Clinical “cytokine storm” as revealed by monocyte intracellular flow cytometry: correlation of tumor necrosis factor α with severe gut graft-versus-host.

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Clinical “cytokine storm” as revealed by monocyte intracellular flow cytometry: correlation of tumor necrosis factor α with severe gut graft-versus-host disease Daniel H. Fowler, Jason Foley, Jeannie Whit-shan Hou, Jeanne Odom, Kate Castro, Seth M. Steinberg, Juan Gea-banacloche, Claude Kasten-sportes, Ronald E. Gress, Michael R. Bishop Clinical Gastroenterology and Hepatology Volume 2, Issue 3, Pages 237-245 (March 2004) DOI: 10.1016/S1542-3565(04)00011-4

Figure 1 Monocyte IL-1α and TNF-α production posttransplantation: grade III acute graft-versus-host disease (GVHD). Peripheral-blood mononuclear cells (PBMCs) were obtained from a patient day 17 posttransplantation (no clinical acute GVHD) and day 38 posttransplantation (before grade III acute GVHD onset day 39 posttransplantation). PBMCs were incubated overnight in media containing monensin A. Adherent and nonadherent cells were stained for a monocyte surface marker (CD14-PE), permeabilized, and stained with either IL-1α fluorescein isothiocyanate (FITC), TNF-α FITC, or isotype control FITC. Results are expressed as percentage of CD14+ cells producing a particular cytokine (percentage of CD14+IL-1+ and percentage of CD14+TNF+). Clinical Gastroenterology and Hepatology 2004 2, 237-245DOI: (10.1016/S1542-3565(04)00011-4)

Figure 2 Monocyte intracellular IL-1α and TNF-α production posttransplantation: kinetics of expression and association with graft-versus-host disease (GVHD) severity. (A, B) Paired IL-1 and TNF values were averaged (with SEM shown) for each successive 2-week interval after allogeneic hematopoietic stem-cell transplantation (HSCT). (C) Average IL-1α values for patients in each of 4 GVHD outcomes; there was no trend for increased monocyte IL-1α production with increased GVHD severity (P = 1.00). (D) Average TNF-α values for patients in each of 4 GVHD outcomes; there was a trend toward increased monocyte TNF-α production with increased GVHD severity (P = 0.0041). Clinical Gastroenterology and Hepatology 2004 2, 237-245DOI: (10.1016/S1542-3565(04)00011-4)

Figure 3 Temporal association of peak monocyte TNF-α value with graft-versus-host disease (GVHD) onset. The percentage of monocytes producing TNF-α was determined for each patient at various times posttransplantation. Patients were grouped according to GVHD outcome (stage 2–3 gut GVHD, stage 1 gut GVHD, skin-only GVHD, or no acute GVHD). Individual patient values can be identified by their assignment (1, 2, 3, 4, or 5) within each GVHD group; these assignments correlate with patient characteristics listed in Table 1. Data points marked by increased font size indicate the TNF-α value obtained closest to the time of GVHD onset. Clinical Gastroenterology and Hepatology 2004 2, 237-245DOI: (10.1016/S1542-3565(04)00011-4)