临床分离耐药的革兰阴性菌脂多糖脂质体的制备及其免疫保护性作用的初步研究-病原生物学专业论文.docxVIP

天津医科大学硕士学位论文Preparation 天津医科大学硕士学位论文 Preparation of a LiposomaI LipopoIysaccharide of Gram-NegatiVe Batteria ISOIated fr伽CI inic and i ts I mmunoprotect i ve Effects M．S．Candidate：Shangwei Supervisor：Professor Zhong Qiping ABSTRACT Object To prepare fl 1iposomal 1ipopolysaccharide(LPS)of gram-negative bacteria isolated from clinic which were multi—drug resistant．And to observe the immunoprotective effects of the liposomal LPS on mice challenged by various LPS or bacteria．We are looking forward to develop a novel vaccine strategy that can be evaluated for surgical， tumor and other high—risk hOspitalized patients． Methods LPSs were extracted from several multi—drug resistant gram- negative bacteria strains which were isolated from clinic by enzyme extraction， and then incorporated into multi—lamellar liposomes consisting of phosphatidyi choline(PC)，phosphatidylserine(PS) and cholesterol in a 4：l：4 molar ratio by the method of reverse evaporating．The basic LPS／lipid ratio was l：10(wt／wt)．The endotoxic activities of both the LPS—containing liposomes and the free LPS were measured by Lfmulus amoebocyte lysate(LAL)assay and the entrapment efficiency was presumed．The particle size of the 1iposomes were measured by laser light scattering instrument．And the transmission electron microscopy(TEM)was used to examine the morphology of 1 iposomes．D-galactosamine(D-GaIN)一sensitized mice model were established to investigate the immuno—protective effects of the 1 iposomes in yi vo against various LPS or bacteria． Ⅲ 天津医科大学硕士学位论文Result 天津医科大学硕士学位论文 Result The multilamella liposomal LPS。consisting of PC，P6 and cholesterol in a 4：1：4 molar ratio was prepared according to the reverse evaporating method developed by Dijkstra et．a1．with some modifications，and was opalescent at room tempreture．The basic LPS／1ipid ratio was 1：10(wt／,t)．The endotoxic activities of the LPS in liposomes measured by the LAL assay reduced 100 to 1．000一fold compared with its original value which suggested