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Cancer is primarily a disease in which late diagnosis is linked to poor prognosis, and unfortunately, detection and management are still challenging. Circulating tumor cells (CTCs) are a potential resource to address this disease. Cell fusion, an event discovered recently in CTCs expressing carcinoma and leukocyte markers, occurs when ≥2 cells become a single entity (hybrid cell) after the merging of their plasma membranes. Cell fusion is still poorly understood despite continuous evaluations in in vitro/in vivo studies. Blood samples from 14 patients with high-grade serous ovarian cancer (A.C. Camargo Cancer Center, São Paulo, Brazil) were collected with the aim to analyze the CTCs/hybrid cells and their correlation to clinical outcome. The EDTA collected blood (6 mL) from patients was used to isolate/identify CTCs/hybrid cells by ISET. We used markers with possible correlation with the phenomenon of cell fusion, such as MC1-R, EpCAM and CD45, as well as CEN8 expression by CISH analysis. Samples were collected at three timepoints: baseline, after one month (first follow-up) and after three months (second follow-up) of treatment with olaparib (total sample = 38). Fourteen patients were included and in baseline and first follow-up all patients showed at least one CTC. We found expression of MC1-R, EpCAM and CD45 in cells (hybrid) in at least one of the collection moments. Membrane staining with CD45 was found in CTCs from the other cohort, from the other center, evaluated by the CellSearch® system. The presence of circulating tumor microemboli (CTM) in the first follow-up was associated with a poor recurrence-free survival (RFS) (5.2 vs. 12.2 months; p = 0.005). The MC1-R expression in CTM in the first and second follow-ups was associated with a shorter RFS (p = 0.005). CEN8 expression in CTCs was also related to shorter RFS (p = 0.035). Our study identified a high prevalence of CTCs in ovarian cancer patients, as well as hybrid cells. Both cell subtypes demonstrate utility in prognosis and in the assessment of response to treatment. In addition, the expression of MC1-R and EpCAM in hybrid cells brings new perspectives as a possible marker for this phenomenon in ovarian cancer.

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BACKGROUND AIMS: Dendritic cell (DC)-tumor cell hybrids have been used clinically in cancer immunotherapy, but their advantage over the simple mixture of tumor cells and DCs is still a matter of controversy. In this study, we compared DC-tumor cell hybrids with the non-fused mixture of DC and tumor cells directly in their ability to induce a specific immune response. METHODS: Hybrids were obtained by electrofusion of tumor cells and monocyte-derived DCs. Cell phenotype was evaluated by flow cytometry and antigen-presenting ability by co-culture with syngeneic T cells followed by tetramer analysis and interferon (IFN)-γ ELISPOT. RESULTS: Less than half the cells in the mixture expressed DC co-stimulatory molecules. Furthermore, DCs in the mixture had significantly lower expression of MHC class I molecules than DCs in the fusion. Conversely, nearly all CD11c(+)Her2/neu(+) hybrids expressed CD80, CD86, CD83, HLA-DR and MHC class I from both tumor cells and DCs. Using tumor cells constitutively expressing a cytomegalovirus (CMV) antigen, we show that expansion of CMV-specific cytotoxic T lymphocytes (CTLs) restricted by DCs' MHC class I molecules was higher when DC-tumor hybrids were the stimulators. Furthermore, only hybrids stimulated CTLs to produce IFN-γ in response to CMV-positive target cells. CONCLUSIONS: These data show the superiority of DC-tumor cell hybrids over their simple mixture as T-cell stimulators. Hybrids expressed more co-stimulatory and MHC molecules, induced higher antigen-specific T-cell expansion and were the only cells able to induce IFN-γ-producing antigen-specific T cells. Thus, these data offer further support for cancer immunotherapeutic approaches using DC-tumor cell hybrids.

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OBJETIVO: Presentar la filosofía utilizada y como y por qué decidimos proteger el nivel adyacente a una fusión. MÉTODOS: En el criterio de selección de 620 pacientes operados entre enero de 2007 y agosto de 2011 por patología degenerativa, inestabilidad y estenosis del conducto lumbosacro, se seleccionaron 30 pacientes con estadios de Pfirmann 3 y 4, de los cuales seis se perdieron en la consulta postoperatoria y cuatro rechazaron la terapéutica quirúrgica, quedando 20 pacientes que fueron intervenidos quirúrgicamente. La edad promedio de los pacientes fue de 46 años (rango: 22 a 71 años), siendo 11 hombres (55%) y 9 mujeres (45%). RESULTADOS: El seguimiento de los casos es de 6 meses a 2 años y hasta el momento no se evidenció empeoramiento clínicoo radiológico, ni aflojamiento de la instrumentación en ningún caso. CONCLUSIONES: Entendemos que la protección del nivel adyacente mediante el empleo de barras semirrígidas en PEEK sería una buena alternativa de protección debido a que no es necesario abordar el ligamento o los pedículos del nivel adyacente a la fusión.

OBJETIVO: Apresentar a filosofia utilizada e como e por que decidimos proteger o nível adjacente a uma fusão. MÉTODOS: No critério de seleção de 620 pacientes operados entre janeiro de 2007 e agosto de 2011 devido a patologia degenerativa, instabilidade e estenose do canal lombossacral, foram escolhidos 30 pacientes com estágios 3 e 4 de Pfirmann, dos quais seis foram perdidos na consulta pós-operatória e quatro recusaram a cirurgia, restando 20 pacientes que foram submetidos à cirurgia. A idade média dos pacientes era 46 anos (faixa: 22 a 71), sendo 11 homens (55%) e 9 mulheres (45%). RESULTADOS: O acompanhamento dos casos é de 6 meses a 2 anos e, até o momento, não se evidenciou piora clínica ou radiológica, nem afrouxamento da instrumentação em nenhum caso. CONCLUSÕES: Entendemos que a proteção do nível adjacente por meio do emprego de hastes semirrígidas em PEEK seria uma boa alternativa de proteção, devido ao fato de não ser necessário abordar o ligamento ou os pedículos do nível adjacente à fusão.

OBJECTIVE: To present the philosophy used, and demonstrate how and why we decided to protect the level adjacent to a bone union. METHODS: In the selection criteria of 620 patients who had undergone surgery between January 2007 and August 2011 due to degenerative pathology, instability and stenosis of the lumbosacral canal, 30 patients were selected with Pfirmann grades 3 and 4, from which six were lost to follow-up and four refused surgery, leaving 20 patients who underwent surgery. The mean age of the patients was 46 years (range: 22 to 71), with 11 men (55%) and 9 women (45%). RESULTS: The follow-up of the cases was 6 months to 2 years, and so far, no clinical or radiological worsening has been observed, or loosening of the instrumentation in any case. CONCLUSIONS: we understand that protection of the adjacent level through the use of semi-rigid rods in PEEK is a good alternative, as it is not necessary to approach the ligament or pedicles of the level adjacent to the union.

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