Expression of the SART1 tumor rejection antigen in renal cell carcinoma

We have previously described the SART1 gene, which encodes both the SART1₂₅₉ antigen expressed in the cytosol of the majority of squamous cell carcinomas and some adenocarcinomas and the SART1₈₀₀ antigen expressed in the nucleus of the majority of proliferating cells. The SART1₂₅₉ antigen is recognized by HLA-A24 and A26-restricted cytotoxic T lymphocytes (CTLs). The present study investigated the expression of these two antigens in renal cell carcinomas (RCCs) in order to identify an appropriate molecule for use in specific immunotherapy for RCC patients. These two antigens were detected in all RCC cell lines and cells of the primary cultures of the RCCs tested. Further, they were detectable in cells of the primary cultures of non-tumorous kidney tissues. In contrast to these cultured cells, SART1₂₅₉ was detectable in only a few uncultured RCC tissues (5/20, 25%) and was undetectable in non-tumorous kidney tissues. SART1₈₀₀ was also scarcely detectable in uncultured RCC tissues (3/20, 15%) and non-tumorous kidney tissues (4/20, 20%). HLA-A2402-restricted and tumor-specific CTL (KE4-CTL) used for the cloning of the SART1 gene showed significant levels of cytotoxicity to both the cells from the RCC cell line and the cells from the primary cultures of RCC tissues, but did not lyse any normal cells, including cells from the primary cultures of non-tumorous kidney tissues. The SART1-derived peptide at positions 690–698 induced HLA-A24 restricted CTLs cytotoxic to RCC cells from peripheral blood mononuclear cells (PBMCs) of RCC patients. Therefore, the SART1 peptide could be an appropriate molecule for use in peptide-based specific immunotherapy for RCC patients. Received: 22 October 1999 / Accepted: 11 January 2000     

Immunological effects of granulocyte-macrophage colony-stimulating factor and autologous tumor vaccine in patients with renal cell carcinoma

PURPOSE: Biological therapy for renal cell carcinoma (RCC) uses agents that mobilize immune effector cells which are able to recognize and destroy cancer. We evaluated the effects of weekly then monthly autologous tumor vaccine combined with daily granulocyte macrophage-colony stimulating factor (GM-CSF) in patients with RCC as a method of stimulating antigen presenting cells. MATERIALS AND METHODS: Eligible patients with pathological stage II to IV RCC were entered into this pilot study. Autologous tumor vaccine (0.5 to 1 x 107 irradiated tumor cells) admixed with 250 microg GM-CSF per vaccine was given subcutaneously weekly for 4 weeks and then monthly for 4 months. GM-CSF (125 microg/m2) was given subcutaneously for 13 days after vaccine injection 1 and injections 4 to 8. Treatment related tumor specific CD4 and CD8 positive T cell precursors were assessed. RESULTS: A total of 22 patients were entered into this study. Patients were stratified by bulk of disease (group 1, 9 patients with micrometastatic disease, and group 2, 13 patients with macrometastatic disease). In general treatment was well tolerated. Of 9 patients in group 1 7 remained disease-free after nephrectomy. In group 2, 6 patients had stable (46.2%) and 7 patients had progressive disease (53.8%). Statistically significant treatment related increases in CD4 (p = 0.028) and CD8 (p = 0.018) positive tumor specific T cell precursors were observed for the entire group of patients. Changes in CD4 and CD8 positive precursors correlated significantly with each other (p = 0.0001). This correlation was seen in the 2 patient subpopulations as well (group 1 p = 0.003, group 2 p = 0.013). Patients with minimal disease, and with changes in CD4 and CD8 positive tumor specific T cell precursors greater than the median appeared to have an improved time to progression as well as a survival benefit. CONCLUSIONS: GM-CSF and autologous vaccine can be given safely in combination to patients with renal cell cancer. We observed treatment related changes in tumor specific circulating lymphocyte populations.     

Phase I trial of a B7-1 (CD80) gene modified autologous tumor cell vaccine in combination with systemic interleukin-2 in patients with metastatic renal cell carcinoma

PURPOSE: A reason that the immune system may fail to reject tumors is that T cells encounter tumor antigen derived peptides on the surface of tumor cells in a tolerizing rather than activating context since tumor cells do not express T cell co-stimulatory molecules such as B7-1 (CD80). In preclinical models over expression of B7-1 on the surface of tumor cells has been shown to activate T cells which kill tumor cells. We conducted a phase I clinical trial testing this approach in patients with metastatic renal cell carcinoma. MATERIALS AND METHODS: Resected tumors from 15 patients were disaggregated and adapted to tissue culture, transduced with the B7-1 gene and injected subcutaneously as a vaccine. The dose of the vaccine was escalated in 3 separate cohorts of patients, and systemic interleukin-2 (IL-2) was administered as an adjuvant designed to enhance the proliferation of the vaccine activated T cells. RESULTS: Of the 15 patients 9 had measurable disease, 2 had a partial response and 2 had stable disease. Perivascular T cell infiltrates at autologous tumor delayed type hypersensitivity skin test sites developed in 3 of the 4 patients with stable disease or partial response. Although the patients experienced the usual and expected toxicity from the IL-2, there was no significant toxicity observed with the vaccine. CONCLUSIONS: The B7-1 gene modified autologous tumor cell vaccine is safe and can be combined with systemic IL-2 with acceptable toxicity. Immunological and clinical responses were observed in some of the patients. A phase II trial is reasonable to determine the efficacy of this approach.     

Collecting duct renal cell carcinoma: clinical study of a rare tumor.

PURPOSE: Collecting duct carcinoma is a rare type of renal cell carcinoma that affects younger patients, and is associated with aggressive regional and distant spread. The clinical and pathological features of 6 patients with collecting duct carcinoma treated at a single institution are described. MATERIALS AND METHODS: There were 6 patients with collecting duct carcinoma included in the University of California School of Medicine, Los Angeles, Kidney Cancer Database. Demographic, clinical, pathological and survival data were gathered. RESULTS: Average patient age plus or minus standard deviation was 56 +/- 11 years, and 5 of 6 had TNM stage IV disease. The average survival of these patients was 11.5 months (range 7 to 17). There was 1 patient who had TNM stage I disease and survived without evidence of disease at 5 years. Transient response to chemotherapy was seen in 1 patient. CONCLUSIONS: Collecting duct carcinoma is associated with poor prognosis. For the majority of patients surgical treatment will not result in a cure. Previously recommended chemotherapy and/or immunotherapy appears to have a limited role in treatment of this disease, and early detection may be the best method for prolonging patient survival.     

5t4在肾透明细胞癌中的表达及意义

目的探讨滋养层糖蛋白5t4在肾透明细胞癌中的表达及其意义。方法采用免疫组织化学技术检测72例肾透明细胞癌组织标本、17例癌旁肾组织以及14例非癌因素的肾脏组织标本中5t4的表达,并对5t4在肾透明细胞癌的表达与组织学分级的关系进行分析。结果 5t4在肾细胞癌组织中的阳性率为70.8%,癌旁肾组织中的阳性率为41.2%,两者具有差异性(P0.05)。5t4在肾癌组织学分级中低分级与高分级的表达存在显著差异(P0.01)。结论 5t4可作为判断肾透明细胞癌分化程度的标志物。     

Combined small cell carcinoma and sarcomatoid squamous cell carcinoma in the renal pelvis

We report here a case of combined small cell carcinoma and sarcomatoid squamous cell carcinoma in the renal pelvis. A 61-year-old female presented with right flank discomfort, microhematuria and progressive renal dysfunction. Following diagnosis of right renal pelvic carcinoma, radical nephroureterectomy with lymph node dissection was performed through a midline incision. The tumor was pathologically diagnosed to be combined small cell carcinoma and sarcomatoid squamous cell carcinoma in the renal pelvis. The patient had no evidence of recurrence or metastasis, 16 months postoperatively. Small cell carcinoma or sarcomatoid squamous cell carcinoma of the renal pelvis is very rare. We believe this is the first such case to be reported in the world.     

肾癌患者手术及免疫治疗前后的血清蛋白指纹图谱的研究

目的:分析肾癌患者手术前后及免疫治疗前后的血清蛋白指纹图谱,构建并评估血清差异蛋白诊断模型。方法:首先应用弱阳离子磁珠技术富集肾癌患者手术前后、免疫治疗后及健康对照组的血清蛋白,其次用基质辅助激光解离-飞行时间质谱(MALDI-TOF-MS)技术构建肾癌患者不同时期的蛋白指纹图谱,再应用有监督以及无监督的化学计量学模式识别方法对构建的指纹图谱进行效力评价,最后应用受试者工作特征曲线(ROC)寻求区别各个时期肾癌患者血清中的差异蛋白。结果:在无监督模式识别方法(主成分分析,PCA)结果不理想的情况下,对比了不同有监督模式识别方法构建的指纹图谱模型,结果表明遗传算法(GA)对各个时期的肾癌患者均有较高的识别率,达到了97.7%;术前和术后差异蛋白只有一个,而术前和免疫治疗后、术后和免疫治疗后存在较多的差异蛋白。结论:肾癌患者经过免疫治疗后,血清蛋白质和术前及术后存在明显差异,肾癌应用遗传算法构建的血清蛋白指纹图谱可有效识别术前、术后、免疫治疗后以及正常对照组的血清差异蛋白。     

Dendritic cell immunotherapy for patients with metastatic renal cell carcinoma: University of Tokyo experience

BACKGROUND: Dendritic cells (DC) are the most potent antigen-presenting cells and induce host antitumor immunity through the T-cell response. A clinical study of immunotherapy using cultured DC loaded with tumor antigen, for patients with metastatic renal cell carcinoma (RCC) was performed. METHODS: Dendritic cells were generated by culturing monocytes from peripheral blood for 7 days in the presence of granulocyte-macrophage colony-stimulating factor and interleukin-4. On day 6 the DC were pulsed with lysate from autologous tumor as the antigen and with keyhole limpet hemocyanin (KLH) as immunomodulator. The patients were given four doses of lysate-pulsed DC by intradermal injection with a 2-week interval between doses. Clinical effect and immune response were, respectively, evaluated by radiological examination and delayed-type hypersensitivity (DTH) test. RESULTS: Three patients were enrolled and the immunotherapy was well tolerated without significant toxicity. The vaccination induced a positive DTH reaction to tumor lysate in two patients and to KLH in all patients. Clinical responses consisted of one case of no change and two cases of progression of disease. However, we did not see a significant reduction of tumor volume in any case. CONCLUSION: Dendritic cell vaccination can safely induce an immunological response against RCC. Further trials are needed to fully evaluate its efficacy.     

富组蛋白-3在肾透明细胞癌患者中的表达及临床意义

目的利用蛋白质芯片技术筛查肾透明细胞癌患者手术前后血清差异性表达的蛋白质,并探讨其临床意义。 方法选取2015年10月至2016年10月在新疆医科大学第一附属医院泌尿外科手术治疗并经术后病理证实为肾透明细胞癌患者89例为肾癌组,选择同期在我院体检的健康志愿者以及非肾细胞癌患者100例为对照组。肾癌组术后规律随访1年。运用表面增强激光解析电离化飞行时间质谱系统（SELDI-TOF-MS）和CM10弱阳离子交换蛋白芯片技术系统检测肾癌组及对照组血清差异性表达的蛋白质。应用线性支持向量机（SVM）方法建立蛋白质指纹图诊断模型,留一法交叉验证模型判别效果。通过ZUCI-PDAS蛋白质谱数据分析系统软件对血清差异性蛋白质进行分析。 结果质荷比（M/Z）为2745的蛋白质在肾癌组与对照组呈差异性表达,所对应为富组蛋白-3,其表达水平在肾癌术前组较对照组呈低表达,差异有统计学意义（P<0.05）;肾癌组术前与术后1个月、3个月、6个月以及12个月后的表达水平比较,术前表达水平低于术后各随访段水平,差异均有统计学意义（P<0.05）。术后3、6、12个月表达水平与术后1个月相比较,差异有统计学意义（P<0.05）。术后6、12个月与术后3个月表达水平相比较,差异无统计学意义（P>0.05）。依据富组蛋白-3在肾癌组患者与健康人血清中表达量不同,利用ROC曲线法分析,曲线下面积AUC=0.804,灵敏度为86.7％,特异度为66.7％。该蛋白质表达强度的某一界点作为标记,预测诊断肾透明细胞癌的灵敏度为86.3％（76/89）,特异度为88.0％（88/100）。 结论富组蛋白-3在肾癌与健康人群中的表达存在差异,且肾癌术后其表达水平升高,提示富组蛋白-3与肾细胞癌具有相关性,有望成为肾透明细胞癌新的分子标记物。     

NNMT在肾透明细胞癌的表达及对肾癌细胞侵袭能力的影响

目的:研究NNMT在肾透明细胞癌中的表达情况及对肾癌细胞侵袭能力的影响。方法:采用RT-PCR和Western blot方法检测正常肾小管上皮细胞株HKC、肾癌细胞株786-O及30例肾透明细胞癌组织、相应癌旁组织中NNMT的mRNA和蛋白的表达水平,并分析NNMT的mRNA水平与临床病理参数的关系。化学合成针对NNMT特异的siRNA序列,应用脂质体Lipofectamine 2000将其转染进786-O细胞中,利用RT-PCR和Western blot法检测NNMT在786-O细胞中的表达水平,用Transwell小室法检测肾癌细胞786-O侵袭能力的变化。结果:NNMT在肾癌细胞786-O中的mRNA和蛋白表达水平显著高于正常肾小管上皮细胞株HKC(P<0.001);肾透明细胞癌组织和对应的癌旁组织中NNMT的mRNA相对表达量分别为(1.582±0.2145)、(0.1269±0.04279),两组比较P<0.001。NNMT的mRNA水平与肿瘤大小、临床分期有关(P<0.05);Tran-swell法检测结果显示降低NNMT的表达后786-O细胞的侵袭能力明显下降。结论:NNMT在肾透明细胞癌组织和细胞中表达升高,可能在肾癌发生、发展过程中发挥重要作用。     

Telomerase pulsed dendritic cells for immunotherapy for renal cell carcinoma

PURPOSE: Renal cell carcinoma (RCC) is known for its immunological susceptibility. Unfortunately RCC lacks specific tumor antigens for the induction of specific immunotherapy. We investigated the role of telomerase as a tumor antigen and pulsed dendritic cells (DCs) as antigen presenting cells with an immunogenic peptide from telomerase. MATERIAL AND METHODS: DCs and immunological effector cells, that is cytokine induced killer (CIK) cells, from patients with RCC or healthy donors were generated. CIK cells were co-cultured with telomerase peptide pulsed DCs. CIK cells were tested for cytotoxic activity against primary cultures. Using the dimer technique we determined the percent of telomerase specific T cells. Activation status was identified using interferon-gamma secretion assay. RESULTS: After pulsing DCs with telomerase peptide co-cultured CIK cells had a significant increase in cytotoxic activity against tumor cells compared with CIK cells without co-culture, that is 100% at an effector-to-target ratio of 60:1 vs 41.7% (p <0.05). Using a complete autologous model with immunological cells derived from patients with metastatic RCC we were able to induce cytotoxicity against autologous, telomerase positive primary cell cultures. We could detect 2.4% telomerase specific effector cells after co-culture with peptide pulsed DCs, which secreted interferon-gamma after re-stimulation. CONCLUSIONS: Telomerase could serve as a specific tumor associated antigen for RCC. The presentation of telomerase peptide by DCs to lymphocytes allows the generation of antigen specific cytotoxic effector cells.     

Current management and future perspectives of metastatic renal cell carcinoma

Over the last number of years, the treatment of metastatic renal cell cancer has evolved tremendously with the advent of targeted therapy. Previously, immunotherapies, such as interferon alpha and interleukin‐2, were the only treatment options available for this chemoresistant malignancy. Currently, seven additional agents, including sunitinib, sorafenib, axitinib, pazopanib, bevacizumab, everolimus and temsirolimus, have been approved for use in metastatic renal cell cancer, with several more in development. The efficacy of these agents depends primarily on inhibition of the vascular endothelial growth factor and mammalian target of rapamycin pathways, and have drastically improved the outcomes of patients diagnosed with metastatic renal cell cancer. This article reviews the major treatment advances that have occurred for metastatic renal cell cancer with the advent of targeted treatments, summarizes the evidence to support their use and addresses clinical issues that have arisen with them. To help guide clinicians in their decision‐making with these emerging therapeutic choices, the evidence for sequencing and combining these agents, and the need for biomarkers will be addressed. The role of surgical management options, such as cytoreductive nephrectomy and metastectomy, in the era of targeted treatment is also reviewed. Several novel treatments are also on the horizon, which might serve as future avenues for treatment advancement in metastatic renal cell cancer.     

Impact of tumor size on the predictive ability of the pT3a primary tumor classification for renal cell carcinoma

PURPOSE: The accuracy of the pT3a primary tumor classification for renal cell carcinoma has been questioned recently. We investigated the association of perinephric and renal sinus fat invasion with death from renal cell carcinoma independent of tumor size. MATERIALS AND METHODS: We identified 2,165 patients treated with open radical nephrectomy or nephron sparing surgery for clinically localized, sporadic pT1a, pT1b, pT2 or pT3a renal cell carcinoma between 1970 and 2002. Patients with pT3a disease were then subdivided into 3 groups according to tumor size to match the size definitions for the pT1a, pT1b and pT2 tumor classifications. RESULTS: There were 834 patients with pT1a RCC, 674 with pT1b, 494 with pT2 and 163 with pT3a RCC. At last followup 317 patients died of RCC at a median of 3.8 years following surgery. The median followup among the 1,087 patients still alive at last followup was 7.8 years (range 0 to 34). The risk ratios (95% CI) for the association between fat invasion and death from RCC among patients with tumors 4 cm or smaller, 4 to 7 cm and more than 7 cm were 6.15 (1.84-20.50, p = 0.003), 4.12 (2.50-6.78, p <0.001) and 2.13 (1.53-2.97, p <0.001), respectively. These associations remained statistically significant in a multivariate analysis that included nuclear grade and histological coagulative tumor necrosis. CONCLUSIONS: Peripheral perinephric and renal sinus fat invasion was associated with death from RCC independent of tumor size. Our data contradict reports suggesting that pT3a tumors should be reclassified according to tumor size only.     

肉瘤样肾细胞癌的诊疗进展

肉瘤样肾细胞癌(sarcomatoid renal cell carcinoma, SRCC)是一种特殊类型的肾癌,临床少见,恶性程度高,无特异性临床表现,缺乏生物标记物,传统治疗方式疗效不佳,预后极差,是临床上较难诊治的疾病。目前诊断仍首选CT检查,联合CT、MRI、PET/CT及穿刺病理活检等多种方式对于SRCC诊断及鉴别诊断具有重要临床意义。治疗上多采取以根治性肾切除术为主,辅以基于免疫治疗的全身治疗。随着SRCC分子机制研究的不断深入及临床试验的开展,免疫联合靶向治疗显示出广阔应用前景,但现有证据较少,且多数为小样本回顾性研究,存在局限性,仍需进一步前瞻性随机对照研究为SRCC的诊疗提供更高等级的循证医学证据。     

Imaging techniques for the patient with renal cell carcinoma

The imaging of renal cell carcinoma continues to evolve from radiographic tomography to state-of-the-art three-dimensional imagery using computed tomography (CT) or magnetic resonance imaging (MRI). This article reviews the current techniques of imaging the patient with renal cell carcinoma. Careful and accurate imaging of these patients allows for the appropriate diagnosis, treatment planning, and follow-up care. At each point in the care of these patients, imaging plays an important role. In particular, the diagnosis and staging of renal cell carcinoma can be accomplished with CT and MRI, with each modality having strengths and weaknesses that are contrasted. Intraoperative ultrasound is used during laparoscopic or conventional partial nephrectomies, whereas ultrasound, CT, and MRI can be used for guiding ablative technologies. Imaging also plays an important role in the follow-up care of these patients. The particular follow-up care is dependent on the stage and grade of the lesion and the treatment modality used. In summary, this article reviews the current imaging approaches for the diagnosis, staging, treatment, and follow-up care of patients with renal cell carcinoma.     

Dexamethasone and interleukin-2 combination therapy for advanced renal cell carcinoma in a patient with paraneoplastic inflammatory syndrome

Interleukin-2 (IL-2) in combination with dexamethasone was administered to a 48-year-old man with renal cell carcinoma accompanied by paraneoplastic inflammatory syndrome, including progressive multiple lung metastases and inferior vena caval tumor thrombus. Although non-steroidal anti-inflammatory drugs had no apparent antipyretic effect on the systemic inflammatory syndrome, oral administration of dexamethasone achieved complete antipyresis and improved his quality of life. After a 4-week period of IL-2 treatment, regression of metastasized lesions was demonstrated despite concurrent oral administration of dexamethasone. Steroids might reduce the action of immunotherapeutic drugs, but in some cases, combination therapy can achieve both alleviation of the paraneoplastic syndrome and tumor shrinkage.     

Immunotherapy against metastatic renal cell carcinoma with mature dendritic cells

OBJECTIVE: We performed a clinical trial of immunotherapy using autologous mature dendritic cells (DC) pulsed with autologous tumor lysate, for patients with metastatic renal cell carcinoma (RCC). METHODS: Patients with refractory metastatic RCC were enrolled in the study. All of them received interferon (IFN)-alpha treatment after nephrectomy and were followed over 3 months prior to this study. Autologous monocyte-derived immature DC were pulsed with lysate from autologous primary tumor as the antigen and keyhole limpet hemocyanin (KLH) as immunomodulator, and cultured in the presence of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and prostaglandin (PG)E2 to generate mature DC. Mature DC were injected intradermally near bilateral inguinal lymph nodes of the patients. A delayed-type hypersensitivity (DTH) test and enzyme-linked immunospot (ELISPOT) assay were performed to evaluate the immunological response. After 4 months from first injection, the clinical effect was evaluated by diagnostic imaging. RESULTS: The treatments were well tolerated without significant toxicity by the patients who were an average of 65.7 years old and had multiple metastases in the lung and other organs. One of the two patients developed a positive DTH reaction to tumor lysate and the other patient only to KLH. The patient with a positive DTH reaction to tumor lysate had stable disease in the clinical evaluation. CONCLUSIONS: We confirmed the safety of DC therapy in this clinical trial. The DTH test revealed that the DC therapy induced immunological response to RCC. On the other hand, it was necessary to reconsider the patient selection criteria.