Immunomodulatory effects of different Lactobacillus probiotic strains in macrophages challenged by Candida albicans

• Grant number: 16/17166-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2016
• End date: September 30, 2017
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Luciane Dias de Oliveira
• Grantee: Barbara Evelyn Begnini
• Host Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil

Abstract

Lactobacilli strains may have probiotic characteristics, being one of the most significant ones the modulation of the host immune system. Many probiotics are composed of a mixture of lactobacilli, in order to enhance this effect. Thus, the aim of this study is to evaluate the immunomodulatory caused by different combinations of lactobacilli suspensions in macrophages challenged by C. albicans, through the analysis of pro-inflammatory cytokines (TNF-±, IL-1², IL-6, IL -17). For this purpose, Lactobacillus rhamnosus ATCC 7469 (LR), L. acidophilus LA 14 (LA), isolated from a commercial product and a clinical strain of L. paracasei (LP) are suspended at a concentration of 2.5 x 107 CFU/mL to obtain 7 final suspension (LR, LA, LP, LR+LA, LR+LP, LA+LP e LR+LA+LP). C. albicans ATCC 18804 will suspended at a concentration of 2 x 106 CFU/mL. Mouse macrophages (RAW 264.7) will be distributed in 24-wells polystyrene microplates (5 x 105 cells/well) and incubated for 24 h (37°C/5% CO2) for cell adhesion. Macrophages will be activated by each probiotic suspension and then challenged by C. albicans. Tests will be conducted in triplicate, with four repetitions in each, resulting 12 samples for each experimental group. After incubation for 16 h at 37ºC and 5% CO2, DMEM with cell supernatant will be collected, aliquoted and frozen in a freezer (-80ºC) for later analysis of cytokines level. It will be evaluated production of pro-inflammatory cytokines (TNF-±, IL-1², IL-6, IL-17) by immunoenzymatic method ELISA. Results will be statistically analyzed in accordance with the normal distribution (ANOVA and Tukey or Kruskal-Wallis and Dunn), with 5% significance level.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DE OLIVEIRA, FELIPE EDUARDO; ROSSONI, RODNEI DENNIS; DE BARROS, PATRICIA PIMENTEL; BEGNINI, BARBARA EVELYN; JUNQUEIRA, JULIANA CAMPOS; CARDOSO JORGE, ANTONIO OLAVO; PEREIRA LEAO, MARIELLA VIEIRA; DE OLIVEIRA, LUCIANE DIAS. Immunomodulatory effects and anti-Candida activity of lactobacilli in macrophages and in invertebrate model of Galleria mellonella. Microbial Pathogenesis, v. 110, p. 603-611, SEP 2017. (16/17166-7)