TY - JOUR
T1 - Extracellular vesicles released from Mycobacterium tuberculosis-Infected neutrophils promote macrophage autophagy and decrease intracellular mycobacterial survival
AU - Alvarez-Jiménez, Violeta D.
AU - Leyva-Paredes, Kahiry
AU - García-Martínez, Mariano
AU - Vázquez-Flores, Luis
AU - García-Paredes, Víctor Gabriel
AU - Campillo-Navarro, Marcia
AU - Romo-Cruz, Israel
AU - Rosales-García, Víctor Hugo
AU - Castañeda-Casimiro, Jessica
AU - González-Pozos, Sirenia
AU - Manuel Hernández, José
AU - Wong-Baeza, Carlos
AU - García-Pérez, Blanca Estela
AU - Ortiz-Navarrete, Vianney
AU - Estrada-Parra, Sergio
AU - Serafín-López, Jeanet
AU - Wong-Baeza, Isabel
AU - Chacón-Salinas, Rommel
AU - Estrada-García, Iris
N1 - Publisher Copyright:
© 2018 Alvarez-Jiménez, Leyva-Paredes, García-Martínez, Vázquez-Flores, García-Paredes, Campillo-Navarro, Romo-Cruz, Rosales-García, Castañeda-Casimiro, González-Pozos, Hernández, Wong-Baeza, García-Pérez, Ortiz-Navarrete, Estrada-Parra, Serafín-López, Wong-Baeza, Chacón-Salinas and Estrada-García.
PY - 2018/2/19
Y1 - 2018/2/19
N2 - Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (Mtb). In the lungs, macrophages and neutrophils are the first immune cells that have contact with the infecting mycobacteria. Neutrophils are phagocytic cells that kill microorganisms through several mechanisms, which include the lytic enzymes and antimicrobial peptides that are found in their lysosomes, and the production of reactive oxygen species. Neutrophils also release extracellular vesicles (EVs) (100-1,000 nm in diameter) to the extracellular milieu; these EVs consist of a lipid bilayer surrounding a hydrophilic core and participate in intercellular communication. We previously demonstrated that human neutrophils infected in vitro with Mtb H37Rv release EVs (EV-TB), but the effect of these EVs on other cells relevant for the control of Mtb infection, such as macrophages, has not been completely analyzed. In this study, we characterized the EVs produced by non-stimulated human neutrophils (EV-NS), and the EVs produced by neutrophils stimulated with an activator (PMA), a peptide derived from bacterial proteins (fMLF) or Mtb, and observed that the four EVs differed in their size. Ligands for toll-like receptor (TLR) 2/6 were detected in EV-TB, and these EVs favored a modest increase in the expression of the co-stimulatory molecules CD80, a higher expression of CD86, and the production of higher amounts of TNF-a and IL-6, and of lower amounts of TGF-β, in autologous human macrophages, compared with the other EVs. EV-TB reduced the amount of intracellular Mtb in macrophages, and increased superoxide anion production in these cells. TLR2/6 ligation and superoxide anion production are known inducers of autophagy; accordingly, we found that EV-TB induced higher expression of the autophagy-related marker LC3-II in macrophages, and the co-localization of LC3-II with Mtb inside infected macrophages. The intracellular mycobacterial load increased when autophagy was inhibited with wortmannin in these cells. In conclusion, our results demonstrate that neutrophils produce different EVs in response to diverse activators, and that EV-TB activate macrophages and promote the clearance of intracellular Mtb through early superoxide anion production and autophagy induction, which is a novel role for neutrophil-derived EVs in the immune response to Mtb.
AB - Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (Mtb). In the lungs, macrophages and neutrophils are the first immune cells that have contact with the infecting mycobacteria. Neutrophils are phagocytic cells that kill microorganisms through several mechanisms, which include the lytic enzymes and antimicrobial peptides that are found in their lysosomes, and the production of reactive oxygen species. Neutrophils also release extracellular vesicles (EVs) (100-1,000 nm in diameter) to the extracellular milieu; these EVs consist of a lipid bilayer surrounding a hydrophilic core and participate in intercellular communication. We previously demonstrated that human neutrophils infected in vitro with Mtb H37Rv release EVs (EV-TB), but the effect of these EVs on other cells relevant for the control of Mtb infection, such as macrophages, has not been completely analyzed. In this study, we characterized the EVs produced by non-stimulated human neutrophils (EV-NS), and the EVs produced by neutrophils stimulated with an activator (PMA), a peptide derived from bacterial proteins (fMLF) or Mtb, and observed that the four EVs differed in their size. Ligands for toll-like receptor (TLR) 2/6 were detected in EV-TB, and these EVs favored a modest increase in the expression of the co-stimulatory molecules CD80, a higher expression of CD86, and the production of higher amounts of TNF-a and IL-6, and of lower amounts of TGF-β, in autologous human macrophages, compared with the other EVs. EV-TB reduced the amount of intracellular Mtb in macrophages, and increased superoxide anion production in these cells. TLR2/6 ligation and superoxide anion production are known inducers of autophagy; accordingly, we found that EV-TB induced higher expression of the autophagy-related marker LC3-II in macrophages, and the co-localization of LC3-II with Mtb inside infected macrophages. The intracellular mycobacterial load increased when autophagy was inhibited with wortmannin in these cells. In conclusion, our results demonstrate that neutrophils produce different EVs in response to diverse activators, and that EV-TB activate macrophages and promote the clearance of intracellular Mtb through early superoxide anion production and autophagy induction, which is a novel role for neutrophil-derived EVs in the immune response to Mtb.
KW - Autophagy
KW - Extracellular vesicles
KW - Macrophage
KW - Neutrophils
KW - Tuberculosis
UR - http://www.scopus.com/inward/record.url?scp=85042196702&partnerID=8YFLogxK
U2 - 10.3389/fimmu.2018.00272
DO - 10.3389/fimmu.2018.00272
M3 - Artículo
C2 - 29520273
SN - 1664-3224
VL - 9
JO - Frontiers in Immunology
JF - Frontiers in Immunology
IS - FEB
M1 - 272
ER -