Relevance of the diversity among members of the Trypanosoma cruzi trans-sialidase family analyzed with camelids single-domain antibodies.
Laura Ratier,Mariela Urrutia,Gastón Paris,Laura Zarebski,Alberto C.C. Frasch,Fernando Alberto Goldbaum +5 more
TLDR
The results suggest that the presence of a large and diverse trans-sialidase family might be required to prevent the inhibitory response against this essential enzyme and might thus constitute a novel strategy of T. cruzi to evade the host immune system.Abstract:
The sialic acid present in the protective surface mucin coat of Trypanosoma cruzi is added by a membrane anchored trans-sialidase (TcTS), a modified sialidase that is expressed from a large gene family. In this work, we analyzed single domain camelid antibodies produced against trans-sialidase. Llamas were immunized with a recombinant trans-sialidase and inhibitory single-domain antibody fragments were obtained by phage display selection, taking advantage of a screening strategy using an inhibition test instead of the classic binding assay. Four single domain antibodies displaying strong trans-sialidase inhibition activity against the recombinant enzyme were identified. They share the same complementarity-determining region 3 length (17 residues) and have very similar sequences. This result indicates that they likely derived from a unique clone. Probably there is only one structural solution for tight binding inhibitory antibodies against the TcTS used for immunization. To our surprise, this single domain antibody that inhibits the recombinant TcTS, failed to inhibit the enzymatic activity present in parasite extracts. Analysis of individual recombinant trans-sialidases showed that enzymes expressed from different genes were inhibited to different extents (from 8 to 98%) by the llama antibodies. Amino acid changes at key positions are likely to be responsible for the differences in inhibition found among the recombinant enzymes. These results suggest that the presence of a large and diverse trans-sialidase family might be required to prevent the inhibitory response against this essential enzyme and might thus constitute a novel strategy of T. cruzi to evade the host immune system.read more
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Single domain antibodies: promising experimental and therapeutic tools in infection and immunity.
Janusz Wesolowski,Vanina Alzogaray,Jan Reyelt,Mandy Unger,Karla Juarez,Karla Juarez,Mariela Urrutia,Ana Cauerhff,Welbeck Danquah,Björn Rissiek,Felix Scheuplein,Nicole Schwarz,Sahil Adriouch,Olivier Boyer,Michel Seman,Alexei Licea,David V. Serreze,Fernando Alberto Goldbaum,Friedrich Haag,Friedrich Koch-Nolte +19 more
TL;DR: The results of several recent proof-of-principle studies are reviewed that open the exciting perspective of using sdAbs for modulating immune functions and for targeting toxins and microbes.
Journal ArticleDOI
Trypanosoma Cruzi ノ培養及其ノ培養基
TL;DR: Metacyclic trypomastigotes (hemoflagellates) are intermittently found in the peripheral blood and are the stage that transmits the infection to vectors or blood recipients.
Journal ArticleDOI
Grading the commercial optical biosensor literature-Class of 2008: 'The Mighty Binders'.
Rebecca L. Rich,David G. Myszka +1 more
TL;DR: This work grades every paper published in 2008 on a scale from A to F and outlines what features make a biosensor article fabulous, middling or abysmal and focuses on a few experimental, analysis and presentation mistakes that are alarmingly common.
Book ChapterDOI
Sialidases in Vertebrates : A Family Of Enzymes Tailored For Several Cell Functions*
Eugenio Monti,Erik J. Bonten,Alessandra d'Azzo,Roberto Bresciani,Bruno Venerando,Giuseppe Borsani,Roland Schauer,Guido Tettamanti +7 more
TL;DR: A review of the recent research development on vertebrate sialidase biology can be found in this article, where the authors summarize the results obtained since 2002, when the last general review on the molecular biology of mammalian Sialidases was written, highlighting the increasing relevance of these enzymes in glycobiology.
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