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Cramoll 1,4 lectin increases ROS production, calcium levels, and cytokine expression in treated spleen cells of rats
Authors:Cristiane Moutinho Lagos de Melo  Bruno Alves Paim  Karina Gotardelo Zecchin  Joseani Morari  Marcos R. Chiaratti  Maria Tereza Santos Correia  Luana Cassandra B. Barroso Coelho  Patrícia Maria Guedes Paiva
Affiliation:1. Departamento de Imunologia, Instituto de Pesquisas Aggeu Magalh?es (IAM/FIOCRUZ), Universidade Federal de Pernambuco (UFPE), Recife, PE, 50670-901, Brazil
7. Departamento de Imunologia, Centro de Pesquisas Aggeu Magalh?es/FIOCRUZ, Cidade Universitária, Av. Moraes Rego s/n, Recife, PE, 50670-420, Brazil
2. Faculdade de Ciências Médicas, Departamento de Patologia Clínica, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, 13083-887, Brazil
3. Faculdade de Odontologia de Piracicaba, Departamento de Diagnóstico Oral, Universidade Estadual de Campinas (UNICAMP), Piracicaba, SP, 13414-018, Brazil
4. Departamento de Clínica Médica, Universidade Estadual de Campinas (UNCAMP), Campinas, SP, 13084-970, Brazil
5. Faculdade de Zootecnia e Engenharia de Alimentos, Departamento de Ciências Básicas, Universidade de S?o Paulo (USP), Pirassununga, SP, 13635-900, Brazil
6. Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco (UFPE), Recife, PE, 50670-901, Brazil
Abstract:This study reports the in vivo stimulatory effects of Cramoll 1,4 on rat spleen lymphocytes as evidenced by an increase in intracellular reactive oxygen species (ROS) production, Ca2+ levels, and interleukin (IL)-1β expression. Cramoll 1,4 extracted from seeds of the Leguminosae Cratylia mollis Mart., is a lectin with antitumor and lymphocyte mitogenic activities. Animals (Nine-week-old male albino Wistar rats, Rattus norvegicus) were treated with intraperitoneal injection of Cramoll 1,4 (235 μg ml?1 single dose) and, 7 days later, spleen lymphocytes were isolated and analyzed for intracellular ROS, cytosolic Ca2+, and IL-6, IL-10, and IL-1 mRNAs. Cell viability was investigated by annexin V-FITC and 7-amino-actinomycin D staining. The data showed that in lymphocytes activated by Cramoll 1,4 the increase in cytosolic and mitochondrial ROS was related to higher cytosolic Ca2+ levels. Apoptosis and necrosis were not detected in statistically significant values and thus the lectin effector activities did not induce lymphocyte death. In vivo Cramoll 1,4 treatment led to a significant increase in IL-1β but IL-6 and -10 levels did not change. Cramoll 1,4 had modulator activities on spleen lymphocytes and stimulated the Th2 response.
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