<i>Trichomonas vaginalis</i> Legumain-2, TvLEGU-2, Is an Immunogenic Cysteine Peptidase Expressed during Trichomonal Infection
Esly Alejandra Euceda-Padilla,
Miriam Guadalupe Mateo-Cruz,
Leticia Ávila-González,
Claudia Ivonne Flores-Pucheta,
Jaime Ortega-López,
Daniel Talamás-Lara,
Beatriz Velazquez-Valassi,
Lidia Jasso-Villazul,
Rossana Arroyo
Affiliations
Esly Alejandra Euceda-Padilla
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Av. IPN 2508, Alcaldía Gustavo A. Madero (GAM), Mexico City 07360, Mexico
Miriam Guadalupe Mateo-Cruz
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Av. IPN 2508, Alcaldía Gustavo A. Madero (GAM), Mexico City 07360, Mexico
Leticia Ávila-González
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Av. IPN 2508, Alcaldía Gustavo A. Madero (GAM), Mexico City 07360, Mexico
Claudia Ivonne Flores-Pucheta
Departamento de Biotecnología y Bioingeniería, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Av. IPN 2508, Alcaldía Gustavo A. Madero (GAM), Mexico City 07360, Mexico
Jaime Ortega-López
Departamento de Biotecnología y Bioingeniería, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Av. IPN 2508, Alcaldía Gustavo A. Madero (GAM), Mexico City 07360, Mexico
Daniel Talamás-Lara
Unidad de Microscopía Electrónica, Laboratorios Nacionales De Servicios Experimentales (LaNSE), Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Av. IPN 2508, Alcaldía Gustavo A. Madero (GAM), Mexico City 07360, Mexico
Beatriz Velazquez-Valassi
Departamento de Vigilancia Epidemiológica, Hospital General de México “Eduardo Liceaga”, Mexico City 06720, Mexico
Lidia Jasso-Villazul
Unidad de Medicina Preventiva, Hospital General de México “Eduardo Liceaga”, Mexico City 06720, Mexico
Rossana Arroyo
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Av. IPN 2508, Alcaldía Gustavo A. Madero (GAM), Mexico City 07360, Mexico
Trichomonas vaginalis is the causative agent of trichomoniasis, the most prevalent nonviral, neglected sexually transmitted disease worldwide. T. vaginalis has one of the largest degradomes among unicellular parasites. Cysteine peptidases (CPs) are the most abundant peptidases, constituting 50% of the degradome. Some CPs are virulence factors recognized by antibodies in trichomoniasis patient sera, and a few are found in vaginal secretions that show fluctuations in glucose concentrations during infection. The CPs of clan CD in T. vaginalis include 10 genes encoding legumain-like peptidases of the C13 family. TvLEGU-2 is one of them and has been identified in multiple proteomes, including the immunoproteome obtained with Tv (+) patient sera. Thus, our goals were to assess the effect of glucose on TvLEGU-2 expression, localization, and in vitro secretion and determine whether TvLEGU-2 is expressed during trichomonal infection. We performed qRT-PCR assays using parasites grown under different glucose conditions. We also generated a specific anti-TvLEGU-2 antibody against a synthetic peptide of the most divergent region of this CP and used it in Western blot (WB) and immunolocalization assays. Additionally, we cloned and expressed the tvlegu-2 gene (TVAG_385340), purified the recombinant TvLEGU-2 protein, and used it as an antigen for immunogenicity assays to test human sera from patients with vaginitis. Our results show that glucose does not affect tvlegu-2 expression but does affect localization in different parasite organelles, such as the plasma membrane, Golgi complex, hydrogenosomes, lysosomes, and secretion vesicles. TvLEGU-2 is secreted in vitro, is present in vaginal secretions, and is immunogenic in sera from Tv (+) patients, suggesting its relevance during trichomonal infection.