Abstract
Physiological cytokine environments arise from factors produced by diverse cell types in coordinated concert. Understanding the contributions of each cell type in the context of cell-cell communication is important for effectively designing disease modifying interventions. Here, we present multi-plexed measurement of 48 cytokines from a coculture system of primary human CD4+ T cells and monocytes across a spectrum of stimuli and for a range of relative T cell/monocyte compositions, coupled with corresponding measurements from PBMCs and plasma from the same donors. Computational analysis of the resulting data-sets elucidated communication-independent and communication-dependent contributions, including both positive and negative synergies. We find that cytokines in cell supernatants were uncorrelated to those found in plasma. Additionally, as an example of positive synergy, production levels of CXCR3 cytokines IP-10 and MIG, depend non-linearly on both IFNγ and TNFα levels in cross-talk between T cells and monocytes. Overall, this work demonstrates that communication between cell types can significantly impact the consequent cytokine environment, emphasizing the value of mixed cell population studies.
Highlights
It is possible to identify changes in circulating cytokines relevant to inflammation in serum samples, it is difficult to derive mechanistic information about overall change in immune activation from these measurements
For each stimulation condition and cell type, we establish that isolated cell populations respond when stimulated with the relevant stimuli– we observe cytokines typically associated with T cells treated when adding T-cell receptor stimulus (TCR) stimulus or phorbol 12-myristate 13-acetate/ionomycin (PI) to isolated CD4+T cells (i.e. IL-2, IL-4, IL-13, GM-CSF, IFN-γ)and monocyte-specific cytokines when monocytes were treated with LPS (i.e. GRO-α, IL-1β, G-CSF)
We applied hierarchical clustering to determine that CD4+T cells stimulated with their relevant stimuli (PI and TCR) cluster separately from resting CD4+T cells and those stimulated with LPS
Summary
Coculture environment yields insights into divergent cellular behavior due to immune cell communication. The PBMCs cluster together with the stimulated CD4+T cells when stimulated with TCR/PI and with the monocytes when stimulated with LPS This multidimensional dataset presents opportunities to examine relationships between commonly studied plasma and cell supernatants, as well as uncover cytokines regulated by cellular communication. HGF, was detected in plasma but not in cultured cell supernatants under any stimuli tested (Fig. 3A) As many of these cytokines were present only in stimulated cells, which may explain their absence in plasma, we further looked at only those cytokines secreted by each cell population under resting conditions. In this case, five cytokines (MIG, IFNα2, HGF, SCF, and SCGFb) are not represented by cellular supernatants (Fig. 3B). Collecting cytokine secretion upon stimulation of isolated cell types is not sufficient to predict the cytokine environment in a multicellular environment such as that found in PBMCs; including interaction data improves the accuracy of the model
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