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1.
BMC Pediatr ; 24(1): 587, 2024 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-39285316

RESUMO

BACKGROUND: Congenital cytomegalovirus (cCMV) infection can lead to a range of adverse outcomes. The majority of cCMV neonates with clinical symptoms are infected postnatally; however, established cases of intrauterine infection are uncommon, resulting in a paucity of reports on clinical findings and lymphocytes expression in CMV-infected neonates. CASE PRESENTATION: We followed a neonate with cCMV infection from the onset of hospitalization to several months of follow-up. This infant was intrauterine CMV-positive in the amniotic fluid of the mother at 21 weeks' gestation and received intravenous ganciclovir infusion and sequential oral valganciclovir after birth. The typical clinical signs manifested in the nervous system, liver, and peripheral blood and were documented during the hospitalizaion period and up to the follow-up visit. Flow cytometry was employed to examine the expression of T cells, their subsets, and the associated cytokines in peripheral blood samples at various time points. The flow data for the cCMV neonate were compared with those of the controls at each time point. Following treatment, clinical symptoms improved and the infant became CMV negative. However, developmental delays occurred later in life. The proportion of CD8+CD28- Tregs in the peripheral blood of the neonate with congenital CMV infection was higher than that in the controls at the three time points. The expression levels of perforin and granzyme B secreted by γδ T cells (Vδ1 and Vδ2 T cells), increased during the course of hospitalization until follow-up and were higher than those in the controls at the three time points. CONCLUSIONS: Despite the alleviation of clinical symptoms, developmental delay in later life remains inevitable in this intrauterine cCMV neonate. CD8+CD28- Tregs and Vδ1 and Vδ2 T cells secreting perforin and granzyme B may be involved in congenital CMV infection, although this hypothesis requires validation in a larger study. This report may contribute to our understanding of the effect of current treatment and the immune status of intrauterine cCMV-infected neonates.


Assuntos
Infecções por Citomegalovirus , Linfócitos T Reguladores , Humanos , Infecções por Citomegalovirus/imunologia , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/tratamento farmacológico , Recém-Nascido , Feminino , Linfócitos T Reguladores/imunologia , Gravidez , Linfócitos T CD8-Positivos/imunologia , Antígenos CD28 , Complicações Infecciosas na Gravidez , Perforina/metabolismo , Antivirais/uso terapêutico , Masculino , Ganciclovir/uso terapêutico , Granzimas/metabolismo
2.
Biomolecules ; 14(8)2024 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-39199299

RESUMO

PRF1 (perforin 1) is a key cytotoxic molecule that plays a crucial role in the killing function of natural killer (NK) cells and cytotoxic T lymphocytes (CTLs). Recent studies have focused on PRF1's role in cancer development, progression, and prognosis. Studies have shown that aberrant PRF1 expression has a significant role to play in cancer development and progression. In some cancers, high expression of the PRF1 gene is associated with a better prognosis for patients, possibly because it helps enhance the body's immune response to tumors. However, some studies have also shown that the absence of PRF1 may make it easier for tumors to evade the body's immune surveillance, thus affecting patient survival. Furthermore, recent studies have explored therapeutic strategies based on PRF1, such as enhancing the ability of immune cells to kill cancer cells by boosting PRF1 activity. In addition, they have improved the efficacy of immunotherapy by modulating its expression to enhance the effectiveness of the treatment. Based on these findings, PRF1 may be a valuable biomarker both for the treatment of cancer and for its prognosis in the future. To conclude, PRF1 has an important biological function and has clinical potential for the treatment of cancer, which indicates that it deserves more research and development in the future.


Assuntos
Neoplasias , Perforina , Humanos , Perforina/metabolismo , Perforina/genética , Neoplasias/terapia , Neoplasias/imunologia , Neoplasias/genética , Neoplasias/metabolismo , Imunoterapia , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Animais , Prognóstico , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Linfócitos T Citotóxicos/imunologia , Regulação Neoplásica da Expressão Gênica
3.
Cells ; 13(16)2024 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-39195200

RESUMO

BACKGROUND: Large granular lymphocyte leukemias (LGLLs) are rare lymphoproliferative malignancies caused by clonal expansion of granular lymphocytes. T-cell LGLL and natural killer (NK) cell LGLL are defined based on their cellular origin. Their clinical manifestation and pathophysiology vary depending on the subtype and include, e.g., neutropenia, anemia, recurrent infections, and autoimmunity. A limited number of available patient-derived cell lines are considered valuable tools to study the biology of these malignancies. They differ in the expression of lineage-specific surface markers, but generally contain cytotoxic effector molecules in characteristic granules. METHODS: We investigated the presence and release of lysosome-associated effector proteins in patient-derived LGLL cell lines by flow and imaging cytometry, by Western blotting and by bottom-up proteomics profiling. RESULTS: The tested cell lines did not express FasL (CD178), but did express CD26/DPP4+. Intracellularly, we detected major differences in the abundance and subcellular distribution of granzymes, perforin, and granulysin. Similar differences were seen in enriched lysosome-related effector vesicles (LREVs). The proteomics profiling of enriched EVs from an NK-LGLL line (NKL) and a T-LGLL line (MOTN-1), confirmed individual profiles of effector molecules. CONCLUSION: Our analyses underscore the individual distribution of effector proteins but also open new routes to define the role of intra- and extracellular granules in the disease manifestation or pathology of LGLLs.


Assuntos
Vesículas Extracelulares , Leucemia Linfocítica Granular Grande , Humanos , Leucemia Linfocítica Granular Grande/patologia , Leucemia Linfocítica Granular Grande/metabolismo , Vesículas Extracelulares/metabolismo , Linhagem Celular Tumoral , Grânulos Citoplasmáticos/metabolismo , Lisossomos/metabolismo , Proteômica , Células Matadoras Naturais/metabolismo , Perforina/metabolismo , Granzimas/metabolismo , Antígenos de Diferenciação de Linfócitos T
4.
Elife ; 132024 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-39133873

RESUMO

Group 1 innate lymphoid cells (ILCs) comprise conventional natural killer (cNK) cells and type 1 innate lymphoid cells (ILC1s). The main functions of liver cNK cells and ILC1s not only include directly killing target cells but also regulating local immune microenvironment of the liver through the secretion of cytokines. Uncovering the intricate mechanisms by which transcriptional factors regulate and influence the functions of liver cNK cells and ILC1s, particularly within the context of liver tumors, presents a significant opportunity to amplify the effectiveness of immunotherapies against liver malignancies. Using Ncr1-drived conditional knockout mouse model, our study reveals the regulatory role of Prdm1 in shaping the composition and maturation of cNK cells. Although Prdm1 did not affect the killing function of cNK cells in an in vivo cytotoxicity model, a significant increase in cancer metastasis was observed in Prdm1 knockout mice. Interferon-gamma (IFN-γ), granzyme B, and perforin secretion decreased significantly in Prdm1-deficient cNK cells and liver ILC1s. Single-cell RNA sequencing (scRNA-seq) data also provided evidences that Prdm1 maintains functional subsets of cNK cells and liver ILC1s and facilitates communications between cNK cells, liver ILC1s, and macrophages. The present study unveiled a novel regulatory mechanism of Prdm1 in cNK cells and liver ILC1s, showing promising potential for developing innovative immune therapy strategies against liver cancer.


Assuntos
Neoplasias Hepáticas , Camundongos Knockout , Fator 1 de Ligação ao Domínio I Regulador Positivo , Animais , Camundongos , Fator 1 de Ligação ao Domínio I Regulador Positivo/genética , Fator 1 de Ligação ao Domínio I Regulador Positivo/metabolismo , Neoplasias Hepáticas/imunologia , Neoplasias Hepáticas/genética , Células Matadoras Naturais/imunologia , Interferon gama/metabolismo , Imunidade Inata , Linfócitos/imunologia , Vigilância Imunológica , Granzimas/metabolismo , Granzimas/genética , Receptor 1 Desencadeador da Citotoxicidade Natural/metabolismo , Receptor 1 Desencadeador da Citotoxicidade Natural/genética , Perforina/metabolismo , Perforina/genética , Fígado/imunologia , Fígado/metabolismo , Camundongos Endogâmicos C57BL , Microambiente Tumoral/imunologia , Antígenos Ly
5.
Front Immunol ; 15: 1434011, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39144143

RESUMO

Background: Gamma-delta (γδ) T cells are a major immune cell subset in pigs. Approximately 50% of circulating T cells are γδ T cells in young pigs and up to 30% in adult sows. Despite this abundance, the functions of porcine γδ T cells are mostly unidentified. In humans and mice, activated γδ T cells exhibit broad innate cytotoxic activity against a wide variety of stressed, infected, and cancerous cells through death receptor/ligand-dependent and perforin/granzyme-dependent pathways. However, so far, it is unknown whether porcine γδ T cells have the ability to perform cytotoxic functions. Methods: In this study, we conducted a comprehensive phenotypic characterization of porcine γδ T cells isolated from blood, lung, and nasal mucosa. To further analyze the cytolytic potential of γδ T cells, in vitro cytotoxicity assays were performed using purified γδ T cells as effector cells and virus-exposed or mock-treated primary porcine alveolar macrophages as target cells. Results: Our results show that only CD2+ γδ T cells express cytotoxic markers (CD16, NKp46, perforin) with higher perforin and NKp46 expression in γδ T cells isolated from lung and nasal mucosa. Moreover, we found that γδ T cells can exhibit cytotoxic functions in a cell-cell contact and degranulation-dependent manner. However, porcine γδ T cells did not seem to specifically target Porcine Reproductive and Respiratory Syndrome Virus or swine Influenza A Virus-infected macrophages, which may be due to viral escape mechanisms. Conclusion: Porcine γδ T cells express cytotoxic markers and can exhibit cytotoxic activity in vitro. The specific mechanisms by which porcine γδ T cells recognize target cells are not fully understood but may involve the detection of cellular stress signals.


Assuntos
Citotoxicidade Imunológica , Vírus da Síndrome Respiratória e Reprodutiva Suína , Animais , Suínos , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Macrófagos Alveolares/imunologia , Macrófagos Alveolares/virologia , Síndrome Respiratória e Reprodutiva Suína/imunologia , Linfócitos T Citotóxicos/imunologia , Biomarcadores , Infecções por Orthomyxoviridae/imunologia , Perforina/metabolismo , Perforina/imunologia , Linfócitos Intraepiteliais/imunologia , Células Cultivadas
6.
Adv Exp Med Biol ; 1448: 129-144, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39117812

RESUMO

Familial forms of hemophagocytic lymphohistiocytosis (HLH) are caused by loss-of-function mutations in genes encoding perforin as well as those required for release of perforin-containing cytotoxic granule constituent. Perforin is expressed by subsets of CD8+ T cells and NK cells, representing lymphocytes that share mechanism of target cell killing yet display distinct modes of target cell recognition. Here, we highlight recent findings concerning the genetics of familial HLH that implicate CD8+ T cells in the pathogenesis of HLH and discuss mechanistic insights from animal models as well as patients that reveal how CD8+ T cells may contribute to or drive disease, at least in part through release of IFN-γ. Intriguingly, CD8+ T cells and NK cells may act differentially in severe hyperinflammatory diseases such as HLH. We also discuss how CD8+ T cells may promote or drive pathology in other cytokine release syndromes (CSS). Moreover, we review the molecular mechanisms underpinning CD8+ T cell-mediated lymphocyte cytotoxicity, key to the development of familial HLH. Together, recent insights to the pathophysiology of CSS in general and HLH in particular are providing promising new therapeutic targets.


Assuntos
Linfócitos T CD8-Positivos , Síndrome da Liberação de Citocina , Linfo-Histiocitose Hemofagocítica , Humanos , Linfócitos T CD8-Positivos/imunologia , Linfo-Histiocitose Hemofagocítica/imunologia , Linfo-Histiocitose Hemofagocítica/genética , Animais , Síndrome da Liberação de Citocina/imunologia , Síndrome da Liberação de Citocina/genética , Células Matadoras Naturais/imunologia , Perforina/genética , Perforina/metabolismo , Citotoxicidade Imunológica/genética , Interferon gama/imunologia , Interferon gama/genética , Interferon gama/metabolismo
7.
Nat Commun ; 15(1): 6443, 2024 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-39085199

RESUMO

Birdshot chorioretinopathy is an inflammatory eye condition strongly associated with MHC-I allele HLA-A29. The striking association with MHC-I suggests involvement of T cells, whereas natural killer (NK) cell involvement remains largely unstudied. Here we show that HLA-A29-positive birdshot chorioretinopathy patients have a skewed NK cell pool containing expanded CD16 positive NK cells which produce more proinflammatory cytokines. These NK cells contain populations that express CD8A which is involved in MHC-I recognition on target cells, display gene signatures indicative of high cytotoxic activity (GZMB, PRF1 and ISG15), and signaling through NK cell receptor CD244 (SH2D1B). Long-term monitoring of a cohort of birdshot chorioretinopathy patients with active disease identifies a population of CD8bright CD244bright NK cells, which rapidly declines to normal levels upon clinical remission following successful treatment. Collectively, these studies implicate CD8bright CD244bright NK cells in birdshot chorioretinopathy.


Assuntos
Coriorretinopatia de Birdshot , Antígenos HLA-A , Células Matadoras Naturais , Família de Moléculas de Sinalização da Ativação Linfocitária , Análise de Célula Única , Humanos , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Coriorretinopatia de Birdshot/imunologia , Coriorretinopatia de Birdshot/metabolismo , Antígenos HLA-A/genética , Antígenos HLA-A/metabolismo , Antígenos HLA-A/imunologia , Análise de Célula Única/métodos , Família de Moléculas de Sinalização da Ativação Linfocitária/metabolismo , Família de Moléculas de Sinalização da Ativação Linfocitária/genética , Antígenos CD8/metabolismo , Antígenos CD8/genética , Coriorretinite/imunologia , Coriorretinite/genética , Feminino , Receptores de IgG/metabolismo , Receptores de IgG/genética , Masculino , Citocinas/metabolismo , Adulto , Proteínas Ligadas por GPI/metabolismo , Proteínas Ligadas por GPI/genética , Pessoa de Meia-Idade , Perforina
8.
Immunol Lett ; 269: 106900, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39032911

RESUMO

Pemphigus vulgaris (PV) is a rare autoimmune disorder where autoantibodies target the desmosomal proteins resulting in blistering of oral mucosa and skin. While the pathogenesis of PV is mainly mediated by the adaptive immune system, key players of innate immunity are also emerging. This study outlines the phenotypic as well as functional attributes of NK cells in PV. Through in-depth analysis using flow cytometry we identified an increase in the frequency of CD56+ CD3- NK cells and their subtypes in periphery. Along with this there is an increased frequency of IFNγ+ CD56bright CD16dim NK cells. mRNA expression of sorted NK cells for differentially expressed genes, particularly key transcription factors such as T-bet and EOMES, as well as surface receptors like NKG2D and KIR2D, and the cytokine IFNγ, displayed significant upregulation. A significant activation of NK cells was seen in the disease state. The levels of perforin and IFNγ were significantly elevated in the culture supernatants of patients. Additionally, a significantly higher cytotoxicity of NK cells in PV was observed. In lesioned tissues of PV, NK related markers were significantly increased. Lastly, we observed NK cells using confocal microscopy in the tissue biopsies of patients which showed significant infiltration of CD56+ CD3- NK cells at the lesional sites. This study aimed to shed light on the pivotal role of NK cells in the immunopathology of PV, offering a thorough understanding of their behaviour and changes in expression which might help in contributing to the development of novel therapeutics.


Assuntos
Células Matadoras Naturais , Pênfigo , Humanos , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Pênfigo/imunologia , Pênfigo/terapia , Pênfigo/metabolismo , Feminino , Masculino , Interferon gama/metabolismo , Pessoa de Meia-Idade , Adulto , Perforina/metabolismo , Imunofenotipagem , Citotoxicidade Imunológica , Ativação Linfocitária/imunologia , Idoso , Contagem de Linfócitos
9.
J Immunol ; 213(5): 753-762, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-38995175

RESUMO

The role of ICOS in antitumor T cell responses and overall tumor progression has been controversial. In this study, we compared tumor progression in mice lacking ICOS selectively in regulatory T (Treg) cells or in all T cells. Using an experimental melanoma lung metastasis model, we found that Treg cell-specific ICOS knockout reduces the overall tumor burden compared with Cre control mice, with increased CD4+-to-Treg cell and CD8+-to-Treg cell ratios in the tumor. In contrast, there was no difference in the tumor burden in mice lacking ICOS in all of the T cell compartments. This suggests a dual role of ICOS costimulation in promoting protumor and antitumor T cell responses. Consistent with reduced tumor burden, we found that Treg cell-specific deletion of ICOS leads to an increase of CD8+ CTLs that express high levels of granzyme B and perforin. Moreover, single-cell transcriptome analysis revealed an increase of Ly108+Eomeshi CD8+ T cells at the cost of the Ly108+T-bethi subset in Treg cell-specific knockout mice. These results suggest that ICOS-expressing Treg cells suppress the CTL maturation process at the level of Eomes upregulation, a critical step known to drive perforin expression and cytotoxicity. Collectively, our data imply that cancer immunotherapies using ICOS agonist Abs may work better in Treg cell-low tumors or when they are combined with regimens that deplete tumor-infiltrating Treg cells.


Assuntos
Proteína Coestimuladora de Linfócitos T Induzíveis , Melanoma Experimental , Camundongos Knockout , Linfócitos T Citotóxicos , Linfócitos T Reguladores , Animais , Proteína Coestimuladora de Linfócitos T Induzíveis/genética , Linfócitos T Reguladores/imunologia , Camundongos , Linfócitos T Citotóxicos/imunologia , Melanoma Experimental/imunologia , Perforina/metabolismo , Camundongos Endogâmicos C57BL , Granzimas/metabolismo , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/patologia , Proteínas Citotóxicas Formadoras de Poros
10.
Ann Hematol ; 103(9): 3775-3782, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39046509

RESUMO

Hemophagocytic Lymphohistiocytosis (HLH) is a rare disorder of immune dysregulation characterized by fever, cytopenias, and splenomegaly. Its primary form poses a therapeutic challenge due to its high fatality when left untreated. We retrospectively analyzed 28 patients who underwent related-donor allogeneic stem cell transplant for primary HLH from 2010 to 2021. Among them were 10 cases of familial HLH, 8 cases of Griscelli syndrome type 2, and 1 case each with PRF1 and STX11 mutations. All the patients underwent transplants with reduced-intensity or myeloablative conditioning and 26 of them achieved neutrophil engraftment at a median of day + 14. The donors were either fully matched (68%) or haploidentical (32%). With a median follow-up of 1 year, overall survival was 68% (n = 19) and disease-free survival was 64.4% (n = 18). OS was better in patients transplanted with a sibling donor (compared to parent donor), who achieved complete donor chimerism, and those transplanted early in the course of the disease (diagnosis to transplant duration less than 6 months).


Assuntos
Transplante de Células-Tronco Hematopoéticas , Linfo-Histiocitose Hemofagocítica , Humanos , Linfo-Histiocitose Hemofagocítica/terapia , Linfo-Histiocitose Hemofagocítica/mortalidade , Masculino , Feminino , Estudos Retrospectivos , Adolescente , Criança , Pré-Escolar , Adulto , Lactente , Condicionamento Pré-Transplante/métodos , Transplante Homólogo , Aloenxertos , Adulto Jovem , Piebaldismo/terapia , Pessoa de Meia-Idade , Taxa de Sobrevida , Intervalo Livre de Doença , Seguimentos , Doenças da Imunodeficiência Primária , Perforina
11.
J Pediatr Hematol Oncol ; 46(6): e393-e401, 2024 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-38968556

RESUMO

OBJECTIVE: Neonatal sepsis and familial hemophagocytic lymphohistiocytosis (fHLH) have similar clinical and laboratory symptoms and the possibility of overlooking fHLH diagnosis is high in newborns with sepsis. History of consanguineous marriage and/or sibling death, hepatomegaly/splenomegaly, and hyperferritinemia (>500 ng/mL) are likely to support fHLH in newborns with sepsis. Therefore, in newborns with sepsis in whom at least 2 of these 3 criteria were detected, genetic variants was investigated for the definitive diagnosed of fHLH. According to the results of genetic examination, we investigated whether these criteria supporting fHLH could be used as a screening test in fHLH. MATERIALS AND METHODS: fHLH-associated genetic variants were investigated in 22 patients diagnosed with neonatal sepsis who fulfilled at least 2 of the following criteria (1) history of consanguineous marriage and/or sibling death, (2) hepatomegaly/splenomegaly, and (3) hyperferritinemia (>500 ng/mL). RESULTS: Heterozygous variants were determined in 6 patients (27.2%): 3 STXBP2 , 1 STX11 , 1 UNC13D , and 1 PRF1 . Polymorphisms associated with the clinical symptoms and signs of HLH were determined in 5 patients (22.7%): 4 UNC13D , 1 PRF1 . Two patients were in the heterozygous variants and polymorphism associated with the clinical symptoms and signs of HLH groups. In 12 patients, benign polymorphisms were detected in STXBP2 and UNC13D genes. No change in fHLH associated genes were found in 1 patient. CONCLUSION: Some variants and/or polymorphisms identified in our patients have been previously reported in patients with HLH. Therefore, we recommend further investigation of fHLH in patients with neonatal sepsis who fulfill at least 2 of the above 3 criteria.


Assuntos
Linfo-Histiocitose Hemofagocítica , Sepse Neonatal , Humanos , Linfo-Histiocitose Hemofagocítica/genética , Linfo-Histiocitose Hemofagocítica/diagnóstico , Recém-Nascido , Masculino , Feminino , Sepse Neonatal/diagnóstico , Sepse Neonatal/genética , Perforina/genética , Proteínas Qa-SNARE/genética , Proteínas de Membrana/genética , Testes Genéticos/métodos
12.
Cell Physiol Biochem ; 58(4): 322-335, 2024 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-39074350

RESUMO

BACKGROUND/AIMS: Immune cells are reported to upregulate CD47 during infection, however, the role of CD47 in innate and adaptive immune cells remains unclear. METHODS: To bridge this knowledge gap, we analysed our single cell (sc)-RNA dataset along with other publicly available sc-RNA datasets from healthy controls, people with HIV-1 (PWH) and COVID-19 patients. We characterized each immune cell based on low, intermediate, and high expression of CD47 . RESULTS: Our analyses revealed that CD47 high pDCs and monocytes exhibited relatively higher expression of IFN-α regulatory genes, antiviral interferon-stimulated genes (ISGs) and MHC-I associated genes compared to CD47 inter. and CD47 low cells. Furthermore, CD47 high NK and CD8+ T cells showed higher expression of antiviral ISGs, as well as genes encoding for cytotoxic markers like granzyme B, perforin, granulysin, interferon gamma and NKG7. Additionally, CD47 high CD8+ T cells expressed higher levels of PD-1 and LAG-3 genes. Lastly, we found that CD47 high B cells had enriched expression of genes involved in cell activation and humoral responses. CONCLUSION: Overall, our analyses revealed that innate and adaptive immune cells expressing elevated activation and functional gene signatures also express higher CD47 levels.


Assuntos
Antígeno CD47 , Linfócitos T CD8-Positivos , Granzimas , HIV-1 , Células Matadoras Naturais , Perforina , Receptor de Morte Celular Programada 1 , RNA Mensageiro , Análise de Célula Única , Humanos , Antígeno CD47/metabolismo , Antígeno CD47/genética , Linfócitos T CD8-Positivos/metabolismo , Linfócitos T CD8-Positivos/imunologia , Granzimas/metabolismo , Granzimas/genética , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Perforina/metabolismo , Perforina/genética , HIV-1/imunologia , RNA Mensageiro/metabolismo , RNA Mensageiro/genética , Receptor de Morte Celular Programada 1/metabolismo , Receptor de Morte Celular Programada 1/genética , COVID-19/imunologia , COVID-19/virologia , COVID-19/genética , Infecções por HIV/imunologia , Infecções por HIV/virologia , Infecções por HIV/genética , Proteína do Gene 3 de Ativação de Linfócitos , Antígenos de Diferenciação de Linfócitos T/metabolismo , Antígenos de Diferenciação de Linfócitos T/genética , SARS-CoV-2/imunologia , Interferon gama/metabolismo , Interferon gama/genética , Monócitos/metabolismo , Monócitos/imunologia , Antígenos CD/metabolismo , Antígenos CD/genética , Linfócitos B/metabolismo , Linfócitos B/imunologia , Imunidade Inata
13.
Nature ; 632(8023): 174-181, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38987594

RESUMO

Changes in the gut microbiome have pivotal roles in the pathogenesis of acute graft-versus-host disease (aGVHD) after allogenic haematopoietic cell transplantation (allo-HCT)1-6. However, effective methods for safely resolving gut dysbiosis have not yet been established. An expansion of the pathogen Enterococcus faecalis in the intestine, associated with dysbiosis, has been shown to be a risk factor for aGVHD7-10. Here we analyse the intestinal microbiome of patients with allo-HCT, and find that E. faecalis escapes elimination and proliferates in the intestine by forming biofilms, rather than by acquiring drug-resistance genes. We isolated cytolysin-positive highly pathogenic E. faecalis from faecal samples and identified an anti-E. faecalis enzyme derived from E. faecalis-specific bacteriophages by analysing bacterial whole-genome sequencing data. The antibacterial enzyme had lytic activity against the biofilm of E. faecalis in vitro and in vivo. Furthermore, in aGVHD-induced gnotobiotic mice that were colonized with E. faecalis or with patient faecal samples characterized by the domination of Enterococcus, levels of intestinal cytolysin-positive E. faecalis were decreased and survival was significantly increased in the group that was treated with the E. faecalis-specific enzyme, compared with controls. Thus, administration of a phage-derived antibacterial enzyme that is specific to biofilm-forming pathogenic E. faecalis-which is difficult to eliminate with existing antibiotics-might provide an approach to protect against aGVHD.


Assuntos
Bacteriófagos , Enterococcus faecalis , Microbioma Gastrointestinal , Doença Enxerto-Hospedeiro , Adulto , Idoso , Animais , Feminino , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Adulto Jovem , Bacteriófagos/enzimologia , Bacteriófagos/genética , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Disbiose/complicações , Disbiose/microbiologia , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/genética , Enterococcus faecalis/crescimento & desenvolvimento , Enterococcus faecalis/metabolismo , Enterococcus faecalis/virologia , Fezes/microbiologia , Vida Livre de Germes , Doença Enxerto-Hospedeiro/complicações , Doença Enxerto-Hospedeiro/microbiologia , Doença Enxerto-Hospedeiro/prevenção & controle , Doença Enxerto-Hospedeiro/terapia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Técnicas In Vitro , Intestinos/efeitos dos fármacos , Intestinos/microbiologia , Perforina/metabolismo , Fatores de Risco , Transplante Homólogo/efeitos adversos , Sequenciamento Completo do Genoma , Farmacorresistência Bacteriana/efeitos dos fármacos , Antibacterianos/farmacologia
14.
Int J Mol Sci ; 25(13)2024 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-39000234

RESUMO

Juvenile Dermatomyositis (JDM) is the most common inflammatory myopathy in pediatrics. This study evaluates the role of Natural Killer (NK) cells in Juvenile Dermatomyositis (JDM) pathophysiology. The study included 133 untreated JDM children with an NK cell count evaluation before treatment. NK cell subsets (CD56low/dim vs. CD 56bright) were examined in 9 untreated children. CD56 and perforin were evaluated in situ in six untreated JDM and three orthopedic, pediatric controls. 56% of treatment-naive JDM had reduced circulating NK cell counts, designated "low NK cell". This low NK group had more active muscle disease compared to the normal NK cell group. The percentage of circulating CD56low/dim NK cells was significantly lower in the NK low group than in controls (0.55% vs. 4.6% p < 0.001). Examination of the untreated JDM diagnostic muscle biopsy documented an increased infiltration of CD56 and perforin-positive cells (p = 0.023, p = 0.038, respectively). Treatment-naive JDM with reduced circulating NK cell counts exhibited more muscle weakness and higher levels of serum muscle enzymes. Muscle biopsies from treatment-naive JDM displayed increased NK cell infiltration, with increased CD56 and perforin-positive cells.


Assuntos
Antígeno CD56 , Dermatomiosite , Células Matadoras Naturais , Debilidade Muscular , Humanos , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Dermatomiosite/imunologia , Dermatomiosite/sangue , Dermatomiosite/patologia , Masculino , Criança , Debilidade Muscular/sangue , Feminino , Antígeno CD56/metabolismo , Pré-Escolar , Perforina/metabolismo , Adolescente , Contagem de Linfócitos
15.
Sci Rep ; 14(1): 15511, 2024 07 05.
Artigo em Inglês | MEDLINE | ID: mdl-38969707

RESUMO

Anti-citrullinated protein autoantibodies (ACPA) are diagnostic for rheumatoid arthritis (RA). The antigens recognized by these autoantibodies are produced by protein arginine deiminases (PADs), particularly PAD4. However, it remains unknown why and how PAD4 causes this aberrant citrullination in RA. Here, we report that poly-perforin pores are present on freshly isolated neutrophils from RA patients, but not on healthy donor neutrophils. Neutrophils with perforin pores also contained intracellular citrullinated proteins in the region adjacent to the pores. This response was replicated in vitro by treating neutrophils with purified perforin, which generated intense dots of anti-perforin immunofluorescence, calcium influx, and intracellular citrullination. Extensive neutrophil killing in Felty's syndrome, an aggressive form of RA, correlated with particularly high ACPA, and PAD4 autoantibodies. In contrast, other forms of death, including NETosis, apoptosis, and pyroptosis, produced minimal citrullination. We conclude that neutrophil targeting by perforin leading to intracellular citrullination takes place in patients with RA.


Assuntos
Anticorpos Antiproteína Citrulinada , Artrite Reumatoide , Citrulinação , Neutrófilos , Perforina , Proteína-Arginina Desiminase do Tipo 4 , Humanos , Neutrófilos/metabolismo , Neutrófilos/imunologia , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Artrite Reumatoide/imunologia , Proteína-Arginina Desiminase do Tipo 4/metabolismo , Anticorpos Antiproteína Citrulinada/metabolismo , Anticorpos Antiproteína Citrulinada/imunologia , Perforina/metabolismo , Feminino , Masculino , Pessoa de Meia-Idade , Autoanticorpos/imunologia , Desiminases de Arginina em Proteínas/metabolismo , Adulto , Síndrome de Felty/metabolismo , Síndrome de Felty/patologia , Armadilhas Extracelulares/metabolismo , Citrulina/metabolismo , Idoso
16.
Proc Natl Acad Sci U S A ; 121(29): e2401420121, 2024 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-38995966

RESUMO

Cerebral (Aß) plaque and (pTau) tangle deposition are hallmarks of Alzheimer's disease (AD), yet are insufficient to confer complete AD-like neurodegeneration experimentally. Factors acting upstream of Aß/pTau in AD remain unknown, but their identification could enable earlier diagnosis and more effective treatments. T cell abnormalities are emerging AD hallmarks, and CD8 T cells were recently found to mediate neurodegeneration downstream of tangle deposition in hereditary neurodegeneration models. The precise impact of T cells downstream of Aß/pTau, however, appears to vary depending on the animal model. Our prior work suggested that antigen-specific memory CD8 T ("hiT") cells act upstream of Aß/pTau after brain injury. Here, we examine whether hiT cells influence sporadic AD-like pathophysiology upstream of Aß/pTau. Examining neuropathology, gene expression, and behavior in our hiT mouse model we show that CD8 T cells induce plaque and tangle-like deposition, modulate AD-related genes, and ultimately result in progressive neurodegeneration with both gross and fine features of sporadic human AD. T cells required Perforin to initiate this pathophysiology, and IFNγ for most gene expression changes and progression to more widespread neurodegenerative disease. Analogous antigen-specific memory CD8 T cells were significantly elevated in the brains of human AD patients, and their loss from blood corresponded to sporadic AD and related cognitive decline better than plasma pTau-217, a promising AD biomarker candidate. We identify an age-related factor acting upstream of Aß/pTau to initiate AD-like pathophysiology, the mechanisms promoting its pathogenicity, and its relevance to human sporadic AD.


Assuntos
Doença de Alzheimer , Linfócitos T CD8-Positivos , Modelos Animais de Doenças , Doença de Alzheimer/imunologia , Doença de Alzheimer/patologia , Doença de Alzheimer/genética , Animais , Linfócitos T CD8-Positivos/imunologia , Camundongos , Humanos , Placa Amiloide/patologia , Placa Amiloide/imunologia , Peptídeos beta-Amiloides/metabolismo , Camundongos Transgênicos , Encéfalo/patologia , Encéfalo/imunologia , Masculino , Interferon gama/metabolismo , Interferon gama/imunologia , Envelhecimento/imunologia , Memória Imunológica , Células T de Memória/imunologia , Perforina/metabolismo , Perforina/genética , Feminino
17.
Front Cell Infect Microbiol ; 14: 1410015, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38957797

RESUMO

Background: Tuberculosis (TB) persists as a global health challenge, with its treatment hampered by the side effects of long-term combination drug therapies and the growing issue of drug resistance. Therefore, the development of novel therapeutic strategies is critical. This study focuses on the role of immune checkpoint molecules (ICs) and functions of CD8+ T cells in the search for new potential targets against TB. Methods: We conducted differential expression genes analysis and CD8+ T cell functional gene analysis on 92 TB samples and 61 healthy individual (HI) samples from TB database GSE83456, which contains data on 34,603 genes. The GSE54992 dataset was used to validated the findings. Additionally, a cluster analysis on single-cell data from primates infected with mycobacterium tuberculosis and those vaccinated with BCG was performed. Results: The overexpression of LAG-3 gene was found as a potentially important characteristic of both pulmonary TB (PTB) and extrapulmonary TB (EPTB). Further correlation analysis showed that LAG-3 gene was correlated with GZMB, perforin, IL-2 and IL-12. A significant temporal and spatial variation in LAG-3 expression was observed in T cells and macrophages during TB infection and after BCG vaccination. Conclusion: LAG-3 was overexpressed in TB samples. Targeting LAG-3 may represent a potential therapeutic target for tuberculosis.


Assuntos
Antígenos CD , Linfócitos T CD8-Positivos , Proteína do Gene 3 de Ativação de Linfócitos , Mycobacterium tuberculosis , Tuberculose , Humanos , Mycobacterium tuberculosis/imunologia , Mycobacterium tuberculosis/genética , Linfócitos T CD8-Positivos/imunologia , Tuberculose/imunologia , Tuberculose/microbiologia , Animais , Antígenos CD/genética , Vacina BCG/imunologia , Macrófagos/imunologia , Macrófagos/microbiologia , Interleucina-2/metabolismo , Interleucina-2/genética , Perfilação da Expressão Gênica , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/microbiologia , Interleucina-12/genética , Interleucina-12/metabolismo , Perforina/genética , Perforina/metabolismo , Masculino
18.
Zhonghua Yi Xue Za Zhi ; 104(23): 2160-2166, 2024 Jun 18.
Artigo em Chinês | MEDLINE | ID: mdl-38871474

RESUMO

Objective: To investigate the clinical and genetic mutation characteristics of patients with primary hemophagocytic lymphohistiocytosis (HLH) and their impact on prognosis. Methods: Sixty-three primary HLH patients with complete medical records admitted and diagnosed at Beijing Friendship Hospital of Capital Medical University from January 2013 to December 2022 were selected. The patients' clinical and laboratory features, genetic and rapid immunological indicator characteristics, treatment outcomes and prognosis were retrospectively analyzed. Follow-up was up to June 30, 2023, with a median follow-up time [M (Q1, Q3)] of 47 (21, 76) months. Overall survival was analyzed using Kaplan-Meier survival curve, and prognostic factors were analyzed using Cox proportional hazards regression model. Results: Sixty-three primary HLH patients included 35 males and 28 females, with a median age [M (Q1, Q3)] of 17 (7, 27) years. Clinical manifestations at the initial diagnosis mainly included fever (93.7%, 59/63), splenomegaly (87.3%, 55/63), hemophagocytosis (65.1%, 41/63), hepatomegaly (52.4%, 33/63) and central nervous system (CNS) involvement (38.1%, 24/63). A total of 39 patients (61.9%) were diagnosed with EB virus (EBV) infection at initial diagnosis.PRF1 and UNC13D gene mutations were the most common mutations, and the highest frequency mutation site in the PRF1 gene was c.1349C>T, and that of UNC13D gene was c.2588G>A. A total of 76.2% (48/63) of patients had reduced activity of natural killer (NK) cells. Cytotoxic cell degranulation function was impaired or absent in 52.7% (29/55) of patients, of which 79.2% (19/24) of patients with primary HLH with defects in degranulation-related genes had impaired degranulation function. The 1-year and 3-year overall survival rates were 74.8% and 66.7%, respectively. Cox multivariate analysis suggested that peripheral blood EBV≥10 000 copies/ml (HR=3.523, 95%CI: 1.418-8.757, P=0.007) was the risk factor for prognosis. Conclusions: The main clinical manifestations of primary HLH patients at the initial diagnosis include fever, splenomegaly, hemophagocytosis, hepatomegaly, and CNS involvement. PRF1 and UNC13D are the most commonly mutated genes. High copy number EBV infection in peripheral blood is the risk factor for prognosis.


Assuntos
Linfo-Histiocitose Hemofagocítica , Mutação , Humanos , Linfo-Histiocitose Hemofagocítica/genética , Linfo-Histiocitose Hemofagocítica/diagnóstico , Masculino , Prognóstico , Feminino , Estudos Retrospectivos , Adolescente , Criança , Adulto , Adulto Jovem , Perforina/genética
19.
Biosens Bioelectron ; 261: 116512, 2024 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-38908292

RESUMO

Natural killer (NK) cells are a crucial component of the innate immune system. This study introduces Cellytics NK, a novel platform for rapid and precise measurement of NK cell activity. This platform combines an NK-specific activation stimulator cocktail (ASC) and lens-free shadow imaging technology (LSIT), using optoelectronic components. LSIT captures digital hologram images of resting and ASC-activated NK cells, while an algorithm evaluates cell size and cytoplasmic complexity using shadow parameters. The combined shadow parameter derived from the peak-to-peak distance and width standard deviation rapidly distinguishes active NK cells from inactive NK cells at the single-cell level within 30 s. Here, the feasibility of the system was demonstrated by assessing NK cells from healthy donors and immunocompromised cancer patients, demonstrating a significant difference in the innate immunity index (I3). Cancer patients showed a lower I3 value (161%) than healthy donors (326%). I3 was strongly correlated with NK cell activity measured using various markers such as interferon-gamma, tumor necrosis factor-alpha, perforin, granzyme B, and CD107a. This technology holds promise for advancing immune functional assays, offering rapid and accurate on-site analysis of NK cells, a crucial innate immune cell, with its compact and cost-effective optoelectronic setup, especially in the post-COVID-19 era.


Assuntos
Técnicas Biossensoriais , Células Matadoras Naturais , Humanos , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/citologia , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Imunidade Inata , COVID-19/imunologia , COVID-19/virologia , Holografia/métodos , Holografia/instrumentação , Ativação Linfocitária , Interferon gama/análise , SARS-CoV-2/imunologia , SARS-CoV-2/isolamento & purificação , Neoplasias/imunologia , Neoplasias/diagnóstico por imagem , Granzimas , Fator de Necrose Tumoral alfa , Perforina/metabolismo
20.
Sci Rep ; 14(1): 14586, 2024 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-38918457

RESUMO

Natural killer (NK) cells play a key role in defense against Salmonella infections during the early phase of infection. Our previous work showed that the excretory/secretory products of Ascaris suum repressed NK activity in vitro. Here, we asked if NK cell functionality was influenced in domestic pigs during coinfection with Ascaris and Salmonella enterica serotype Typhimurium. Ascaris coinfection completely abolished the IL-12 and IL-18 driven elevation of IFN-γ production seen in CD16 + CD8α + perforin + NK cells of Salmonella single-infected pigs. Furthermore, Ascaris coinfection prohibited the Salmonella-driven rise in NK perforin levels and CD107a surface expression. In line with impaired effector functions, NK cells from Ascaris-single and coinfected pigs displayed elevated expression of the inhibitory KLRA1 and NKG2A receptors genes, contrasting with the higher expression of the activating NKp46 and NKp30 receptors in NK cells during Salmonella single infection. These differences were accompanied by the highly significant upregulation of T-bet protein expression in NK cells from Ascaris-single and Ascaris/Salmonella coinfected pigs. Together, our data strongly indicate a profound repression of NK functionality by an Ascaris infection which may hinder infected individuals from adequately responding to a concurrent bacterial infection.


Assuntos
Ascaríase , Coinfecção , Células Matadoras Naturais , Doenças dos Suínos , Animais , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Ascaríase/imunologia , Ascaríase/veterinária , Ascaríase/parasitologia , Coinfecção/imunologia , Coinfecção/microbiologia , Coinfecção/parasitologia , Suínos , Doenças dos Suínos/parasitologia , Doenças dos Suínos/imunologia , Doenças dos Suínos/microbiologia , Salmonelose Animal/imunologia , Salmonella typhimurium/imunologia , Salmonella typhimurium/patogenicidade , Ascaris suum/imunologia , Interferon gama/metabolismo , Perforina/metabolismo , Interleucina-12/metabolismo , Proteínas com Domínio T/metabolismo , Proteínas com Domínio T/genética , Interleucina-18/metabolismo
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