Manusama E R, Nooijen P T, Stavast J, de Wilt J H, Marquet R L, Eggermont A M
Department of Surgery, University Hospital Rotterdam-Dr Daniel den Hoed Cancer Center, Rotterdam, The Netherlands.
J Surg Res. 1998 Aug;78(2):169-75. doi: 10.1006/jsre.1997.5256.
Isolated limb perfusion (ILP) with TNFalpha in combination with melphalan and IFNgamma has resulted in an immediate and dramatic tumor response in patients. Such an effect was also noted following ILP in a rat sarcoma model. This model enables us to investigate several factors responsible for the TNFalpha-induced tumor responses. We applied total body irradiation (TBI) to reduce white blood cell count, to investigate the contribution of leukocytes to the anti-tumor effect of TNFalpha.
Small fragments of the nonimmunogenic BN 175 sarcoma were implanted sc in the lower hind leg. A 5 Gy TBI was performed before ILP at a tumor diameter of approximately 15 mm. The hind limbs of 63 rats were perfused and were divided into 6 groups: group 1, sham perfusion, n = 9; group 2, TBI + sham perfusion, n = 6; group 3, TNFalpha 50 microgram, n = 9; group 4, melphalan 40 microgram, n = 9; group 5, TNFalpha 50 microgram + melphalan 40 microgram, n = 22; group 6, TBI + TNFalpha + melphalan ILP, n = 8. In addition, 10 rats were perfused for histological analysis at 24 h post-ILP.
We observed in Group 1: 9/9 progressive disease (PD); Group 2: 6/6 PD; Group 3: 9/9 PD; Group 4: 9/9 no change (NC) of tumor diameter for at least 4 days; Group 5: 6/22 NC, 16/22 complete remission (CR), 12/16 of which showed skin necrosis at the tumor site; and Group 6: 7/8 NC and 1/8 CR (without skin necrosis). After TBI, WBC reduction of 80-95% was observed, while the number of platelets was not significantly reduced and platelet aggregation was maintained at 72 %. Histological analysis revealed decreased hemorrhagic necrosis associated with the absence of PMN infiltration at the tumor margins in the TBI rats.
TBI and the associated reduction in WBC count decreased the tumor response by TNFalpha and melphalan significantly and abrogated the immediate response of skin necrosis at the tumor site, as found in rats treated with TNFalpha and melphalan without TBI. These data strongly suggest that leukocytes play an important role in the hemorrhagic effects of TNFalpha.
在患者中，使用肿瘤坏死因子α（TNFα）联合美法仑和干扰素γ进行肢体隔离灌注（ILP）已产生即时且显著的肿瘤反应。在大鼠肉瘤模型的ILP后也观察到了这种效果。该模型使我们能够研究导致TNFα诱导肿瘤反应的几个因素。我们应用全身照射（TBI）来降低白细胞计数，以研究白细胞对TNFα抗肿瘤作用的贡献。
将非免疫原性的BN 175肉瘤小片段皮下植入大鼠后肢下部。在肿瘤直径约为15 mm时，于ILP前进行5 Gy的TBI。对63只大鼠的后肢进行灌注，并分为6组：第1组，假灌注，n = 9；第2组，TBI + 假灌注，n = 6；第3组，TNFα 50微克，n = 9；第4组，美法仑40微克，n = 9；第5组，TNFα 50微克 + 美法仑40微克，n = 22；第6组，TBI + TNFα + 美法仑ILP，n = 8。此外，10只大鼠在ILP后24小时进行灌注以进行组织学分析。
我们在第1组观察到：9/9进展性疾病（PD）；第2组：6/6 PD；第3组：9/9 PD；第4组：9/9肿瘤直径至少4天无变化（NC）；第5组：6/22 NC，16/22完全缓解（CR），其中12/16在肿瘤部位出现皮肤坏死；第6组：7/8 NC和1/8 CR（无皮肤坏死）。TBI后，观察到白细胞减少80 - 95%，而血小板数量未显著减少，血小板聚集维持在72%。组织学分析显示，TBI大鼠肿瘤边缘出血性坏死减少，且无中性粒细胞浸润。
TBI及相关的白细胞计数降低显著降低了TNFα和美法仑的肿瘤反应，并消除了在未进行TBI的TNFα和美法仑治疗大鼠中观察到的肿瘤部位皮肤即时坏死反应。这些数据强烈表明白细胞在TNFα的出血效应中起重要作用。
