Application of CyTOF based high dimensional single cell analysis to characterize the peripheral blood profile in acute infectious disease.

MALEKI KIMIA; KLINGSTROM JONAS; GARCÍA MARINA; MARTINEZ VALERIA; SOLA RIERA CARLES; SCHIERLOH PABLO

Ciudad Autónoma de Buenos Aires

Simposio; Frontiers in Bioscience 3; 2018

Max Planck Institute

Hantaviruses are enveloped negative single stranded RNA viruses belonging to the Hantaviridae family. The Old World hantaviruses cause Haemorrhagic Fever with Renal Syndrome (HFRS) in Europe and Asia, whereas the New World Hantaviruses cause Hantavirus Pulmonary Syndrome in the Americas (HPS), including Andes virus (ANDV) that causes HPS in Argentina. We have previously reported a massive plasmablast response in circulation of HPS patients from Argentina, characterized by flow cytometry [1]. Herein we aimed to study the peripheral blood mononuclear cells (PBMC) of hantavirus infected patients at high dimensional level (n>50 parameters/cell), by means of single cell mass cytometry (CyTOF) technology [2].PBMC from hantavirus-infected patients were assayed using designed panels of 28 heavy metal labelled monoclonal antibodies. T-distributed Stochastic Neighbour Embedding (t-SNE) machine learning algorithm was applied for dimensional reduction during broad exploration of the data. 2D-viSNE plots can be used to reveal defined clusters of cells that vary in proportion over time, allowing for a quick screening and detection of altered cell populations with respect to normality. This allowed for identification of several altered populations within the lymphocyte lineage during the acute phase of disease, as previously described in literature by means of multiple independent lineage panels and flow cytometry. The combined use of CyTOF with a dimensionality reduction algorithm can be very useful to pinpoint altered cellular compartments in patient blood samples, which can then be further assessed for multiple parameters. Another benefit from mass cytometry data is that it does not require compensation for spillover between channels, simplifying analysis in great extent. Furthermore, given the critical hemodynamic state in which many hantavirus-infected patients are, limited blood sample can be withdrawn for research purposes. In this sense, CyTOF can be a very useful approach for maximizing the data to be extracted from these precious human samples.