Thomas Chandan, Rawat Amit, Bai Shuhua, Ahsan Fakhrul
Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center, 1300 Coulter, Amarillo, TX 79106, USA.
J Pharm Sci. 2008 Mar;97(3):1213-23. doi: 10.1002/jps.21069.
This study was designed to test the hypothesis that formulation of hepatitis B vaccine with tetradecyl-beta-maltoside (TDM) enhances the immune response after pulmonary administration in a rodent model. Commercially available recombinant hepatitis B vaccine (rHBV) was formulated with varying concentrations of TDM and administered intratracheally to anesthetized male Sprague-Dawley rats. rHBV administered intramuscularly at doses of 2 and 4 microg served as positive controls. All formulations were administered on days 0 and 14 and the immune response was evaluated for 28 days. Specific antibodies generated to HBsAg were analyzed by ELISA. Safety studies were carried out by measuring the levels of alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and tumor necrosis factor alpha (TNF-alpha) in bronchoalveolar lavage (BAL) fluid. There was a significant increase in the immune response when the vaccine was administered intramuscularly at a dose of 4 microg. Only a modest increase in the immune response was observed when plain rHBV was administered intratracheally at the same dose. However, a pulmonary formulation of 4 microg rHBV plus 0.5% TDM produced a fourfold increase in the immune response compared to plain rHBV administered via the pulmonary route. No increase in immune response was observed for formulations containing rHBV plus 0.125% or 0.25% TDM. The levels of ALP and LDH in the BAL fluid suggest that the hepatitis B vaccine plus TDM formulations cause some injury to the lungs after the first intratracheal instillation of the formulation; however, the enzyme levels tended to be lower after the second instillation. The level of TNF-alpha in the BAL fluid of TDM-treated rats was substantially lower than that in rats treated with the positive control substance, sodium dodecyl sulfate. Overall, rHBV formulated with TDM increases the immune response after pulmonary administration, and pulmonary formulation of rHBV plus TDM could be used as an alternative to needle-based delivery of hepatitis B vaccine.
在啮齿动物模型中，用十四烷基-β-麦芽糖苷（TDM）配制乙肝疫苗可增强经肺部给药后的免疫反应。将市售重组乙肝疫苗（rHBV）与不同浓度的TDM混合配制，并经气管内给予麻醉的雄性Sprague-Dawley大鼠。以2微克和4微克剂量肌肉注射rHBV作为阳性对照。所有制剂均在第0天和第14天给药，并在28天内评估免疫反应。通过酶联免疫吸附测定（ELISA）分析针对乙肝表面抗原（HBsAg）产生的特异性抗体。通过测量支气管肺泡灌洗（BAL）液中碱性磷酸酶（ALP）、乳酸脱氢酶（LDH）和肿瘤坏死因子α（TNF-α）的水平进行安全性研究。当以4微克剂量肌肉注射疫苗时，免疫反应显著增强。当以相同剂量经气管内给予纯rHBV时，仅观察到免疫反应有适度增强。然而，与经肺部途径给予的纯rHBV相比，4微克rHBV加0.5%TDM的肺部制剂使免疫反应提高了四倍。对于含有rHBV加0.125%或0.25%TDM的制剂，未观察到免疫反应增强。BAL液中ALP和LDH的水平表明，乙肝疫苗加TDM制剂在首次经气管内滴注后会对肺部造成一定损伤；然而，第二次滴注后酶水平趋于降低。TDM处理大鼠的BAL液中TNF-α水平明显低于用阳性对照物质十二烷基硫酸钠处理的大鼠。总体而言，用TDM配制的rHBV可增强经肺部给药后的免疫反应，rHBV加TDM的肺部制剂可作为基于针头注射的乙肝疫苗的替代方法。
