目的 重復(fù)給予食蟹猴治療用乙型肝炎腺病毒注射液（TC007），觀察其免疫原性、免疫毒性、生物分布。方法 普通級(jí)食蟹猴隨機(jī)分為溶媒對(duì)照組，TC007低、中、高劑量（1.0×10⁹、1.0×10¹⁰、1.0×10¹¹ VP/只）組，Ad5-Null（1.0×10¹¹ VP/只，空載對(duì)照）組，每組10只，背部肩胛區(qū)sc給藥，每周給藥1次，共7次，停藥恢復(fù)28 d。應(yīng)用流式細(xì)胞儀進(jìn)行外周血中淋巴細(xì)胞分群檢測(cè)，檢測(cè)經(jīng)TC007刺激后分泌干擾素-γ（IFN-γ）的特異T細(xì)胞水平；免疫組化檢測(cè)肝臟IFN-γ、腫瘤壞死因子-α（TNF-α）、白細(xì)胞介素-2（IL-2）表達(dá)；試劑盒法測(cè)定血清中抗Ad5抗體的中和活性，補(bǔ)體成分C3、C4及免疫復(fù)合物（CIC）水平；給藥期及恢復(fù)期結(jié)束，進(jìn)行多個(gè)組織取材，HE染色后進(jìn)行病理學(xué)閱片，TaqMan探針qPCR法進(jìn)行Ad5分布檢測(cè)。結(jié)果 與溶媒對(duì)照組比較，各組動(dòng)物血漿中CD3⁺CD8⁺T細(xì)胞、CD4⁺/CD8⁺、CD3^-CD56⁺NK細(xì)胞、CD3⁺CD56⁺NKT細(xì)胞、巨噬細(xì)胞CD45⁺CD14⁺百分率均未見明顯異常；Ad5-Null組特異T細(xì)胞均未見明顯改變，TC007各劑量組針對(duì)HBV 3種抗原（Core、Env、Pol）分泌IFN-γ的特異性T細(xì)胞水平顯著增加（P<0.05、0.01），停藥恢復(fù)末期（d68），效應(yīng)值仍略高；TC007各劑量組和Ad5-Null組食蟹猴肝臟組織中IFN-γ、TNF-α、IL-2均高于溶媒對(duì)照組，且呈劑量相關(guān)性，但TC007各劑量組與Ad5-Null組比較未見明顯差異；TC007各劑量、Ad5-Null組動(dòng)物均產(chǎn)生抗Ad5抗體，39/40例具有中和活性；TC007各劑量組補(bǔ)體C3、C4未見明顯異常，高劑量組CIC輕微升高（P<0.05）。Ad5主要分布于給藥局部，在腋下淋巴結(jié)有少量分布，其他組織未見分布。TC007及Ad5-Null組均可見給藥局部輕微至輕度皮下組織炎性細(xì)胞浸潤(rùn)，停藥恢復(fù)28 d，病變可見明顯恢復(fù)，其他組織未見病理學(xué)改變。結(jié)論 TC007在食蟹猴體內(nèi)具有一定免疫原性，抗Ad5抗體具有中和活性，高劑量組可見輕微免疫復(fù)合物形成，未見免疫器官損傷，生物分布主要集中于給藥部位，可產(chǎn)生特異性T細(xì)胞。

Objective In this study, 6-week repeated sc injection to cynomolgus monkey with TC007 to assess the immunogenicity, immunotoxicity, and vivo biodistribution.Methods Ordinary grade cynomolgus monkeys were randomly divided into solvent control group, TC007 low, medium, and high doses (1.0×10⁹, 1.0×10¹⁰, 1.0×10¹¹ VP/rat) group, Ad5-Null (1.0×10¹¹ VP/rat, no-load control) group, 10 monkeys in each group, in the back scapular region was sc given once a week for 7 weeks, and the withdrawal was resumed for 28 d. Flow cytometry was used to detect the lymphocyte population in peripheral blood and the level of IFN-γ- secreting T cells stimulated by TC007; immunohistochemistry was used to detect the expression of IFN-γ/TNF-α/IL-2 in liver; kit method was used to determine the neutralization activity of anti-Ad5 antibody in serum and the levels of complement components C3, C4 and immune complex (CIC). At the end of convalescence, multiple tissue samples were taken, histopathological examination was performed after HE staining, and the distribution of Ad5 was detected by TaqMan probe qPCR.Results Compared with solvent control group, the percentage of CD3⁺ CD8⁺ T cells, CD4⁺/CD8⁺, CD3^-CD56⁺ NK cells, CD3⁺ CD56⁺ NKT cells and macrophage CD45⁺CD14⁺ in plasma of all groups were not significantly abnormal; the specific T cells of Ad5-Null group were not significantly changed, and the level of specific T cells targeted the three antigens of HBV (Core, Env, and Pol) secreting IFN-γ increased significantly in TC007 group, and the effect remained slightly higher after 4 weeks of withdrawal and recovery. The levels of IFN-γ/TNF-α/IL-2 in liver tissues of cynomolgus monkeys in TC007 and Ad5-Null groups were higher than those in the solvent control group in a dose-dependent manner; But there was no significant difference between TC007 and Ad5-Null groups. Anti-Ad5 antibodies were produced in all the animals in Ad 5-Null group, and 39 of 40 cases had neutralization activity. Complement C3 and C4 were not abnormal in TC007 different dose groups, but CIC was slightly elevated in high dose group (P<0.05). Ad5 was mainly distributed in the local administration, but there was a little distribution in the axillary lymph nodes with no distribution in other tissues. In TC007 and Ad5-Null groups, mild subcutaneous inflammatory cell infiltration was stopped for 28 days, and the pathological changes were obviously recovered. No pathological changes were found in other tissues. Conclusion TC007 has a certain immunogenicity in cynomolgus monkeys and anti-Ad5 antibody has neutralizing activity. In high-dose group, slight immunocomplex formation can be found. There is no damage to immune organs. The biodistribution is mainly concentrated at the site of administration. Specific T cells can be found in high dose group.