Human blood monocytes can be broadly divided into two distinct subsets: CD14+CD16- and CD14+/lowCD16+ subsets. Perturbation in their proportions in the blood has been observed in several disease conditions. Although numerous phenotypic and functional differences between the two subsets have already been described, the roles contributed by each subset during homeostasis or disease conditions are still largely unclear. To uncover novel differences to aid in elucidating their functions, we perform a global analysis of the two subsets utilizing both proteomics and transcriptomics approaches. From the proteomics and transcriptomics data, the expression of 613 genes by the two subsets is detected at both the protein and mRNA levels. These 613 genes are assessed for up-regulation in each subset at the protein and mRNA levels using a cutoff fold change of > or =|1.5| between subsets. Proteins and mRNAs up-regulated in each subset are then mapped in silico into biological functions. This mapping reveals copious functional differences between the subsets, many of which are seen at both protein and mRNA levels. For instance, expression of genes involved in F(CY) receptor-mediated phagocytosis are up-regulated in the CD14+/lowCD16+ subset, while those involved in antimicrobial function are up-regulated in the CD14+CD16- subset. We uncover novel functional differences between the monocyte subsets from differences in gene expression at the protein and mRNA levels. These functional differences would provide new insights into the different roles of the two monocyte subsets in regulating innate and adaptive immune responses.

High macrophage infiltration into tumours often correlates with poor prognoses; in colorectal, stomach and skin cancers, however, the opposite is observed but the mechanisms behind this phenomenon remain unclear. Here, we sought to understand how tumour-associated macrophages (TAMs) in colorectal cancer execute tumoursuppressive roles. We found that TAMs in a colorectal cancer model were pro-inflammatory and inhibited the proliferation of tumour cells. TAMs also produced chemokines that attract T cells, stimulated proliferation of allogeneic T cells and activated type-1 T cells associated with anti-tumour immune responses. Using colorectal tumour tissues, we verified that TAMs in vivo were indeed pro-inflammatory. Furthermore, the number of tumour-infiltrating T cells correlated with the number of TAMs, suggesting that TAMs could attract T cells; and indeed, type-1 T cells were present in the tumour tissues. Patient clinical data suggested that TAMs exerted tumour-suppressive effects with the help of T cells. Hence, the tumour-suppressive mechanisms of TAMs in colorectal cancer involve the inhibition of tumour cell proliferation alongside the production of pro-inflammatory cytokines, chemokines and promoting type-1 T-cell responses. These new findings would contribute to the development of future cancer immunotherapies based on enhancing the tumour-suppressive properties of TAMs to boost anti-tumour immune responses. To elucidate the roles of TAMs, we first used an in vitro model known as the multi-cellular tumour spheroid (MCTS) model. This model has been proven to exhibit micro-environmental heterogeneity comparable to that of tumours in vivo, in terms of oxygen, nutrient, catabolite and metabolite gradients, resulting in sub-populations of proliferative and necrotic tumour cells typical of non-vascular tumour micro-regions [9,10]. Compared with using animal models, this MCTS model offers the advantages of studying the interactions between tumour cells and TAMs without confounding factors from other cell types, and in a 'human' microenvironment. In this study, we used colorectal cancer as a model to study the mechanisms underlying the tumour-suppressive effects of TAMs. We co-cultured primary human monocytes with human colorectal tumour cells for 8 days as MCTSs, during which time the monocytes would differentiate into TAMs. We performed global gene expression profiling to obtain an overview of the biological functions of TAMs, followed by validation with functional assays. Subsequently, we verified the in vitro findings with tumour tissues from colorectal cancer patients.The TAMs in the colorectal cancer model were pro-inflammatory and inhibited the proliferation of tumour cells. The TAMs also secreted chemokines that attract T cells and expressed surface molecules for antigen presentation and T-cell co-stimulation. In a mixed lymphocyte reaction (MLR) assay, the TAMs stimulated proliferation of allogeneic T cells and activated type-1 T cells, which are associated with anti-tumour immune responses [11]. To co...