人免疫重建荷人人乳头瘤病毒16阳性宫颈癌-严重联合免疫缺陷鼠模型的建立及其免疫学特性研究

Development of HPV16 positive cervical cancer model in the hu-PBL-SCID mouse and its immunological features

目的 建立人免疫重建荷人人乳头瘤病毒 (HPV) 16阳性宫颈癌 严重联合免疫缺陷(SCID)小鼠模型并研究其免疫学特性。方法 对SCID鼠腹腔注射人外周血淋巴细胞 (hu PBL)后2 4h ,皮下接种HPV16阳性的人宫颈癌细胞株SiHa细胞建立人免疫重建荷人HPV16阳性宫颈癌SCID鼠模型 ,观察荷瘤鼠的一般生物学特性、异种移植物抗宿主病 (XGVHD)情况 ,检测血清中人IgG含量、外周血和脾中人CD3+ 、CD4 + 和CD8+ T细胞、脾重、肿瘤浸润淋巴细胞 (TIL)以及脾细胞毒杀伤功能。结果 免疫重建的小鼠成瘤率为 10 0 %。未发现异种移植物抗宿主反应。荷瘤第 5天 ,血浆中人IgG水平人化组 (0 98μg/ml± 0 2 0 μg/ml)、人化荷瘤组 (1 39μg/ml± 0 2 5 μg/ml)均显著高于空白组 (0 2 0 μg/ml± 0 11μg/ml) (t=7 6 5 5、9 937,均P =0 0 0 0 ) ,人化荷瘤组显著高于人化组 (t=3 2 0 0 ,P =0 0 0 6 ) ,荷瘤组与空白组比较差异无显著意义 (t=0 0 90 ,P =0 930 )。 30d内免疫重建组血浆中人IgG水平随重建时间延长而升高 ,与未人化组比显著升高 (P <0 0 5 )。外周血和脾中人CD3+ ,CD4 + 和CD8+ T细胞均显著升高 (P <0 0 5 ) ,脾重显著增加 (P <0 0 5 )。组织学观察到移植瘤局部TIL浸润并经免疫组织化学检测到人CD4 +

Objective To develop a HPV16 positive cervical cancer model in the hu-PBL-SCID mouse and investigate its immunological features. Methods Thirty-two CB17SCID mice were randomly divided into 4 groups: group A (5 mice) subcutaneously injected with phosphate-buffered saline, group B (5 mice) intraperitoneally injected with human peripheral blood lymphocyte(PBL) for immune reconstruction, group C(11 mice) subcutaneously injected with human cervical carcinoma cell line SiHa, and group D (11 mice) intraperitoneally injected with PBL and subcutaneously injected with SiHa cells after 24 hours of PBL transplantation. The tumor growth, behaviors and status of xenogeneic graft versus host disease(XGVHD) were observed. Human immunoglobulins G(IgG) in mouse serum,the percentage of human CD3+,CD4+ and CD8+ T cells in peripheral blood and spleen, spleen weight, tumor infiltrating lymphocytes and human CD4+ T cells, and cytotoxicity test of spleen cells were detected. Results The rate of successful tumor transplantation was 100%. XGVHD was not found . On the 5th day ,human IgG level in the group B (0.98 μg/ml±0.20 μg/ml)and group D (1.39 μg/ml±0.25 μg/ml) was significantly higher than that in the group A (t=7.655,9.937,both P=0.000). Human IgG level in group D was significantly higher than that in the group B (t=3.200,P=0.006). Only very low levels of human serumal IgG were detected in the group C and group A with no significantly difference. The level of human serumal IgG was gradually elevated in all the humanized SCID mice as the the time after PBL transplantation went on, and was significantly higher than that in non-humanized mice (P<0.05). The percentage of human CD3+,CD4+ and CD8+ T cells was significantly increased in the peripheral blood and spleen of immunoreconstituted SCID mice. The weight of spleen was markedly increased in the group D. TIL infiltrating in the tumor were remarkable and human CD4+ T cells was detected by immunohistochemistry in the group D but not in the group C. The spleen cells in the group D displayed stronger cytotoxicity to the target cells (P<0.05). Conclusion Human immune function can be successfully reconstructed in SCID mouse via intraperitoneally injecting with human PBL, and induce anti-tumor immune response to the transplantated tumor of HPV16 positive cervical cancer.