Generation of potent cytotoxic T lymphocytes against in male patients with non-muscle invasive bladder cancer by dendritic cells loaded with dying T24 bladder cancer cells

ABSTRACT Background In order to induce a potent cytotoxic T lymphocyte (CTL) response in dendritic cell (DC)-based immunotherapy for bladder cancer, various tumor antigens can be loaded onto DCs. Objective The aim of this study was to establish a method of immunotherapy for male patients with non-muscle invasive bladder cancer (NMIBC), using bladder cancer-specific CTLs generated in vitro by DCs. Materials and Methods Monocyte-derived DCs from bladder cancer patients were induced to mature in a standard cytokine cocktail (IL-1β, TNF-α, IL-6, and PGE2: standard DCs, sDCs) or anα-type 1-polarized DC (αDC1) cocktail (IL-1β, TNF-α, IFN-α, IFN-γ, and polyinosinic:polycytidylic acid) and loaded with the UVB-irradiated bladder cancer cell line, T24. Antigen-loaded αDC1s were evaluated by morphological and functional assays, and the bladder cancer-specific CTL response was analyzed by cytotoxic assay. Results The αDC1s significantly increased the expression of several molecules pertaining to DC maturation, regardless of whether or not the αDC1s were loaded with tumor antigens, relative to sDCs. The αDC1s demonstrated increased production of interleukin-12 both during maturation and after subsequent stimulation with CD40L that was not significantly affected by loading with tumor antigens as compared to that of sDCs. Bladder cancer-specific CTLs targeting autologous bladder cancer cells were successfully induced by αDC1s loaded with dying T24 cells. Conclusion Autologous αDC1s loaded with an allogeneic bladder cancer cell line resulted in increased bladder cancer-specific CTL responses as compared to that with sDCs, and therefore, may provide a novel source of DC-based vaccines that canbe used in immunotherapy for male patients with NMIBC.

CTLA-4 Exon one +49 A/G gene variants in patients with superficial and invasive bladder cancer: a study in southern Iran

Cytotoxic T-cell lymphocyte antigen 4 [CTLA-4] is a member of the superfamily of immunoglobulins that are mainly expressed by activated T cells. It is established that blockade of CTLA-4 receptors leads to the enhancement of an immune response. Different polymorphisms of the CTLA-4 gene have been described which cause increased susceptibility to various malignancies such as lymphoma or multiple myeloma. Considering that bladder cancer is one of the most prevalent cancers worldwide, we have evaluated the role of CTLA-4 gene polymorphism at position +49 A/G in the formation or progression of bladder cancer in southern Iran. A total of 226 individuals between February 2005 and June 2006 were included and placed into two subgroups: patients diagnosed with bladder cancer and a control group. Demographic data and risk factors were collected from both groups. The DNA of all subjects was extracted from their blood samples. Different genotypes of the CTLA-4 gene were determined using the restriction fragment length polymorphism [RFLP] technique and data were compared in both groups by using Pearson's chi-square test. The prevalence of AA, AG and GG genotypes at position 49, according to the PCR-RFLP method, were 57.5%, 37.2% and 5.3% in the control group, respectively. In the patient group, the prevalence of these genotypes was: AA in 57.5%, AG in 32.7% and GG in 9.8%. Statistical analysis of data showed no significant difference in both groups [P value=0.40]. Also there was no correlation between different genotypes of the CTLA-4 gene and invasiveness of the disease in our cases. Although polymorphism of the CTLA-4 gene at position 49 of exon-1 increases susceptibility to several malignancies, our study showed no relationship between polymorphism at this position and genetic susceptibility to the development of bladder cancer, nor was there any association with disease progression

Costimulatory molecule B7-H1 on the immune escape of bladder cancer and its clinical significance / 华中科技大学学报（医学）（英德文版）

B7-H1, a recently described member of the B7 family of costimulatory molecules, is thought to be involved in tumor immune escape by inducing T-cell apoptosis. In order to investigate the relationship between B7-H1 and immune escape of bladder cancer, B7-H1 expression in 50 cases of bladder cancer was detected by using immunohistochemical method. Survival curves were constructed using the Kaplan-Meier method and independent prognostic factors were evaluated using the Cox regression model. Our results showed that the positive rate of B7-H1 immunostaining in normal bladder tissue and bladder cancer was 0 and 72% respectively. The expression of B7-H1 was strongly associated with the pathological grade, clinical stage and recurrence (P<0.05). The survival rate was significantly lower in patients with B7-H1 positive group than in those with B7-H1 negative group and multi-variable analysis revealed that B7-H1 could be regarded as an independent factor in evaluating the prognosis of bladder cancer. It is concluded that the expression of B7-H1 is strongly associated with neoplastic progression and prognosis of bladder cancer. The manipulation of B7-H1 may become a beneficial target for immunotherapy in human bladder cancer.

Immunoreactivity of proliferating cell nuclear antigen in transitional cell carcinoma of urinary bladder

This is a descriptive study designed to evaluate Proliferating cell nuclear antigen [PCNA] in transitional cell carcinoma. A total of fifty per urethra resected bladder tumour samples [TUR BT] were collected from Mayo Hospital and Services Hospital, Lahore and processed for H and E and PCNA stain. The grading of tumours were made on H and E stain. While Proliferating cell nuclear antigen labeling index was recorded for each case. The mean PCNA labeling index was significantly higher in grade III when compared with tumours of grade II. [p < 0.001] carcinoma. Similarly mean PCNA labeling index was significantly higher [p < 0.05] in patients having duration of symptoms up to 3 month when compared with the patients having longer duration of symptoms. The mean PCNA labeling index had significantly higher value in high grade tumours as compared to low grade tumours [p < 0.001]. Although determination of PCNA labeling index is costly yet it has significant role in tumour grading

El bacilo de Calmette-Guérin como inmunomodulador en el cáncer de vejiga / The bacillus Calmette-Guérin as immunomodulator in bladder cancer

The bacillus Calmette-Guérin (BCG) is regarded as the most successful immunotherapy against superficial bladder carcinoma recurrences to date. BCG intravesical therapy for superficial bladder cancer has shown its efficacy and advantage over classical therapeutic strategies. This efficacy is based on complex and long lasting immune activation. The initial step is the binding of mycobacteria to the urothelial lining, which depends on the interaction of a fibronectin attachment protein on the bacteria surface with fibronectin in the bladder wall. Granulocytes and other immunocompetent mononuclear cells became attracted to the bladder wall and a cascade of proinflammatory cytokines sustains the immune response. In the bladder wall a largely TH1 based cytokine milieu and granuloma-like cellular foci are established. Within this scenario, the most important effector mechanisms might be the direct antitumor activity of interferons and the cytotoxic activity of NK cells. Current treatment consists of an induction phase of 6 weeks and a maintenance dose schedule of 3 weeks every three months up to 36. The majority of patients present adverse events related to dose administration due to bladder inflammatory response and on only a few ocassions, there are mayor complications like granulomatous prostatitis. Among all the neoplasms only in superficial bladder cancer the BCG is proved to be effective.

El bacilo de Calmette-Guérin (BCG) es considerado como la inmunoterapia más exitosa en contra del carcinoma de vejiga superficial recidivante hasta la fecha. La terapia intravesical con el BCG para el cáncer superficial de vejiga ha mostrado su eficacia y ventaja sobre otras estrategias terapéuticas; esta eficacia está basada en una compleja y larga duración de la activación inmunológica. El paso inicial es la unión de la micobacteria al urotelio, la cual depende de la interacción con la fibronectina de la bacteria con la fibronectina del urotelio. Los granulocitos y otras células mononucleares inmunocompetentes son atraídos hacia la pared vesical, así activando una cascada inmunológica a través de secreción de diversas citocinas, quienes estimulan a las células asesinas naturales (NK) activadas por el BCG, las cuales son capaces de diferenciar células neoplásicas y del epitelio urinario normal. En la pared vesical se encuentra un medio ambiente de citocinas principalmente del tipo TH1 y se forman focos celulares similares a granulomas. Dentro de este escenario los mecanismos efectores más importantes parecen ser la actividad antitumoral directa de los interferones y la actividad citotóxicas de las células NK. El tratamiento actual consiste en la administración intravesical del bacilo en una primera fase de inducción de seis semanas y posteriormente dosis de mantenimiento cada tres meses hasta los 36 meses. La mayoría de los pacientes presentan efectos adversos locales secundarios a la reacción inflamatoria y en un porcentaje muy pequeño se presentan complicaciones mayores como prostatitis y orquiepididimitis granulomatosa. De entre todas estas neoplasias sólo en el cáncer superficial de vejiga se han demostrado resultados satisfactorios con el empleo del BCG.

Bladder preserving approach for muscle invasive bladder cancer--role of mycobacterium w.

Mycobacterium w (immuvac), a new potent immunomodulator was evaluated as concomitant therapy in the management of muscle invasive bladder cancer along with external beam radiation therapy. There was no residual tumour in all the five patients after two months of therapy. All patients remained disease free for an observation period of more than 2 years.

Bacilo Calmette-Guérin intravesical em pacientes com carcinoma superficial de bexiga: atualizaçäo / Intravesical bacillus Calmette-Guérin in patients with superficial bladder carcinoma: up to date

O carcinoma de células transitional de bexiga superficial é uma doença multipotencial e a sua evoluçäo natural algumas vezes näo pode ser prevista precisamente. Tumor, grau e estádio da doença fornecem as mais importantes informaçöes até hoje. Há uma grande pesquisa e um enorme desafio nesta área, como os biomarcadores moleculares para indicar os fatores prognósticos nos pacientes de alto risco. As principais metas no tratamento do câncer superficial da bexiga säo: erradicaçäo da doença existente, profilaxia contra novas recorrências e prevençäo da progressäo da doença. O melhor tratamento é a imunoterapia com BCG liofilizado e uma resposta duradoura pode ser alcançada quando aplicada a manutençäo da terapia, mas há alguns tumores agressivos que devem ser tratados com cistectomia radical precoce. Os efeitos antitumorais do BCG intravesical contra o câncer urotelial parecem estar ligados com mecanismos efetores imunológicos, com aumento dos linfócitos T por infiltraçäo na lâmina própria da bexiga.

Experiencia conjunta del uso de dosis bajas de BCG cepa danesa en el tratamiento y la profilaxis del cáncer superficial de vejiga / Joined experience with the use of low doses of Danish strain of BCG in the treatment and prophilaxis of bladder superficial cancer

Se presenta el resultado de ocho años de experiencia con la utilización del bacilo de Calmette-Guérin (BCG) de la cepa danesa, a 35 mg por dosis, de los servicios de urología de las instituciones Centro Médico Nacional "20 de Noviembre" del ISSSTE y Hospital Central Sur de Alta Especialidad de PEMEX. Se ha comprobado en el pasado decenio la eficacia del BCG para el tratamiento del cáncer de células transicionales en etapas superficiales; el resultado puede ser el mejor de los obtenidos con los agentes antitumorales utilizados, pero los pacientes han tenido que pagar un precio muy alto a causa de las reacciones adversas que se presentan con la instilación intravesical de bacilos vivos; incluso se han informado casos fatales en la bibliografía. Se trataron 112 pacientes con BCG, durante un periodo de ocho años, con dos esquemas de tratamiento: 100 se trataron con 35 mg por dosis durante seis semanas, dos pacientes con esquema prolongado de tratamiento a dos años, y diez pacientes se sometieron a resección como único tratamiento. A continuación se compararon los resultados. De los 100 pacientes tratados con esquema corto, se obtuvo una reacción adecuada en 83 por ciento, con una proporción de recidivas de 17 por ciento. Ocurrieron reacciones adversas en 34 por ciento de los casos. Hubo un caso de sepsis del sistema nervioso central. Los dos pacientes tratados con esquema largo se encuentran libres de la enfermedad. Entre los pacientes tratados con resección nada más hubo 60 por ciento de recidivas y 20 por ciento de progreso tumoral

Inmunoterapia intravesical con BCG en el carcinoma superficial de células transicionales de la vejiga / Intravesical immunotherapy with BCG in bladder superficial transitional cell carcinoma

Se realizó un estudio prospectivo para conocer la eficacia del BCG intravesical como tratamiento en el carcinoma superficial del la vejiga de células transicionales y para prevenir la recurrencia de esta clase de tumores. Se trataron siete pacientes (seis varones y una mujer) con carcinoma superficial de vejiga de células transicionales estadio A, grado II, con promedio de edad de 61.2 años. Se utilizó la cepa danesa 1331 a dosis de 50 mg en 50 ml, equivalente a 500 y 1 000 millones de BCG. El esquema inicial consistió en la aplicación de 50 mg por semana durante seis semanas. Los pacientes con mayor riesgo de recurrencia se sometieron a esquema mensual de sostén. El seguimiento fue de 7.6 meses, a partir de la primera dosis, mediante vigilancia trimestral con cistoscopia, examen general de orina, urocultivo y citologías urinarias. La respuesta es de 100 por ciento, según lo establecido por el protocolo de los autores. Un paciente presentó recidiva después de haber recibido el ciclo inicial, y se sometió a un esquema de reinducción previa y RTU de tumor. Actualmente se encuentra libre de actividad tumoral. Este protocolo muestra prácticamente los mismos resultados de comparación con otros estudios sobre BCG sólo que con diferentes dosis y esquema de seguimiento

Primary adenocarcinoma of the urinary bladder--a case report with review of literature.

Primary adenocarcinoma of the urinary bladder is a rare neoplasm. We present the clinicopathological and immunohistochemical analysis of a case in a 50 year old male. The histogenesis of the tumour and a review of literature is also discussed.

Analysis of the immunologic mechanism of intravesical bacillus Calmette-Guerin therapy for superficial bladder tumors: distribution and function of immune cells

Intravesical bacillus Calmette-Guerin (BCG) administration has been used as an adjuvant therapy after transurethral resection for superficial bladder cancer, but the exact mechanisms of its antitumor activity are not yet known. The aim of this study was to characterize the immunologic aspects of antitumor activity of BCG using an animal model. C3H/He inbred mice and murine bladder tumor cell line, MBT-2 were used. The changes in immune cells such as helper T cells, suppressor T cells, macrophages and natural killer cells in the bladder and spleen were analysed by immunohistochemical method in intravesical BCG instilled in normal bladder, MBT-2 implanted after electrocauterization of the bladder mucosa and MBT-2 implanted and intravesical BCG treated group. The changes in natural killer cell activity of the splenocytes and peritoneal lymphocytes were evaluated using 51chromium release assay at regular time intervals following intraperitoneal BCG instillation. The prophylactic anticancer effect was evaluated by observing the tumor growth in the intravesically BCG treated group after intravesical MBT-2 implantation. In immunohistochemical examination, a remarkable infiltration of macrophage and helper T cell was observed in the lamina propria of the bladder, and the helper and suppressor T cells ratio (Th/Ts ratio) was increased after intravesical BCG therapy. In 51chromium release assay, enhanced natural killer cell activity of the splenocytes and peritoneal lymphocytes was observed after intraperitoneal BCG inoculation. The growth of implanted tumor was suppressed following intravesical instillation of BCG. These results suggest that the antitumor activity of BCG is not related to the simple inflammatory reaction but to the local and systemic immune response in which helper T lymphocytes and mononuclear cells play an important role.

Reduction of interleukin 1 and interleukin 2 production by patients with advanced cancer

The concentrations of interleukin 1 (IL-1) and interleukin 2 (IL-2) produced in the supernatants of peripheral blood mononuclear cell cultures from patients with advanced cancer were measured to identify some of the causes of the immunological impairment characteristic of malignant disease. Mononuclear cells obtained from 19 cancer patients were stimulated to produce IL-1 and IL-2 and compared with those of healthy controls. A severe reduction of both IL-1 and IL-2 activity was observed. There was no correlation between the lower number of OKT4+ cells observed in these patients and the levels of IL-2 production. The removal of monocytes did not bring IL-2 levels to normal. Impaired IL-2 production could not be restored to normal by addition of IL-1. These results suggest that exogenous IL-01 and IL-2 may be useful in cancer immunotherapy

Expressäo de antígenos do grupo sanguíneo ABO(H), antígeno T, antígeno carcinoembriônico e gonadotrofina coriônica humana subunidade beta em carcinoma de células transicionais de bexiga / ABO (H) and T blood group antigen, CEA and Beta-hCG expression in transitional cell carcinoma of bladder

O carcinoma de células transicionais é um grupo heterogênio de tumores que variam consideravelmente em sua história natural. A graduaçäo histológica, associada ou näo ao estadiamento, tem valor limitado na previsäo de quais os pacientes com CCT que säo de maior risco para recorrência/progressäo da doença. Vários outros parâmetros têm sido estudados para avaliar o comportamento biológico dos CCT, estando entre os mais importantes as anomalias cromossômicas e a expressäo de marcadores tumorais pelas células neoplásicas. O presente trabalho teve como principal objetivo estabelecer parâmetros fidedignos que pudessem prever quais os CCT que evoluíram com progressäo e/ou recorrência. Dentre esses parâmetros foram estudados: grau histológico e estádio dos CCT no momento do diagnóstico inicial e expressäo de marcadores tumorais (antígenos ABO(H), antígeno T, antígeno carcinoembriônico (CEA) e gonadotrofina coriônica humana sybunidade beta (hCG), em tecido, detectados por método de avidina-biotina-peroxidase. A análise da expressäo desses marcadores foi feita individualmente e em conjunto. Associando-se ao método da imunoperoxidase, utilizaram-se como reagentes específicos, para detecçäo dos antígenos, anticorpos monoclonais e lectina do Ulex europaeus para antígenos ABO(H), lectina do amendoim (Arachis hypogaea) para antígeno T, anticorpo monoclonal para antígeno carcinoembriônico e policlonal para gonadotrofina coriônica humana subunidade beta. Para análise dos CCT quanto ao grau histológico baixo e estádio superficial), G2 (grau histológico baixo e estádio superficial), G4 (grau histológico alto e estádio invasivo). Os principais resultados encontrados no presente trabalho foram: - G4 foi o grupo que mais se correlacionou com progressäo tumoral, principalmente quando comparado com G1. Os marcadores antígeno T, CEA e hCG apresentaram maior expressäo nos CCT do grupo G4, näo apresentando os antígenos ABO (H) diferença de expressäo nos grupos G1, G2, G3 e G4. A expressäo do conjunto dos marcadores näo mostrou diferença entre esses grupos de CCT. A hCG foi o único marcador tumoral que teve forte correlaçäo com progressäo. O antígeno críptico T foi o único marcador a ter maior expressäo nos CCT com recorrências. A expressäo do conjunto dos marcadores näo teve correlaçäo com progressäo e recorrência. Esses dados mostram que apesar da forte correlaçäo entre o grupo G4 e progressäo, os grupos G2 e G3 ficaram em posiçöes intermediárias...

Immunological reaction to bilharzial carcinoma of urinary bladder

Operative specimens taken from the urinary bladder of 50 patients formed the material of the present work. Immunohistochemical study of paraffin sections stained for subpopulations of lymphocytes using monoclonal antibodies and the immunoperoxidase procedure. Lymphocytic infiltration was seen in all studied cases of bilharzial carcinoma of the bladder.In transitional cell variety more than other cell types and in squamous Cell variety more than adenocarcinoma and undifferentiated carcinoma. In most cases lymphocytic infiltration was heavier in well differentiated tumour than in poorly differentiated and undifferentiated tumours. This study suggests that the natural host defence mediated through lymphocytes play a crucial role in determining the outcome of bilharzial carcinoma of the urinary bladder

Avaliaçäo prognóstica de pacientes com câncer de bexiga através da pesquisa de antígenos de superfície ABH / Prognosis evaluation of patients with bladder neoplasms by research of ABH surface antigens

A expressäo dos antígenos de superfície ABH tem sido estudada clinicamente em câncer de bexiga no sentido de se avaliar o potencial biológico da neoplasia. Com este objetivo, desenvolveu-se um estudo no qual se correlacionou a ocorrência de antígenos ABH na lesäo primária com a evoluçäo de 35 pacientes portadores de carcinoma de células transicionais de bexiga. Para a pesquisa dos antígenos ABH utilizou-se uma nova técnica de imunoperoxidase envolvendo o emprego consecutivo de lectina purificada do Ulex europaeus, anticorpo biotinado anti-Ulex obtido em carneiro e o complexo avidina-biotina-peroxidase. De acordo com este protocolo, observou-se que a expressäo dos antígenos de superfície ABH correlacionou-se de forma significativa com os índices de progressäo subseqüente da neoplasia vesical. Progressäo ocorreu em 13% dos pacientes com antígenos ABH presentes na lesäo inicial e em 75% dos casos com estes antígenos ausentes na neoplasia primária. Ademais, a expressäo dos antígenos de superfície ABH sugeriu a evoluçäo dos pacientes com lesöes de mesmo grau histológico. Nestes casos, progressäo da neoplasia ocorreu de forma mais freqüente quando os antígenos ABH estavam ausentes na lesäo inicial. Os resultados do presente estudo indicam que a expressäo dos antígenos ABH, avaliada pela técnica de imunoperoxidase descrita, reflete o potencial biológico das neoplasias vesicais, servindo para definir a propensäo de progressäo das mesmas. Desta forma, o estudo destes antígenos deve ser incorporado à rotina clínica, de forma a auxiliar na elaboraçäo da estratégia terapêutica de pacientes portadores de carcinoma de células transicionais de bexiga

Detecçäo imuno-histoquímica de antígenos de surperfície ABH em carcinoma de células transicionais da bexiga / Immunohistochemical detection of ABH surface antigens in transitional cell carcinoma of the bladder

Perda dos antígenos de superfície ABH é relatada em diversas neoplasias malignas, incluindo-se os carcinomas de bexiga. Estudos empregando técnica de aderência de hemácias (SRCA) sugeram que esta perda é mais significativa nas neoplasias menos diferenciadas, com pior prognóstico. Entretanto, o impacto destes achados em termos práticos foi atenuado por limitaçöes metodológicas do SRCA. Sabendo da elevada eficácia dos métodos imuno-histoquímicos mais modernos, recorremos à lectina do Ulex europaeus para a detecçäo dos antígenos ABH, mediante amplificaçäo com o complexo avidina-biotina-peroxidase, pesquisando correlaçäo com a graduaçäo histológica, critério clássico na avaliaçäo prognóstica. Dos 66 estudados, 39 (59,1%) apresentavam manutençäo dos antígenos em suas membranas. Nos casos grau I a positividade atingiu 85,0%, sendo quantificada como "intensa" em 69,2%. Nos casos graus II a positividade foi 82,7%, mas só em 58,6% mostrou-se "intensa". No outro extremo, apenas 17,0% dos casos grau III mostraram positividade, que foi "intensa" em 12,5%. Concluímos, entäo, que este método imuno-histoquímico é de fácil execuçäo e aplicaçäo a amostras fixadas em formol e incluídas em parafina, podendo vir a ser útil na rotina urológica com vistas à avaliaçäo prognóstica

Comportamento dos antígenos de superfície ABH em neoplasias vesicais superficiais e infiltrativas: inferências sobre os mecanismos de progressäo tumoral / Behavior of ABH surface antigens in superficial and invasive bladder neoplasms: interferences on tumoral progression mechanisms

A expressäo dos antígenos de superfície ABH relaciona-se com o comportamento biológico dos tumores vesicais. Estes antígenos estäo presentes em neoplasias superficiais e ausentes em neoplasias infiltrativas de bexiga. No sentido de elucidar os mecanismos de progressäo destas lesöes, foram estudados em dez pacientes com carcinoma de células transicionais de bexiga, o perfil antigênico ABH de tumores com componentes invasivo e näo invasivo concomitante. Os antígenos ABH, avaliados por um método de imunoperoxidase associando lectina de Ulex europaeus e o complexo avidina-biotina-peroxidase, estavam presentes em dez dos dez componentes näo-invasivos (100%) e em nenhum dos dez componentes invasivos (0%). Estes dados indicam que os tumores invasivos e näo-invasivos originar-se-iam de diferentes populaçöes celulares ou, alternativamente, teriam mesma origem clonal, associando-se nesta última situaçäo, em fenômeno de desdiferenciaçäo de parte das células, com conseqüente perda dos antígenos ABH e desenvolvimento do componente invasivo

Isolation of carcinoembryonic antigen from human bladder carcinoma

Carcinoembryonic antigen [CEA] was isolated from human bladder carcinoma for the first time. The obtained CEA was purified by a conventional procedure involving perchloric acid extraction followed by ethanol precipitation of tumor tissue and fractionation of this extract by gel filtration on Sepharose 4B and Sephadex G-200. The isolated purified CEA have been determined by radioimmuno-assay [RIA] and immunoelectrophoresis [IEP]. The CEA isolation procedure, which was described, saved a considerable amount of time and leads to a remarkable purification of CEA from bladder carcinoma