圆PLoS|oNE A Pre-Clinical Safety Evaluation of SBP(HBsAg-Binding Protein) a x 200(HE staining).(a)Ns group, 3 days after the last injection, normal tissue. (b)V+SBP(high dose)group, 3 days after t necrosis of myofiber(focal). (c)V+SBP (high dose)group, 3 days after the last injection, mesenchyme inflammatory cell int macrophage aggregation(multi-focus).(d)V+SBP(high dose)group, at the end of the recovery period, mesenchyme inflammatory cell infiltration and macrophage aggregation(multi-focus) doi: 10. 1371/journal pone. 0170313.g003 decreased in male rats from the SBP(1.5 mL)and SBP-adjuvanted vaccine(0.5 mL) groups ( data not shown). Although these changes were statistically significant, there was no clear cor relation with injection dosage or time. It is likely that these data do not have toxicological SBP and high-dosage SBP-adjuvanted vaccine could cause allergic reaction in guinea pigs The allergenicity of SBP and SBP-adjuvanted vaccine was evaluated in guinea pigs. No abnor malities were observed during the sensitization period. After the antigens were administered, different levels of active systemic allergic reaction were observed in both SBP groups. Further- more, the higher dosage of SBP-adjuvanted vaccine also caused active systemic allergic reac- tions ( table 2 SBP could induce the production of SBP-specific antibodies in rats As a foreign protein for rats, SBP could induce antibody production after injection. In both SBP groups, SBP-specific antibodies were detected on day 21, after 2 injections, and declined 7 PLOS ONE DO1: 10. 1371/journal pone. 0170313 Januarydecreased in male rats from the SBP (1.5 mL) and SBP-adjuvanted vaccine (0.5 mL) groups (data not shown). Although these changes were statistically significant, there was no clear cor￾relation with injection dosage or time. It is likely that these data do not have toxicological meaning. SBP and high-dosage SBP-adjuvanted vaccine could cause allergic reaction in guinea pigs The allergenicity of SBP and SBP-adjuvanted vaccine was evaluated in guinea pigs. No abnor￾malities were observed during the sensitization period. After the antigens were administered, different levels of active systemic allergic reaction were observed in both SBP groups. Further￾more, the higher dosage of SBP-adjuvanted vaccine also caused active systemic allergic reac￾tions (Table 2). SBP could induce the production of SBP-specific antibodies in rats As a foreign protein for rats, SBP could induce antibody production after injection. In both SBP groups, SBP-specific antibodies were detected on day 21, after 2 injections, and declined 7 Fig 3. SBP could induce local irritation reaction. Representative photomicrographs of treated site are shown with original magnification: × 200 (HE staining). (a) NS group, 3 days after the last injection, normal tissue. (b) V+SBP (high dose) group, 3 days after the last injection, necrosis of myofiber (focal). (c) V+SBP (high dose) group, 3 days after the last injection, mesenchyme inflammatory cell infiltration and macrophage aggregation (multi-focus). (d) V+SBP (high dose) group, at the end of the recovery period, mesenchyme inflammatory cell infiltration and macrophage aggregation (multi-focus). doi:10.1371/journal.pone.0170313.g003 A Pre-Clinical Safety Evaluation of SBP (HBsAg-Binding Protein) PLOS ONE | DOI:10.1371/journal.pone.0170313 January 19, 2017 6 / 11