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1.
J Immunol ; 195(10): 4555-63, 2015 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-26546687

RESUMO

CD (cluster of differentiation) Ags are cell surface molecules expressed on leukocytes and other cells relevant for the immune system. CD nomenclature has been universally adopted by the scientific community and is officially approved by the International Union of Immunological Societies and sanctioned by the World Health Organization. It provides a unified designation system for mAbs, as well as for the cell surface molecules that they recognize. This nomenclature was established by the Human Leukocyte Differentiation Antigens Workshops. In addition to defining the CD nomenclature, these workshops have been instrumental in identifying and determining the expression and function of cell surface molecules. Over the past 30 y, the data generated by the 10 Human Leukocyte Differentiation Antigens Workshops have led to the characterization and formal designation of more than 400 molecules. CD molecules are commonly used as cell markers, allowing the identification and isolation of leukocyte populations, subsets, and differentiation stages. mAbs against these molecules have proven to be essential for biomedical research and diagnosis, as well as in biotechnology. More recently, they have been recognized as invaluable tools for the treatment of several malignancies and autoimmune diseases. In this article, we describe how the CD nomenclature was established, present the official updated list of CD molecules, and provide a rationale for their usefulness in the 21st century.


Assuntos
Anticorpos Monoclonais/imunologia , Antígenos CD/classificação , Terminologia como Assunto , Antígenos CD/imunologia , Biomarcadores , Humanos
2.
Am J Physiol Heart Circ Physiol ; 309(8): H1303-13, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26371161

RESUMO

A growing number of studies have suggested microRNAs (miRNAs) are involved in the modulation of myocardial ischemia-reperfusion (MI/R) injury; however, the role of endogenous miRNAs targeting endothelial cells (ECs) and its interaction with ICAM-1 in the setting of MI/R remain poorly understood. Our microarray results showed that miR-146a, miR-146b-5p, miR-155*, miR-155, miR-497, and miR-451 were significantly upregulated, whereas, miR-141 and miR-564 were significantly downregulated in the ECs challenged with TNF-α for 6 h. Real-time PCR analyses additionally validated that the expression levels of miR-146a, miR-155*, and miR-141 were consistent with the microarray results. Then, ICAM-1 was identified as a novel target of miR-141 by Target Scan software and the reporter gene system. Further functional experiments showed that elevated levels of miR-141 inhibited ICAM-1 expression and diminished leukocytes adhesion to ECs in vitro. In an in vivo murine model of MI/R injury, pretreatment with miR-141 mimics through the tail vein downregulated the expression level of ICAM-1 in heart and attenuated MI/R injury as evidenced by decreased infarct size and decline of serum cardial troponin I (cTnI) and lactate dehydrogenase (LDH) concentration. The cardioprotective effects of miR-141 mimics may be attributed to the decreased infiltration of CD11b(+) cells and F4/80(+) macrophages into ischemic myocardium tissue. In conclusion, our results demonstrate that miR-141, as a novel repressor of ICAM-1, is involved in the attenuation of MI/R injury via antithetical regulation of ICAM-1 and inflammatory cells infiltration. Thus miR-141 may constitute a new therapeutic target in the setting of ischemic heart disease.


Assuntos
Células Endoteliais/metabolismo , Terapia Genética/métodos , Molécula 1 de Adesão Intercelular/metabolismo , MicroRNAs/metabolismo , Infarto do Miocárdio/prevenção & controle , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miocárdio/metabolismo , Regiões 3' não Traduzidas , Animais , Adesão Celular , Técnicas de Cocultura , Modelos Animais de Doenças , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/patologia , Feminino , Regulação da Expressão Gênica , Células HL-60 , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Molécula 1 de Adesão Intercelular/genética , Leucócitos/metabolismo , Macrófagos/metabolismo , Camundongos Endogâmicos BALB C , MicroRNAs/genética , Infarto do Miocárdio/genética , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/genética , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Miocárdio/patologia , RNA Mensageiro/metabolismo , Fatores de Tempo , Transfecção , Fator de Necrose Tumoral alfa/farmacologia
3.
Int Immunol ; 26(5): 269-81, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24355664

RESUMO

CD4(+) T cells play critical roles in orchestrating adaptive immune responses. Their activation and proliferation are critical steps that occur before they execute their biological functions. Despite the important role of this process, the underlying molecular events are not fully understood. MicroRNAs (miRNAs) have been shown to play important roles in lymphocyte development and function. However, the miRNAs that regulate T-cell differentiation, activation and proliferation are still largely unknown. In our previous study, using a miRNA array, we found that several miRNAs (including miR-202, 33b, 181c, 568 and 576) are differentially expressed between resting and activated CD4(+) T cells. In this study, we focused on the function of miR-568 during CD4(+) T-cell activation. We showed that the expression level of miR-568 decreased during the activation of T cells, including Jurkat cells and human peripheral blood CD4(+) T cells. When Jurkat or human peripheral blood CD4(+) T cells were transfected with miR-568 mimics, cell activation was significantly inhibited, as shown by the inhibited expression of activation markers such as CD25, CD69 and CD154; decreased IL-2 production; and inhibited cell proliferation. Using software predictions and confirmatory experiments, we demonstrated that nuclear factor of activated T cells 5 (NFAT5) is a target of miR-568. Treg cells are an important CD4(+) T-cell subpopulation, so we also evaluated the function of miR-568 in Treg-cell activation and differentiation. We showed that the miR-568 level decreased, while the NFAT5 protein level increased during CD4(+)CD25(+) Treg-cell activation, and the transfection of miR-568 mimics inhibited the NFAT5 expression, inhibited the production of both TGF-ß and IL-10 and also inhibited the proliferation of Treg cells. Our further study showed that over-expression of miR-568 can inhibit Treg-cell differentiation and can inhibit the suppressive effect of these cells on effector cells. In addition, inhibition of NFAT5 by siRNA-mediated knockdown can inhibit the activation and differentiation of Treg cells. These findings reveal that miR-568 can inhibit the activation and function of both CD4(+) T cells and Treg cells by targeting NFAT5. Since miR-568 plays an important role in both CD4(+) T cells and Treg cells, these findings may provide leads for the development of novel treatments for human inflammatory and autoimmune diseases.


Assuntos
Linfócitos T CD4-Positivos/imunologia , MicroRNAs/imunologia , Linfócitos T Reguladores/imunologia , Fatores de Transcrição/imunologia , Regiões 3' não Traduzidas/genética , Western Blotting , Linfócitos T CD4-Positivos/metabolismo , Diferenciação Celular/genética , Diferenciação Celular/imunologia , Proliferação de Células , Células Cultivadas , Citometria de Fluxo , Expressão Gênica/imunologia , Células HEK293 , Humanos , Interleucina-2/imunologia , Interleucina-2/metabolismo , Células Jurkat , Luciferases/genética , Luciferases/metabolismo , Ativação Linfocitária/genética , Ativação Linfocitária/imunologia , MicroRNAs/genética , Mutação , Interferência de RNA , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Linfócitos T Reguladores/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
4.
Gastroenterology ; 141(6): 2076-2087.e6, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21925125

RESUMO

BACKGROUND & AIMS: Human epidermal growth factor receptor 2 (HER2) (neu/ERBB2) is overexpressed on many types of cancer cells, including gastric cancer cells; HER2 overexpression has been associated with metastasis and poor prognosis. We investigated the mechanisms by which HER2 regulates cell migration and invasion. METHODS: HER2 expression or activity was reduced in gastric cancer cell lines using small interfering RNAs or the monoclonal antibody, trastuzumab. We identified proteins that interact with HER2 or microRNAs (miRNAs) involved in HER2 signaling. We used various software programs to identify miRNAs that regulate factors in the HER2 signaling pathway. We analyzed expression patterns of these miRNAs in gastric cancer cell lines and tumor samples from patients. RESULTS: We found that CD44 binds directly to HER2, which up-regulates the expression of metastasis-associated protein-1, induces deacetylation of histone H3 lysine 9, and suppresses transcription of microRNA139 (miR-139) to inhibit expression of its target gene, C-X-C chemokine receptor type 4 (CXCR4). Knockdown of HER2 and CD44 reduced invasive activity of cultured gastric cancer cells and suppressed tumor growth in nude mice. Lymph node metastasis was associated with high levels of HER2, CD44, and CXCR4, and reduced levels of miR-139 in human metastatic gastric tumors. Cultures of different types of metastatic cancer cells with histone deacetylase inhibitors and/or DNA methyltransferase resulted in up-regulation of miR-139. CONCLUSIONS: HER2 interaction with CD44 up-regulates CXCR4 by inhibiting expression of miR-139, at the epigenetic level, in gastric cancer cells. These findings indicate how HER2 signaling might promote gastric tumor progression and metastasis.


Assuntos
Epigênese Genética/genética , Receptores de Hialuronatos/metabolismo , MicroRNAs/genética , Receptor ErbB-2/metabolismo , Receptores CXCR4/metabolismo , Neoplasias Gástricas/genética , Animais , Northern Blotting , Movimento Celular , Primers do DNA/química , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Humanos , Camundongos , Camundongos Nus , Metástase Neoplásica , Técnicas de Amplificação de Ácido Nucleico , Células Tumorais Cultivadas , Regulação para Cima
5.
Res Pract Thromb Haemost ; 6(7): e12813, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36246479

RESUMO

Background: Hemorrhagic fever with renal syndrome (HFRS) induced by Hantaan virus infection and heparin-induced thrombocytopenia (HIT) are associated with symptoms such as thrombocytopenia and thrombosis. However, related molecules, such as anti-platelet factor 4 (PF4)/heparin antibodies, in patients with HFRS have not been evaluated. Objectives: To test plasma levels of anti-PF4/heparin antibodies and study the possible role of these antibodies in HFRS pathogenesis. Methods: Indirect ELISA was used to determine plasma levels of anti-PF4/heparin antibodies in 75 patients with HFRS and 20 normal controls. The 4Ts (thrombocytopenia, timing of platelet count fall, thrombosis or other sequelae, and other causes of thrombocytopenia) scoring system was used to determine the probability of HIT occurrence. A PF4-enhanced platelet activation assay was used to detect the pathological effects of anti-PF4/heparin antibodies. The laboratory/clinical features and viral load of all the patients were also assessed. Results: Of the 75 patients with HFRS enrolled in this study, 69 had thrombocytopenia. Platelet count was negatively correlated with Hantaan viral load. Moreover, the optical density (OD) values of plasma antibodies against PF4/heparin in normal controls were less than 0.65, 4 patients tested strongly positive for anti-PF4/heparin antibodies (OD values, 1.51-3.87), 21 patients were weakly positive (OD values, 0.66-0.74), and 50 patients were negative (OD values, 0.16-0.65). Moreover, all 4 patients who tested strongly positive for anti-PF4/heparin antibodies showed a low probability of HIT (4Ts score of 3 or less) and had negative results in the PF4-enhanced platelet activation assay. Conclusions: Hantaan virus infection produces nonpathogenic antibodies against PF4/heparin; however, the generation mechanism of these antibodies requires further study.

6.
Arch Biochem Biophys ; 499(1-2): 49-55, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20460098

RESUMO

HER2 overexpression is associated with metastasis-the main cause of death in individuals with gastric cancer. In this study, we demonstrated that vector-based shRNA significantly knocked down the expression of HER2 and considerably inhibited both the migration and invasion of gastric cancer cells. HER2 knockdown resulted in the downregulation of the expression of MMP-1, while HER2 overexpression improved the transcription of MMP-1 through the activation of an MMP-1 promoter. The promoter region of MMP-1 between -2500 and -2000 bp was found to be crucial for the upregulation of HER2-mediated transcription. Furthermore, a truncated promoter (-70 to+63) did not display any transcriptional activity. Cell invasion activity was almost completely inhibited when MMP-1 was knocked down. Conversely, the overexpression of MMP-1 partly rescued the invasion ability of cell strains with knocked-down HER2. These findings help further understanding of the molecular mechanisms through which HER2 promotes malignancy, and suggest that targeting both HER2 and MMP-1 may be required to effectively block HER2 signaling in gastric cancer therapy.


Assuntos
Metaloproteinase 1 da Matriz/metabolismo , Receptor ErbB-2/metabolismo , Neoplasias Gástricas/metabolismo , Sequência de Bases , Linhagem Celular , Linhagem Celular Tumoral , Primers do DNA/genética , Técnicas de Silenciamento de Genes , Genes erbB-2 , Humanos , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Invasividade Neoplásica/genética , Invasividade Neoplásica/fisiopatologia , Análise de Sequência com Séries de Oligonucleotídeos , Regiões Promotoras Genéticas , RNA Interferente Pequeno/genética , Receptor ErbB-2/genética , Neoplasias Gástricas/genética , Neoplasias Gástricas/patologia , Regulação para Cima
7.
Int J Cancer ; 124(12): 2893-8, 2009 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-19296537

RESUMO

CT45 is a cancer/testis gene that we previously identified by massively parallel signature sequencing. Encoded by a multigene family on chromosome X, CT45 showed restricted mRNA expression to normal testis and various cancers. In this study, monoclonal antibodies were generated against recombinant CT45 protein, and CT45 protein expression in normal and tumor tissues was evaluated by immunohistochemical analysis. In adult normal tissue, CT45 expression was restricted to testicular germ cells, detected as a nuclear protein mainly at the stage of primary spermatocytes. In tumors, CT45 protein expression correlated with the mRNA levels detected by quantitative RT-PCR, and most lung cancer and ovarian cancers with CT45 mRNA at levels >1% of testicular expression were CT45 protein-positive. In positive cases, CT45 showed expression patterns that ranged from diffuse strong staining to heterogeneous and patchy expression. In lung cancer, CT45 expression was least frequent in adenocarcinoma, more frequent in squamous cell carcinoma and neuroendocrine tumors. Using tissue microarrays, 376 lung cancer, 219 ovarian cancer and 155 breast cancer were evaluated for CT45 protein expression. The expression frequency was highest in ovarian cancer (37%), followed by lung cancer (13%) and lowest in breast cancer (<5%). Given the focal nature of CT45 expression in many cases, these numbers represented the minimal frequency of expression in these tumor types. In summary, the expression frequency and characteristics of CT45 expression are similar to other CT cancer vaccine targets currently in clinical trials, e.g., NY-ESO-1 and MAGE-A, suggesting CT45 as a potentially useful cancer target.


Assuntos
Antígenos de Neoplasias/genética , Antígenos de Neoplasias/metabolismo , Neoplasias da Mama/metabolismo , Neoplasias Pulmonares/metabolismo , Neoplasias Ovarianas/metabolismo , RNA Mensageiro/metabolismo , Neoplasias Testiculares/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/secundário , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/metabolismo , Antígenos de Neoplasias/imunologia , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/secundário , Feminino , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Prognóstico , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias Testiculares/genética , Neoplasias Testiculares/patologia , Análise Serial de Tecidos
8.
Zhonghua Wai Ke Za Zhi ; 47(14): 1070-4, 2009 Jul 15.
Artigo em Zh | MEDLINE | ID: mdl-19781272

RESUMO

OBJECTIVE: To investigate the correlation between major histocompatibility complex (MHC) class I chain-related gene A (MICA) gene alleles matching rates and graft rejection in small intestine, liver and kidney transplantation. METHODS: Genome DNA were extracted from blood samples or pathological sections collected from donors and recipients of living-related transplantation, included 4 cases of small bowel transplantation, 5 cases of liver transplantation and 6 cases of kidney transplantation. The correlation between MICA alleles matching rates and acute graft rejection was analyzed following 13 MICA alleles determination by polymerase chain reaction based on sequence-specific primers (PCR-SSP). RESULTS: HLA zygosity of all donors and recipients was confirmed to be half-matching. The recipients displaying higher matching rates of MICA alleles with donors showed lighter clinical and pathological rejection and longer survival time. On the contrary, recipients with lower matching rates of MICA alleles with donors showed severer clinical and pathological rejection and shorter survival time relatively. CONCLUSION: Matching rates of MICA alleles has negative relevance to acute rejection, and positive relevance to survival time of recipients in small bowel, liver, and kidney transplantation.


Assuntos
Rejeição de Enxerto/genética , Antígenos de Histocompatibilidade Classe I/genética , Doadores Vivos , Transplante de Órgãos , Alelos , Rejeição de Enxerto/imunologia , Humanos , Intestino Delgado/transplante , Transplante de Rim/imunologia , Transplante de Fígado/imunologia
9.
Appl Microbiol Biotechnol ; 81(1): 183-9, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18818915

RESUMO

Based on a series of mAbs against four frequently used tags--the human Ig Fc fragment, GST, maltose-binding protein, and thioredoxin--we developed corresponding sandwich enzyme-linked immunosorbent assay (ELISA) to detect these tag fusion proteins. As a supplement for Western blot, the successfully established ELISA was specific, sensitive, quantitative, easy to perform, time-saving, and last but not least, suitable for high-throughput screening of tag fusion proteins. Determination of soluble tag fusion proteins expressed by various systems with the sandwich ELISA developed in the present study could be a valuable and promising tool for the wide application of tag-protein fusion systems in the rapidly growing field of proteomics research.


Assuntos
Ensaio de Imunoadsorção Enzimática/métodos , Proteínas Recombinantes de Fusão/análise , Animais , Anticorpos Monoclonais/análise , Western Blotting , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo
10.
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue ; 20(9): 516-9, 2008 Sep.
Artigo em Zh | MEDLINE | ID: mdl-18786305

RESUMO

OBJECTIVE: To investigate the relationship between dysfunction of cell-mediated immunity and prognosis in severely burned patients. METHODS: Twenty-six patients with total burn surface area larger than 70% total body surface area (TBSA) were included in the present study, and they were divided into non-survival group (14 cases) and survival group (12 cases). Eleven healthy volunteers served as controls. The blood samples were collected on days 1, 3, 7, 14 postburn, the human leukocyte antigen-DR (HLA-DR) bound on CD14+ mononuclear cell surface of burned patients was quantified by flow cytometry (using monoclonal antibody, QuantiBRITE anti-HLA-DR PE/anti-monocyte PerCP-Cy5.5). The ability of T lymphocyte proliferation was determined by thiazolyl blue (MTT) method, and interleukin-2 (IL-2) release was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared to the survival group, the T lymphocyte proliferative activity (0.739+/-0.299 vs. 0.320+/-0.237) and IL-2 release [(11.02+/-2.50) ng/L vs. (8.21+/-2.63) ng/L] in non-survival group were significantly decreased on postburn day 14 (P < 0.01 and P < 0.05). The expression of HLA-DR on CD14+ mononuclear cell surface was persistently decreased in non-survivors, while it markedly elevated in survivors after 14 days postburn (P < 0.01). CONCLUSION: The cellular immune function is consistently suppressed in severely burned patients. Sequential monitoring of T lymphocyte proliferation, IL-2 release, and the amount of HLA-DR on CD14+ mononuclear cell surface might be of clinical prognostic significance in patients with massive burn injury.


Assuntos
Queimaduras/imunologia , Imunidade Celular , Adulto , Estudos de Casos e Controles , Feminino , Antígenos HLA-DR/metabolismo , Humanos , Interleucina-2/metabolismo , Leucócitos Mononucleares/imunologia , Receptores de Lipopolissacarídeos/metabolismo , Masculino , Pessoa de Meia-Idade , Prognóstico , Linfócitos T/imunologia , Adulto Jovem
11.
Zhonghua Wai Ke Za Zhi ; 46(10): 759-62, 2008 May 15.
Artigo em Zh | MEDLINE | ID: mdl-18953932

RESUMO

OBJECTIVE: To investigate the change in T cell-mediated immunity and its relationship with plasma high mobility group box-1 protein (HMGB1) levels in severely burned patients. METHODS: Thirty-five extensively burned patients (> 30% total body surface area) were included in this study, and were divided into MODS group (n = 13) and non-MODS group (n = 22). The blood samples were collected on post burn days 1, 3, 5, 7, 14, 21 and 28. The plasma levels of HMGB1 were measured by using ELISA, and T lymphocyte proliferation response and its IL-2 production ability in peripheral blood were determined too. In addition, the ratio of CD4+/CD8+ T cells were detected by using flow cytometry. RESULTS: Plasma HMGB1 levels were markedly elevated on post burn day 1 in severely burned patients, and HMGB1 level was significantly higher in MODS group than in non-MODS group (P < 0.05). Lymph proliferation response and IL-2 production of T cells in peripheral blood, and the ratio of CD4+/CD8+ T cells in MODS group were markedly lower than those in non-MODS group on post burn days 1, 14, 21 and 28 (all P < 0.05). It indicated that plasma HMGB1 levels were negatively correlated to T cellular immune function parameters, including lymphocyte proliferation response, IL-2 production, and the ratio of CD4+/ CD8+ T cells in extensively burned patients (all P < 0.05). CONCLUSIONS: Extensive burns could lead to T cellular immune dysfunction, which appears to be associated with the development of MODS. HMGB1, as an important late mediators of inflammation, might be involved in the pathogenesis of suppression of T cell-mediated immunity in these patients.


Assuntos
Queimaduras/imunologia , Proteína HMGB1/sangue , Linfócitos T/imunologia , Adolescente , Adulto , Queimaduras/sangue , Queimaduras/complicações , Feminino , Humanos , Imunidade Celular/imunologia , Masculino , Pessoa de Meia-Idade , Insuficiência de Múltiplos Órgãos/etiologia , Insuficiência de Múltiplos Órgãos/imunologia
12.
J Immunol Methods ; 319(1-2): 1-5, 2007 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-17174972

RESUMO

The Human Leucocyte Differentiation Antigens Workshops (HLDA) have since 1984 provided a forum for the characterization and study of leucocyte surface molecules and antibodies against them. HLDA devised the CD nomenclature, which is sanctioned by IUIS. The HLDA Council reviewed and modified the objectives of HLDA in 2004, and changed the name of the organization to Human Cell Differentiation Molecules (HCDM) to reflect the broader objectives. Workshop studies under the HCDM banner proceeded during 2005 and early 2006, culminating in a meeting in May 2006. At that meeting the Council, acting as Nomenclature Committee, approved a number of new CD designations and changes to some pre-existing CD designations, which are summarized in this report.


Assuntos
Antígenos CD/classificação , Diferenciação Celular/imunologia , Terminologia como Assunto , Antígenos CD/genética , Antígenos CD/imunologia , Diferenciação Celular/genética , Humanos , Cooperação Internacional
13.
Cell Res ; 15(6): 430-8, 2005 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15987601

RESUMO

TRAIL, tumor necrosis factor-related apoptosis-inducing ligand, is a member of the TNF family of proteins. Tumour cells were initially found to have increased sensitivity to TRAIL compared with normal cells, raising hopes that TRAIL would prove useful as an anti-tumor agent. The production of reliable monoclonal antibodies against TRAIL and its receptors that can stain fixed specimens will allow a thorough analysis of their expression on normal and malignant tissues. Here we report the generation of monoclonal antibodies against TRAIL and its four membrane-bound receptors (TR1-4), which have been used to stain a range of normal and malignant cells, as routinely fixed specimens. Low levels of TRAIL expression were found to be limited mostly to smooth muscle in lung and spleen as well as glial cells in the cerebellum and follicular cells in the thyroid. Expression of the TRAIL decoy receptors (TR3 and 4) was not as widespread as indicated by Northern blotting, suggesting that they may be less important for the control of TRAIL cytotoxicity than previously thought. TR1 and TR2 expression increases significantly in a number of malignant tissues, but in some common malignancies their expression was low, or patchy, which may limit the therapeutic role of TRAIL. Taken together, we have a panel of monoclonal antibodies that will allow a better assessment of the normal role of TRAIL and allow assessment of biopsy material, possibly allowing the identification of tumors that may be amenable to TRAIL therapy.


Assuntos
Glicoproteínas de Membrana/biossíntese , Neoplasias/fisiopatologia , Receptores do Fator de Necrose Tumoral/biossíntese , Fator de Necrose Tumoral alfa/biossíntese , Anticorpos Monoclonais/biossíntese , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose , Feminino , Proteínas Ligadas por GPI , Humanos , Imuno-Histoquímica/métodos , Células Jurkat , Masculino , Glicoproteínas de Membrana/imunologia , Neoplasias/patologia , Receptores do Ligante Indutor de Apoptose Relacionado a TNF , Receptores do Fator de Necrose Tumoral/imunologia , Membro 10c de Receptores do Fator de Necrose Tumoral , Coloração e Rotulagem/métodos , Ligante Indutor de Apoptose Relacionado a TNF , Distribuição Tecidual , Receptores Chamariz do Fator de Necrose Tumoral , Fator de Necrose Tumoral alfa/imunologia
14.
World J Gastroenterol ; 11(3): 344-7, 2005 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-15637741

RESUMO

AIM: To investigate the expression pattern of epithelial cell adhesion molecule (Ep-CAM) on normal and malignant colon tissues to evaluate its diagnostic and therapeutic significance. METHODS: cDNA encoding Ep-CAM extracellular domain was cloned by reverse transcription-polymerase chain reaction (RT-PCR) from excised malignant colon tissues and inserted into a glutathione S-transferase (GST)-tagged vector. Ep-CAM-GST fusion protein was induced by isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified with glutathione-sepharose. The Ep-CAM-GST fusion protein was mixed with Freund's adjuvant and Balb/c mice were immunized with it. Sp2/0 myeloma cells were fused with the spleen cells of the immunized mice. After having selected by indirect ELISA, the anti-Ep-CAM monoclonal antibodies (MAbs) were generated and the corresponding ascites were obtained. Finally, the human colon carcinoma tissue array prepared from seventy individual patients was stained with the anti-Ep-CAM MAbs. RESULTS: The isolated Ep-CAM cDNA sequence was identical to the data in GenBank. The expressed fusion protein was almost soluble and had a molecular weight (MW) of 53 ku. Four MAbs against Ep-CAM were obtained and designated as FMU-Ep1, FMU-Ep2, FMU-Ep3 and FMU-Ep4 respectively. Among them, FMU-Ep4 could recognize the natural Ep-CAM on Colo205 and SW480 cells, and all of them could be used for immunohistochemical staining of tissue sections. It was found that Ep-CAM was distributed differently in normal and various malignant colon tissues, including squamous cell carcinoma, signet-ring cell carcinoma and adenocarcinoma. In normal colon gland epithelia, Ep-CAM antigen was mainly distributed on the basolateral membrane and in the region between the basolateral membrane and the cytoplastic part near the nuclei, whereas the expression pattern of colon malignancies was mainly on the whole surface of epithelia and the expression was much higher than the normal colon tissues. The staining pattern of tissue array showed in adenocarcinoma and papillary adenocarcinoma, and the expression of Ep-CAM was increased from grade I to grade III. CONCLUSION: MAbs against Ep-CAM might be useful for research on the structure and function of Ep-CAM and may have diagnostic and therapeutic value to various colon carcinomas.


Assuntos
Antígenos de Neoplasias/metabolismo , Moléculas de Adesão Celular/metabolismo , Colo/metabolismo , Neoplasias do Colo/metabolismo , Animais , Linhagem Celular Tumoral , Molécula de Adesão da Célula Epitelial , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Distribuição Tecidual
15.
Zhonghua Yi Xue Za Zhi ; 85(3): 198-202, 2005 Jan 19.
Artigo em Zh | MEDLINE | ID: mdl-15854468

RESUMO

OBJECTIVE: To investigate the effect of specific small interfering RNA (siRNA) targeting against bcr-abl chimeric gene on the biological traits of chronic myelogenous leukemia (CML) cells. METHODS: CML cells of the line K561 transcribing a type of b3: a2 mRNA of bcr-abl chimeric gene were cultured. A 21nt siRNA targeting against the chimeric location of the b3: a2 mRNA of bcr-abl chimeric gene was designed, synthesized, and transfected into the K562 cells as RNA interference group. Another K562 cells were transfected with fluorescein enzyme gene specific siRNA as indifferent controls, or with lipid alone as blank vector controls. Some K562 cells without treatment were used as normal controls. 48 hours after the transfection Western blotting was used to detect the expression of P210bcr-abl fusion protein. 3H-TdR incorporation was used to detect the proliferation activity of K562. Annexin V-fluorescencein isothiocyanate (FITC)/phosphatidylinositol (PI) staining was used to detect the apoptosis of K2562 cells. Flow cytometry was used to observe the cell cycle of K562 cells. Benzidine staining was used to detect the differentiation of K562 cells towards erythrocytic series. Western blotting was used further to detect the expression of apoptosis-related protein Bcl-xL/Bax. RESULTS: (1) In contrast with the control groups, the expression level of bcr-abl chimeric gene was much lower in the RNAi group. (2) (3)H-TdR incorporation test showed time-dependent inhibition of proliferation of K562 cells, reflected in decrease of counts per minute (CPM) value in RNAi group 24 h, 48 h, 72 h, and 96 h after siRNA transfection by 33.06%, 52.25%, 57.64%, and 70.87% respectively (F=17.7, P < 0.01). (3) About 43.2% of K562 cells in the RNAi group were apoptotic 48 h after siRNA transfection (F=13.6, P < 0.01). (4) In contrast with the control groups, the expression of apoptosis-associated protein Bcl-xL was greatly down-regulated; however, the expression of Bax protein showed little change. (5) The percentage of benzidine-positive cells in the RNAi group was 23.5% +/- 3.2%, significantly higher than those in the indifferent control group, blank vector group, and normal control group (2.4% +/- 0.3%, 4.5% +/- 0.5%, and 3.6% +/- 0.2% respectively, all P < 0.01), which meant that part of the K562 cells differentiated towards erythrocytic series. (6) The percentage of G1 phase of K561 cells in the RNA1 group was significantly higher than those of the other groups (F = 6.2, P < 0.05), showing a capture in G1-phase of cell cycle. CONCLUSION: The specific siRNA distinctly inhibits the expression of bcr-abl chimeric gene and influences essential biological traits of K562 cells, which will ultimately result in differentiation or apoptosis of K562 cells.


Assuntos
Proteínas de Fusão bcr-abl/fisiologia , Leucemia Mielogênica Crônica BCR-ABL Positiva , RNA Interferente Pequeno/farmacologia , Relação Dose-Resposta a Droga , Proteínas de Fusão bcr-abl/genética , Genes abl , Humanos , Células K562/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , RNA Mensageiro/análise , Transfecção
16.
Mol Cancer Ther ; 2(3): 301-6, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12657725

RESUMO

Phage display technology is now well established as an important experimental approach in designing new reagents for diagnosis of diseases and development of novel vaccines. Aiming at identifying possible immunogenic mimotopes of gastric cancer-associated antigen, we used a monoclonal antibody against gastric cancer, named monoclonal antibody MG7, to screen two phage-displayed nanopeptide libraries. All selected phages were used to immunize BALB/c mice separately. By immunohistochemical staining the tissue sections with the immunized mice sera, we identified one phage that induced gastric cancer-specific antibody response. Consistent with our reported results before, we demonstrate that specific immunogenic epitopes can be isolated by screening phage-displayed library even when natural antigens are not readily available.


Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Antineoplásicos/imunologia , Antígenos de Neoplasias/imunologia , Epitopos de Linfócito B/imunologia , Mimetismo Molecular , Neoplasias Gástricas/imunologia , Animais , Anticorpos Monoclonais/genética , Anticorpos Antineoplásicos/genética , Bacteriófagos/imunologia , DNA/análise , Ensaio de Imunoadsorção Enzimática/métodos , Escherichia coli/virologia , Feminino , Imunofluorescência , Imunização , Camundongos , Camundongos Endogâmicos BALB C , Biblioteca de Peptídeos
17.
Vaccine ; 33(29): 3367-76, 2015 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-26027907

RESUMO

BACKGROUND: Hantaviral diseases can have a high case fatality rate within the absence of broadly effective antiviral treatments or vaccines. We developed a DNA vaccine targeting the Hantavirus glycoprotein N-terminal (Gn) to major histocompatibility complex class II compartment by fusing the antigen with lysosome-associated membrane protein 1 (LAMP1), which altered antigen presenting pathway and activated the CD4+ T cells. METHODS: The segments of Gn and LAMP1 were cloned into vector pVAX1, and recombinant plasmid was constructed by inserting Gn sequence into LAMP1, between luminal and the transmembrane/cytoplasmic domains. Subsequently, the protein expression was identified through immunoprecipitation, western blot and Immunofluorescent assay. Adaptive immune responses were assessed by the presence of specific and neutralizing antibodies, interferon (ELISpot results, and cytotoxic T-lymphocyte (CTL) cytotoxicity. Epitope mapping was performed to study the T-cell epitopes. Protective immunity in vivo was evaluated using a novel HTNV-challenging model, and safety evaluation was based on histological and behavioral observations. RESULTS: Native or LAMP1 targeting HTNV Gn was successfully identified. Humoral immune responses were enhanced, featuring with satisfying titers of specific and neutralizing antibody production. The boosted activities of IFN-γ and CTL cytotoxicity witnessed enhanced cellular immune responses. Effective protection against HTNV in vivo was conferred in all three vaccine groups by the challenge model. Safety was confirmed and one dominant T-cell epitope screened from immunized mice overlapped the specific T-cell hot spot in HFRS patients. CONCLUSION: LAMP1 targeting strategy successfully enhanced the efficacy of HTNV Gn-based vaccine, which is highly immunogenic and safe, showing promise for immunoprophylaxis against HFRS. Further investigations are warranted in the future.


Assuntos
Vacinas de DNA/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Testes Imunológicos de Citotoxicidade , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , ELISPOT , Feminino , Glicoproteínas/genética , Glicoproteínas/imunologia , Infecções por Hantavirus/imunologia , Infecções por Hantavirus/prevenção & controle , Interferons/metabolismo , Proteínas de Membrana Lisossomal/genética , Proteínas de Membrana Lisossomal/imunologia , Camundongos Endogâmicos BALB C , Testes de Neutralização , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Análise de Sobrevida , Vacinas de DNA/administração & dosagem , Vacinas de DNA/efeitos adversos , Vacinas de DNA/genética , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/efeitos adversos , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Proteínas Virais/genética , Proteínas Virais/imunologia , Vacinas Virais/administração & dosagem , Vacinas Virais/efeitos adversos , Vacinas Virais/genética
18.
World J Gastroenterol ; 9(2): 334-7, 2003 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12532461

RESUMO

AIM: To observe the localization of TRAIL/TRAIR (DR4, DR5, DcR1, DcR2) in the fetal pancreas. METHODS: Fetal pancreas of 32 weeks of pregnancy were obtained from induced abortions, embedded in paraffin, and 4-microm sections were prepared. The localization of TRAIL/TRAILR in fetal pancreas was investigated by fluorescence immunohistochemical method combined with laser scanning confocal microscopy. RESULTS: TRAIL immunoreactive cells were mainly located on the periphery of the pancreas islets. There were a few DcR1 and DcR2 positive cells whereas there were no immunoreactive cells of DR4 and DR5 in the pancreas islets. In the acini and the ducts of the exocrine pancreas there were no TRAIL/TRAILR immunoreactive cells. CONCLUSION: This study not only describes the distribution of TRAIL/TRAILR in the fetal pancreas, but also provides a morphological basis for deducing the function of TRAIL/TRAILR in pancreas, suggesting that in normal pancreatic islets, the pancreatic cells are resistant towards apoptosis too.


Assuntos
Glicoproteínas de Membrana/metabolismo , Pâncreas/embriologia , Receptores de Superfície Celular/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Proteínas Reguladoras de Apoptose , Feto/metabolismo , Humanos , Ligante Indutor de Apoptose Relacionado a TNF , Distribuição Tecidual
19.
World J Gastroenterol ; 8(1): 108-13, 2002 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11833083

RESUMO

AIM: To confirm the existence of CD226 ligand and its distribution, which is a novel molecule cloned in 1996. METHODS: The mRNA was extracted from TPA activated Jurkat cells and used as a template for reverse-transcription. After PCR amplification, the fragment including CD226 extracellular region and the splice donor sequence "ACTTACCTGT" was obtained and cloned into fusion expression vector pIG. The recombinant vector pCD226/Ig was transfected in COS-7 cells by DEAE-Dextran method, the secreting fusion protein was identified by Sandwich ELISA, and was purified by anti-CD226 affinity chromatography. This fusion protein was used as a probe in the investigation of CD226 ligand by immunohistochemistry. Existence of CD226 ligand was further identified by adhesion experiment. RESULTS: Expression of a secreting fusion protein was identified by sandwich ELISA,indicating that both CD226 extracellular domain and IgGFc domain could be recognized respectively by anti-CD226 and anti-hIgFc mAb. About 130g CD226/Ig fusion protein could be obtained from 100mL COS-7 culture supernatants by anti-CD226 affinity chromatography purification. SDS-PAGE showed that this fusion protein has a molecular mass of 83ku. It was confirmed by immunohistochemistry that CD226 ligand expressed on the Colo205 cells, but not on Jurkat cell, U937 cell and mixed lymphocyte culture cells. In adhesive assay, resting Jurkat cells did not have significant adhesion to Colo205 cells. In contrast, activated Jurkat cells could bind to colon carcinoma Colo205 cells and this adhesive reaction could be blocked by CD226/Ig fusion protein or anti-CD226 mAb. Immunochemical experiment showed that Colo205 cells could be specifically stained by CD226/Ig, indicating that CD226 ligand exists on the surface of Colo205 cells. CONCLUSION: Existence of CD226 ligand on the surface of Colo205 cells was identified by immunohistochemistry and adhesion blocking experiment. In addition, the secreting CD226/Ig fusion protein prepared in this study will be a potential tool for further investigation of CD226 ligand.


Assuntos
Antígenos de Diferenciação de Linfócitos T/genética , Antígenos de Superfície/genética , Neoplasias do Colo , Animais , Antígenos de Diferenciação de Linfócitos T/análise , Antígenos de Superfície/análise , Células COS , Primers do DNA , Expressão Gênica , Humanos , Imuno-Histoquímica , Células Jurkat , Plasmídeos , RNA Mensageiro/análise , Proteínas Recombinantes de Fusão/genética
20.
Sheng Li Xue Bao ; 55(3): 355-9, 2003 Jun 25.
Artigo em Zh | MEDLINE | ID: mdl-12817306

RESUMO

In order to study the possible mechanism of CD226 monoclonal antibody (mAb)-mediated intracellular message transduction in human umbilical vein endothelial cells (HUVECs), the influence of CD226 mAb and its cross-linking by secondary antibody (II Ab) on the concentration changes in [Ca(2+)](i) in the HUVECs under different conditions were determined by confocal laser scanning microscopy. The main results are as follows. (1) When the culture medium was balanced by Hanks Buffer, [Ca(2+)](i) in HUVECs increased slowly after stimulation by CD226 mAb, whereas [Ca(2+)](i) increase was accompanied by [Ca(2+)](o) decrease after the mAb was cross-linked by goat anti-mouse IgG. Then [Ca(2+)](i) and [Ca(2+)](o) all returned to the normal level. (2) When the culture medium was balanced by D-Hanks buffer, [Ca(2+)](i) in HUVECs showed little variation when the cells were stimulated by CD226 mAb, but [Ca(2+)](i) decreased markedly after cross-linking. (3) When HUVECs were pretreated with EGTA, there was no variation in [Ca(2+)](i) of HUVECs after CD226 mAb stimulation alone or cross-linking of the mAb. Our results suggest that stimulation by CD226 mAb and cross-linking by goat anti-mouse IgG induce the variation of [Ca(2+)](i) in HUVECs under different conditions and the variation of [Ca(2+)](i) in HUVECs may play an important role in many physiological and pathological processes.


Assuntos
Anticorpos Monoclonais/farmacologia , Antígenos de Diferenciação de Linfócitos T/imunologia , Cálcio/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Células Endoteliais da Veia Umbilical Humana/fisiologia , Células Cultivadas , Humanos
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