Single-Cell Network Profiling of Peripheral Blood Mononuclear Cells from Healthy Donors Reveals Age- and Race-Associated Differences in Immune Signaling Pathway Activation

Abstract

A greater understanding of the function of the human immune system at the single-cell level in healthy individuals is critical for discerning aberrant cellular behavior that occurs in settings such as autoimmunity, immunosenescence, and cancer. To achieve this goal, a systems-level approach capable of capturing the response of the interdependent immune cell types to external stimuli is required. In this study, an extensive characterization of signaling responses in multiple immune cell subpopulations within PBMCs from a cohort of 60 healthy donors was performed using single-cell network profiling (SCNP). SCNP is a multiparametric flow cytometry-based approach that enables the simultaneous measurement of basal and evoked signaling in multiple cell subsets within heterogeneous populations. In addition to establishing the interindividual degree of variation within a broad panel of immune signaling responses, the possible association of any observed variation with demographic variables including age and race was investigated. Using half of the donors as a training set, multiple age- and race-associated variations in signaling responses in discrete cell subsets were identified, and several were subsequently confirmed in the remaining samples (test set). Such associations may provide insight into age-related immune alterations associated with high infection rates and diminished protection following vaccination and into the basis for ethnic differences in autoimmune disease incidence and treatment response. SCNP allowed for the generation of a functional map of healthy immune cell signaling responses that can provide clinically relevant information regarding both the mechanisms underlying immune pathological conditions and the selection and effect of therapeutics.

Causes, consequences, and reversal of immune system aging

“…As previously discussed, class switch recombination is impaired in aging murine and human B cells due to decreased expression of enzymes such as AID (53,84). Aberrant T cell signaling has also been associated with aging (48,85), and the list of dysregulated genes continues to grow, as indicated by several recent reports (86)(87)(88), two of which focused on altered expression of dual specific phosphatases (DUSPs). DUSPs deactivate target kinases, including those in the MAPK pathway whose activity is critical for T cell activation, differentiation, and cytokine production.…”

Aquaporins in Infection and Inflammation

“…The differences observed for the different MOIs could in part be explained by the difference in concentrations of AHL. Donor-specific variations of the macrophages response capabilities, including differences in migration and phagocytosis could also influence the response [241,242].…”

Association between B-cell receptor responsiveness and disease progression in B-cell chronic lymphocytic leukemia: results from single cell network profiling studies

“…[14][15][16] The association of SCNP data with in vivo acute myeloid leukemia (AML) chemosensitivity/chemoresistance 17 as well as FLT3R signaling deregulation 18 and applications in immunology 19,20 have been previously reported. Here the SCNP assay was first applied in an exploratory study to functionally characterize elements of the BCR signaling network and to assess their association with clinical and prognostic parameters with the goal of ultimately developing a clinically useful tool for the prediction of B-CLL progression and early (<3 years) TTFT in patients with early stage B-CLL.…”