Gating strategy for the detection of different immune cell subsets from malignant ascites of ovarian cancer patients.

Cells collected from ascites were stained with a 9-color flow cytometry panel and analyzed in an X-20 Fortessa flow cytometer. (A) The gating strategy is depicted starting with the detection of lymphocytes, followed by live cells and CD3+ T cells that are separated according to CD4 and CD8 expression. (B) Fluorescence-minus-one (FMO) staining is shown for the different markers analyzed on CD4+ and CD8+ T cells. (PDF)</p

Ascitic fluid cell CDR3β motifs associated with prognosis.

(A) Examples of CD4+ T cell receptor (TCR) motifs associated with different prognoses. (B) Examples of CD8+ TCR motifs associated with different prognoses. The figures show the logo and phylogenetic tree of the clustered CDR3β peptides for each motif. The phylogenetic tree also includes peptides found as the best hit from McPAS; the complete set of motifs associated with excellent or poor/worst prognosis is shown in S7A Table. (C) Association networks constructed to form the cluster of motifs associated with response to platinum therapy. Nodes in the networks indicate individual patients; the color of the node indicates prognosis. Edges in the networks indicate an association between a pair of samples if they share GLIPH specificity groups associated with prognosis. The networks validated the statistical algorithm used to identify prognosis-associated specificity groups.</p

Difference in T cell receptor (TCR) characteristics between ascites of patients grouped in Cluster A and ascites of patients grouped in the remaining clusters (others).

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Increased T cell receptor productive rearrangements in ascites of patients with <i>BRCA1/2</i> tumor mutations.

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Difference in flow subset (expressed as a percentage of CD3+ live lymphocytes) between ascites of patients grouped in Cluster A and ascites of patients grouped in the remaining clusters (others).

(PDF)</p