摘要
目的观察HBsAg P24亚基结合紧密程度对其抗原性的影响。方法3批重组酵母表达纯化的P24亚基松弛结合型HBsAg经半转型和转型处理后，成为混合和紧密结合型HBsAg，用HPLC法检测P24亚基结合紧密程度，松弛型和紧密型HBsAg免疫小鼠后，检测NK细胞杀伤活性及抗原特异性脾淋巴细胞增殖活性。将HBsAg吸附于佐剂Al（OH）3凝胶，免疫小鼠后用半数有效剂量法（ED50）检测所诱导的体液免疫应答，用体外相对效力法（RP）检测抗原的反应原性。结果3批松弛型、混合型和紧密型共9组HBsAg P24亚基紧密结合型比率分别在23．00％～32．00％、38．43％～50．98％和39．65％～52．76％之间，混合型组P24亚基紧密结合程度接近紧密型组；3批紧密型HBsAg免疫小鼠后可比松弛型HBsAg诱导更强的NK细胞活性，松弛型HBsAg免后4周内诱导抗原特异性脾淋巴细胞活性高于紧密型HBsAg，但总体上HBsAg仅诱导较弱的特异性脾淋巴细胞应答；紧密型HBsAg组比松弛型组诱导较强的体液免疫应答，1批混合型抗原体液免疫应答低于同批松弛型和紧密型组，2批松弛型组和其同批紧密型组的体液免疫应答程度接近；混合型HBsAg RP平均值显着高于松弛型组（P〈0．01），紧密型组RP平均值小于混合型组。结论重组酵母表达P24亚基结合程度不同的HBsAg均保持一定程度的抗原性。紧密型HBsAg免疫可诱导较强的非特异性NK细胞和体液免疫应答，混合型HBsAg的免疫反应原性较高。
Objective To identify the impact of the HBsAg P24 unit conjugating extent on the antigenicity of the recombinant yeast expressed HBsAg. Methods Three lots of purified HBsAg derived from recombinant yeast were treated with or without thiocyanate to prepare non-transformed, semi-transformed and transformed HBsAg, the P24 unit is theoretically loose-conjugated, semi-fight-conjugated and fight-conjugated. The tight-conjugation of polypepfide P24 unit for HBsAg was detected by HPLC. The BALB/e mice NK activities and HBsAg specific spleen lymphecytes proliferation immunized with the loose-conjugated and tight-conjugated HBsAg were tested. Results The average fight-conjugated P24 percentages of the three lots loose-eonjugated, semi-tight-conjugated and tight-conjugated HBsAg are 23.00%-32.00%, 38.43%-50.98% and 39.65%-52.76%, respectively. The extent of tight-conjugated P24 semi-transformed HBsAg is similar to transformed ones. The NK activities induced by tight-conjugated HBsAg were higher than that induced by loose-conjugated antigen, the HBsAg specific spleen lymphocytes proliferation induced by loose-conjugated antigen was higher than that of tight-conjugated antigen in 4 weeks. All loose-conjugated, semi-tight-conjugated and tight-conjugated HBsAg effectively induced humoral immunoresponse. Although all vaccines showed effective immunogenicity, the semi-tight-conjugate vaccines are signifi- cantly more effective than that of loose-conjugated （ P 〈 0.05）. Both the median effective dose （EDs0） and in vitro relative potencies （RP） of the loose-conjugated and the semi-tight-conjugated trial hepatitis B vaccines meet the requirement of the recombinant yeast derived hepatitis B vaccine. Conclusion Various P24 unit conjugated degree HBsAg maintains antigenicity. The tight-conjugated HBsAg could induce stronger NK activities than loosed HBsAg. The semi-transform treatment may improve the immunegenicity of HBsAg.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第2期132-136,共5页
Chinese Journal of Microbiology and Immunology
