Molecular profiling of CD3- CD4+ T-cells from patients with the lymphocytic variant of hypereosinophilic syndrome

The clonal CD3- CD4+ Th2 cell population characterizing some hypereosinophilic syndrome patients stably endures for years provoking a chronic inflammatory skin disease, with a subgroup of patients ultimately progressing to T-cell lymphoma. The aim of this study is the identification of the molecular changes (1) associated with the persistence of the pre-malignant clone (2) associated with the activation of co-stimulatory receptors and (3) associated with the emergence of malignant T-cell subclones. (1) Molecular changes associated with the persistence of the pre-malignant clone. The gene expression profile of CD3- CD4+ T-cells isolated at diagnosis of three LV-HES patients (chronic disease phase) was compared to the gene expression profile of CD4+ T-cells isolated from four healthy controls. Patients samples were P1 (yr.0), P2 (yr.0) and P3 (yr.0). For P1 (yr.0), three experimental replicates were processed: P1.a (yr.0), P1.b (yr.0) and P1.c (yr.0). For P2 (yr.0), two experimental replicates were processed: P2.a (yr.0) and P2.b (yr.0). For P3 (yr.0), two experimental replicates were processed: P3.a (yr.0) and P3.b (yr.0). Healthy controls were D1, D2, D3 and D4. (2) Molecular changes associated with the activation of co-stimulatory receptors. The gene expression profile of CD3- CD4+ T-cells isolated at diagnosis from three LV-HES patients (chronic disease phase) and activated in vitrowith a combination of rhIL2 and anti-CD2/CD28 antibodies was compared to the gene expression profile of the same CD3- CD4+ T-cell populations, cultured without activation. In vitro stimulated patients samples were P1 (yr.0) Stim, P2 (yr.0) Stim and P3 (yr.0) Stim. For P1 (yr.0) Stim, three experimental replicates were processed: P1.a (yr.0) Stim, P1.b (yr.0) Stim and P1.c (yr.0) Stim. For P2 (yr.0) Stim, two experimental replicates were processed: P2.a (yr.0) Stim and P2.b (yr.0) Stim. For P3 (yr.0) Stim, two experimental replicates were processed: P3.a (yr.0) Stim and P3.b (yr.0) Stim. Unstimulated patients samples were P1 (yr.0), P2 (yr.0) and P3 (yr.0). For P1 (yr.0), three experimental replicates were processed: P1.a (yr.0), P1.b (yr.0) and P1.c (yr.0). For P2 (yr.0), two experimental replicates were processed: P2.a (yr.0) and P2.b (yr.0). For P3 (yr.0), two experimental replicates were processed: P3.a (yr.0) and P3.b (yr.0). (3) Molecular changes associated with the emergence of malignant T-cell subclones. The gene expression profiles of CD3- CD4+ T-cells isolated at three stages in patient 1’s clinical progression were assessed: two chronic disease stages were at diagnosis P1 (yr.0) and follow up P1 (yr.+4) and one T-lymphoma stage was at follow up P1 (yr.+6). For P1 (yr.0), three experimental replicates were processed: P1.a (yr.0), P1.b (yr.0) and P1.c (yr.0). For P1 (yr.+4), three experimental replicates were processed: P1.a (yr.+4), P1.b (yr.+4) and P1.c (yr.+4). For P1 (yr.+6), three experimental replicates were processed: P1.a (yr.+6), P1.b (yr.+6) and P1.c (yr.+6).