory disorders in patients. This review will summarize our current understanding of CARD9 signaling in the innate immune system, its physiological and pathological functions and their implications for human immune-mediated diseases.


Assuntos
Proteínas Adaptadoras de Sinalização CARD , Imunidade Inata , Proteínas Adaptadoras de Sinalização CARD/genética , Humanos , Sistema Imunitário , Lectinas Tipo C , Receptores de Reconhecimento de Padrão/genética , Transdução de Sinais
10.
Anticancer Res ; 40(4): 1953-1962, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32234884

RESUMO

BACKGROUND/AIM: Caspase recruitment domain family, member 14 (CARD14) is a member of the CARD family of proteins, which play an important role in immune and inflammatory response, and cell survival and proliferation. Here, we identified the role of CARD14 in human breast cancer. MATERIALS AND METHODS: Immunohistochemistry was performed to evaluate CARD14 expression in breast cancer. Using CARD14 knockdown cells by small interfering RNA, colony formation and MTT assays, flow cytometry analyses, and migration assays were performed to evaluate the proliferation, cell cycle distribution, apoptosis, and migration ability of MCF7 and SK-BR-3 cells. RESULTS: CARD14 expression was significantly higher in breast cancer samples than in normal breast samples. CARD14 knockdown inhibited cell proliferation and migration, caused cell cycle arrest at the G1/S boundary, and promoted apoptosis. CONCLUSION: CARD14 regulates the proliferation and migration of MCF7 and SK-BR-3 cells; it is thus, a novel potential therapeutic target in breast cancer.


Assuntos
Neoplasias da Mama/genética , Proteínas Adaptadoras de Sinalização CARD/genética , Movimento Celular/genética , Proliferação de Células/genética , Guanilato Ciclase/genética , Proteínas de Membrana/genética , Apoptose/genética , Neoplasias da Mama/patologia , Ciclo Celular/genética , Pontos de Checagem do Ciclo Celular/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Técnicas de Silenciamento de Genes , Humanos , Células MCF-7 , RNA Interferente Pequeno/genética
11.
Fish Shellfish Immunol ; 100: 324-333, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32198069

RESUMO

Apoptosis-associated speck-like protein containing a C-terminal caspase recruit domain (ASC) is an important adapter protein in the inflammasome complex that mediates inflammatory caspase activation and host innate immunity in mammals. However, the function of inflammasome components in lower vertebrate remains poorly understood. In this study, full length of SmASC was cloned from turbot (Scophthalmus maximus). Through bioinformatic analysis, we found that SmASC shares relatively high identity with ASC in bony fish. Furthermore, we found that the intact SmASC can form an oligomeric speck-like structure, while the PYD segment of SmASC can form the filamentous structure. Moreover, expression of SmASC was induced after intraperitoneal injection of Edwardsiella piscicida (E. piscicida) in vivo. To further explore the role of SmASC during infection, we constructed SmASC knockdown and overexpression models by administration of siRNA and overexpression plasmids in vivo, respectively. Expression of SmASC decreased the propagation of E. piscicida in different immune organs. In summary, our results characterize the function of SmASC in S. maximus, suggesting that the SmASC plays a critical role in turbot immune responses.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/genética , Infecções por Enterobacteriaceae/veterinária , Proteínas de Peixes/genética , Linguados/genética , Linguados/imunologia , Inflamassomos/genética , Animais , Clonagem Molecular , Biologia Computacional , Edwardsiella/imunologia , Infecções por Enterobacteriaceae/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Perfilação da Expressão Gênica , Técnicas de Silenciamento de Genes , Imunidade Inata , Inflamassomos/imunologia , RNA Mensageiro , RNA Interferente Pequeno
12.
Comp Immunol Microbiol Infect Dis ; 70: 101417, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32113042

RESUMO

Caspase-associated recruitment domain-containing protein 9 (CARD9) deficiency is an autosomal-recessive primary immunodeficiency characterized by susceptibility to recurrent Candida infections, and its diagnosis and treatment is challenging. The present study aims to investigate the genetic characteristic and treatment strategy of a Chinese pediatric patient with CARD9 deficiency. In the present study, whole-exome sequencing (WES) was performed to screen the causal variants in a Chinese pediatric patient who exhibited an invasive Candida infection in the abdominal cavity and central nervous system. After the disease-causing gene being confirmed, the patient was treated with a combination of G-CSF and antifungal agents. DNA sequencing revealed a homozygous insertion mutation (c.819-820insG) in exon 6 of the CARD9 gene, which led to downstream amino acids conversion on codon 274 (p.D274fsX60). Th17 cell populations and cytokine levels showed decreased levels. The treatment regimen successfully resolved the patient's symptoms, and he remained symptom-free after more than 1 year of follow-up. This study described an invasive Candida infection in a pediatric patient and WES identified an insertion variant of the CARD9 gene. A combination of G-CSF and antifungal agents was highly effective in treating the invasive fungal infection accompanied by CARD9-induced immunodeficiency.


Assuntos
Antifúngicos/uso terapêutico , Proteínas Adaptadoras de Sinalização CARD/genética , Candidíase Mucocutânea Crônica/complicações , Candidíase Invasiva/tratamento farmacológico , Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Cavidade Abdominal/diagnóstico por imagem , Cavidade Abdominal/microbiologia , Encéfalo/diagnóstico por imagem , Encéfalo/microbiologia , Candidíase Mucocutânea Crônica/microbiologia , Criança , Humanos , Imagem por Ressonância Magnética , Mutação , Indução de Remissão , Células Th17 , Tomografia Computadorizada por Raios X , Resultado do Tratamento , Sequenciamento Completo do Exoma
13.
Immunogenetics ; 72(4): 217-224, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32020248

RESUMO

Systemic lupus erythematosus (SLE) is a complex autoimmune disorder involving heterogeneous clinical manifestations and numerous susceptibility genes. Several findings evidence the critical role of inflammasomes in the predisposition to autoimmune diseases and in SLE. We investigated whether inflammasome polymorphins could affect susceptibility to develop and/or severity SLE. Moreover, differences in inflammasome activation in peripheral blood were also evaluated in SLE patients and controls. The distribution of 13 SNPs in eight inflammasome genes was evaluated. To assess inflammasome priming in peripheral blood monocytes of SLE and controls, differential expression of selected inflammasome genes and IL-1ß production was analyzed in resting condition as well as after LPS and ATP stimulation. Results showed that the gain-of-function variant rs10754558 (NLRP3) was significantly more frequent in SLE patients with nephritis, reinforcing the concept of a key role of NLRP3 inflammasome not only in SLE but also especially in kidney disease. SLE monocytes in resting condition showed a higher level of IL-1ß expression and produced higher levels of IL-1ß when stimulated with LPS+ATP comparing to controls. The stimulation induced a significant expression of NLRP1, AIM2, CASP1, and IL1B genes, suggesting that the NLRP1 inflammasome is responsible for the IL-1ß production observed in monocytes. These data emphasized once more the important contribution of inflammasome in SLE-associated inflammation.


Assuntos
Inflamassomos/genética , Lúpus Eritematoso Sistêmico/genética , Polimorfismo de Nucleotídeo Único , Proteínas Adaptadoras de Transdução de Sinal/genética , Adulto , Proteínas Reguladoras de Apoptose/genética , Proteínas Adaptadoras de Sinalização CARD/genética , Proteínas de Ligação ao Cálcio/genética , Estudos de Casos e Controles , Caspase 1/genética , Proteínas de Ligação a DNA/genética , Feminino , Expressão Gênica , Humanos , Interleucina-1beta/genética , Lúpus Eritematoso Sistêmico/complicações , Masculino , Pessoa de Meia-Idade , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteínas de Neoplasias/genética , Nefrite/genética
14.
Cell ; 180(5): 878-894.e19, 2020 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-32059783

RESUMO

Pathogenic autoantibodies arise in many autoimmune diseases, but it is not understood how the cells making them evade immune checkpoints. Here, single-cell multi-omics analysis demonstrates a shared mechanism with lymphoid malignancy in the formation of public rheumatoid factor autoantibodies responsible for mixed cryoglobulinemic vasculitis. By combining single-cell DNA and RNA sequencing with serum antibody peptide sequencing and antibody synthesis, rare circulating B lymphocytes making pathogenic autoantibodies were found to comprise clonal trees accumulating mutations. Lymphoma driver mutations in genes regulating B cell proliferation and V(D)J mutation (CARD11, TNFAIP3, CCND3, ID3, BTG2, and KLHL6) were present in rogue B cells producing the pathogenic autoantibody. Antibody V(D)J mutations conferred pathogenicity by causing the antigen-bound autoantibodies to undergo phase transition to insoluble aggregates at lower temperatures. These results reveal a pre-neoplastic stage in human lymphomagenesis and a cascade of somatic mutations leading to an iconic pathogenic autoantibody.


Assuntos
Autoanticorpos/genética , Doenças Autoimunes/genética , Linfócitos B/imunologia , Linfoma/genética , Animais , Autoanticorpos/imunologia , Doenças Autoimunes/imunologia , Doenças Autoimunes/patologia , Linfócitos B/patologia , Proteínas Adaptadoras de Sinalização CARD/genética , Proteínas de Transporte/genética , Evolução Clonal/genética , Evolução Clonal/imunologia , Ciclina D3/genética , Guanilato Ciclase/genética , Humanos , Proteínas Imediatamente Precoces/genética , Região Variável de Imunoglobulina/genética , Região Variável de Imunoglobulina/imunologia , Proteínas Inibidoras de Diferenciação/genética , Linfoma/imunologia , Linfoma/patologia , Camundongos , Mutação/genética , Mutação/imunologia , Proteínas de Neoplasias/genética , Análise de Sequência de DNA/métodos , Análise de Sequência de RNA/métodos , Análise de Célula Única/métodos , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/genética , Proteínas Supressoras de Tumor/genética , Recombinação V(D)J/genética
15.
Biochem Biophys Res Commun ; 525(2): 412-417, 2020 04 30.
Artigo em Inglês | MEDLINE | ID: mdl-32102753

RESUMO

Caspase recruitment domain 6 (CARD6) was initially implicated in the immune system and oncogenesis, which has also been emerged to play an important role in cardio-metabolic diseases. Nevertheless, the potential role of CARD6 on macrophage activation remains unknown. In the present study, we observed a decreased CARD6 expression in bone marrow derived macrophages (BMDMs) and mouse peritoneal macrophages (MPMs) isolated from ApoE deficiency mice and administrated with OX-LDL, which were tested by RT-PCR and western bolt analysis. Moreover, the immunofluorescence co-staining revealed that a weaker immunoreactivity of CARD6 was found and primary located in cytoplasm of macrophages induced by OX-LDL. Phenotypically, loss-of-function of CARD6 dramatically increased pro-inflammatory M1 macrophage but decreased resolving M2 macrophage markers expression. Additionally, CARD6 knockdown significantly promoted cholesterol uptake but attenuated cholesterol efflux, which lead to increased foam cell formation. Mechanistically, a downregulated AMP-activated protein kinase (AMPK) expression was required for the promoted effect of CARD6 knockdown on macrophage activation. Taken together, these results suggest that CARD6 protects against macrophage activation partially through activation of AMPK-dependent mechanism.


Assuntos
Proteínas Quinases Ativadas por AMP/imunologia , Proteínas Adaptadoras de Sinalização CARD/imunologia , Inflamação/imunologia , Ativação de Macrófagos , Proteínas Quinases Ativadas por AMP/genética , Animais , Proteínas Adaptadoras de Sinalização CARD/genética , Células Cultivadas , Regulação para Baixo , Técnicas de Silenciamento de Genes , Inflamação/genética , Lipoproteínas LDL/imunologia , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos
16.
Blood ; 135(16): 1332-1343, 2020 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-32107531

RESUMO

Hemophagocytic lymphohistiocytosis (HLH) is a syndrome describing patients with severe systemic hyperinflammation. Characteristic features include unremitting fever, cytopenias, hepatosplenomegaly, and elevation of typical HLH biomarkers. Patients can develop hepatitis, coagulopathy, liver failure, central nervous system involvement, multiorgan failure, and other manifestations. The syndrome has a high mortality rate. More and more, it is recognized that while HLH can be appropriately used as a broad summary diagnosis, many pediatric patients actually suffer from an expanding spectrum of genetic diseases that can be complicated by the syndrome of HLH. Classic genetic diseases in which HLH is a typical and common manifestation include pathogenic changes in familial HLH genes (PRF1, UNC13D, STXBP2, and STX11), several granule/pigment abnormality genes (RAB27A, LYST, and AP3B1), X-linked lymphoproliferative disease genes (SH2D1A and XIAP), and others such as NLRC4, CDC42, and the Epstein-Barr virus susceptibility diseases. There are many other genetic diseases in which HLH is an infrequent complication of the disorder as opposed to a prominent manifestation of the disease caused directly by the genetic defect, including other primary immune deficiencies and inborn errors of metabolism. HLH can also occur in patients with underlying rheumatologic or autoinflammatory disorders and is usually designated macrophage activation syndrome in those settings. Additionally, HLH can develop in patients during infections or malignancies without a known (or as-yet-identified) genetic predisposition. This article will attempt to summarize current concepts in the pediatric HLH field as well as offer a practical diagnostic and treatment overview.


Assuntos
Linfo-Histiocitose Hemofagocítica/diagnóstico , Linfo-Histiocitose Hemofagocítica/terapia , Animais , Proteínas Adaptadoras de Sinalização CARD/genética , Proteínas de Ligação ao Cálcio/genética , Criança , Gerenciamento Clínico , Infecções por Vírus Epstein-Barr/complicações , Proteína 7 com Repetições F-Box-WD/genética , Predisposição Genética para Doença , Variação Genética , Humanos , Linfo-Histiocitose Hemofagocítica/genética , Linfo-Histiocitose Hemofagocítica/fisiopatologia , Proteína Associada à Molécula de Sinalização da Ativação Linfocitária/genética , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/genética
17.
Biomed Res Int ; 2020: 2867505, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32104685

RESUMO

Caspase recruitment domain family member 8 (CARD8) is an adaptor molecule that negatively regulates nuclear factor-κB (NF-κB) activation, interleukin (IL)-1ß secretion, and apoptosis. These play important roles in the pathogenesis of psoriasis. Genetic variants of CARD8 have been associated with an increased risk of several inflammatory diseases and psoriasis in Europe. However, nothing is known about the association of the polymorphisms of CARD8 and psoriasis vulgaris (PsV) in the Han population of northeastern China. To investigate the potential association between them, we designed a case-control study to genotype four selected single nucleotide polymorphisms (SNPs) using the improved multiplex ligation reaction (iMLDR) method. Model-based single SNP frequentist-test and haplotype association studies were performed to assess the association between SNPs and PsV. The results showed that the intron SNP rs10403848 was significantly associated with PsV (additive model p=0.0418, p'=0.0411, and statistical power 0.1902; heterozygous model p=0.0418, p'=0.0164, and statistical power 0.9406). A potential risk locus of nonsynonymous SNP rs2043211 found in the European population did not show a significant association in our study. We found that the polymorphism rs10403848 in CARD8 is significantly associated with PsV risk in the Han population of northeastern China. CARD8 may be involved in PsV in this population, as in the European population, but a different genetic process should be considered for the heterogeneity of risk loci.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/genética , Proteínas de Neoplasias/genética , Polimorfismo de Nucleotídeo Único , Psoríase/genética , Adulto , China/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Psoríase/epidemiologia , Fatores de Risco
18.
Med Sci Monit ; 26: e916935, 2020 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-32080163

RESUMO

BACKGROUND The aim of this case-control study was to evaluate the correlation of caspase recruitment domain-containing protein 8 (CARD8) gene rs2043211 (exon) and rs7253718 (intron) polymorphisms with the susceptibility of ankylosing spondylitis (AS) in the Chinese Han population. MATERIAL AND METHODS CARD8 polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 118 AS patients and 122 healthy persons. Linkage disequilibrium (LD) and haplotype analysis were carried out using Haploview software. Distribution differences of genotypes, alleles and haplotypes between the case and control groups were tested by chi-square test. Relative risk of AS was expressed by odds ratios (ORs) and 95% confidence intervals (CIs). Logistic regression analysis was used to adjust the results of association by clinical parameters. RESULTS For rs2043211, distribution of variant allele T was obviously different between AS patients and healthy controls (P=0.046). It indicated that T allele might increase the susceptibility of AS (OR=1.441, 95%CI=1.006-2.065). Adjusted by clinical characteristics, the significance of difference was slightly decreased (P=0.050, OR=1.439, 95%CI=0.999-2.072). Strong LD existed between rs2043211 and rs7253718 polymorphisms, and rs2043211T-rs7253718G haplotype was significantly associated with increased AS susceptibility (OR=1.787, 95%CI=1.165-2.740). In subgroup analysis, we found that the TT genotype and T allele of rs2043211 significantly increased the risk of AS in males (TT versus AA: OR=2.554, 95%CI=1.079-6.049; T versus A: OR=1.661, 95%CI=1.067-2.586), but not females. CONCLUSIONS CARD8 polymorphisms are likely to be associated with the elevated susceptibility of AS. Present results should be confirmed in the future studies.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/genética , Predisposição Genética para Doença , Proteínas de Neoplasias/genética , Polimorfismo de Nucleotídeo Único/genética , Espondilite Anquilosante/genética , Adolescente , Adulto , Idoso , Alelos , Estudos de Casos e Controles , Feminino , Haplótipos/genética , Humanos , Desequilíbrio de Ligação/genética , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Adulto Jovem
20.
Sci Rep ; 10(1): 198, 2020 01 13.
Artigo em Inglês | MEDLINE | ID: mdl-31932628

RESUMO

NOD-like receptors (NLRs) localize in the cytosol to recognize intracellular pathogen products and initialize the innate immune response. However, the ligands and ligand specificity of many NLRs remain unclear. One such NLR, NLRP6, plays an important role in maintaining intestinal homeostasis and protecting against various intestinal diseases such as colitis and intestinal tumorigenesis. Here, we show that the major component of the outer membrane of gram-negative bacteria, lipopolysaccharide (LPS), binds NLRP6 directly and induces global conformational change and dimerization. Following stimulation by ATP, the NLRP6 homodimer can further assemble into a linear molecular platform, and ASC is recruited to form higher molecular structures, indicative of a step-by-step activation mechanism. Our study sheds light on the mystery of LPS-induced inflammasome initiation, reveals the architecture and structural basis of potential pre-inflammasome, and suggests a novel molecular assembly pattern for immune receptors.


Assuntos
Trifosfato de Adenosina/metabolismo , Inflamassomos/metabolismo , Inflamação/patologia , Peptídeos e Proteínas de Sinalização Intracelular/química , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Lipopolissacarídeos/toxicidade , Proteínas Adaptadoras de Sinalização CARD/genética , Proteínas Adaptadoras de Sinalização CARD/metabolismo , Homeostase , Humanos , Imunidade Inata , Inflamassomos/efeitos dos fármacos , Inflamação/induzido quimicamente , Inflamação/imunologia , Inflamação/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/genética , Multimerização Proteica , Transdução de Sinais
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