TLR4-mediated immunomodulatory properties of the bacterial metalloprotease arazyme in preclinical tumor models.
Despite the recent approval of new agents for metastatic melanoma, its treatment remains challenging. Moreover, few available immunotherapies induce a strong cellular immune response, and selection of the correct immunoadjuvant is crucial for overcoming this obstacle. Here, we studied the immunomodulatory properties of arazyme, a bacterial metalloprotease, which was previously shown to control metastasis in a murine melanoma B16F10-Nex2 model. The antitumor activity of arazyme was independent of its proteolytic activity, since heat-inactivated protease showed comparable properties to the active enzyme; however, the effect was dependent on an intact immune system, as antitumor properties were lost in immunodeficient mice. The protective response was IFNγ-dependent, and CD8(+) T lymphocytes were the main effector antitumor population, although B and CD4(+) T lymphocytes were also induced. Macrophages and dendritic cells were involved in the induction of the antitumor response, as arazyme activation of these cells increased both the expression of surface activation markers and proinflammatory cytokine secretion through TLR4-MyD88-TRIF-dependent, but also MAPK-dependent pathways. Arazyme was also effective in the murine breast adenocarcinoma 4T1 model, reducing primary and metastatic tumor development, and prolonging survival. To our knowledge, this is the first report of a bacterial metalloprotease interaction with TLR4 and subsequent receptor activation that promotes a proinflammatory and tumor protective response. Our results show that arazyme has immunomodulatory properties, and could be a promising novel alternative for metastatic melanoma treatment.
Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP), São Paulo, Brazil; Department of Immunology, Instituto de Ciências Biomédicas (ICB), University of São Paulo (USP), São Paulo, Brazil; HIV and Malaria Vaccine Program, Aaron Diamond AIDS Research Center (ADARC), Rockefeller University, NY, USA.
Felipe V
FV
Pereira
Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP), São Paulo, Brazil; Department of Immunology, Instituto de Ciências Biomédicas (ICB), University of São Paulo (USP), São Paulo, Brazil.
Amanda C L
AC
Melo
Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP) , São Paulo, Brazil.
Filipe M
FM
de Melo
Immunobiology Laboratory, Cancer Research UK, London Research Institute , London, UK.
Diego
D
Mourão-Sá
Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP) , São Paulo, Brazil.
Priscila
P
Silva
Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP) , São Paulo, Brazil.
Rodrigo
R
Berzaghi
Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP) , São Paulo, Brazil.
Carolina C A
CC
Herbozo
HIV and Malaria Vaccine Program, Aaron Diamond AIDS Research Center (ADARC), Rockefeller University, NY, USA; Rene Rachou Research Center, Oswaldo Cruz Foundation, FIOCRUZ, Minas Gerais, Brazil.
Jordana
J
Coelho-Dos-Reis
Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP) , São Paulo, Brazil.
Jorge A
JA
Scutti
Department of Immunology, Instituto de Ciências Biomédicas (ICB), University of São Paulo (USP) , São Paulo, Brazil.
Clarice S T
CS
Origassa
Department of Immunology, Instituto de Ciências Biomédicas (ICB), University of São Paulo (USP) , São Paulo, Brazil.
Rosana M
RM
Pereira
Department of Biophysics, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP) , São Paulo, Brazil.
Luis
L
Juliano
Department of Biophysics, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP) , São Paulo, Brazil.
Maria Aparecida
MA
Juliano
Department of Biophysics, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP) , São Paulo, Brazil.
Adriana K
AK
Carmona
Department of Immunology, Instituto de Ciências Biomédicas (ICB), University of São Paulo (USP) , São Paulo, Brazil.
Niels O S
NO
Câmara
HIV and Malaria Vaccine Program, Aaron Diamond AIDS Research Center (ADARC), Rockefeller University , NY, USA.
Moriya
M
Tsuji
Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP) , São Paulo, Brazil.
Luiz R
LR
Travassos
Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina (EPM), Federal University of São Paulo (UNIFESP) , São Paulo, Brazil.
Elaine G
EG
Rodrigues
4T1; arazyme; B16F10; TLR4; bacterial protease; breast adenocarcinoma; immunoadjuvant; melanoma