Department of Urology, University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA.
J Urol. 2010 Jan;183(1):333-8. doi: 10.1016/j.juro.2009.08.110.
The immunological consequences of cryoablation for renal cell carcinoma are largely unknown. Cryoablation is an attractive therapeutic option for tumors due to its minimally invasive nature. Cryoablation is also potentially immunogenic. We describe the development of an animal model to deliver in vivo renal cryotherapy to orthotopically implanted renal cell carcinoma and the results of multiple immunological interrogations after cryoablation.
Four to 6-week-old female Balb/c mice (Jackson Laboratories, Bar Harbor, Maine) underwent renal subcapsular implantation of the syngeneic murine renal cell carcinoma Renca. Two weeks later contact cryoablation was done in tumor bearing kidneys. Another group of animals underwent cryoablation of normal kidneys. Animals were sacrificed 2 weeks after tumor injection or 1 and 2 weeks after cryoablation, respectively. Kidneys, spleens and draining lymph nodes were harvested. Evaluation consisted of immunohistochemistry, immunofluorescence and gene expression profiling using reverse-transcriptase polymerase chain reaction.
Subcapsular tumor implantation was successful in all cases and confirmed histologically. No significant lymphocytic infiltrate was seen in tumor only animals but those treated with cryoablation (tumor and nontumor bearing) had a significant inflammatory response primarily in sublethal tissue injury and perivascular areas. After cryoablation most infiltrating cells were neutrophils, macrophages and T cells. Polymerase chain reaction showed increased interferon-gamma production in kidneys after cryoablation.
This study shows the potential feasibility of this animal model for studying cryo-immunology. We confirm the absence of any significant immune cell infiltration in tumor bearing kidneys and report a significant inflammatory infiltrate after cryoablation, consisting primarily of neutrophils, macrophages, and CD4+ and CD8+ T cells with an increase in the T helper type 1/2 ratio. This orthotopic murine model can form the basis of future studies of additional immunological aspects of renal cryoablation.
冷冻消融治疗肾细胞癌的免疫学后果在很大程度上尚不清楚。由于其微创的性质，冷冻消融是一种有吸引力的治疗选择。冷冻消融也具有潜在的免疫原性。我们描述了一种动物模型的开发，用于将体内肾冷冻消融应用于原位植入的肾细胞癌，并描述了冷冻消融后的多次免疫学研究结果。
4-6 周龄雌性 Balb/c 小鼠（杰克逊实验室，缅因州巴港）接受同基因鼠肾细胞癌 Renca 的肾被膜下植入。2 周后，在肿瘤部位进行接触冷冻消融。另一组动物接受了正常肾脏的冷冻消融。分别在肿瘤注射后 2 周或冷冻消融后 1 周和 2 周处死动物。采集肾脏、脾脏和引流淋巴结。评估包括免疫组织化学、免疫荧光和逆转录聚合酶链反应的基因表达谱。
所有情况下被膜下肿瘤植入均成功，并通过组织学确认。仅肿瘤动物未见明显淋巴细胞浸润，但接受冷冻消融治疗（肿瘤和非肿瘤部位）的动物则表现出明显的炎症反应，主要发生在亚致死性组织损伤和血管周围区域。冷冻消融后，大多数浸润细胞为中性粒细胞、巨噬细胞和 T 细胞。聚合酶链反应显示冷冻消融后肾脏中干扰素-γ的产生增加。
本研究表明，该动物模型用于研究冷冻免疫学具有潜在的可行性。我们证实了肿瘤部位肾脏中不存在任何明显的免疫细胞浸润，并报告了冷冻消融后的显著炎症浸润，主要由中性粒细胞、巨噬细胞、CD4+和 CD8+T 细胞组成，辅助性 T 细胞 1/2 比值增加。这种原位鼠模型可以作为未来研究肾冷冻消融的其他免疫学方面的基础。
