EFFECT OF SUPERNATANT OF CERVICAL CANCER CELLS IN THE POLARIZATION OF THP-1 MACROPHAGE TOWARD M2 SUPPRESOR MACROPHAGE BY INDUCING EXPRESSION OF SOCS1 AND SOCS3
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1
CENTRO DE INVESTIGACION BIOMEDICA DE OCCIDENTE (CIBO), INMUNOLOGIA, Mexico
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2
CENTRO UNIVERSITARIO DE CIENCIAS DE LA SALUD UDG, DOCTORADO EN CIENCIAS BIOMEDICAS, Mexico
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3
UNIVERSIDAD AUTONOMA DE GUADALAJARA, FACULTAD DE MEDICINA, Mexico
Background
Cervical cancer is the fourth cancer that more affect a women in worldwide, this is a multistep process characterized by dysplastic lesions developed after a persistent infection with some subtype of high risk of human papillomaviruses (HPV) such as HPV-16 or HPV-18.
The macrophages are part of innate immune system and important immune effector cells against pathogens and tumors cells; can be differentiated into two phenotypes, identified as M1 macrophage (classically activated), and M2 macrophage (alternatively activated).
Classics activators from macrophage are Interferon-gamma (IFN-γ) and Lipopolysaccharides (LPS), that polarize the macrophage toward the M1 phenotype, characterized by their ability to release proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), Interleukin (IL)-1β, IL-6, IL-12 as well as reactive nitrogen and oxygen intermediates. While that TH2 cytokines such as IL-4, IL-13, and IL-10, as well as glucocorticoids, give rise to M2 macrophages, a phenotype characterized by low expression of IL-12 and high expression of IL-10, transforming growth factor-beta (TGF-β), and scavenger and mannose receptors as CD163 and CD206; respectively.
The polarization toward any specific phenotype is mainly mediated by JAK/STAT as well as NF-kB signaling pathway. NF-kB and STAT1 on macrophages are involved in the expression of many inflammatory genes including iNOS, CCL2, CCL5 and others as IFN-γ and IFN-α and –β. On the other hand STAT3 and STAT6 are transcription factors that mediate the activation of M2 macrophages.
The suppressor of cytokines signaling (SOCS) proteins are regulators of signaling pathway of JAK/STAT, principally SOCS1 and SOCS3. Previous reports have described the importance of SOCS1 and SOCS3 proteins in the regulation of inflammation state and in the polarization of the macrophage phenotype.
The role of tumor associated macrophages (TAM) remains controversial. However, many observations indicate that TAM express several functions associated with the M2 macrophage phenotype, including promotion of angiogenesis, matrix remodeling, and suppression of adaptative immunity.
Objectives
To determinate effect of the factors secreted by cervical cancer cell lines on SOCS1 and SOCS3 proteins in the polarization of THP-1-derived macrophages toward M2 suppressor macrophage.
Methods
Cervical cancer cell lines HeLa, SiHa, C-33A (1 x 105) were grown on 2 mL of DMEM. After 5 day of culture, the supernatant of these cell lines was collected for use in the culture of THP-1-derived macrophages in the corresponding experimental groups.
THP-1 cell lines (1 × 106 cells) were differentiated into macrophages in the presence 200 nM of phorbol myristate acetate (PMA) for 3 days. After incubation, adherent cells were washed with PBS, after were treated with the supernatant of HeLa, SiHa, C-33A cells and incubated for 7 days or 3 hours. The macrophages cultured 7 days with the supernatant were stained for assessment of CD163, CD14, CD86, CD206, HLA-DR and Arginase 1 by Flow cytometry analysis. On the other hand, the macrophages treated with the supernatant for 3 hours were washed with PBS and collected by RNA extraction for assessment the gene expression of IL-1β, TGF-β, IL-10, CD80, CD86, SOCS1 and SOCS3.
Results
We observed that cervical cancer cell lines HeLa, SiHa and C-33A induced a significant increase in the CD163 and CD206 also a decrease in HLA-DR expression of THP-1-derived macrophages.
The supernatant of cervical cancer cells induced a decrease in the CD80 gene expression and an augment in the gene expression of SOCS1 and SOCS3 in the THP-1-derived macrophages treated 3 hours with the supernatants.
Conclusions
Our results indicated that THP-1-derived macrophages treated with the supernatants of HeLa, SiHa, and C-33A polarized the macrophages toward M2 macrophages characterized by CD163 and CD206 high expression, however not decrease the CD86 expression was observed. On the other hand, this THP-1-derived macrophages treated with the supernatants of HeLa, SiHa, and C-33A induce a significant decrease of CD80 and a significant increase of SOCS1 and SOCS3 in the gene expression, that is related with the polarization toward M2 suppressor macrophages.
Keywords:
cervical cancer,
Macrophages,
socs3,
SOCS1,
M1 macrophages,
M2 macrophages,
macrophages polarization,
THP-1
Conference:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología, Medellin, Colombia, 13 Oct - 16 Oct, 2015.
Presentation Type:
Poster Presentation
Topic:
Innate Immunity
Citation:
PEDRAZA BRINDIS
EJ,
ORTIZ LAZARENO
PC,
BRAVO CUELLAR
A,
JAVE SUAREZ
LF,
HERNANDEZ FLORES
G,
AGUILAR LEMARROY
A,
Reyes
KS and
LOPEZ LOPEZ
B
(2015). EFFECT OF SUPERNATANT OF CERVICAL CANCER CELLS IN THE POLARIZATION OF THP-1 MACROPHAGE TOWARD M2 SUPPRESOR MACROPHAGE BY INDUCING EXPRESSION OF SOCS1 AND SOCS3.
Front. Immunol.
Conference Abstract:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología.
doi: 10.3389/conf.fimmu.2015.05.00018
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Received:
02 Jun 2015;
Published Online:
14 Sep 2015.
*
Correspondence:
PhD. PABLO C ORTIZ LAZARENO, CENTRO DE INVESTIGACION BIOMEDICA DE OCCIDENTE (CIBO), INMUNOLOGIA, GUADALAJARA, JALISCO, 44340, Mexico, pablolazareno@gmail.com