Kateete Dp, Alezuyo C, Nanteza A, Asiimwe C, Lubega Gw
School of Biomedical Sciences, Makerere University College of Health Sciences, Kampala, Uganda ; College of Veterinary Medicine, Animal Resources & Biosecurity, Makerere University, Kampala, Uganda.
Iran J Parasitol. 2012;7(3):54-63.
There are only four drugs for treating African trypanosomiasis, a devastating disease in sub-Saharan Africa. With slow discovery of better drugs, vaccination is viewed as the best method of control. We previously showed that antibodies to native Trypanosoma brucei brucei tubulin inhibit the growth of trypanosomes in culture. Here, we aimed to determine the effect of antibodies to bacterially expressed trypanosome tubulin on T. brucei brucei growth.
T. brucei brucei alpha and beta tubulin genes were individually expressed in Escherichia coli under the tryptophan promoter. Monoclonal tubulin antibodies reacted specifically with the expressed tubulins with no cross-reaction with the opposite tubulin. Rabbits were immunized with 450µg each of the concentrated recombinant tubulin, and production of antibodies assessed by ELISA and Western blotting. The effect of polyclonal antibodies on trypanosome growth was determined by culturing bloodstream T. brucei brucei in up to 25% of antisera.
Low antisera dilutions (25%) from the immunized rabbits inhibited trypanosome growth. The most cytotoxic antisera were from one rabbit immunized with a mixture of both alpha and beta tubulins. However, the result was not reproduced in other rabbits and there was no apparent effect on growth at higher antisera dilutions.
Antibodies to bacterially expressed trypanosome tubulin are not effective at killing cultured bloodstream trypanosomes.
治疗非洲锥虫病仅有四种药物，而非洲锥虫病是撒哈拉以南非洲地区一种极具破坏性的疾病。由于新型药物研发进展缓慢，疫苗接种被视为最佳防控方法。我们之前的研究表明，针对布氏锥虫天然微管蛋白的抗体可抑制锥虫在培养物中的生长。在此，我们旨在确定针对细菌表达的锥虫微管蛋白的抗体对布氏锥虫生长的影响。
布氏锥虫的α和β微管蛋白基因分别在大肠杆菌中色氨酸启动子的控制下表达。单克隆微管蛋白抗体与表达的微管蛋白特异性反应，与另一种微管蛋白无交叉反应。用450μg每种浓缩的重组微管蛋白免疫兔子，通过ELISA和蛋白质印迹法评估抗体产生情况。通过在高达25%的抗血清中培养布氏锥虫血流形式来确定多克隆抗体对锥虫生长的影响。
免疫兔子的低抗血清稀释度（25%）可抑制锥虫生长。细胞毒性最强的抗血清来自一只用α和β微管蛋白混合物免疫的兔子。然而，其他兔子未重现该结果，且在较高抗血清稀释度下对生长无明显影响。
针对细菌表达的锥虫微管蛋白的抗体对培养的布氏锥虫血流形式无杀伤作用。
