CoChat
Sign Up
Cited by 7Trypanosoma species research and implicationsMonoclonal and Polyclonal Antibodies ResearchResearch on Leishmaniasis Studies

Engineering a single-chain antibody against Trypanosoma cruzi metacyclic trypomastigotes to block cell invasion

Open access
Demeu, Lara Maria Kalempa, Soares, Rodrigo Jahn, Miranda, Juliana Severo, Pacheco-Lugo, Lisandro A., Oliveira, Kelin Gonçalves, Cortez Plaza, Cristian Andrés, Billiald, Philippe, Ferreira de Moura, Juliana +3 more·2019·PLOS ONE
DOI 10.1371/journal.pone.0223773PMC PMC6795462
Open this paper in CoChat — free accountOpen PDF

Abstract

Trypanosoma cruzi is a flagellate protozoan pathogen that causes Chagas disease. Currently there is no preventive treatment and the efficiency of the two drugs available is limited to the acute phase. Therefore, there is an unmet need for innovative tools to block transmission in endemic areas. In this study, we engineered a novel recombinant molecule able to adhere to the T. cruzi surface, termed scFv-10D8, that consists of a single-chain variable fragment (scFv) derived from mAb-10D8 that targets gp35/50. The synthetic gene encoding scFv-10D8 was cloned and fused to a 6×His tag and expressed in a prokaryotic expression system. Total periplasmic or 6xHis tag affinity-purified fractions of scFv-10D8 retained the capacity to bind to gp35/50, as shown by Western blot analyses. Pre-incubation of metacyclic trypomastigotes with scFv-10D8 showed a remarkable reduction in cell invasion capacity. Our results suggest that scFv-10D8 can be used in a paratransgenic approach to target parasites in insect vectors, avoiding dissemination of infective forms. Such advances in the development of this functional molecule will surely prompt the improvement of alternative strategies to control Chagas disease by targeting mammalian host stages.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1371/journal.pone.0223773

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • The 22 amino acids long PelB signal sequence from Erwinia carotovora is the most frequently used for transportation of Fabs and scFvs.

    “The 22 amino acids long PelB (pectate lyase B) signal sequence from Erwinia carotovora [157] is the most frequently used for transportation of the Fabs [88,96] and scFvs [30,101,158,159].”

    From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [158]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

No references could be resolved yet.

Research works printed but not linked: 48

  1. [1]Tibayrenc M, Barnabé C, Telleria J. Reticulate Evolution in: Medical and Epidemiological Implications In: Telleria J, Tibayrenc M, editors. American trypanosomiasis: Chagas disease One hundred years of research Burlington: Elsevier; 2010 475– 488.
  2. [3]Silveira AC, Dias JCP. O controle da transmissão vetorial. Rev Soc Bras Med Trop 44 Supplement. 2011; 2: 52– 63. 10.1590/S0037-86822011000800009 21584358 · DOI
  3. [4]Coura JR. The main sceneries of Chagas disease transmission. The vectors, blood and oral transmissions: a comprehensive review. Mem Inst Oswaldo Cruz. 2015; 110: 277– 282. 10.1590/0074-0276140362 25466622 PMC4489464 · DOI
  4. [5]Browne AJ, Guerra CA, Alves RV, da Costa VM, Wilson AL, Pigott DM, et al The contemporary distribution of Trypanosoma cruzi infection in humans, alternative hosts and vectors. Sci Data. 2017; 4: 170050 10.1038/sdata.2017.50 28398292 PMC5387921 · DOI
  5. [6]World Health Organization (2015) Chagas disease in Latin America: an epidemiological update based on 2010 estimates. Wkly Epidemiol Rec Feb 90: 33– 43 25671846
  6. [7]Gurgel-Gonçalves R, Galvão C, Costa J, Peterson AT. Geographic distribution of Chagas disease vectors in brazil based on ecological niche modeling. J Trop Med. 2012;:Article ID 705326. 10.1155/2012/705326 22523500 PMC3317230 · DOI
  7. [8]Vinhaes MC, de Oliveira SV, Reis PO, de Lacerda Sousa AC, Silva RA, Obara MT, et al Assessing the vulnerability of Brazilian municipalities to the vectorial transmission of Trypanosoma cruzi using multi-criteria decision analysis. Acta Trop. 2014; 137: 105– 110. 10.1016/j.actatropica.2014.05.007 24857942 · DOI
  8. [10](2016) Brazilian consensus on Chagas disease. Epidemiol Serv Saúde, Brasília 25(núm. esp.): 7– 86 27869914
  9. [11]Mougabure-Cueto G, Picollo MI. Insecticide resistance in vector Chagas disease: evolution, mechanisms and management. Acta Trop 2015; 149: 70– 85. 10.1016/j.actatropica.2015.05.014 26003952 · DOI
  10. [12]Urbina JA, Docampo R. Specific chemotherapy of Chagas disease: controversies and advances. Trends Parasitol. 2003; 19: 495– 501. 10.1016/j.pt.2003.09.001 14580960 · DOI
  11. [13]Filardi LS, Brener Z. Susceptibility and natural resistance of Trypanosoma cruzi strains to drugs used clinically in Chagas disease. Trans R Soc Trop Med Hyg. 1987; 81: 755– 759. 10.1016/0035-9203(87)90020-4 3130683 · DOI
  12. [14]Teston AP, Monteiro WM, Reis D, Bossolani GD, Gomes ML, De Araújo SM, et al In vivo susceptibility to Benznidazole of Trypanosoma cruzi strains from the western Brazilian Amazon. Trop Med Int Health. 2013; 18: 85– 95. 10.1111/tmi.12014 23130989 · DOI
  13. [15]Baral TN, Magez S, Stijlemans B, Conrath K, Vanhollebeke B, Pays E, et al Experimental therapy of African trypanosomiasis with a nanobody-conjugated human trypanolytic factor. Nat Med. 2016; 12: 580– 584. 10.1038/nm1395 16604085 · DOI
  14. [16]Arias JL, Unciti-Broceta JD, Maceira J, Del Castillo T, Hernández-Quero J, Magez S, et al Nanobody conjugated PLGA nanoparticles for active targeting of African trypanosomiasis. J Control Release. 2014; 197 10: 190– 198. 10.1016/j.jconrel.2014.11.002 25445702 · DOI · Paper page
  15. [17]Unciti-Broceta JD, Arias JL, Maceira J, Soriano M, Ortiz-González M, Hernández-Quero J, et al Specific cell targeting therapy bypasses drug resistance mechanisms in African trypanosomiasis. PLoS Pathog. 2015; 25: e1004942(6). 10.1371/journal.ppat.1004942 26110623 PMC4482409 · DOI
  16. [18]Stijlemans B, Caljon G, Natesan SK, Saerens D, Conrath K, Pérez-Morga D, et al High affinity nanobodies against the Trypanosome brucei VSG are potent trypanolytic agents that block endocytosis. PLoS Pathog. 2011; 7: e1002072 10.1371/journal.ppat.1002072 21698216 PMC3116811 · DOI
  17. [19]Berasategui A, Shukla S, Salem H, Kaltenpoth M. Potential applications of insect symbionts in biotechnology. Appl Microbiol Biotechnol. 2016; 100: 1567– 1577. 10.1007/s00253-015-7186-9 26659224 PMC4737797 · DOI
  18. [20]Hurwitz I, Fieck A, Read A, Hillesland H, Klein N, Kang A, et al Paratransgenic control of vector borne diseases. Int J Biol Sci. 2011; 7: 1334– 1344. 10.7150/ijbs.7.1334 22110385 PMC3221369 · DOI
  19. [21]Durvasula RV, Sundaram RK, Kirsch P, Hurwitz I, Crawford CV, Dotson E, et al Genetic transformation of a Corynebacterial symbiont from the Chagas disease vector Triatoma infestans. Exp Parasitol. 2018; 119: 94– 98. 10.1016/j.exppara.2007.12.020 18331732 PMC2635415 · DOI
  20. [22]Matthews S, Rao VS, Durvasula RV. Modeling horizontal gene transfer (HGT) in the gut of the Chagas disease vector Rhodnius prolixus. Parasites Vectors. 2011; 4: 77 10.1186/1756-3305-4-77 21569540 PMC3117810 · DOI
  21. [23]De Vooght CG, De Ridder K, Van Den Abbeele J. Delivery of a functional anti-trypanosome Nanobody in different tsetse fly tissues via a bacterial symbiont, Sodalis glossinidius. Microb Cell Factories. 2014; 13: 156 10.1186/s12934-014-0156-6 25376234 PMC4230353 · DOI
  22. [24]Yoshida N, Mortara RA, Araguth MF, Gonzalez JC, Russo M. Metacyclic neutralizing effect of monoclonal antibody 10D8 directed to the 35- and 50-kilodalton surface glycoconjugates of Trypanosoma cruzi. Infect Immun. 1989; 57: 1663– 1667 2656520 PMC313336
  23. [25]Mortara RA, Da Silva S, Araguth MF, Blanco SA, Yoshida N. Polymorphism of the 35- and 50-kilodalton surface glycoconjugates of Trypanosoma cruzi metacyclic trypomastigotes. Infect Immun. 1992; 60: 4673– 4678 1328061 PMC258217
  24. [26]Urban I, Santurio LB, Chidichimo A, Yu H, Chen X, Mucci J, et l. Molecular diversity of the Trypanosoma cruzi TcSMUG family of mucin genes and proteins. Biochem J. 2011; 438: 303– 313. 10.1042/BJ20110683 21651499 · DOI
  25. [27]Yoshida N. Molecular basis of mammalian cell invasion by Trypanosoma cruzi. An Acad Bras Cienc. 2006; 78: 87– 111. doi: /S0001-37652006000100010 16532210
  26. [28]Jones C, Todeschini AR, Agrellos OA, Previato JO, Mendonça-Previato L. Heterogeneity in the biosynthesis of mucin O-glycans from Trypanosoma cruzi tulahuen strain with the expression of novel galactofuranosyl-containing oligosaccharides. Biochemistry. 2004/ 43(37): 11889– 97. 10.1021/bi048942u 15362875 · DOI
  27. [29]Milenic DE, Yokota T, Filpula DR, Finkelman MA, Dodd SW, Wood JF, et al Construction, binding properties, metabolism, and tumor targeting of a single-chain Fv derived from the pancarcinoma monoclonal antibody CC49. Cancer Res. 1991; 51: 6363– 71 1933899
  28. [30]Kuan CT, Srivastava N, Mclendon RE, Marasco WA, Zalutsky MR, Bigner DD. Recombinant single-chain variable fragment antibodies against extracellular epitopes of human multidrug resistance protein MRP3 for targeting malignant gliomas. Int J Cancer. 2010; 127: 598– 611. 10.1002/ijc.25062 19937796 PMC2878888 · DOI
  29. [31]Crivianu-Gaita V, Thompson M (2016) Aptamers, antibody scFv, and antibody Fab' fragments: an overview and comparison of three of the most versatile biosensor biorecognition elements. Biosens Bioelectron. 2016; 85: 32– 45. 10.1016/j.bios.2016.04.091 27155114 · DOI
  30. [32]Fields C, O’Connell D, Xiao S, Lee GU, Billiald P, Muzard J. Creation of recombinant antigen-binding molecules derived from hybridomas secreting specific antibodies. Nat Protoc. 2013; 8: 1125– 1148. 10.1038/nprot.2013.057 23680984 · DOI
  31. [33]Lyskov S, Chou FC, Conchúir SÓ, Der BS, Drew K, Kuroda D, et al Serverification of molecular modeling applications: the Rosetta online server that includes everyone (ROSIE). PLoS One. 2013 8: e63906 10.1371/journal.pone.0063906 23717507 PMC3661552 · DOI
  32. [34]Ghoorah AW, Devignes MD, Smaïl-Tabbone M, Ritchie DW. Protein docking using case-based reasoning. Proteins. 2013; 81: 2150– 2158. 10.1002/prot.24433 24123156 · DOI
  33. [35]Mcguffin LJ, Atkins JD, Salehe BR, Shuid AN, Roche DB. IntFOLD: an integrated server for modelling protein structures and functions from amino acid sequences. Nucleic Acids Res. 2015; 43: W169– W173. 10.1093/nar/gkv236 25820431 PMC4489238 · DOI
  34. [36]Karim-Silva S, Moura Jd, Noiray M, Minozzo JC, Aubrey N, Alvarenga LM, et al Generation of recombinant antibody fragments with toxin-neutralizing potential in loxoscelism. Immunol Lett. 2016; 176: 90– 6. 10.1016/j.imlet.2016.05.019 27288291 · DOI
  35. [37]Zingales B, Andrade SG, Briones MRS, Campbell DA, Chiari E, Fernandes O, et al A new consensus for Typanosoma cruzi intraespecific nomenclature: second revision meeting recommends TcI to TcVI. Mem Inst Oswaldo Cruz. 2009; 104: 1051– 1054. 10.1590/s0074-02762009000700021 20027478 · DOI
  36. [38]Contreras VT, Araujo-Jorge TC, Bonaldo MC, Thomaz N, Barbosa HS, Meirelles N, et al Biological aspect of the Dm28c clone of Trypanosoma cruzi after metacyclogenesis in chemically defined media. Mem Inst Oswaldo Cruz. 1988; 83: 123– 133. 10.1590/s0074-02761988000100016 3074237 · DOI
  37. [39]Teixeira MMG, Yoshida N. Stage-specific surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi identified by monoclonal antibodies. Mol Biochem Parasitol. 1986; 18: 271– 282. 10.1016/0166-6851(86)90085-x 3515178 · DOI
  38. [40]Serrano AA, Schenkman S, Yoshida N, Mehlert A, Richardson JM, Ferguson MA. The lipid structure of the glycosylphosphatidylinositol-anchored mucin-like sialic acid acceptors of Trypanosoma cruzi changes during parasite differentiation from epimastigotes to infective metacyclic trypomastigote forms. J Biol Chem. 1995; 270: 27244– 27253. 10.1074/jbc.270.45.27244 7592983 · DOI
  39. [41]Mcgwire BS, Olson CL, Tack BF, Engman DM. Killing of African trypanosomes by antimicrobial peptides. J Infect Dis. 2003; 188: 146– 152. 10.1086/375747 12825184 · DOI
  40. [42]Smulski C, Labovsky V, Levy G, Hontebeyrie M, Hoebeke J, Levin MJ. Structural basis of the cross-reaction between an antibody to the Trypanosoma cruzi ribosomal P2β protein and the human β1 adrenergic receptor. FASEB J. 2006; 20: 1396– 1406. 10.1096/fj.05-5699com 16816115 · DOI
  41. [43]Ayub MJ, Nyambega B, Simonetti L, Duffy T, Longhi SA, Gómez KA, et al Selective blockade of trypanosomatid protein synthesis by a recombinant antibody anti-Trypanosoma cruzi P2β protein. PLOS ONE. 2012: 7:e36233. 10.1371/journal.pone.0036233 22570698 PMC3343115 · DOI
  42. [44]Mendonça-Previato L, Penha L, Garcez TC, Jones C, Previato JO. Addition of alpha-O-GlcNAc to threonine residues define the post-translational modification of mucin-like molecules in Trypanosoma cruzi. Glycoconj J. 2013; 30: 659– 666. 10.1007/s10719-013-9469-7 23430107 PMC3769586 · DOI
  43. [45]Ossysek K, Uchański T, Kulesza M, Bzowska M, Klaus T, Woś K, et al A New expression vector facilitating production and functional analysis of scFv antibody fragments selected from Tomlinson I+J phagemid libraries. Immunol Lett. 2015; 167: 95– 102. 10.1016/j.imlet.2015.07.005 26219832 · DOI
  44. [46]Lim KP, Li H, Nathan S. Expression and purification of a recombinant scFv towards the exotoxin of the pathogen, Burkholderia pseudomallei. J Microbiol. 2004; 42: 126– 132 15357306
  45. [47]Yusakul G, Sakamoto S, Nuntawong P, Tanaka H, Morimoto S. Different expression systems resulted in varied binding properties of anti–paclitaxel single–chain variable fragment antibody clone 1C2. J Nat Med. 2018; 72: 310– 316. 10.1007/s11418-017-1136-z 29027080 · DOI
  46. [48]Yoshida N, Dorta ML, Ferreira AT, Oshiro ME, Mortara RA, Acosta-Serrano A, et l. Removal of sialic acid from mucin-like surface molecules of Trypanosoma cruzi metacyclic trypomastigotes enhances parasite-host cell interaction. Mol Biochem Parasitol. 1997; 84: 57– 67. 10.1016/s0166-6851(96)02783-1 9041521 · DOI
  47. [49]Lebozec K, Jandrot-Perrus M, Avenard G, Favre-Bulle O, Billiald P. Quality and cost assessment of a recombinant antibody fragment produced from mammalian, yeast and prokaryotic host cells: A case study prior to pharmaceutical development. N Biotechnol. 2018; 44: 31– 40. 10.1016/j.nbt.2018.04.006 29689305 · DOI
  48. [50]Cámara MLM, Balouz V, Centeno Cameán C, Cori CR, Kashiwagi GA, Gil SA, et al Trypanosoma cruzi surface mucins are involved in the attachment to the Triatoma infestans rectal ampoule. PLoS Negl Trop Dis. 2019; 13(5): e0007418 10.1371/journal.pntd.0007418 31107901 PMC6544316 · DOI

Cited by

Sorted by citations

Authors
Demeu, L. M. K., Soares, R. J., Miranda, J. S., Pacheco-Lugo, L. A., Oliveira, K. G., Cortez Plaza, C. A., Billiald, P., Ferreira de Moura, J., Yoshida, N., Alvarenga, L. M., & DaRocha, W. D.
Affiliations
  • Universidade Federal do Paraná (BR)
  • Universidad Simón Bolívar (CO)
  • Université Paris-Sud (FR)
  • Universidade Federal de São Paulo (BR)
Corresponding author
  • Demeu, Lara Maria Kalempa
  • Soares, Rodrigo Jahn
  • Miranda, Juliana Severo
  • Pacheco-Lugo, Lisandro A.
  • Oliveira, Kelin Gonçalves
  • Billiald, Philippe
  • Yoshida, Nobuko
  • Alvarenga, Larissa Magalhães
  • DaRocha, Wanderson Duarte
ORCID
Venue
PLOS ONE
Publisher
Public Library of Science (PLoS)
Published
2019-10-16
Volume
14
Issue
10
Pages
e0223773
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0223773
PMC
PMC6795462
OpenAlex
W2981142738
Cited by
7
References
0
Citations by year
2021: 1 · 2020: 5 · 2017: 1
FWCI
1.14
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • HAL (submittedVersion, other-oa)
  • LA Referencia (Red Federada de Repositorios Institucionales de Publicaciones Científicas) (submittedVersion, cc-by-nc-nd)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Figshare (submittedVersion, cc-by)
Funders
  • Fundação Araucária (48.018)
  • Conselho Nacional de Desenvolvimento Científico e Tecnológico (401355/2014-4, 303733/2016-0, 311376/2017-7)
Topics
  • Health Sciences › Medicine › Epidemiology › Trypanosoma species research and implications
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
  • Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Research on Leishmaniasis Studies
Keywords
Trypanosoma cruzi, Chagas disease, scFv, metacyclic trypomastigotes, cell invasion, paratransgenesis, insect vectors, recombinant antibodies, Chagas disease prevention
MeSH terms
Antibodies, Protozoan, Antigens, Protozoan, Cell Line, HeLa Cells, Humans, Recombinant Proteins, Trypanosoma cruzi, Chagas Disease, Protein Engineering, Single-Chain Antibodies

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review