Response of peripheral blood mononuclear cells to conditioned medium from cultured oral squamous cell carcinomas

Abstract

Response of peripheral blood mononuclear cells to conditioned medium from cultured oral squamous cell carcinomas Abstract: The current study investigated the capacity for tumor factors secreted by head and neck squamous cell carcinoma (HNSCC) cell lines, KB, KB16, and HEP, to induce the secretion of various cytokines from peripheral blood mononuclear cells (PBMCs). PBMCs were isolated from blood samples collected from six healthy volunteers and these cells were incubated for 6, 24, 48, or 72 hours in the presence of 50% conditioned medium collected from cultured cell lines pretreated with, or without, stimulants such as phytohemagglutinin (PHA) or lipopolysaccharides (LPS). Aliquots of each supernatant were then assayed for levels of IFN-γ, vascular endothelial growth factor (VEGF), TNF-α, and IL-4 using enzyme linked immunosorbent assays (ELISAs). Data collected were analyzed using Student's t-test, an ANOVA test followed by Tukey's test, and tests of Pearson's Correlation. PBMCs cultured with KB16-conditioned medium produced the highest levels of IFN-γ. VEGF was also detected in conditioned media collected from all of the squamous cell carcinoma (SCC) cell lines used, and a significant difference in VEGF levels between control and KB-or KB16-conditioned media was observed. TNF-α was secreted by all PBMC groups within 6 hours of receiving conditioned media, and these levels increased up to the 24 hour timepoint, after which levels of TNF-α stabilized. In contrast, none of the supernatant samples contained detectable levels of IL-4. In combination, these data suggest that direct contact between fresh human PBMCs and conditioned media from tumor cells induces the secretion of TNF-α and VEGF by PBMCs, and this represents an initial angiogenic response.

A mammary adenocarcinoma murine model suitable for the study of cancer immunoediting

“…The higher proliferation of M-406 cells observed when cultured with CMO from both Naïve CBi/L and CBi mice indicates that these media would contain factor/s like IL-2, IL-4, INF-γ, IL-10, TNF-α and many others, yet to be identified, capable to stimulate or inhibit tumor growth. This observation agrees with the results obtained by Franca [24] who found that direct contact between fresh human mononuclear cells and conditioned media from tumor cells induces the secretion of TNF-α and VEGF, factors which are clearly involved in tumor growth.…”

IRF5 is a novel regulator of CXCL13 expression in breast cancer that regulates CXCR5+ B‐ and T‐cell trafficking to tumor‐conditioned media

“…Upregulation of this gene cluster caused increased recruitment of lymphocytes to the site of injury 29 . In addition, several of the immune‐stimulating cytokines/chemokines identified to be potential IRF5 target genes by RT‐qPCR array (Table 1) have been shown to be expressed in normal head and neck squamous epithelial cells, fallopian tubes, vaginal, corneal, stomach, lung, salivary, colon and pancreatic epithelial cells as well 29 , 31 , 36 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 . These data suggest that IRF5 may have an important and more global role in the regulation of epithelial‐mediated inflammation.…”

“…Data obtained from the focused RT‐qPCR array offer significant insight into the complexity of the tumor immune microenvironment; every ligand and receptor listed has the ability to shape an immune response. For instance, the release of IL‐33 from necrotic cells, such as the necrotic and hypoxic cells in the tumor center, along with the chemokine CXCL5, can cause a rapid influx of macrophages and thus a rapid inflammatory response 30 , 31 , 32 , 37 , 38 , 39 , 54 , 55 , 62 , 63 , 64 . Interferon‐γ has been shown to inhibit breast cancer cell growth, 65 whereas CXCR2 () allows for sustained senescence in otherwise neoplastic breast tumors 66 .…”