Individuals differ in many respects including their response to stress and infection. Before trying to identify both the genetic and environmental reasons for those individual differences we first need to define and characterize different immune responses – immune phenotypes. By characterizing the response to stress and infection we will create reference values ranges for a number of immune phenotypes for a healthy population.
We are characterizing the major cell populations in the circulation of healthy donors through multi-parameter flow cytometry analysis. The numbers of these cells can be influenced by factors such as age, gender, lifestyle, previous infections as well as by genetic factors.
The major response to infection or stress is the production of effector proteins by cells of the immune system. To characterize the induced immune response to stimulation we have performed 32 whole blood assays (against a range of stimuli) across our 1000 healthy donor cohort. Samples have been biobanked and are in the process of being analyzed by protein immunoassays (Luminex based multi-analyte profiling) to quantify the secretion of the major cytokines and chemokines.
Transcriptome (mRNA, microRNA)
To identify and characterize the pathways behind the secreted protein response to the whole blood stimulations we will perform RNA-Seq, high throughput hybridization assays, and multiplex qPCR approaches. This will enable identification and quantification of mRNA and microRNAs altered by stress, infection, and stimulation.
Fibroblast and EBV cell lines have been created from our healthy donor populations. Experimental assays are being developed that will enable the assessment of different phenotypic traits associated with these cell types.