title: Altering the motility of Trypanosoma cruzi with rabbit polyclonal anti-peptide antibodies reduces infection to susceptible mammalian cells.
abstract: Trypanosoma cruzi's trypomastigotes are highly active and their incessant motility seems to be important for mammalian host cell infection. The kinetoplastid membrane protein-11 (KMP-11) is a protein expressed in all parasite stages, which induces a cellular and humoral immune response in the infected host, and is hypothesized to participate in the parasite's motility. An N-terminal peptide from KMP-11, termed K1 or TcTLE, induced polyclonal antibodies that inhibit parasitic invasion of Vero cells. The goal of this study was to evaluate the motility and infectivity of T. cruzi when exposed to polyclonal anti-TcTLE antibodies. Rabbits were immunized with TcTLE peptide along with FIS peptide as an immunomodulator. ELISA assay results showed that post-immunization sera contained high titers of polyclonal anti-TcTLE antibodies, which were also reactive against the native KMP-11 protein and live parasites as detected by immunofluorescence and flow cytometry assays. Trypomastigotes of T. cruzi were incubated with pre- or post-immunization sera, and infectivity to human astrocytes was assessed by Giemsa staining/light microscope and flow cytometry using carboxyfluorescein diacetate succinimidyl ester (CFSE) labeled parasites. T. cruzi infection in astrocytes decreased approximately by 30% upon incubation with post-immunization sera compared with pre-immunization sera. Furthermore, trypomastigotes were recorded by video microscopy and the parasite's flagellar speed was calculated by tracking the flagella. Trypomastigotes exposed to post-immunization sera had qualitative alterations in motility and significantly slower flagella (45.5 µm/s), compared with those exposed to pre-immunization sera (69.2 µm/s). In summary, polyclonal anti-TcTLE serum significantly reduced the parasite's flagellar speed and cell infectivity. These findings support that KMP-11 could be important for parasite motility, and that by targeting its N-terminal peptide infectivity can be reduced.
authors: Grupo de Ciencias Básicas, Facultad de Medicina, Universidad de Los Andes, Bogotá, DC, Colombia.
authors: Eli J
authors: EJ
authors: Finkelsztein
authors: Grupo de Ciencias Básicas, Facultad de Medicina, Universidad de Los Andes, Bogotá, DC, Colombia.
authors: Juan C
authors: JC
authors: Diaz-Soto
authors: Grupo de Ciencias Básicas, Facultad de Medicina, Universidad de Los Andes, Bogotá, DC, Colombia.
authors: Juan C
authors: JC
authors: Vargas-Zambrano
authors: Grupo de Biofísica, Departamento de Física, Universidad de los Andes, Bogotá, DC, Colombia.
authors: Elizabeth
authors: E
authors: Suesca
authors: Núcleo de Biotecnología Curauma, Pontificia Universidad Católica Valparaíso, Valparaíso, Chile.
authors: Fanny
authors: F
authors: Guzmán
authors: Instituto de Parasitología y Biomedicina López Neyra, Consejo Superior de Investigaciones Científicas (IPBLN-CSIC) P.T. de Ciencias de la Salud, Granada, Spain.
authors: Manuel C
authors: MC
authors: López
authors: Instituto de Parasitología y Biomedicina López Neyra, Consejo Superior de Investigaciones Científicas (IPBLN-CSIC) P.T. de Ciencias de la Salud, Granada, Spain.
authors: M Carmen
authors: MC
authors: Thomas
authors: Grupo de Biofísica, Departamento de Física, Universidad de los Andes, Bogotá, DC, Colombia.
authors: Manu
authors: M
authors: Forero-Shelton
authors: Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, DC, Colombia.
authors: Adriana
authors: A
authors: Cuellar
authors: Laboratorio de Parasitología Molecular, Departamento de microbiología, Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, DC, Colombia.
authors: Concepción J
authors: CJ
authors: Puerta
authors: Grupo de Ciencias Básicas, Facultad de Medicina, Universidad de Los Andes, Bogotá, DC, Colombia. Electronic address: johgonza@uniandes.edu.co.
authors: John M
authors: JM
authors: González
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology*; D000953:Antigens, Protozoan / Q000276:immunology; D001253:Astrocytes / Q000469:parasitology*; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D008297:Male; D018715:Microscopy, Video; D009068:Movement; D015800:Protozoan Proteins / Q000276:immunology*; D011817:Rabbits; D014349:Trypanosoma cruzi / Q000276:immunology / Q000502:physiology*
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D015800:Protozoan Proteins
keywords: Anti-peptide antibodies; Chagas disease; Flow cytometry; Trypanosoma cruzi