Ruiz DH Andy¹, Juárez Esmeralda¹, González Yolanda¹, Favela-Hernández Juan Manuel² and Guerrero G Gloria³*

¹Instituto Nacional de Enfermedades Respiratorias (INER), Departamento de Microbiología, México

*Corresponding Author: Guerrero G Gloria, Universidad Autónoma de Zacatecas, Unidad Académica de Ciencias Biológicas, Lab de Imunobiologia, Campus II, Zacatecas, Zac, México.

Received: November 07, 2022; Published: November 18, 2022

Abstract

Autophagy is a highly conserved degradative and recycling program to maintain homeostasis. In particular, it plays an important role in the innate immune response against intracellular pathogens. Several studies have shown that BCG and/or M. tuberculosis block autophagosome formation, inhibiting thus, activation of the autophagy machinery, and survival of mycobacteria. Human macrophages preparation from Peripheral blood mononuclear cells (buffy coats of the healthy donor) (blood bank of INER. MX). Monocytes were further isolated by CD14 positive selection and differentiated into monocyte-derived macrophages. Infection of macrophages with M. bovis BCG vaccine (ATCC, Manassas, VA, US) at MOI 1:5. Stimulation with Cry1A-Cr2A (5 mg/ml Rapamycin (250 mg/ml) and/or Wortmannin (100 nM). Autophagy detection and fluorescence microscopy were made in the uninfected well and the infected-stimulated cells were stained with rabbit anti-LC3B coupled to Alexa Fluor 488. The percentage of cells with more than 5 LC3+ puncta (autophagosomes) was calculated as well as the percentage of bacteria co-localizing with LC3. The percentage of mycobacteria BCG co-localizing with light chain 3 (LC3-II) in human macrophages is greater in infected and stimulated human macrophages with the commercial preparation based on Cry1A-Cry2A Bt proteins versus rapamycin and/or Wortmannin. The commercial preparation of Cry1A-Cry2A in combination with the BCG vaccine represents a potential alternative to enhance the autophagy-mediated elimination of intracellular pathogens such as M. tuberculosis.

Keywords: Bacillus thuringiensis; Cry1A Toxins; Autophagy; Macrophages; Microtubule Light Chain 3 (LC3); Tuberculosis

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Citation

Citation: Guerrero G Gloria., et al. “Autophagy Inducing Capacities of a Commercial Preparation Based on of Bacillus thuringiensis Cry 1A-2A Toxins in Human Macrophages: Implication Against Mycobacterial Infections". Acta Scientific Microbiology 5.12 (2022): 62-69.

Copyright

Copyright: © 2022 Guerrero G Gloria., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.