Strontium (Sr2+) är det aktiva ämnet i läkemedel som används för att reducera frakturrisken hos patienter som lider av osteoporos. På senare tid har Sr2+ kombinerats med olika biomaterial i syfte att gynna benbildning, emellertid är ämnet vagt studerat avseende effekten på parodontala vävnader. Trots omfattande användning, är verkningsmekanismer av Sr2+ inte helt klarlagda. Denna studie syftar därmed till att utvärdera effekten av Sr2+ på humana parodontalligament-celler (PDL-celler) och osteoblaster avseende proliferation och osteogen aktivitet. Odlade humana PDL celler och osteoblastcellinjerna MG63 och hFOB 1.19 behandlades med SrCl2 (0,1-10 mM) eller vehikel under 72 h. Manuell cellräkning utfördes med en Bürker kammare. Det totala proteininnehållet fastställdes med en kolorimetrisk mätning genom användning av Bio-Rad proteinanalys. Aktiviteten av alkaliskt fosfatas bestämdes enzymatiskt och normaliserades till totalt proteininnehåll. SrCl2 hade ingen signifikant effekt på PDL celler (p> 0.05), däremot observerades en tendens till inducerade osteoblastiska egenskaper. I motsats till det, ökade 5 mM SrCl2 den totala proteinhalten i MG63 celler med 37% (p <0,01) jämfört med vehikel, medan en lägre koncentration (0,1 mM) inte hade någon påverkan. 5 mM SrCl2 ökade MG63 cellantalet med 38% (p <0,001), medan en högre koncentration (10 mM) inte uppvisade en signifikant ökad effekt jämfört med 5 mM (+54%, jämfört med vehikel, p<0.05). Resultaten visar att 72 h administrering av ≥ 5 mM SrCl2 ger en pro-proliferativ effekt på humana osteoblastliknande MG63 celler och uppvisar en tendens till att stimulera osteogena egenskaper hos primära humana PDL-celler. / Strontium (Sr2+) is the active substance of pharmaceuticals used for reducing fracture risk in osteoporotic patients. Lately, Sr2+ is combined with biomaterials to enhance osteogenesis, which has been vaguely studied considering periodontal tissue regeneration. Despite extensive use, the mechanisms of action of Sr2+ are not fully understood. The present study assesses the impact of Sr2+ on primary human periodontal ligament cells (PDL cells) and human osteoblasts in regard to proliferation and pro-osteogenic activity. Cultured human PDL cells and osteoblast cell lines MG63 and hFOB 1.19 were treated with SrCl2 (0.1-10 mM) or vehicle for 72 h. Cells were counted manually using a Bürker chamber. Total protein content was determined by colorimetric analysis using Bio-Rad protein assay. Alkaline phosphatase activity was determined enzymatically and normalized to total protein content. SrCl2 had no significant effect on PDL cells (p>0.05), but a tendency towards induced osteogenic characteristics was observed. In contrast, 5 mM SrCl2 enhanced total MG63 cell protein content by 37% (p<0.01), compared to vehicle, whereas a lower concentration (0.1 mM) did not. 5 mM SrCl2 increased MG63 cell number by 38% (p<0.001), while a higher concentration (10 mM) did not have a significant additional effect over the 5 mM (+54%, compared to vehicle, p<0.05). The results demonstrate that 72 h administration of ≥ 5 mM SrCl2 exerts a pro-proliferative effect on human osteoblast-like MG63 cells and display a tendency to induce osteogenic characteristics in primary human PDL cells.

Yes / The Mg/Ca ratios of biogenic calcite is commonly seen as a valuable palaeo-proxy for reconstructing past ocean temperatures. The temperature dependence of Mg/Ca ratios in bivalve calcite has been the subject of contradictory observations. The palaeoceanographic use of a geochemical proxy is dependent on initial, rigorous calibration and validation of relationships between the proxy and the ambient environmental variable to be reconstructed. Shell Mg/Ca ratio data are reported for the calcite of two bivalve species, Mytilus edulis (common mussel) and Pecten maximus (king scallop), which were grown in laboratory culturing experiments at controlled and constant aquarium seawater temperatures over a range from 10 to 20 C. Furthermore, Mg/Ca ratio data of laboratory- and fieldgrown M. edulis specimens were compared. Only a weak, albeit significant, shell Mg/Ca ratio¿temperature relationship was observed in the two bivalve species: M. edulis (r2=0.37, p<0.001 for laboratory-cultured specimens and r2=0.50, p<0.001 for field-cultured specimens) and P. maximus (r2=0.21, p<0.001 for laboratory-cultured specimens only). In the two species, shell Mg/Ca ratios were not found to be controlled by shell growth rate or salinity. The Mg/Ca ratios in the shells exhibited a large degree of variability among and within species and individuals. The results suggest that the use of bivalve calcite Mg/Ca ratios as a temperature proxy is limited, at least in the species studied to date. Such limitations are most likely due to the presence of physiological effects on Mg incorporation in bivalve calcite. The utilization is further limited by the great variability both within and among shells of the same species that were precipitated under the same ambient conditions

INTRODUCTION. La guérison rapide des sites donneurs des greffes cutanées favorise la survie des victimes de brûlures graves (>50 % de superficie brûlée). La mortalité élevée de ces patients est attribuable au fait que la superficie des brûlures excède celle de la peau saine. Des cultures épithéliales autologues (CEA) sont des feuillets de kératinocytes produits en culture à partir de la peau du patient. Cette étude a évalué l’effet des CEA sur l'épithélialisation des sites donneurs chez les grands brûlés. MÉTHODES. Tous les patients recevant des CEA ont été prospectivement inclus. Les plaies des sites donneurs ont été recouvertes de CEA, sauf pour une région contrôle randomisée de 7 x 7 cm. Des biopsies faites sur la greffe de peau ont permis de contrôler la profondeur des plaies sur les sites donneurs. Il y avait deux types de contrôles, avec gaze non adhérente trempée dans le milieu de culture ou dans le salin. L’épithélialisation était quantifiée globalement (% d’épithélialisation par photographie) et histologiquement (par biopsie au poinçon) à simple insu. La guérison des zones de contrôle et CEA était comparée par analyse de variance et par le test de Student. RÉSULTATS. Entre 2008 et 2009, 6 patients furent recrutés avec un total de 11 sites donneurs. Ces patients avaient en moyenne 43.5 ans, 56 % de superficie brûlée, 45% de brûlure pleine épaisseur, 66% avaient une brûlure d’inhalation, 75 jours de séjour. Il n’y a aucune corrélation entre le pourcentage d’épithélialisation et l’épaisseur du prélèvement des greffes (Pearson 0.19). Le score photographique est significativement influencé par le traitement (CEA vs Contrôle; p = 0,039) et par le jour postopératoire (p < 0,001). Le temps moyen pour atteindre un score photographique de guérison pour les zones contrôles fut de 10.2 jours contre 8.6 jours pour le CEA (p = 0,021). A l’évaluation histologique, les sites donneurs traités par le milieu de culture ont évolué aussi favorablement que ceux traités par des feuillets de CEA. CONCLUSION. L’utilisation de CEA sur les sites donneurs semble accélérer leur épithélialisation chez les victimes de brûlures graves. Cet effet est probablement le résultat d’une stimulation de la réépithélialisation innée de la plaie, plutôt que par une adhérence des feuillets de kératinocytes cultivés à la surface de la plaie. / RATIONALE: Prompt healing of split thickness skin graft donor sites is primordial to the survival of severely burned patients. Increased mortality of patients with >50 % TBSA is attributable to the limited availability of donor sites. This study evaluated the effect of Cultured Epithelial Autograft (CEA) application on skin graft donor site healing. METHODS: All burn patients receiving CEA were prospectively included. Donor site wounds were covered with CEA except a randomly designated 7x7 cm control region. Autograft biopsies were taken to document graft harvest thickness. One half of the controls were covered with non-adherent gauze soaked in the culture media and the other controls only received a non-adherent gauze dressing. Epithelialization was objectively evaluated by scoring blinded photographs with an analogue scale. Punch biopsies of the donor sites were evaluated histologically. Repeated measures ANOVA and T-test were used. RESULTS: Between 2008 and 2009, 6 patients were enrolled for a total of 11 donor sites. The patients averaged 43.5 years, 56 % TBSA, 45 % FT-TBSA, 66 % had inhalation injury and mean length of stay was 75 days. As expected, dermatome settings and autograft thickness measured by microscope did not correlate (Pearson 0.19). There was no correlation between the percentage of epithelialization of the punch biopsies of the donor sites and the thickness of the harvest. Photographic score was significantly influenced by its treatment CEA vs Control (p=0.039) and by postoperative day (p<0.001). Mean time to healing was 8.6 days for CEA compared to 10.2 days for controls (p=0.021). Infection was noted on only one donor site. On histologic analysis, the control sites dressed with gauze soaked in the culture media healed as nicely and promptly as the CEA sheet treated region. CONCLUSION: Use of CEA on donor sites appears to stimulate epithelialization. This effect is probably mediated by stimulation of local wound healing processes rather than by engraftment of keratinocytes from the CEA sheets.

O diferencial da cultura de células em 3-dimensões é permitir que as células explorem as 3-dimensões do espaço, aumentando assim as interações com o ambiente e entre as células. Em estudos relacionados à biologia do câncer de mama, vem ganhando espaço a utilização de esferóides para estudos que visam à compreensão da morfogênese do espaço luminal. Neste trabalho foi mostrado que as células MCF-7 reorganizam-se em estruturas tubulares e acinares. Em ambas as situações, a formação do lúmen veio acompanhada pelo estabelecimento de uma camada de células polarizadas, arranjo este muito semelhante ao encontrado em glândulas mamárias. Os resultados apresentados apontam para a existência de uma população de células na linhagem MCF-7 que não estão totalmente comprometidas ao fenótipo tumoral. Mantidos diferenciados, os esferóides de células MCF-7 apontam como um novo modelo para estudos relacionados à formação do lúmen, permitindo assim explorar o papel de diferentes vias como as relacionadas a apoptose, autofagia, diferenciação e sobrevivência celular. / As a particularity, a 3D cell culture permits cells to explore the three dimensions of the space thereby increasing cell-cell interactions, as well as interaction with the environment. In studies related to breast cancer biology, spheroids are becoming widely used in the aim to comprehend luminal space morphogenesis. We showed that MCF-7 cells reorganize themselves in tubular and acinar structures. In both situations, lumen formation was accompanied by the establishment of a layer of polarized cells, an arrangement that is very similar to that of breast glands. The presented results suggest the existence of an MCF cell line population not completely committed to the tumor phenotype. When maintained as differentiated, MCF-7 cell spheroids can be a new model for studies regarding lumen formation, thereby exploring the role of diiferent pathways, such as those related to cell apoptosis, autophagy, differentiation and survival.

A própolis, um produto natural derivado de resinas de plantas coletadas por abelhas, foi usada por milhares de anos na medicina tradicional pelo mundo inteiro. Neste estudo, investigamos o efeito de uma fração da própolis vermelha brasileira (BRVD), no crescimento das células de melanoma murino (B16F10), das linhagens hematológicas humanas (HL-60 e K562), e de fibroblastos humanos (MCR-5 e FP). Após a análise preliminar de várias frações da própolis BRV, encontramos que a Fração BRVD inibiu fortemente o crescimento das células de uma maneira dose-tempo dependente pela necrose. Os resultados mostraram que essa fração induz eficazmente um efeito citotóxico em todas as linhagens estudadas, com média da IC50 em torno de 30 g/mL em 24 h de exposição. Estes resultados sugerem que a atividade antitumor da fração BRVD ocorre com a indução de necrose e os compostos dessa fração podem ser úteis como um agente contra o câncer / Propolis, a natural product derived from plant resins collected by honeybees, has been used for thousands of years in traditional medicine all over the world. In this study we have investigated the effect of some fractions from red Brazilian propolis on the growth of murine melanoma cell ( B16/F10), human hematological cells ( HL-60 and K562), and human fibroblasts cells (MCR-5 and FP). We found that BRVD strongly inhibited the growth of the cells in a dose- and time-dependent through induction of necrosis. Our results showed that BRVD effectively induced a cytotoxic effect on all cell lines studied, with IC50 average about 30 g/mL for 24 h of exposition. These results suggest that the antitumor activity of BRVD from red Brazilian propolis occurs through the induction of necrosis and its compounds may be useful as a anticancer agent

No câncer de mama, o metabolismo tumoral, ação dos moduladores de estrógenos são fatores que podem influenciar as células dendríticas (DCs). Neste trabalho avaliou o fenótipo de DCs em amostras tumorais, a diferenciação de DCs a partir de células mononucleares do sangue periférico (PBMCs) das pacientes e comparou com voluntárias saudáveis. Os resultados mostraram que há alteração na capacidade de geração, no fenótipo, na capacidade aloestimuladora, maior produção de interleucina 10 e expressão de HSP27 nas DCs de pacientes, comparadas com as DCs de voluntárias saudáveis que produzem mais Interferon-gama. A via p38MAPK parece ser importante na diferenciação de PBMCs em DCs, entretanto, estímulos estressantes podem ativar esta via e induzir a síntese de HSP27 inibindo este processo. O tratamento com tamoxifeno parece modular a expressão de algumas moléculas de membrana. Desta forma, os resultados sugerem que as DCs diferenciadas de pacientes com câncer de mama apresentam alterações fenotípicas e funcionais causadas pelo microambiente tumoral. / In breast cancer, the tumor metabolism, the action of estrogens antagonists can influence dendritic cells (DC) generation. The aim of this work was to evaluate the frequency of DCs in tumor tissue and the differentiation of DCs derived from peripheral blood mononuclear cells (PBMCs) and compared the phenotypic and functionally of these cells from healthy individuals and breast cancer patients. The results showed that patients PBMCs were unable to generate phenotypicaly, functionally mature DCs and presented larger production of interleukin 10 and higher expression of HSP27 when compared with healthy volunteers\' DCs, presented higher production of Interferon-gamma. The p38 MAPK signaling pathway seems to be important in PBMCs differentiation into DCs, and its activation by stress can induce the synthesis of HSP27, that inhibits DC generation. The tamoxifen treatment caused modulation of membrane DC markers expression. Therefore, these results show that patients\' DCs present phenotypic and functional alterations which can be caused by tumor microenvironment.

A profunda perda de tecido adiposo é considerada um marcador na caquexia associada ao câncer. O objetivo do estudo foi avaliar os efeitos da caquexia associada ao câncer em componentes da matriz extracelular do tecido adiposo subcutâneo (TAS) de pacientes. Pacientes do Hospital Universitário (HU) foram divididos em dois grupos: portadores de tumor com caquexia (TC) e controles (C). Amostras de TAS foram analisadas quanto aos aspectos morfológicos, morfométricos, ultraestruturais, moleculares por RT-PCR em tempo real para os genes: COL1A1, COL3A1, COL6A1, FN1 e MMP2, e por imunohistoquímica para colágeno (III, VI), fibronectina e metaloproteinase 2 (MMP2). O presente estudo relata alterações das características morfológicas dos adipócitos, bem como na expressão gênica do COL6A1, FN1 e MMP2 no TC. A imunopositividade observada estava modificada para colágeno III, VI, fibronectina e na MMP2. Conclusão: A caquexia associada ao câncer afeta profundamente o tecido adiposo conduzindo à fibrose tecidual. / Profound loss of adipose tissue is a hallmark of cancer cachexia. Nevertheless, the changes caused by cancer cachexia regarding the adipose tissue extracellular matrix have not yet been fully described. The aim of the study was to evaluate the effects of cancer cachexia upon extracellular matrix components of the subcutaneous adipose tissue (TAS) of cancer patients. Patients of the Hospital University (HU) were divided into two groups: tumour cachexia (TC) and control (C). Samples were analysed for morphological aspects, ultrastructurals, morphometric, molecular analyses by real time RT-PCR for gene COL3A1, COL1A1, COL6A1, FN1 and MMP2, and immunohistochemistry for collagen (III, VI), fibronectin and metalloproteinase 2 (MMP2). This study shows modifications of the morphological characteristics of the adipocytes as well as in gene expression of COL6A1, FN1 and MMP2 in TC. The imunopositivity also was modified to collagen III, VI, fibronectin and MMP2. Conclusion: cancer cachexia affects deeply the adipose tissue, leading to the emergence of tissue fibrosis.

As células dendríticas (DCs) são as mais eficazes células apresentadoras de antígenos. Mesmo com a possibilidade da geração de DCs in vitro, que permitiu a criação de protocolos de vacinação antitumoral, mecanismos de tolerância periférica, mediados por células T reguladoras, impedem uma resposta imune antitumoral eficaz. O presente estudo visou avaliar, in vitro, a geração de linfócitos T reguladores por células dendríticas derivadas de pacientes portadoras de câncer de mama. Para tanto, DCs foram diferenciadas a partir de monócitos do sangue periférico de pacientes com câncer, por sete dias, na presença de GM-CSF e IL-4 (DCs imaturas - iDCs), e ativadas, por adição de TNF-<font face=\"Symbol\">a no dia cinco de cultura (DCs maduras - mDCs). As DCs foram caracterizadas, por citometria de fluxo, quanto à: expressão de CD1a, CD11c, CD14, CD80, CD86, CD83, CD123, PD-L1, HLA-ABC e HLA-DR; produção de IL-10 e TGF-beta1, por ELISA; e ainda em ensaio funcional, que se deu pela co-cultura das DCs com linfócitos T (CD3+, CD3+CD25neg ou CD4+CD25neg), isolados por microsferas imunomagnéticas. Após co-cultura, a expressão de CD25, a proliferação (diluição de CFSE), a produção de citocinas (IFN-<font face=\"Symbol\">g, IL-10, TGF-beta1) e a geração de células Tregs foram analisadas. As células foram caracterizadas como Tregs por seu fenótipo (CD4+CD25+CD127lowCTLA-4+Foxp3+) e sua capacidade supressora sobre linfócitos alogeneicos. iDCs de pacientes apresentaram aumento da expressão de CD86 (com duas subpopulações: CD86High e CD86Low) e CD123 além de produção elevada de IL-10 e TGF-beta1 bioativo. Co-culturas com DCs de pacientes apresentaram níveis altos de TGF-beta1 bioativo (298,08 pg/ml x ctrl: 57,63 pg/ml) e induziram um alta freqüência de Tregs (iDCs: 57% ± 4,1; mDCs: 48% ± 5,0 x ctrl: 2,5% ± 0,7) a partir de precursores CD25negFoxp3neg, que foram capazes de suprimir a proliferação de linfócitos alogeneicos. O bloqueio de TGF-beta nas co-culturas reduziu parcialmente a freqüência de Treg geradas por DCs de pacientes. Esses achados são condizentes com a alta freqüência de Tregs no sangue periférico dessas mesmas pacientes (19,5% ± 2,3 x 8% ± 2,3) e com a presença de células com fenótipo de DCs no sangue, apresentando marcação semelhante a iDCs geradas in vitro. Por outro lado, iDCs provenientes de doadoras saudáveis induziram estimulação linfocitária mais intensa (35,7% ± 7,9 x 11,8 ± 5,9% CD25+), intensa proliferação de linfócitos CD4+ (82,7% x 29,4%) e CD8+ (73,8% x 21%) e alta produção de IFN-<font face=\"Symbol\">g (109,85 pg/ml x 7,86 pg/ml) nas co-culturas. Estes dados indicam que DCs derivadas de monócitos de pacientes com câncer de mama apresentam um viés imunossupressor que não é estritamente dependente do seu status de maturação ou de TGF-beta. Esses achados além de contribuir para a compreensão das interações entre o sistema imune e as neoplasias, devem ser considerados no delineamento de protocolos imunoterapêuticos baseados em DCs. / Dendritic cells (DCs) are the most effective professional antigen-presenting cells. Even considering the possibility of generating DCs in vitro, which allowed the design of antitumor vaccination protocols, mechanisms of peripheral tolerance mediated by regulatory T cells prevent an effective antitumor immune response. The aim of our study was evaluate, in vitro, the induction of regulatory T cells by dendritic cells derived from breast cancer patients.DCs were differentiated from breast cancer patients blood monocytes, for seven days, in the presence of GM-CSF and IL-4 (immature DCs- iDCs) and activated by TNF-<font face=\"Symbol\">a on day five of culture (mature DCs- mDCs). DCs were characterized by flow cytometry to CD1a, CD11c, CD14, CD80, CD86, CD83, CD123, PD-L1, HLA-ABC and HLA-DR expression; the cytokine secretion to IL-10 and bioactive TGF-beta1, by ELISA; and in functional assay by co-culturing DCs with T lymphocytes (CD3+, CD3+CD25neg or CD4+CD25neg) isolated by microbeads. Cell activation (CD25 expression), proliferation (CFSE dilution), cytokine production (IFN-gamma, IL-10 and TGF-beta1) and de novo regulatory T cells (Tregs) generation, were analyzed in these co-cultures after 5 or 6 days. Tregs were characterized by their phenotype (CD4+CD25+CD127LowCTLA-4+Foxp3+) and suppressive capability on allogeneic T cell proliferation. Patients iDCs showed a higher expression of CD86 (two subpopulation: CD86High and CD86Low) and CD123 beyond the elevated production of IL-10 and bioactive TGF-beta1. Co-cultures using patients DCs presented high levels of bioactive TGF-beta1 (298.08 pg/ml x ctrl: 57.63 pg/ml) and induced elevated frequency of Tregs (iDCs: 57% ± 4.1; mDCs: 48% ± 5.0 x ctrl: 2.5% ± 0.7) from CD25neg Foxp3neg precursors, which were able to suppress the allogeneic lymphocyte proliferation. The TGF-beta blocking partially reduced the frequency of induced Tregs by patients DCs. These findings are consistent with the higher frequency of Tregs on peripheral blood of those patients (19.5% ± 2.3 x ctrl 8% ± 2.3) and the presence of DCs also on the blood, showing similar markings with iDCs generated in vitro. Contrastingly, iDCs from healthy donors were better stimulator cells, leading to a higher CD25+ cell frequency (ctrl 35.7% ± 7.9 x 11.8 ± 5.9% CD25+), more intense proliferation of CD4+ (82.7% x 29.4%) and CD8+ (73.8% x 21%) cells and higher production of IFN-gamma (109.85 pg/ml x 7.86 pg/ml) on co-cultures. These data indicate that DCs derived from breast cancer patients show an immunosuppressive bias that is not strictly dependent on DCs maturation status or TGF-beta. Finally, these observations call to caution in the use of patients monocytes for the generation of DC-based vaccines and also contribute to the comprehension of the interactions between the immune system and cancer.

Introdução: O carcinoma de células renais metastático (CCRm) é um tumor altamente agressivo e resistente. Seu tratamento é baseado em terapia alvo molecular e citocinas. Avaliamos os efeitos antineoplásicos do Amblyomin-X, sobre CCR. Métodos: Avaliadas culturas de CCR RENCA e fibroblastos normais NIH/3T3 tratadas ou não com Amblyomin-X. Realizados ensaios de viabilidade celular por MTT e determinação, por citometria de fluxo, da proporção de células em apoptose/necrose; expressão da P-gp; Bad; Bax; Bcl-2; ciclina D1; caspase 3; Ki-67; p53; VEGFR1; citocromo c; análise das fase do ciclo celular; e atividade do proteassoma. Analisamos as populações celulares por microscopia eletrônica de varredura. Empregados testes T e One-way ANOVA para análise estatística. Resultados: O Amblyomin-X demonstrou citotoxicidade em células RENCA por indução de apoptose, diminuição de proliferação celular, inibição do proteassoma e modulação do ciclo celular em G0/G1. Em fibroblastos normais não houve citotoxicidade Conclusão: O Amblyomin-X apresentou efeito antineoplásico em CCR e não exerceu efeito citotóxico em células normais, demonstrando um possível potencial terapêutico no tratamento do CCRm. Estudos futuros deverão confirmar estes resultados. / Introduction: Metastatic renal cell carcinoma (mRCC) is a highly agressive and resistant tumour. Its treatment is based on targeted therapies and cytokines. We have evaluated the antineoplasic effects of Amblyomin-X on RCC. Methods: RCC (RENCA) and fibroblasts (NIH/3T3) cell cultures treated or not with Amblyomin-X were evaluated. MTT assay was performed to determine cell viability. Apoptosis/necrosis ratio; expression of P-gp; Bad; Bax; Bcl-2; cyclin D1; caspase-3; Ki-67; p53; VEGFR1; cytochrome c; cell cycle analysis and proteasome activity were obtained by flow cytometry. Cellular populations were analised by Scanning Electron Microscopy. Statistical analyses was performed using T-Tests and One-way ANOVA. Results: Amblyomin-X showed cytotoxic activity on RENCA tumor cells. It has induced apoptosis, decreased tumor cell proliferation, targeted the ubiquitinproteasome system and modulated genes related to cell cycle in G0/G1. There was no toxicity on fibroblasts. Conclusion: Amblyomin-X showed antineoplasic effects on RCC cells preserved normal fibroblast cells. There is a potential role of its therapeutic use in mRCC treatment. Future studies should confirm our initial results.

Acredita-se que as células dendríticas (DCs) sejam as mais eficientes na ativação de linfócitos T (LT) naive. Com a possibilidade de geração in vitro de DCs, muitos protocolos explorando este potencial vêm sendo desenvolvidos, principalmente em abordagens imunoterapêuticas para o câncer. Todavia, em situações fisiológicas a apresentação antigênica dificilmente ocorre por um tipo celular único. Deste modo, este trabalho investigou os padrões de reposta de LT induzidos por DCs maduras (mDCs) em combinação com diferentes APCs, incluindo linfócitos B, monócitos, macrófagos e DCs imaturas. Os padrões de resposta gerados pelos LT foram analisados por citometria de fluxo, ELISA e BIOPLEX. A estimulação dos LT pelas mDCs isoladas apresentaram o maior poder de estimulação, seguidas pelas outras APCs. Todas as combinações diminuiram a resposta induzida pelas mDCs, principalmente de LT CD4+. Deste modo, os dados confirmam o efeito das interações de APCs na estimulação de LT provendo ferramentas para o refinamento de abordagens imunoterapêuticas. / Dendritic cells (DC) are the main APC able to activate T cells (TC). Since they may be generated in vitro, many protocols based on their immunostimulatory potential are currently underway, mainly in immunotherapeutic approaches for cancer. In physiological conditions, however, other APC may participate in antigen presentation, thus influencing the immune response pattern developed. Therefore, the aim of this study was to evaluate the TC response patterns induced in vitro by mature DC (mDC) combined with different APC, including B cells, monocytes, macrophages and immature DC. The response patterns were analyzed flow cytometry, ELISA and Multiplex flow immunoassay. The TC stimulation by isolated mDC presented the highest capacity of stimulation, followed by the other APC. All combinations decreased the response induced by mDC, mainly CD4+ T cells. These data confirm the effects of APC interactions upon TC stimulation and may provide a tool for fine-tuning of immune response induction in immunotherapeutic approaches.