戊二醛脱毒前后的百日咳杆菌外膜蛋白及其组分的毒性与免疫原性

TOXICITY AND ANTIGENICITY OF THE OUTER MEMBRANE PROTEIN AND ITS FRACTIONS OF BORDETELLA PERTUSSIS BEFORE AND AFTER GLUTARALDEHYDE DETOXIFICATION

应用TritonX-100抽提和Sephadex G-100层析获得百日咳杆菌外膜蛋白(OMP)及其5个组分(OMP_1～OMP_5).OMP_1～OMP_4均具有促白细胞增多、组胺敏感和胰岛活化蛋白的活性.OMP及其各组分对中国仓鼠卵巢细胞(CHO细胞)具有毒性作用,可使该细胞聚集生长、~3H-TdR掺入量下降,及核糖体大量脱落和膜旁颗粒减少等现象.OMP组分OMP_1～OMP_4经0.3%戊二醛处理后,上述生物学活性均明显下降,CHO细胞不出现聚集生长,~3H-TdR掺入量明显高于未脱毒的组分,与正常对照组比较无显著性差异(P>0.05).脱毒前后的OMP各组分对小鼠的主动保护作用未见明显变化(P>0.05).实验结果表明,戊二醛能有效地降低OMP_1～OMP_4的毒性,但脱毒后的OMP_1～OMP_4仍具备较强的免疫原性和保护力.因此,OMP及其组分经戊二醛处理后,有希望成为百日咳杆茵的无细胞疫苗.

The outer membrane protein (OMP) and its fractions (OMP_1～OMP_5) of B. pertussis were obtained through Triton X-100 treatment and Sephadex G-100 column chromatography. OMP_1～OMP_4 had LP, HS and IAP activities. The OMP and its fractions exhibited cytotoxity to CHO cells such as clustered growth, inhibition of ~3H-TdR incorporation, falling of the ribosomes and reduction of the juxtamembranous particles. The activities of LP, HS, IAP and CHO cell clustered growth decreased obviously after glutaraldehyde detoxification. The ~3H-TdR incorporation of CHO cells in the presence of the detoxified fractions was much higher than that of the non-detoxified ones, and not differed significantly from the black controls (P>0.05). There were ? significant differences among the actively protective activities in mice of the non-detoxified and the detoxified fractions (P>0.05). The results showed that the toxicities of the fractions (OMP_1～OMP_4) declined obviously after treatment with 0.3% glutaraldehyde while the detoxified fractions still possessed strong antigenicities and protective activities. Thus, the OMP and its fractions after glutaraldehyde detoxification might be adopted as acellular vaccine of B. pertussis.