In Vitro Trypanocidal Activity of Antibodies to Bacterially Expressed Trypanosoma brucei Tubulin
Background: 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...
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Tehran University of Medical Sciences
2012-09-01
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doaj-5902391c57ec4b6194797d86cf347f8a2021-03-02T10:15:39ZengTehran University of Medical SciencesIranian Journal of Parasitology1735-70202008-238X2012-09-01735463In Vitro Trypanocidal Activity of Antibodies to Bacterially Expressed Trypanosoma brucei TubulinGW LubegaC AsiimweA NantezaC AlezuyoDP KateeteBackground: 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. Methods: 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. Results: 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. Conclusion: Antibodies to bacterially expressed trypanosome tubulin are not effective at killing cul­tured bloodstream trypanosomes.http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/22125.pdf&manuscript_id=22125Alpha tubulinBeta tubulinTrypanosoma bruceiProtein expressionGene cloning |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
GW Lubega C Asiimwe A Nanteza C Alezuyo DP Kateete |
spellingShingle |
GW Lubega C Asiimwe A Nanteza C Alezuyo DP Kateete In Vitro Trypanocidal Activity of Antibodies to Bacterially Expressed Trypanosoma brucei Tubulin Iranian Journal of Parasitology Alpha tubulin Beta tubulin Trypanosoma brucei Protein expression Gene cloning |
author_facet |
GW Lubega C Asiimwe A Nanteza C Alezuyo DP Kateete |
author_sort |
GW Lubega |
title |
In Vitro Trypanocidal Activity of Antibodies to Bacterially Expressed Trypanosoma brucei Tubulin |
title_short |
In Vitro Trypanocidal Activity of Antibodies to Bacterially Expressed Trypanosoma brucei Tubulin |
title_full |
In Vitro Trypanocidal Activity of Antibodies to Bacterially Expressed Trypanosoma brucei Tubulin |
title_fullStr |
In Vitro Trypanocidal Activity of Antibodies to Bacterially Expressed Trypanosoma brucei Tubulin |
title_full_unstemmed |
In Vitro Trypanocidal Activity of Antibodies to Bacterially Expressed Trypanosoma brucei Tubulin |
title_sort |
in vitro trypanocidal activity of antibodies to bacterially expressed trypanosoma brucei tubulin |
publisher |
Tehran University of Medical Sciences |
series |
Iranian Journal of Parasitology |
issn |
1735-7020 2008-238X |
publishDate |
2012-09-01 |
description |
Background: 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. Methods: 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. Results: 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. Conclusion: Antibodies to bacterially expressed trypanosome tubulin are not effective at killing cul­tured bloodstream trypanosomes. |
topic |
Alpha tubulin Beta tubulin Trypanosoma brucei Protein expression Gene cloning |
url |
http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/22125.pdf&manuscript_id=22125 |
work_keys_str_mv |
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