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PURPOSE: Information on incentives for COVID-19 testing is needed to understand effective practices that encourage testing uptake. We describe characteristics of those who received an incentive after performing a rapid antigen test. DESIGN: Cross-sectional descriptive analysis of survey data. SETTING: During April 29-May 9, 2021, COVID-19 rapid antigen testing was offered in 2 Maryland cities. SAMPLE: Convenience sample of 553 adults (≥18 years) who tested and received an incentive; 93% consented to survey. MEASURES: Survey questions assessed reasons for testing, testing history, barriers, and demographics. ANALYSIS: Robust Poisson regressions were used to determine characteristic differences based on testing history and between participants who would re-test in the future without an incentive vs participants who would not. RESULTS: The most common reasons for testing were the desire to be tested (n = 280; 54%) and convenience of location (n = 146; 28%). Those motivated by an incentive to test (n = 110; 21%) were 5.83 times as likely to state they would not test again without an incentive, compared to those with other reasons for testing (95% CI: 2.67-12.72, P < .001). CRITICAL LIMITATIONS: No comparative study group. CONCLUSION: Results indicate internal motivation and convenience were prominent factors supporting testing uptake. Incentives may increase community testing participation, particularly among people who have never tested. Keywords COVID-19, pandemic, incentives, health behavior, community testing.
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COVID-19 , Motivação , Adulto , Humanos , Maryland , Teste para COVID-19 , Estudos Transversais , COVID-19/diagnósticoRESUMO
Lower levels of physical activity among children in the United States can be attributed in part to the lack of access to safe, low-cost recreational facilities. Shared use, or a partnership allowing the community to use school recreational facilities outside of normal hours, has received increased attention. The objective of this study was to determine the extent of knowledge among school district decision makers about a law passed clarifying liability for school shared use in Minnesota and to understand perceptions held by school decision makers regarding shared use of recreational facilities. A survey of Minnesota school superintendents and other decision makers (N = 182) was conducted to understand the issues relevant to sharing school recreational facilities with the public. The majority (90%) of respondents indicated concern about liability for injury on school property outside of normal hours, and that insurance and contracts provided the most protection from liability. Most respondents indicated they were not familiar with the Minnesota shared use legislation and its provisions (61.4%, n = 108). Findings suggest the importance of education and training to further school superintendents' knowledge of Minnesota shared use legislation, legal and policy issues relevant to shared use, and issues related to the implementation of shared use within their districts.
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The multifaceted immunomodulatory activity of DNA hypomethylating agents improves immunogenicity and immune recognition of neoplastic cells; thus, we predicted they could be utilized to design new immunotherapeutic combinations in cancer. Testing this hypothesis, the antitumor efficacy of the DNA hypomethylating agent 5-aza-2'-deoxycytidine (5-AZA-CdR) combined with the anti-CTLA-4 monoclonal antibody (mAb) 9H10 in syngeneic transplantable murine models was investigated. Murine mammary carcinoma TS/A or mesothelioma AB1 cells were injected in BALB/c, athymic nude, and SCID/Beige mice that were treated with 5-AZA-CdR, mAb 9H10, or their combination. Tumor volumes were captured at different time-points; molecular and immunohistochemical assays investigated changes in neoplastic and normal tissues. A significant antitumor effect of 5-AZA-CdR combined with mAb 9H10 was found: compared to controls, a 77% (p < 0.01), 54% (p < 0.01) and 33% (p = 0.2) decrease in TS/A tumor growth was induced by 5-AZA-CdR combined with mAb 9H10, 5-AZA-CdR or mAb 9H10, respectively. These antitumor activities were confirmed utilizing the AB1 model. 5-AZA-CdR-based regimens induced a promoter-demethylation-sustained tumor expression of cancer testis antigens. MHC class I expression was up-regulated by 5-AZA-CdR. Antitumor efficacy of 5-AZA-CdR in athymic nude and SCID/Beige mice was not increased by mAb 9H10. In BALB/c mice, combined treatment induced the highest tumor infiltration by CD3+ lymphocytes, which included both CD8+ and CD4+ T cells; no such infiltrates were observed in normal tissues. This significant immune-related antitumor activity of 5-AZA-CdR combined with CTLA-4 blockade, demonstrated in highly aggressive mouse tumor models, provides a strong scientific rationale to implement epigenetically-based immunotherapies in cancer patients.
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During the last decade, debate about the nation's ailing healthcare system has moved to the forefront. In 2010, the Patient Protection and Affordable Care Act (PPACA) was signed into law. This groundbreaking piece of legislation impacts every aspect of the health industry, affecting everyone from doctors and health-care facilities to insurers and benefits consultants to business owners and patients. The ultimate goal of PPACA is to decrease the number of uninsured Americans and reduce the overall costs of healthcare.
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BACKGROUND: Epigenetic remodelling of cancer cells is an attractive therapeutic strategy and distinct DNA hypomethylating agents (DHA) are being actively evaluated in patients with hemopoietic or solid tumours. However, no studies have investigated the modulation of gene expression profiles (GEP) induced by DHA in transformed and benign tissues. Such information is mandatory to clarify the fine molecular mechanism(s) underlying the clinical efficacy of DHA, to identify appropriate therapeutic combinations, and to address safety issues related to their demethylating potential in normal tissues. Thus, utilising a syngeneic mouse model, we investigated the remodelling of GEP of neoplastic and normal tissues induced by systemic administration of DHA. METHODS: The murine mammary carcinoma cells TS/A were injected s.c. into female BALB/c mice that were treated i.p. with four cycles of the DHA 5-aza-2'-deoxycytidine (5-AZA-CdR) at a fractioned daily dose of 0.75 mg kg(-1) (q8 h × 3 days, every week). Whole mouse transcriptomes were analysed by microarrays in neoplastic and normal tissues from control and treated mice. Results were processed by bioinformatic analyses. RESULTS: In all, 332 genes were significantly (P ≤ 0.05; FC ≥ 4) modulated (294 up and 38 downregulated) in neoplastic tissues from 5-AZA-CdR-treated mice compared with controls. In decreasing order of magnitude, changes in GEP significantly (P ≤ 0.05) affected immunologic, transport, signal transduction, spermatogenesis, and G-protein-coupled receptor protein signalling pathways. Epigenetic remodelling was essentially restricted to tumour tissues, leaving substantially unaltered normal ones. CONCLUSION: The ability of 5-AZA-CdR to selectively target tumour GEP and its major impact on immune-related genes, strongly support the clinical use of DHA alone or combined with immunotherapeutic agents.
Assuntos
Azacitidina/análogos & derivados , Transformação Celular Neoplásica/efeitos dos fármacos , Transformação Celular Neoplásica/genética , Imunoterapia/métodos , Neoplasias Mamárias Experimentais/genética , Neoplasias Mamárias Experimentais/terapia , Animais , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/imunologia , Azacitidina/farmacologia , Linhagem Celular Tumoral , Transformação Celular Neoplásica/imunologia , Metilação de DNA , Decitabina , Epigênese Genética , Epigenômica/métodos , Feminino , Perfilação da Expressão Gênica/métodos , Regulação Neoplásica da Expressão Gênica , Neoplasias Mamárias Experimentais/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Regiões Promotoras Genéticas , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/imunologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Transdução de Sinais/imunologiaAssuntos
Melanoma/diagnóstico , Melanoma/genética , Mutação de Sentido Incorreto , Proteínas Proto-Oncogênicas B-raf/genética , Seleção Genética/fisiologia , Neoplasias Cutâneas/diagnóstico , Neoplasias Cutâneas/genética , Substituição de Aminoácidos/genética , Progressão da Doença , Ácido Glutâmico/genética , Humanos , Melanoma/patologia , Melanoma/terapia , Mutação de Sentido Incorreto/fisiologia , Metástase Neoplásica , Prognóstico , Proteínas Proto-Oncogênicas B-raf/metabolismo , Neoplasias Cutâneas/patologia , Neoplasias Cutâneas/terapia , Resultado do Tratamento , Valina/genéticaRESUMO
BACKGROUND: Osteoarthritis (OA) affects an estimated 49 million adults in North America, or nearly 1 of every 6 adults. More than 8 million North Americans have limited mobility to some extent because of OA. By 2030, an estimated 71 million North Americans will be diagnosed with OA, an increase of 45% over current figures. For one group-model health maintenance organization (HMO), the average cost of care for patients with OA was $543 per member, a total annual cost to the HMO of $4,728,425. Of this total amount, 46% was for inpatient care, 32% was for medication, and 22% was for ambulatory care. OBJECTIVE: To determine the impact of OA on managed care and discuss treatment options available to those with OA, particularly of the knee. SUMMARY: OA represents an advanced stage of an active, progressive disease process. We know from medical research that OA is the endpoint of a progression in tissue degradation that results in loss of cartilage structure and function. Relief of pain and preservation of joint tissue must evolve to encompass treatments that interfere with cartilage-degrading mechanisms that follow acute or chronic injury, restore normal cartilage and joint homeostasis, and arrest the progression of disease. Optimal future treatments will also reverse existing damage and restore normal cartilage structure and function. Viscosupplementation with an elastoviscous fluid containing polymers of hylan derivatives of the natural glycosaminoglycan hyaluronan is indicated for treating pain of OA of the knee that has not responded to or is contraindicated for conservative nonpharmacologic therapy and traditional analgesics. These analgesics include acetaminophen, nonsteroidal anti-inflammatory drugs (NSAIDs), and cyclooxygenase-2 (COX-2) inhibitors. Clinicians in the managed care setting may consider using viscosupplementation in patients (1) who have persistent pain despite their use of conservative nonpharmacologic and pharmacologic therapy (e.g., exercise, weight loss, physician therapy, bracing/orthotics, NSAIDs, COX-2 inhibitors, and intra-articular glucocorticoids); (2) who have compromised gastrointestinal (GI) function or who are at risk for GI bleeding due to the adverse events of NSAIDs; (3) who are taking concomitant anticoagulant therapy for any condition; (4) who have cardiovascular or renal risk factors that preclude use of COX-2 inhibitors; and (5) for whom surgery is not appropriate. Further study should be conducted with larger numbers of patients to help identify a subgroup of patients with OA in whom viscosupplementation may have even greater effects. Additional research should also concentrate on assessing the risks and benefits of extended treatments, because limited data are available concerning the effectiveness of multiple courses of therapy. CONCLUSION: OA is an important public health issue as the leading cause of disability in North America. As populations age, socioeconomic costs of OA will dramatically increase. Among available treatment options, viscosupplementation is a valuable alternative to more conservative therapy and has the benefit of circumventing the possible side effects of systemically administered pharmacologic agents. Viscosupplementation demonstrated efficacy in OA of the knee, and its use in the managed care arena may generate savings in hospitalizations and other costs.
Assuntos
Programas de Assistência Gerenciada/economia , Osteoartrite do Joelho/tratamento farmacológico , Viscossuplementos/administração & dosagem , Adulto , Progressão da Doença , Custos de Cuidados de Saúde , Humanos , Ácido Hialurônico/administração & dosagem , Ácido Hialurônico/química , Ácido Hialurônico/economia , América do Norte/epidemiologia , Osteoartrite do Joelho/economia , Osteoartrite do Joelho/epidemiologia , Osteoartrite do Joelho/fisiopatologia , Seleção de Pacientes , Viscossuplementação/métodos , Viscossuplementos/economiaRESUMO
Recent evidences suggest that malignant mesothelioma may be sensitive to immunotherapy; however, little is known about malignant mesothelioma-associated tumour antigens. Focusing on cancer/testis antigens, the expression of well-characterised immunogenic tumour-associated antigens was investigated in malignant mesothelioma cells. At variance with MAGE-4 and NY-ESO-1, malignant mesothelioma cells frequently expressed MAGE-1, -2 and -3, GAGE 1-2, GAGE 1-6, SSX-2 and SSX 1-5, and distinct malignant mesothelioma cells concomitantly expressed at least four cancer/testis antigens. Additionally, the tumour-associated antigens RAGE-1 was expressed at high levels in both benign and malignant mesothelial cells. Lastly, treatment with the DNA hypomethylating agent 5-aza-2'-deoxycytidine induced and up-regulated the expression of the cancer/testis antigen examined in malignant mesothelioma cells. Overall, these findings strongly suggest that cancer/testis antigens-based immunotherapy may represent a suitable therapeutic approach to malignant mesothelioma, and foresee the clinical use of 5-aza-2'-deoxycytidine to design new chemo-immunotherapeutic strategies in malignant mesothelioma patients.
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Antígenos de Neoplasias/análise , Metilação de DNA , Proteínas de Membrana , Mesotelioma/imunologia , Testículo/imunologia , Animais , Azacitidina/farmacologia , Feminino , Humanos , Imunoterapia , Masculino , Antígenos Específicos de Melanoma , Mesotelioma/terapia , Proteínas de Neoplasias/análise , Proteínas/análise , Coelhos , Proteínas Repressoras/análiseRESUMO
Endoglin (CD105) is a cell membrane glycoprotein over-expressed on highly proliferating endothelial cells in culture, and on endothelial cells of angiogenetic blood vessels within benign and malignant tissues. CD105 binds several factors of the Transforming Growth Factor (TGF)-beta superfamily, and its over-expression modulates cellular responses to TGF-beta1. The complex of experimental findings accumulated in the last few years strongly indicate that CD105 is a powerful marker of angiogenesis, and that it might play a critical role in the pathogenesis of vascular diseases and in tumor progression. In this paper, we will review the structural, biological and functional features of CD105, as well as its distribution within normal and neoplastic tissues, emphasizing its foreseeable role as a molecular target for new diagnostic and bioimmunotherapeutic approaches in human malignancies.
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Neoplasias/metabolismo , Neovascularização Patológica/fisiopatologia , Fator de Crescimento Transformador beta/metabolismo , Molécula 1 de Adesão de Célula Vascular/metabolismo , Animais , Antígenos CD , Biomarcadores Tumorais/metabolismo , Endoglina , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Humanos , Substâncias Macromoleculares , Neoplasias/irrigação sanguínea , Neoplasias/patologia , Neoplasias/terapia , Neovascularização Fisiológica/fisiologia , Receptores de Superfície Celular , Fator de Crescimento Transformador beta/genética , Molécula 1 de Adesão de Célula Vascular/química , Molécula 1 de Adesão de Célula Vascular/genéticaRESUMO
Conflicting evidences suggested that levels of HLA-A and -B antigens expressed on normal and neoplastic cells of given individuals are genetically predetermined, or, on the other hand, regulated by molecular mechanisms generating the down-regulated expression of HLA-B antigens frequently observed on melanoma cells. In our study, we quantitated, both at the protein and mRNA level, the amounts of HLA-A and -B antigens constitutively expressed on 23 primary cultures of metastatic melanomas and on autologous peripheral blood mononuclear cells (PBMC). Flow cytometric analyses identified a significantly (p < 0.01) lower expression of HLA-B antigens on melanoma cell cultures but not on autologous PBMC. Consistently, lower amounts of HLA-B antigens mRNA were detected by RNase protection assay exclusively in neoplastic cells. This unbalanced expression of HLA-A and -B antigens was readily reverted by interferon (IFN)-gamma but not by the DNA hypomethylating agent 5-aza-2'-deoxycytidine in 4 melanoma cell cultures investigated. Significantly (p < 0.05) lower levels of HLA-B antigens were also detected on cells from solid malignancies of different histotypes but not on neoplastic cells from hemopoietic neoplasms; levels of HLA-B antigens were rapidly up-regulated by IFN-gamma exclusively on non-hemopoietic transformed cells. Together, these data strongly argue against a genetic predetermination of the amounts of HLA-A and -B antigens expressed on normal and neoplastic cells of distinct melanoma patients and suggest that constitutively low levels of HLA-B antigens are a specific feature of non-hemopoietic transformed cells that is controlled by common regulatory mechanism(s) and that is possibly shared by non-hemopoietic normal cells.
Assuntos
Azacitidina/análogos & derivados , Antígenos HLA-A/biossíntese , Antígenos HLA-B/biossíntese , Interferon gama/uso terapêutico , Melanoma/metabolismo , Antimetabólitos Antineoplásicos/uso terapêutico , Antineoplásicos/uso terapêutico , Azacitidina/uso terapêutico , Sequência de Bases , Células Cultivadas , DNA Complementar/metabolismo , Decitabina , Regulação para Baixo , Citometria de Fluxo , Antígenos HLA-A/genética , Antígenos HLA-B/genética , Humanos , Leucócitos Mononucleares/metabolismo , Melanoma/genética , Dados de Sequência Molecular , Neoplasias/genética , Neoplasias/metabolismo , Fenótipo , RNA Mensageiro/metabolismo , Proteínas Recombinantes/metabolismo , Ribonucleases/metabolismo , Homologia de Sequência do Ácido Nucleico , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/metabolismo , Células Tumorais Cultivadas , Regulação para CimaRESUMO
The clinical efficacy of therapeutic complement (C)-activating monoclonal antibodies (mAb) to melanoma-associated antigens can be impaired by the levels of expression of C-inhibitory molecules on neoplastic cells. Protectin (CD59) is a glycosylphosphatidylinositol (GPI)-anchored cell membrane glycoprotein, acting as terminal regulator of C cascade, which is heterogeneously expressed in melanomas and represents the main restriction factor of C-mediated lysis of melanoma cells. Thus, we investigated whether the overexpression of CD59 could influence the constitutive susceptibility of distinct melanoma cells to homologous C. Infection of CD59-positive Mel 100 and 70-W melanoma cells by a retroviral vector carrying the CD59 cDNA, significantly (P < 0.05) upregulated their constitutive expression of CD59, whereas it did not affect that of additional C-regulatory molecules. Transduced CD59 was entirely GPI-anchored and showed a molecular weight identical to native CD59. Additionally, higher amounts of soluble CD59 were detected in the conditioned media of CD59-transduced melanoma cells compared with parental cells. CD59-transduced melanoma cells, sensitized by the anti-GD3 disialoganglioside mAb R24, were significantly (P < 0.05) less susceptible to homologous C-lysis than were parental cells; this effect was fully reverted by the masking of CD59 with F(ab')(2) fragments of the anti-CD59 mAb YTH53.1. These results provide conclusive evidence demonstrating that absolute levels of CD59 expression regulate the susceptibility to homologous C of specific melanoma cells, and suggest an additional explanation for the poor clinical results obtained with C-activating mAb in the clinical setting.
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Antígenos CD59/genética , Ativação do Complemento/genética , Melanoma/genética , Melanoma/imunologia , Antígenos CD59/biossíntese , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica/imunologia , Vetores Genéticos , Humanos , Retroviridae , Transfecção , Células Tumorais CultivadasRESUMO
Over the past two decades, complement (C)-activating monoclonal antibodies (mAb), directed to specific tumor-associated antigens (TAA), have been extensively utilized for passive immunotherapy of solid tumors of different histology. However, the clinical outcome of this therapeutic approach has been substantially disappointing; antigenic heterogeneity of neoplastic cells and their limited accessibility by therapeutic mAb, have been provided as substantial explanations for the poor clinical results obtained. Nevertheless, in light of the recent advances in the knowledge of the mechanisms regulating C-activity, it begins to be evident that membrane and soluble C-inhibitory proteins play a key role in the protection of neoplastic cells from C-attack, providing additional insights on biological features of transformed cells that may hamper the clinical efficacy of humoral immunotherapy. Among C-regulatory proteins investigated, this review will focus on protectin (CD59) that represents the main restriction factor of C-susceptibility of neoplastic cells from solid malignancies. In view of the functional role of CD59, we will describe its tissue distribution and biological features in malignant neoplasms; major emphasis will be given to cutaneous melanoma, in which the C-regulatory role of CD59 has been extensively investigated, and clinical approaches of humoral immunotherapy have been implemented. According to the available data, the foreseeable strategies to improve the therapeutic efficacy of humoral immunotherapy of solid malignancies will be discussed.
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Antígenos CD59/fisiologia , Proteínas Inativadoras do Complemento/fisiologia , Imunoterapia/métodos , Neoplasias/tratamento farmacológico , Humanos , Neoplasias/genéticaRESUMO
Increasing evidence suggests that endoglin (CD105) is a new powerful marker of neovascularization in solid malignancies; thus, using breast cancer as a model, we investigated whether targeting of CD105 by monoclonal antibody (mAb) MAEND3 can be used for in vivo imaging of solid tumors. Immunohistochemistry and flow cytometry identified differential expression of CD105 on breast cancer and endothelial cells; in fact, neoplastic cells were weakly and rarely stained by mAb MAEND3, which in contrast, strongly and invariably stained blood vessel endothelia within the breast adenocarcinomas investigated and cultured endothelial cells. Moreover, in contrast to CD31, which currently represents the reference marker to assess angiogenetic activity, CD105 expression was highest in semiconfluent and actively proliferating endothelial cells, and it progressively decreased as cells reached tight confluency and low [3H]thymidine uptake. i.v. administration of 18 MBq of 125I-labeled mAb MAEND3 efficiently imaged spontaneous mammary adenocarcinomas in two dogs; the uptake of radiolabeled mAb was rapid and intense because tumor: background ratios of 8.2:1 and 9.3:1 were reached 8 h after mAb administration, in the absence of immediate and/or long-term clinical side effects. Altogether, our present data suggest that targeting of CD105 on tumor-associated blood vessels may represent a new strategy for in vivo imaging of solid malignancies, regardless of their histological origin.
Assuntos
Neoplasias/diagnóstico por imagem , Molécula 1 de Adesão de Célula Vascular/análise , Adenocarcinoma/diagnóstico por imagem , Adenocarcinoma/metabolismo , Animais , Anticorpos Monoclonais/imunologia , Antígenos CD , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Contagem de Células , Divisão Celular , Linhagem Celular , Modelos Animais de Doenças , Cães , Endoglina , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Feminino , Câmaras gama , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Radioisótopos do Iodo , Neoplasias Mamárias Animais/diagnóstico por imagem , Neoplasias Mamárias Animais/metabolismo , Neoplasias/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análise , RNA Neoplásico/genética , RNA Neoplásico/metabolismo , Cintilografia , Receptores de Superfície Celular , Células Tumorais Cultivadas , Molécula 1 de Adesão de Célula Vascular/genética , Molécula 1 de Adesão de Célula Vascular/imunologiaRESUMO
Melanoma cells constitutively release intercellular adhesion molecule 1 (ICAM-1) as soluble ICAM-1 (sICAM-1), and its levels are elevated in melanoma patients and correlate with disease progression. However, this correlation is not absolute, suggesting that specific characteristics of neoplastic cells and/or ICAM-1-positive non-neoplastic cells may influence the amounts of circulating sICAM-1. In this study, we found a weak correlation (r = 0.55; r2 = 0.3) between sICAM-1 release by 40 metastatic melanomas (36 primary cultures and 4 cell lines), and ICAM-1 expression on neoplastic cells. In addition, melanoma-secreted interleukin-1alpha (IL-1alpha) (1/40) but not vascular endothelial growth factor (VEGF) (29/40), significantly (P < 0.05) up-regulated the shedding of sICAM-1 by human umbilical vein endothelial cells (HUVEC). This was completely abolished by IL-1alpha/beta neutralizing antibodies both at the protein and mRNA level. Altogether, our results suggest that (i) the extent of sICAM-1 release is distinctive for individual melanomas and can be independent of ICAM-1 expression; (ii) tumor endothelia may sustain levels of sICAM-1 in selected melanomas; (iii) melanoma-released VEGF does not affect ICAM-1 expression and sICAM-1 release by HUVEC. Melanoma-derived sICAM-1 inhibits cell-mediated cytotoxicity of melanoma cells; therefore, constitutive levels of sICAM-1 release and IL-1alpha secretion by individual melanomas can differentially influence tumor progression and the clinical effectiveness of cytotoxic-cell-based vaccines.
Assuntos
Endotélio Vascular/metabolismo , Molécula 1 de Adesão Intercelular/biossíntese , Melanoma/fisiopatologia , Células Cultivadas , Regulação da Expressão Gênica , Humanos , Molécula 1 de Adesão Intercelular/genética , Interleucina-1/fisiologia , RNA Mensageiro/análise , Células Tumorais CultivadasRESUMO
The immunogenic potential of melanoma cells and their recognition by the host's cytotoxic cells depends on the presence and on the level of expression of human leukocyte antigen (HLA) class I antigens, costimulatory molecules and melanoma-associated antigens (MAA), on neoplastic cells. In this study, we demonstrate that the DNA hypomethylating agent 5-aza-2'-deoxycytidine (5-AZA-CdR), significantly (p < 0.05) enhanced the constitutive expression of HLA class I antigens, HLA-A1 and -A2 alleles, and of the costimulatory molecules intercellular adhesion molecule-1 and lymphocyte function-associated antigen-3, on a panel of 12 melanoma cells. This upregulation peaked at day 4, slowly decreased thereafter, and returned to baseline levels 32 days after the end of treatment. In addition, treatment with 5-AZA-CdR induced a persistent expression of MAGE-1 in Mel 275 melanoma cells; this was still detectable, by reverse transcriptase polymerase chain reaction, 60 days after the end of treatment. In contrast, 5-AZA-CdR did not affect the constitutive expression of the high molecular weight-MAA by the melanoma cells investigated. These observations, together with data obtained comparing the effect of 5-AZA-CdR with that of interferon-gamma, strongly suggest that 5-AZA-CdR may have prospective therapeutic implications in active and/or passive specific immunotherapy for human melanoma.
Assuntos
Azacitidina/análogos & derivados , Antígenos CD58/biossíntese , Antígenos de Histocompatibilidade Classe I/biossíntese , Molécula 1 de Adesão Intercelular/biossíntese , Melanoma/imunologia , Alelos , Antígenos de Neoplasias , Azacitidina/farmacologia , Western Blotting , Decitabina , Citometria de Fluxo , Antígenos de Histocompatibilidade Classe I/genética , Humanos , Antígenos Específicos de Melanoma , Proteínas de Neoplasias/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Células Tumorais Cultivadas , Regulação para Cima/efeitos dos fármacosRESUMO
Protectin (CD59) is a glycosyl-phosphatidylinositol-anchored cell membrane glycoprotein, ubiquitously expressed, though to a different extent, on benign and malignant cells. CD59 inhibits complement (C)-mediated lysis of target cells by preventing the formation of the membrane attack complex, in the terminal step of C-activation. Recent experimental evidence demonstrates that CD59 is the main restriction factor of C-mediated lysis of malignant cells of different histotype. Additionally, a soluble form of CD59, that retains its anchoring ability and functional properties, has been most recently identified in body fluids and in culture supernatants of different malignant cells. In view of its functional role, CD59 may protect neoplastic cells from C-mediated lysis, contributing to their escape from innate C-control and to tumor progression; additionally, the expression of CD59 by neoplastic cells may contribute to impair the therapeutic efficacy of C-activating monoclonal antibodies (mAb) directed to tumor-associated antigens. In the light of the functional role of CD59, this review focuses on the structural features, tissue distribution and regulation of the expression of CD59 in malignant tissues, and on the foreseeable application(s) of CD59 to improve the therapeutic efficacy of clinical approaches of humoral immunotherapy with C-activating mAb in human malignancies.
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Antígenos CD59/fisiologia , Proteínas do Sistema Complemento/fisiologia , Neoplasias/metabolismo , Antígenos CD59/química , Antígenos CD59/metabolismo , Ativação do Complemento/fisiologia , Humanos , Imunoterapia , Neoplasias/imunologia , Neoplasias/terapiaRESUMO
Protectin (CD59), a glycosylphosphatidylinositol-anchored cell membrane glycoprotein, is differentially expressed on melanocytic cells and represents the main restriction factor of C-mediated lysis of melanoma cells. In this study, we report that CD59-positive melanoma cells constitutively release a soluble form of CD59 (sCD59), and that its levels directly correlate (r = 0.926; P < 0.05) with the amount of membrane-bound CD59. SDS-PAGE analysis showed that the molecular components of sCD59 are similar to those of cellular CD59 expressed by melanoma cells. Melanoma-released sCD59 is anchor positive since it inserts into cell membranes of homologous cells that transiently increase their expression of CD59. Moreover, sCD59 is functional: it blocks the binding of the anti-CD59 mAb YTH53.1 to melanoma cells and reverses its effects on C-mediated lysis. In fact, preincubation of mAb YTH53.1 with scalar doses of conditioned media of CD59-positive but not of CD59-negative melanoma cells reduced significantly (P < 0.05), and in a dose-dependent fashion, the enhancement of C-mediated lysis of anti-GD3-sensitized melanoma cells induced by the masking of cellular CD59 by mAb YTH53.1. Altogether, these data demonstrate that CD59-positive human melanoma cells release a soluble form of CD59 that is structurally similar to cellular CD59, retains its anchoring ability, is functional, and may impair the effectiveness of clinical approaches to humoral immunotherapy for human melanoma.
Assuntos
Antígenos CD59/fisiologia , Proteínas do Sistema Complemento/fisiologia , Citotoxicidade Imunológica , Melanoma/imunologia , Animais , Anticorpos Monoclonais/imunologia , Antígenos CD59/análise , Humanos , Melanoma/terapia , Camundongos , Ratos , Células Tumorais CultivadasRESUMO
Triggering of HLA class II antigens by the anti-HLA-DR monoclonal antibody (mAb) L243 significantly (P < 0.05) and differentially enhanced the release of tumor necrosis factor alpha (TNF-alpha) by the non-Hodgkin's lymphoma cells Ri-I, Ci-I, and Sc-I, which are at a distinct stage of B-cell differentiation, and by the more mature Burkitt lymphoma cell Raji; in contrast, it did not induce TNF-alpha release by the pre-B leukemia cells Nalm-6 and BV173. TNF-alpha release peaked at 24 h and decreased thereafter, and it was dose dependent and preceded by an increase of TNF-alpha mRNA detectable after 3 h of stimulation with mAb L243. Secreted TNF-alpha mediated the enhancement of nuclear factor kappa B (NF-kappa B) and activator protein-1 (AP-1) binding activity; in fact, the triggering of HLA-DR antigens in the presence of antihuman TNF-alpha-neutralizing antibodies did not upregulate NF-kappa B and AP-1. In contrast, released TNF-alpha was not responsible for the homotypic aggregation of Ri-I, Ci-I, Sc-I, and Raji cells induced by mAb L243, and it did not affect the proliferation of B cells investigated. Altogether, our data demonstrate that: (a) the ability of B cells to release TNF-alpha after triggering of HLA-DR antigens depends on their stage of differentiation; (b) levels of released TNF-alpha seem to correlate with the stage of B-cell maturation but do not correlate with the amounts of cell surface HLA-DR antigens; (c) secreted TNF-alpha regulates the levels of expression of NF-kappa B and AP-1 by an autocrine loop; and (d) intracellular signals mediating TNF-alpha release by B cells are distinct from those regulating homotypic aggregation and proliferation.
Assuntos
Linfócitos B/imunologia , Antígenos HLA-DR/imunologia , Ativação Linfocitária/imunologia , Fator de Necrose Tumoral alfa/imunologia , Anticorpos Monoclonais/imunologia , Linfócitos B/citologia , Diferenciação Celular , Linhagem Celular , Humanos , NF-kappa B/imunologiaRESUMO
Levels of circulating soluble intercellular adhesion molecule-1 (sICAM-1) are elevated in patients affected by solid malignancies; however, the cellular sources generating high levels of sICAM-1 remain to be characterized. Using conditioned media (CM) from seven ICAM-1-positive or -negative neoplastic cells, we demonstrate that tumour-derived interleukin 1alpha (IL-1alpha) significantly (P < 0.05) up-regulates the release of sICAM-1 by human umbilical vein endothelial cells. The intensity of the effect correlated with the amounts of IL-1alpha detectable in CM. Levels of ICAM-1 mRNA were also up-regulated by tumour-secreted IL-1alpha. The up-regulation of the shedding of sICAM-1 and of its expression at protein and mRNA level were completely reversed by the addition of anti-IL-1alpha neutralizing antibodies. Consistent with the in vitro data, tumour endothelia were strongly stained for ICAM-1 compared with autologous normal tissue endothelia. Taken altogether, our observations reveal an IL-1alpha-mediated tumour-endothelium relationship sustaining the shedding of sICAM-1 by endothelial cells. This is a general phenomenon in solid malignancies that correlates with the ability of neoplastic cells to secrete IL-1alpha rather than with their expression of ICAM-1 and/or histological origin. sICAM-1 has been previously shown to inhibit LFA-1/ICAM-1-mediated cell-cell interactions; therefore, the ability of neoplastic cells to secrete IL-1alpha is likely to represent a mechanism for their escape from immune interaction.
Assuntos
Endotélio Vascular/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Interleucina-1/fisiologia , Neoplasias/imunologia , Células Cultivadas , Meios de Cultivo Condicionados , Humanos , Molécula 1 de Adesão Intercelular/análise , Regulação para Cima , Neoplasias da Bexiga Urinária/químicaRESUMO
Normal and neoplastic cells are protected from autologous complement (C) attack by different cell-surface C-regulatory proteins including CD59 (protectin), CD46 (membrane cofactor protein) and CD55 (decay-accelerating factor). Indirect immunofluorescence (IIF) analysis showed a differential expression of CD59, CD46, and CD55 in nine human melanoma cell lines and that the expression of CD59 was highly heterogeneous compared with that of CD46 and CD55. Levels of cell membrane CD59 were found to regulate the differential sensitivity of melanoma cells investigated to homologous C-mediated lysis; in fact, an inverse correlation (r > 0.7, p < 0.05) was found between levels of cell membrane CD59, but not of CD46 and CD55, and extent of C-mediated lysis of melanoma cells sensitized with scalar concentrations of the anti-GD3 ganglioside mAb R24. Masking of CD59 by 2.5 micrograms/ml of the anti-CD59 mAb YTH53.1 induced or enhanced C-mediated lysis of melanoma cells sensitized with 2.5 micrograms/ml of mAb R24; the latter phenomenon was found to be directly correlated (r > 0.865, p < 0.01) with levels of cell membrane CD59. CD59 is bound to melanoma cells by a glycosylphosphatidylinositol anchor: treatment of C-resistant melanoma cells Mel 97, by increasing doses of phosphatidylinositol-specific phospholipase C (PI-PLC), progressively decreased cell-surface expression of CD59 and increased C-mediated lysis of cells sensitized with mAb R24. Staining of 38 benign and malignant lesions of melanocytic origin by mAb YTH53.1 demonstrated that CD59 is consistently expressed in vivo and confirmed the heterogeneous expression detected in vitro. Our data, altogether, demonstrate that CD59 is the main restriction factor of C-mediated lysis of melanoma cells and that levels of CD59 may account for their differential resistance to C-mediated lysis. The analysis of the levels of CD59 could represent an useful strategy in selecting melanoma patients who may benefit from immunotherapeutic treatment(s) that trigger C activation.