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1.
Exerc Immunol Rev ; 26: 116-131, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32139354

RESUMO

Type 1 diabetes (T1D) is a T cell mediated autoimmune disease that targets and destroys insulin-secreting pancreatic beta cells. Although T cell mediated, a number of other immune cells are also critically involved in coordinating the events leading to T1D. Specifically, innate subsets play an important role in the pathogenesis of T1D. NK cells are one of the first cell types to infiltrate the pancreas, causing damage and release of beta cell antigens. Previous work in our group has shown differential mobilisation of highly differentiated CD8+ T cells during vigorous intensity exercise in T1D compared to a control cohort. Here, we aimed to explore exercise-induced mobilisation of other cell types involved in T1D pathogenesis. In this study, we investigated the effects of a single bout of vigorous (80% predicted VO2max) intensity exercise on innate cell mobilisation in T1D and control participants. T1D (N=12, mean age 33.2yrs, predicted VO2max 32.2 ml.kg.min⁻¹, BMI 25.3 kg.m⁻²) and control (N=12, mean age 29.4yrs, predicted VO2 max 38.5 ml.kg.min⁻¹, BMI 23.7 kg.m⁻² male participants completed a 30-minute bout of cycling at 80% predicted VO2 max in a fasted state. Peripheral blood was collected at baseline, immediately post-exercise, and 1 hour post-exercise. NK cell subsets mobilised during vigorous intensity exercise in both control and T1D participants. However, mature NK cells, defined as the CD56dimCD16bright subset, displayed a lower percentage increase following vigorous intensity exercise in T1D participants (Control: 185.12%, T1D: 97.06%). This blunted mobilisation was specific to early mature NK cells (KIR+) but not later differentiated NK cells (KIR+CD57+). Myeloid lineage subsets mobilised to a similar extent in both control and T1D participants. In conclusion, vigorous exercise mobilises innate immune cells in people with T1D albeit to a different extent to those without T1D. This mobilisation of innate immune cells provides a mechanistic argument to support exercise in people with T1D where it has the potential to improve surveillance for infection and to modulate the autoimmune response to the beta cell.


Assuntos
Diabetes Mellitus Tipo 1/imunologia , Exercício Físico , Células Matadoras Naturais/citologia , Ativação Linfocitária , Adulto , Antígeno CD56 , Proteínas Ligadas por GPI , Humanos , Masculino , Receptores de IgG
2.
Ann Lab Med ; 40(1): 1-6, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31432632

RESUMO

BACKGROUND: JL1, a CD43 epitope and mucin family cell surface glycoprotein, is expressed on leukemic cells. An anti-JL1 antibody combined with a toxic substance can have targeted therapeutic effects against JL1-positive leukemia; however, JL1 expression on bone marrow (BM) lymphoma cells has not been assessed using flow cytometry. We investigated JL1 expression on BM lymphoma cells from patients with non-Hodgkin lymphoma (NHL) to assess the potential of JL1 as a therapeutic target. METHODS: Patients with BM involvement of mature B-cell (N=44) or T- and natural killer (NK)-cell (N=4) lymphomas were enrolled from May 2015 to September 2016. JL1 expression on BM lymphoma cells was investigated using flow cytometry. Clinical, pathological, and cytogenetic characteristics, and treatment responses were compared according to JL1 expression status. RESULTS: Of the patients with NHL and BM involvement, 37.5% (18/48) were JL1-positive. Among mature B-cell lymphomas, 100%, 38.9%, 33.3%, 100%, and 25.0% of Burkitt lymphomas, diffuse large B-cell leukemias, mantle cell leukemias, Waldenstrom macroglobulinemia, and other B-cell lymphomas, respectively, were JL1-positive. Three mature T- and NK-cell NHLs were JL1-positive. JL1 expression was associated with age (P=0.045), complete response (P=0.004), and BM involvement at follow-up (P=0.017), but not with sex, performance status, the B symptoms, packed marrow pattern, cytogenetic abnormalities, or survival. CONCLUSIONS: JL1 positivity was associated with superior complete response and less BM involvement in NHL following chemotherapy.


Assuntos
Antígenos de Diferenciação de Linfócitos T/metabolismo , Células da Medula Óssea/metabolismo , Linfoma de Células B/patologia , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Células da Medula Óssea/citologia , Criança , Pré-Escolar , Feminino , Citometria de Fluxo , Humanos , Células Matadoras Naturais/citologia , Células Matadoras Naturais/metabolismo , Linfoma de Células B/metabolismo , Masculino , Pessoa de Meia-Idade , Linfócitos T/citologia , Linfócitos T/metabolismo , Adulto Jovem
3.
Anticancer Res ; 39(11): 6389-6392, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31704872

RESUMO

BACKGROUND/AIM: Neurofibromatosis type 1 (NF1) is characterized by the occurrence of multisystem tumors, among which the most characteristic are optic pathway gliomas (OPGs) and plexiform neurofibromas (PNFs). With the development of new anticancer drugs targeting the immune system, it is important to examine the immunological status of patients with NF1. Furthermore, the immune system has been suggested as a probable modulator of NF1-associated phenotypes. The objective of this study was the analysis of lymphocyte subset populations with respect to the presence of PNFs and OPGs. PATIENTS AND METHODS: Fifty-three patients with NF1 diagnosed with OPG/PNF were analyzed for lymphocyte subpopulations. RESULTS: Significantly lower levels of B-cells, T-cells and natural killer (NK) cells were observed in the group of patients with PNFs compared to those with OPG. CONCLUSION: Our observation may indicate a correlation between weakened functioning of the immune system and the formation of PNFs.


Assuntos
Subpopulações de Linfócitos B/citologia , Células Matadoras Naturais/citologia , Neurofibroma Plexiforme/imunologia , Neurofibromatose 1/imunologia , Glioma do Nervo Óptico/imunologia , Subpopulações de Linfócitos T/citologia , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Neurofibroma Plexiforme/etiologia , Neurofibromatose 1/complicações , Glioma do Nervo Óptico/etiologia
4.
BMC Infect Dis ; 19(1): 931, 2019 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-31690258

RESUMO

BACKGROUND: The sepsis-induced immunodepression contributes to impaired clinical outcomes of various stress conditions. This syndrome is well documented and characterized by attenuated function of innate and adaptive immune cells. Several pharmacological interventions aimed to restore the immune response are emerging of which interferon-gamma (IFNγ) is one. It is of paramount relevance to obtain clinical information on optimal timing of the IFNγ-treatment, -tolerance, -effectiveness and outcome before performing a RCT. We describe the effects of IFNγ in a cohort of 18 adult and 2 pediatric sepsis patients. METHODS: In this open-label prospective multi-center case-series, IFNγ treatment was initiated in patients selected on clinical and immunological criteria early (< 4 days) or late (> 7 days) following the onset of sepsis. The data collected in 18 adults and 2 liver transplanted pediatric patients were: clinical scores, monocyte expression of HLA-DR (flow cytometry), lymphocyte immune-phenotyping (flow cytometry), IL-6 and IL-10 plasma levels (ELISA), bacterial cultures, disease severity, and mortality. RESULTS: In 15 out of 18 patients IFNγ treatment was associated with an increase of median HLA-DR expression from 2666 [IQ 1547; 4991] to 12,451 [IQ 4166; 19,707], while the absolute number of lymphocyte subpopulations were not affected, except for the decrease number of NK cells 94.5 [23; 136] to 32.5 [13; 90.8] (0.0625)]. Plasma levels of IL-6 464 [201-770] to 108 (89-140) ng/mL (p = 0.04) and IL-10 from IL-10 from 29 [12-59] to 9 [1-15] pg/mL decreased significantly. Three patients who received IFNγ early after ICU admission (<4 days) died. The other patients had a rapid clinical improvement assessed by the SOFA score and bacterial cultures that were repeatedly positive became negative. The 2 pediatric cases improved rapidly, but 1 died for hemorrhagic complication. CONCLUSION: Guided by clinical and immunological monitoring, adjunctive immunotherapy with IFNγ appears well-tolerated in our cases and improves immune host defense in sepsis induced immuno suppression. Randomized clinical studies to assess its potential clinical benefit are warranted.


Assuntos
Tolerância Imunológica , Interferon gama/uso terapêutico , Sepse/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Feminino , Antígenos HLA-DR/metabolismo , Humanos , Lactente , Unidades de Terapia Intensiva , Interleucina-10/sangue , Interleucina-6/sangue , Células Matadoras Naturais/citologia , Células Matadoras Naturais/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/citologia , Monócitos/metabolismo , Estudos Prospectivos , Pseudomonas aeruginosa/isolamento & purificação , Sepse/microbiologia
5.
Chem Asian J ; 14(23): 4268-4273, 2019 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-31591824

RESUMO

The 9-mer peptide MFCH401 (N: 165-173: DTILWKDIF), which is located in the extracellular domain of HER2, has been predicted to be a novel epitope. Self-adjuvanting anti-HER2 vaccine constructs were designed and synthesized via covalently attaching MFCH401 or its linear tandem repeats (2×MFCH401, 3×MFCH401) to a lipopeptide Pam3 CSK4 via iterative condensation reaction. The in vivo results showed the Pam3 CSK4 -MFCH401 vaccine construct can induce higher antibody titers of IgG and IgM than those of other conjugates, and the analysis of changes in plasma cytokines level indicate the activation of Th1 cells and NK cells. In addition, the Pam3 CSK4 -MFCH401 vaccine conjugate induced a specific immune response to HER2-overexpressing human BT474 cells. Our data clearly indicated that MFCH401 is a promising epitope; moreover, its linear tandem repeats were unsuitable for anticancer vaccine design when conjugating with Pam3 CSK4 , which provided useful evidence for developing further anti-HER2 cancer vaccines.


Assuntos
Vacinas Anticâncer/imunologia , Peptídeos/imunologia , Animais , Anticorpos/sangue , Anticorpos/imunologia , Anticorpos/metabolismo , Vacinas Anticâncer/química , Linhagem Celular Tumoral , Citocinas/metabolismo , Humanos , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Camundongos , Peptídeos/síntese química , Peptídeos/química , Receptor ErbB-2/química , Células Th1/citologia , Células Th1/imunologia , Células Th1/metabolismo
6.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(8): 738-743, 2019 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-31638571

RESUMO

Objective To investigate the value of serum interleukin-6 (IL-6) and soluble interleukin-6 receptor (sIL-6R) levels of breast cancer patients in evaluating immune function after radiotherapy. Methods We randomly selected 55 cases of breast cancer patients who had received radiotherapy as an observation group, and 55 cases of normal healthy adults as a control group. ELISA was used to detect serum IL-6 and sIL-6R levels. Flow cytometry was used to detect CD45+CD19+ B lymphocytes, CD45+CD3+CD4+ T lymphocytes, CD45+CD3+CD8+ T lymphocytes and CD45+CD16+CD56+ NK cells in peripheral blood. The correlation of lymphocyte subsets with IL-6 or sIL-6R levels was analyzed. Results Serum IL-6 and sIL-6R levels of the observation group patients after radiotherapy were significantly higher than those before radiotherapy and in the control group, and the IL-6 and sIL-6R levels of patients of III or IV stage breast cancer increased significantly. Flow cytometry showed that the ratios of total T lymphocytes and NK cells in the observation group after radiotherapy were significantly reduced, while the ratio of B lymphocytes was significantly elevated. In addition, the levels of IL-6 and sIL-6R were positively correlated with the ratio of NK cells and negatively correlated with the ratio of B lymphocytes. Conclusion Serum IL-6 and sIL-6R levels in patients with breast cancer can be used as references for assessing the course of breast cancer and immune function after radiotherapy.


Assuntos
Neoplasias da Mama , Interleucina-6 , Células Matadoras Naturais , Subpopulações de Linfócitos , Receptores de Interleucina-6 , Adulto , Neoplasias da Mama/sangue , Neoplasias da Mama/imunologia , Neoplasias da Mama/radioterapia , Estudos de Casos e Controles , Feminino , Humanos , Interleucina-6/sangue , Células Matadoras Naturais/citologia , Subpopulações de Linfócitos/citologia
7.
Mol Med Rep ; 20(5): 4358-4366, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31545423

RESUMO

Natural killer (NK) cells are a group of large granular lymphocytes that play an important regulatory role in innate immunity and adaptive immunity. Immune­related pancytopenia (IRP) is a type of pancytopenia resulting from bone marrow hematopoietic cells that were destroyed or suppressed by auto­antibodies. The specific mechanism of IRP is not clear. In the present study, it was identified that the percentage of NK cells in peripheral blood lymphocytes was decreased in patients with IRP. Subsequently, high purity NK cells were extracted from 6 untreated patients with IRP using the immunomagnetic beads sorting, magnetic­activated cell­sorting method, which were then cultured then in RPMI­1640 medium containing 20% FBS. NK cell expansion agents, with or without recombinant interleukin (IL)­15, were used to amplify high­purity NK cells on the basic of recombinant IL­2. Expression of the activated receptors NKG2­D type II integral membrane protein (NKG2D) and natural killer cell p46­related protein (NKp46), and the inhibitory receptors CD158a and NKG2­A/NKG2­B type II integral membrane protein (NKG2A), in CD56+ NK cells were detected by flow cytometry before and after cell culture. It was observed that treatment with an NK cell expansion agent combined with the stimulation of recombinant IL­2 and recombinant IL­15 could increase the number whilst maintaining the purity of NK cells. There were no significant changes in the expression of NKG2D, NKp46, NKG2A and CD158a in patients with IRP before and after NK cell culture. This new amplification method lays a foundation for clinical NK cell immunotherapy and anti­tumor applications.


Assuntos
Interleucina-15/farmacologia , Interleucina-2/farmacologia , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Proteínas Recombinantes/farmacologia , Adolescente , Adulto , Idoso , Biomarcadores , Técnicas de Cultura de Células , Criança , Citocinas/farmacologia , Feminino , Citometria de Fluxo , Humanos , Imunoterapia Adotiva , Células Matadoras Naturais/citologia , Células Matadoras Naturais/metabolismo , Masculino , Pessoa de Meia-Idade , Pancitopenia/diagnóstico , Pancitopenia/etiologia , Pancitopenia/terapia , Adulto Jovem
8.
Food Chem Toxicol ; 134: 110816, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31518602

RESUMO

Our previous study has demonstrated that Pseudostellaria heterophylla protein hydrolysate (PPH) has immunomodulatory activity on murine spleen lymphocytes. The aim of this study was to investigate the excitation of PPH in RAW264.7 macrophage cells and the protective effect in cyclophosphamide (CTX)-treated mice. The results showed PPH of 50 µg/mL could stimulate macrophages resulting in significant promotions of nitric oxide (NO) production, endocytosis and reactive oxygen species formation. Meanwhile, enzyme-linked immunosorbent assay (ELISA) revealed that the levels of tumor necrosis factor-α and interleukin-10 were significantly upregulated by PPH. Furthermore, 50 mg/kg per day PPH restored the T lymphocyte proliferation and natural killer cell activity, and increased NO production and pinocytosis of peritoneal macrophages in CTX-treated mice. These findings indicate PPH plays a crucial role in RAW264.7 macrophage cells activation and in the protection against immunosuppression in CTX-treated mice and could be used as a potential immunostimulant agent.


Assuntos
Caryophyllaceae/química , Ciclofosfamida/farmacologia , Ativação de Macrófagos/efeitos dos fármacos , Hidrolisados de Proteína/farmacologia , Animais , Citocinas/metabolismo , Suplementos Nutricionais , Alimento Funcional , Imunidade Inata/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Células Matadoras Naturais/citologia , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Macrófagos Peritoneais/citologia , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/imunologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Peso Molecular , Hidrolisados de Proteína/química , Células RAW 264.7 , Linfócitos T/citologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia
9.
BMC Infect Dis ; 19(1): 792, 2019 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-31500589

RESUMO

BACKGROUND: Alveolar echinococcosis (AE) is caused by the larval stage of Echinococcus multilocularis (E. multilocularis), and considered as public health issue. Parasite-host immune interaction is pivotal during infection. As a subset of innate lymphoid cells, NK cells are known to play an important role during virus, bacteria, intra/extracellular parasitic infections and tumor progression. However, the possible role of NK cells in E. multilocularis infection in both human and murine is little known. Herein, the functional alteration of hepatic NK cells and their related molecules in E. multilocularis infected mice were studied. METHODS: 2000 protoscoleces (PSCs) were injected to C57BL/6 mice via the portal vein to establish secondary E. multilocularis infection. NK cells population and their related molecules (CD69, Ly49D, Ly49G2, Ly49H, Ly49I, NKG2A, NKG2D, granzyme B, IFN-γ, TNF-α) were assessed by using fluorescence-activated cell sorter (FACS) techniques and qRT-PCR. NK cell depletion was performed for further understanding the possible function of NK cells during infection. RESULTS: The total frequencies of NK cells and NK-derived IFN-γ production were significantly reduced at designated time points (2, 4, 12 weeks). The liver resident (CD49a+DX5-) NK cells are decreased at 4 weeks after inoculation and which is significantly lower than in control mice. Moreover, in vivo antibody-mediated NK cell depletion increased parasitic load and decreased peri-parasitic fibrosis. Expression of the inhibitory receptor NKG2A was negatively related to NK- derived IFN-γ secretion. CONCLUSIONS: Our study showed down regulates of NK cells and upper regulates of NKG2A expression on NK cells during E. multilocularis infection. Reduction of NK cell frequencies and increased NKG2A might result in low cytotoxic activity through decreased IFN-γ secretion in E. multilocularis infection. This result might be helpful to restore NK cell related immunity against E. multilocularis infection to treat alveolar echinococcosis.


Assuntos
Echinococcus multilocularis/fisiologia , Subfamília C de Receptores Semelhantes a Lectina de Células NK/metabolismo , Animais , Equinococose/microbiologia , Equinococose/patologia , Feminino , Tolerância Imunológica , Integrina alfa1/metabolismo , Interferon gama/metabolismo , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Fígado/imunologia , Fígado/patologia , Camundongos , Camundongos Endogâmicos C57BL , Subfamília C de Receptores Semelhantes a Lectina de Células NK/genética , Regulação para Cima/efeitos dos fármacos
10.
Oncol Rep ; 42(5): 2049-2056, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31485666

RESUMO

Osteosarcoma is a serious malignancy in pediatric patients, which comprises 2.4% of fatal cancer in children and achieves 20% of all primary bone cancers. In the present study, we employed three human osteosarcoma cell lines MG­63, HOS and U2OS for susceptibility to cytolytic activity of freshly isolated healthy donor NK cells. Cells were lysed by NK cells in a dose dependent manner. MG­63 cells exhibited less susceptibility to NK cells than HOS and U2OS cells at all cell ratios. The specific mechanism underlying the effects of NK cells on osteosarcoma cells was determined by antibody blockage experiments. The results revealed that granzyme B was the key factor in the NK cell­induced cytotoxicity of human osteosarcoma cells. To the best of our knowledge, the present study is the first to investigate the expression of PD­L1 in MG­63, HOS and U2OS cells. The relative expression of the PD­L1 gene and protein in MG­63 cell was greater than HOS and U2OS cells. The specific lysis of human osteosarcoma cells induced by NK cells was enhanced when PD­L1/PD­1 was blocked by the PD­L1 antibody. The present study proposed that the PD­L1/PD­1 axis serves an important role in NK cell­induced cytotoxicity in osteosarcoma via granzyme B secretion. Our findings may contribute to the development of precise treatments for osteosarcoma based on the expression profile of PD­L1 in patients with this disease.


Assuntos
Antígeno B7-H1/genética , Neoplasias Ósseas/imunologia , Granzimas/metabolismo , Células Matadoras Naturais/citologia , Osteossarcoma/imunologia , Receptor de Morte Celular Programada 1/genética , Adulto , Apoptose , Antígeno B7-H1/metabolismo , Neoplasias Ósseas/genética , Neoplasias Ósseas/terapia , Linhagem Celular Tumoral , Citotoxicidade Imunológica , Relação Dose-Resposta a Droga , Humanos , Células Matadoras Naturais/imunologia , Masculino , Osteossarcoma/genética , Osteossarcoma/terapia , Receptor de Morte Celular Programada 1/metabolismo , Adulto Jovem
11.
Hum Immunol ; 80(10): 842-847, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31320124

RESUMO

The natural killer group 2 (NKG2) family of receptors, encoded within the NK complex gene region (NKC), modulate the cytotoxic activity of NK cells. Two haplotype blocks throughout the NKC, hb-1 and hb-2 have been associated with different levels of overall natural cytotoxicity. Here, we evaluated allelic and genotype frequencies at rs1049174, rs2617160, rs2617170, rs2617171, rs1983526 (hb-1 haplotype), and rs2255336 and rs2246809 (hb-2 haplotype) in 928 subjects examined from Mexico City. The most frequent alleles and genotypes were as follows: C, CG to rs1049174; G, GG to rs2255336; T, AT to rs2617160; G, GG to rs2246809; C, CT to rs2617170; G, CG to rs2617171; and G, CG to rs1983526. Linkage disequilibrium analysis revealed that rs1049174, rs2617160, rs2617170, and rs2617171 constituted the haplotype block-1 variant (hb-1v) (r2 ≥ 0.89). Two predominant haplotypes of hb-1v were identified based on the allele content and included CTCG and GATC. This study is the first to evaluate the allelic and genotype frequency distribution of rs1049174, rs2255336, rs2617160, rs2246809, rs2617170, rs2617171, and rs1983526 in the population of Mexico City.


Assuntos
Haplótipos/genética , Subfamília C de Receptores Semelhantes a Lectina de Células NK/genética , Polimorfismo de Nucleotídeo Único/genética , Adulto , Alelos , Feminino , Frequência do Gene/genética , Técnicas de Genotipagem/métodos , Heterozigoto , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Células Matadoras Naturais/citologia , Células Matadoras Naturais/fisiologia , Desequilíbrio de Ligação/genética , Masculino , México , Pessoa de Meia-Idade
12.
Immunol Rev ; 290(1): 85-99, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31355489

RESUMO

Adoptive immunotherapy with natural killer cells was pioneered 30 years ago in human clinical trials with the development of cytokine-induced killer cells-unfractionated peripheral blood mononuclear cell (PBMC) populations activated overnight with IL-2. Higher doses were subsequently made possible through the advent of steady-state apheresis, allowing the collection of PBMC numbers equivalent to an entire adult blood volume, and increased purity made feasible through magnetic CD3-depletion and/or CD56-selection methods. Still, these approaches rarely achieved clinical dosing above a single infusion of 108 NK cells/kg, except with substantial donor-recipient size mismatch (eg, parents donating cells to children). To address this shortcoming, leukemia cell lines with NK cell-like function or ex vivo expansion approaches centered on the homeostatic cytokine IL-15 were developed. Here, we describe the development of an ex vivo expansion system based on a feeder cell expressing membrane-bound IL-21 that enables log-phase growth of primary NK cells for many weeks without inducing senescence, and describe the biology, correlative science, and translation to clinical trials for patients with leukemia, brain tumors, and solid tumors.


Assuntos
Terapia Baseada em Transplante de Células e Tecidos , Imunoterapia Adotiva , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Animais , Técnicas de Cultura de Células , Terapia Baseada em Transplante de Células e Tecidos/efeitos adversos , Terapia Baseada em Transplante de Células e Tecidos/métodos , Estudos Clínicos como Assunto , Criopreservação , Citotoxicidade Imunológica , Células Alimentadoras , Humanos , Imunoterapia Adotiva/efeitos adversos , Imunoterapia Adotiva/métodos , Interleucinas/metabolismo , Células Matadoras Naturais/citologia , Modelos Animais , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Resultado do Tratamento
13.
Zhonghua Gan Zang Bing Za Zhi ; 27(6): 436-439, 2019 Jun 20.
Artigo em Chinês | MEDLINE | ID: mdl-31357759

RESUMO

Objective: To evaluate the changes in natural killer cell subsets marked with CD27 and CD11b for HBV carrier mice. Methods: The pAAV-HBVl.2 plasmid was injected into the tail vein of C57BL/6 mice by hydrodynamic injection method to construct HBV-carrier model group and empty vector as the control group. Liver function and virological examination at different time points were used to judge the construction of HBV- plasmid carrier animal model. Flow cytometry was used to detect the frequency of NK cells and CD11b combined with CD27 NK cell subsets in spleen and liver. GraphPad Prism software was used for statistical analysis. Results: HBV-carrier mouse model was successfully constructed. There were no statistically significant difference in NK cell frequencies between spleen and liver of HBV carrier mice (P> 0.05), compared to control group. NK cells were divided into four subsets with in combination to CD27 and CD11b: CD11b(+)CD27(-)(CD11b(+)SP), CD11b(+)CD27(+)(DP), CD11b(-)CD27(+)(CD27(+)SP) and CD11b-CD27-(DN). Furthermore, the spleen of HBV-carrier mice had no statistically significant difference (P> 0.05) with the frequency of the four NK-cell subsets. The frequency of DN NK cell subsets was significantly increased in the liver of HBV carrier mice than control group (P< 0.001); however, the frequency of CD11b(+)SP cell subsets was significantly decreased (P < 0.05).There were no statistical significance in the frequency comparison between NK subgroups of DP and CD27(+)SP NK cell subsets (P> 0.05). Conclusion: HBV-carrier mice with abnormal distribution of hepatic NK cell subsets significantly increased and decreased the frequency of DN NK cell subsets and CD11b(+)SP cell subsets.


Assuntos
Antígeno CD11b , Vírus da Hepatite B , Células Matadoras Naturais , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral , Animais , Antígeno CD11b/metabolismo , Modelos Animais de Doenças , Citometria de Fluxo , Células Matadoras Naturais/citologia , Camundongos , Camundongos Endogâmicos C57BL
14.
Cells ; 8(6)2019 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-31181729

RESUMO

Hepatocellular carcinoma (HCC) is a primary malignancy of the liver, and currently the second most common cause of cancer-related deaths worldwide with increasing incidence and poor prognosis. Exosomes are now considered as important mediators of host anti-tumor immune response as well as tumor cell immune escape. HCC-derived exosomes have been shown to attenuate the cytotoxicity of T-cells and NK cells, and promote the immuno-suppressive M2 macrophages, N2 neutrophils, and Bregs. These exosomes harbor several immune-related non-coding RNAs and proteins that drive immune-escape and tumor progression, and thus may serve as potential diagnostic biomarkers and therapeutic targets for HCC. In a previous study, we identified miR146a as an exosomal factor that promotes M2-polarization and suppresses the anti-HCC function of T-cells. In this review, we summarized the role of tumor-derived exosomes and their key components in mediating tumor immune escape during HCC development.


Assuntos
Carcinoma Hepatocelular/patologia , Exossomos/metabolismo , Neoplasias Hepáticas/patologia , Linfócitos B/citologia , Linfócitos B/imunologia , Linfócitos B/metabolismo , Carcinoma Hepatocelular/imunologia , Carcinoma Hepatocelular/metabolismo , Células Dendríticas/citologia , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Humanos , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Neoplasias Hepáticas/imunologia , Neoplasias Hepáticas/metabolismo , MicroRNAs/metabolismo , Monócitos/citologia , Monócitos/imunologia , Monócitos/metabolismo , Linfócitos T/citologia , Linfócitos T/imunologia , Linfócitos T/metabolismo
15.
Bull Exp Biol Med ; 167(1): 169-176, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31183653

RESUMO

We studied changes in angiogenesis during contact interaction of natural killer cells and endothelial cells in the presence of secretory products of trophoblast cells activated by various cytokines. Activated trophoblast regulates angiogenesis by producing soluble factors that affect endothelial cells either directly or indirectly through activation of proangiogenic activity of natural killer cells. A stimulating effect of the trophoblast supernatants activated by IL-1ß and an inhibitory effect of trophoblast supernatants activated by IL-6 and TGFß for the formation of tube-like structures by endothelial cells were revealed. During contact culturing, natural killer cells increased the length of tube-like structures formed by endothelial cells. The trophoblast activated by IL-1ß affects angiogenesis both directly through the production of proangiogenic factors and indirectly through activation of the proangiogenic potential of natural killer cells. Trophoblast activated by IFNγ affects angiogenesis only by stimulating the proangiogenic potential of natural killer cells. Under conditions of contact interaction of natural killer cells and endothelial cells, soluble factors of trophoblast activated by IL-6 or TGFß attenuated the angiogenesis-stimulating effect of natural killer cells.


Assuntos
Células Endoteliais/citologia , Células Endoteliais/metabolismo , Células Matadoras Naturais/citologia , Células Matadoras Naturais/metabolismo , Trofoblastos/citologia , Trofoblastos/metabolismo , Comunicação Celular/fisiologia , Linhagem Celular , Linhagem Celular Tumoral , Feminino , Humanos , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Neovascularização Patológica/patologia , Fator de Crescimento Transformador beta/metabolismo
16.
Pharmacol Res Perspect ; 7(3): e00481, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31143450

RESUMO

Augmentation of natural killer (NK) cell cytotoxicity is one of the greatest challenges for cancer immunotherapy. Although histidine-rich glycoprotein (HRG), a 75-kDa glycoprotein with various immunomodulatory activities, reportedly elicits antitumor immunity, its effect on NK cell cytotoxicity is unclear. We assessed NK cell cytotoxicity against K562 cells. We also measured concentrations of cytokines and granzyme B in the cell supernatant. The proportion of CD56bright NK cells and NK cell surface PD-1 expression was assessed with flow cytometry. The neutralizing effects of anti-C-type lectin-like receptor (CLEC) 1B against HRG were also measured. NK cell morphological changes were analyzed via confocal microscopy. HRG significantly increased NK cell cytotoxicity against K562 cell lines. HRG also increased the release of granzyme B and the proportion of CD56bright NK cells. Further, HRG was able to decrease NK cell surface PD-1 expression. The effects of HRG on NK cells were reversed with anti-CLEC-1B antibodies. Additionally, we confirmed NK cell nuclear morphology and F-actin distribution, which are involved in the regulation of cytotoxic granule secretion. Because both PD-1 and CLEC-1B are associated with prognosis during malignancy, HRG incorporates these molecules to exert the antitumor immunity role. These facts indicate the potential of HRG to be a new target for cancer immunotherapy.


Assuntos
Células Matadoras Naturais/citologia , Lectinas Tipo C/metabolismo , Neoplasias/imunologia , Receptor de Morte Celular Programada 1/metabolismo , Proteínas/metabolismo , Actinas/metabolismo , Antígeno CD56/metabolismo , Núcleo Celular/metabolismo , Sobrevivência Celular , Células Cultivadas , Granzimas/metabolismo , Humanos , Imunoterapia , Células K562 , Células Matadoras Naturais/imunologia , Neoplasias/terapia
17.
Lab Chip ; 19(11): 2009-2018, 2019 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-31065640

RESUMO

Cytotoxicity exerted by cytotoxic lymphocytes against cancer cells is an essential cellular function for successful cancer immunotherapy. Standard cytotoxicity assays mostly provide population level information, whereas live cell imaging-based cytotoxicity assays can assess single cell level heterogeneity. However, long term tracking of individual cytotoxic lymphocyte-hematological cancer cell interactions is technically challenging because both cells can float around and form multi-cellular aggregates. To overcome this limitation, single hematological cancer cell arrays with immobilized hematological cancer cells are fabricated using microwell arrays. Using this new platform, single cell level natural killer (NK) cell cytotoxicity against leukemic cells is quantitatively assessed. Depending on microwell surface adhesiveness and inter-microwell distances, distinct modes of NK-leukemic cell interactions that result in different NK cell cytotoxicity are observed. For microwell arrays coated with bovine serum albumin, which prevents cell adhesion, NK cells stably contacted cancer cells with rounded morphologies, whereas for microwell arrays coated with fibronectin (FN), which triggers integrin signals, NK cells contacting cancer cells exhibited dynamic behaviors with elongated morphologies and constantly explored extracellular spaces by membrane extension. In addition, FN on extracellular spaces facilitate NK cell detachment from leukemic cells after killing by providing anchorage for force transmission, and promote cytotoxicity and serial killing. Single hematologic cell arrays are not only an efficient method for lymphocyte cytotoxicity analysis but also a useful tool to study the role of signaling molecules in extracellular spaces on lymphocyte cytotoxicity.


Assuntos
Espaço Extracelular/metabolismo , Neoplasias Hematológicas/imunologia , Neoplasias Hematológicas/patologia , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Análise de Célula Única/instrumentação , Análise Serial de Tecidos/instrumentação , Adesão Celular , Linhagem Celular Tumoral , Humanos
19.
Cells ; 8(5)2019 05 16.
Artigo em Inglês | MEDLINE | ID: mdl-31100864

RESUMO

The immune system is a fundamental part of the tumor microenvironment. In particular, cytotoxic lymphocytes, such as cytolytic T cells and natural killer cells, control tumor growth and disease progression by interacting and eliminating tumor cells. The actin cytoskeleton of cytotoxic lymphocytes engaged in an immunological synapse has received considerable research attention. It has been recognized as a central mediator of the formation and maturation of the immunological synapse, and its signaling and cytolytic activities. In comparison, fewer studies have explored the organization and function of actin filaments on the target cancer cell side of the immunological synapse. However, there is growing evidence that the actin cytoskeleton of cancer cells also undergoes extensive remodeling upon cytotoxic lymphocyte attack, and that such remodeling can alter physical and functional interactions at the immunological synapse. In this article, we review the current knowledge of actin organization and functions at both sides of the immunological synapse between cytotoxic lymphocytes and cancer cells, with particular focus on synapse formation, signaling and cytolytic activity, and immune evasion.


Assuntos
Citoesqueleto de Actina/metabolismo , Citoesqueleto de Actina/fisiologia , Sinapses Imunológicas/imunologia , Células Matadoras Naturais/imunologia , Neoplasias/imunologia , Linfócitos T Citotóxicos/imunologia , Microambiente Tumoral/imunologia , Animais , Linhagem Celular , Humanos , Evasão da Resposta Imune , Células Matadoras Naturais/citologia , Camundongos , Linfócitos T Citotóxicos/citologia
20.
Int J Biol Macromol ; 133: 202-213, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-30991069

RESUMO

Setaria equina heat shock protein (SeqHSP) 70 gene was characterized, cloned and expressed to recombinant protein (rSeqHSP70). The protein was tested for binding with an anti-filarial drug "diethylcarbamazine citrate (DEC)" by equilibrium dialysis method. Molecular docking was also used to determine the binding sites and residues of binding with DEC. The mice were immunized with the protein alone or bound to DEC. Serum IFN-γ levels in the immunized group with protein-drug complex were significantly higher (P < 0.05) than the protein-immunized group. Mouse anti-SeqHSP70 polyclonal IgG recognized 2 bands at 70 and 75 kDa in S. equina adult worm and human cancer cell lines (HepG2 and MCF-7) extracts. The proliferation assay for mice splenocytes revealed a potentiation and down-regulating effects in non-immunized and immunized groups, respectively with the drug-protein complex. The proliferation and IFN-γ assays for purified human NK cells indicated a potentiating effect of the drug-protein complex (DEC concentration is 50 µM) comparable to the protein. DEC at lower concentration (25 mM) could also show a significant increase (P < 0.05) in IFN-γ. From the results, DEC was postulated to induce conformational changes in the protein exposing more epitopes for NK cell binding and activation.


Assuntos
Dietilcarbamazina/metabolismo , Filarioidea/genética , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/metabolismo , Adjuvantes Imunológicos/farmacologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Proliferação de Células/efeitos dos fármacos , Clonagem Molecular , Reações Cruzadas , Expressão Gênica , Proteínas de Choque Térmico HSP70/química , Proteínas de Choque Térmico HSP70/farmacologia , Proteínas de Helminto/química , Proteínas de Helminto/farmacologia , Humanos , Interferon gama/metabolismo , Células Matadoras Naturais/citologia , Células Matadoras Naturais/efeitos dos fármacos , Camundongos , Simulação de Acoplamento Molecular , Conformação Proteica , Análise de Sequência , Baço/imunologia
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