ovarian cancer-specific vaccine therapy by carrier cell

gfp引入了感染溶瘤腺病毒AdE3-IAI的非小细胞肺癌A549细胞。将3B注射到小鼠皮下肿瘤中，检测GFP和AdE3-IAI的DNA含量。肿瘤中的3B在一天后达到峰值，两周后消失。以A549细胞为基础，通过限制稀释建立EHK-2细胞，其抗肿瘤效果比原A549细胞高10倍。皮下注射AdE3-IAI后，40%小鼠的OVHM皮下肿瘤形成被拒绝。3b感染的EHK-2载体细胞。这表明在注入载体细胞后引入了异种抗肿瘤免疫。载体细胞在感染次数为200MOI、感染时间为33 h时的抗肿瘤效果最好。未感染的载体细胞形态未发生改变，但AdE3-IAI发生改变。乙型肝炎病毒感染的载体细胞表现为核膜分叶。载体细胞冻融后，分叶核膜破裂，腺病毒颗粒从细胞核释放到细胞质中。在这些感染条件下，体外抗肿瘤效果提高了20%，预先免疫腺病毒后90%的小鼠肿瘤完全缩小。在OVHM卵巢癌细胞腹腔播散小鼠模型中，腹腔注射AdE3-IAI。3b感染的载体细胞不能治愈任何小鼠，但Ad-mGM-CSF和AdE3-IAI感染的载体细胞不能治愈任何小鼠。3B显示，在先前的腺病毒免疫后，所有治疗小鼠的肿瘤完全减少。经载体细胞治疗的完全减瘤小鼠，OVHM卵巢癌细胞的肿瘤形成均被排斥，并引入自身抗肿瘤免疫。

GFP-introduced non small cell lung cancer A549 cells infected with oncolytic adenovirus, AdE3-IAI.3B was injected into the mouse subcutaneous tumors and DNA contents of GFP and AdE3-IAI.3B in the tumors peaked one day later and disappeared two weeks later. EHK-2 cell was established from A549 cells by limiting dilution and showed the 10 time higher antitumor effect than the original A549 cells. OVHM subcutaneous tumor formation was rejected in 40% of mice after the subcutaneous injection of AdE3-IAI.3B-infected EHK-2 carrier cells. This indicated that xenogenic antitumor immunity was introduced after the injection of carrier cells. The infection condition of carrier cells at 200MOI (multiplicity of infection) and 33 hours showed the best antitumor effect. The morphology of non-infected carrier cells was not changed but AdE3-IAI.3B-infected carrier cells showed the nuclear membrane lobulation. After the freeze and thaw of carrier cells, the lobulated nuclear membrane was ruptured and the adenovirus particles were released from the nucleus into the cytoplasm. Under these infection conditions, the in vitro antitumor effect increased by 20% and 90% of mouse showed the complete tumor reduction after the prior immunization of adenovirus. In the mouse model of intraperitoneal dissemination of OVHM ovarian cancer cells, the intraperitoneal injection of AdE3-IAI.3B-infected carrier cells did not cure any mice but the carrier cells infected with Ad-mGM-CSF and AdE3-IAI.3B showed the complete tumor reduction in all treated mice after the prior adenoviral immunization. The tumor formation of OVHM ovarian cancer cells was rejected in all complete tumor reduced mice treated with carrier cells and self-antitumor immunity was introduced in these mice.