Journal: Viruses
Article Title: Characterization of Natural Killer Cell Profile in a Cohort of Infected Pregnant Women and Their Babies and Its Relation to CMV Transmission
doi: 10.3390/v16050780
Figure Lengend Snippet: Comparative analysis of NK cell profile in CMV-transmitting vs. non-transmitting mothers. ( A,B ) Scatter plots with bar (mean ± SD) depict frequency distribution, as in B,C. Unpaired t test and Mann–Whitney test were used to assess differences in cell frequencies between transmitting (red dots, n = 8) and non-transmitting (blue dots, n = 9) mothers. Significance was set at p < 0.05. ( C ) tSNE algorithm was configured to distribute data combined from transmitting and non-transmitting mothers’ samples according to the expression of NK cell markers CD56, CD16, NKG2C, NKG2A, CD57, NKG2D, DNAM-1, KIRs, and PD-1. Multigraph histogram overlays with geomean values were generated in a combined FCS file obtained by concatenating 2000 events in NK cell down-sample (identified as CD3-CD19-CD14- live lymphocytes) of transmitting (red, n = 7) and non-transmitting (blue, n = 8) mothers. ( D ) By using the same combined FCS file, single parameter heatmaps were obtained for transmitting (upper panels) and non-transmitting (lower panels) groups. Outline population (black circle) is specific to non-transmitting mothers and only the expressed markers are reported. TD NK: terminally differentiated NK; ML NK: memory-like NK.
Article Snippet: The “DownSample” FlowJo plugin was run on NK cells in order to reduce and make uniform the population sizes for the further concatenation of samples from different groups into a single FCS file. tSNE was performed on the concatenated file using the “TSNE” plugin and the maps generated using data from the following compensated parameters as inputs: CD56, CD16, NKG2C, NKG2A, NKG2D, DNAM-1, CD57, KIR2DL1/S1/S3/S5, KIR2DL2/L3, and PD-1 for the phenotype analysis and CD56, CD16, NKG2C, DNAM-1, CD57, NKp46, PD-1, and CD107a for the degranulation analysis; under the following tSNE settings: iteration 1000, perplexity 30, learning rate (Eta) 910–2100, the ANNOY algorithm as the k nearest neighbors (KNN) algorithm and the fast Fourier transform (FTT) interpolation as the gradient algorithm, resulting in tSNE plots with less than 5 million events ( ).
Techniques: MANN-WHITNEY, Expressing, Generated