经济动物学报

以人参茎叶总皂苷（GSLS）以及人参皂苷单体Rb1作为免疫增强剂，制备IBR灭活疫苗，探究该疫苗在小鼠体内的免疫原性。首先增殖IBRV NM8株病毒并测定TCID50，以BEI为灭活剂进行病毒灭活，从而得到用于疫苗制备的抗原。随后，按照预先设定的比例，把GSLS与Rb1加入抗原之中，乳化后分别制备含有高剂量（0.5 mg/mL）、中剂量（0.25 mg/mL）、低剂量（0.05 mg/mL）GSLS和人参皂苷Rb1的IBR灭活疫苗。将制备的各组疫苗分别以皮下注射的方式接种小鼠，每只小鼠接种0.2 mL，同时以不含人参皂苷的ISA206佐剂疫苗和只含有灭活IBRV的病毒液作为对照疫苗，首免后14 d以相同途径和剂量进行二次免疫。分别于首免后14 d和28 d取小鼠的脾脏和血液，进行淋巴细胞增殖试验和细胞因子、抗体水平检测。增殖后IBRV NM8株的TCID50为108.0TCID50/mL，当NM8株感染60 h时，病毒含量达峰值；经研究确定，BEI的最佳灭活浓度和时间分别为3 mmol/L ，20～24 h制备的灭活疫苗物理性状和安全性检验等均合格。GSLS和人参单体皂苷Rb1作为免疫增强剂可以提高IBRV灭活疫苗免疫小鼠产生特异性抗体IgG1、IgG2的水平，提高IL-4、IL-1β和IFN-γ等细胞因子表达水平；可促进小鼠脾脏淋巴细胞的增殖。其中单体皂苷Rb1低剂量组效果最为明显。人参茎叶总皂苷以及人参皂苷单体Rb1，均对IBR灭活疫苗呈现出显著的免疫增强作用，为IBR疫苗的后续研发提供了极具价值的参考，也为新型疫苗免疫增强剂的探索开拓了新方向。

An inactivated IBR vaccine was prepared using total ginseng stem and leaf saponins as well as ginsenoside monomer Rb1 as immune enhancers to investigate the immunogenicity of the vaccine in mice. First， IBRV NM8 strain virus was multiplied and TCID50 was measured， and the virus was inactivated using BEI as an inactivating agent to obtain the antigen for vaccine preparation. Subsequently， GSLS and Rb1 were added to the antigen according to a predetermined ratio， and IBR inactivated vaccines containing high-dose (0.5 mg/mL)， medium-dose (0.25 mg/mL)， and low-dose (0.05 mg/mL) GSLS and ginsenoside Rb1 were emulsified， and the physical properties and safety of the vaccines were examined. The prepared groups of vaccines were inoculated into mice by subcutaneous injection of 0.2 mL each， while the ISA206 adjuvanted vaccine without ginsenoside and the viral solution containing only inactivated IBRV were used as control vaccines， and the second immunization was performed 14 d after the first immunization by the same route and dose. The spleens and blood of mice were taken at 14 and 28 d after the first immunization， respectively， and tested for lymphocyte proliferation assay and cytokine and antibody levels. The TCID50 of the proliferated IBRV NM8 strain was 108.0 TCID50/mL， and the virus content peaked when the NM8 strain was infected for 60 h. It was determined that the optimal inactivation concentration and time of BEI were 3 mmol/L and 20-24 h， respectively.The prepared inactivated vaccine passed the physical properties and safety tests， etc. GSLS and ginsenoside Rb1 as immune-enhancing agents GSLS and ginseng monosaponin Rb1 were used as immune enhancers， which could increase the level of specific antibodies IgG1 and IgG2 produced by mice immunized with IBRV inactivated vaccine， and the expression level of cytokines such as IL-4， IL-1β， and IFN-γ; and they could promote the proliferation of splenic lymphocytes in mice. Among them， the effect of monosaponin Rb1 low dose group was most obvious. Ginseng stem and leaf total saponins， as well as ginsenoside monomer Rb1， both showed significant immune-enhancing effects on IBR inactivated vaccines， providing a highly valuable reference for the subsequent development of IBR vaccines and opening up a new direction for the exploration of novel vaccine immune enhancers.