Figure 2
Figure 2. Cytokine and chemokine levels relative to timing of blinatumomab and tocilizumab. Cytokine and chemokine levels were measured 4 days after treatment with blinatumomab and 1 day prior to tocilizumab (toci) (red lines) and repeated 3 days later (2 days after tocilizumab) (blue lines), using Luminex bead array technology and kits purchased from Life Technologies (Invitrogen 30-plex; Carlsbad, CA). Assays were performed according to the manufacturer’s protocol with the 9-point standard curve generated using a 3-fold dilution series. Each sample was evaluated in duplicate at 1:3 dilution; calculated percentage (%) of coefficient of variation for the duplicate measures was in most cases less than 5% and always less than 15%. Data were acquired on a FlexMAP-3D and analyzed using XPonent 4.0 software and 5-parameter logistic regression analysis. Standard curve quantification ranges were determined by the 80% to 120% (observed/expected value) range. IL-2R, IL-6, IL-8, IL-10, monocyte chemoattractant protein (MCP)-1, macrophage-inflammatory protein (MIP)1B, and INF-γ were elevated, and IL-1B, IL-4, IL-5, IL-7, IL-12, IL-13, IL-17, tumor necrosis factor (TNF)-a, and granulocyte macrophage–colony-stimulating factor (GM-CSF) were normal. This cytokine prolife is identical to other published work investigating cytokines in children with HLH.7,8,14,15 IL-2 was normal in this patient. IL-2 can be normal or elevated in HLH; however, IL2R is universally elevated in HLH, as was found in this patient.7,8,14,15 The top half of the figure (above the black horizontal line) indicate cytokines expected to be elevated in HLH. The bottom half of the figure (below the black horizontal line) indicate cytokines expected to be normal in HLH. X-axis is cytokine level in pg/mL. Numbers beside red/blue line pair represent fold changes in cytokine level pre- and post-tocilizumab. A number of additional cytokines that have not been studied in the published HLH literature were also measured. A complete list of all cytokines tested with absolute values before and after tocilizumab treatment is included, which is described in supplemental Table 1.

Cytokine and chemokine levels relative to timing of blinatumomab and tocilizumab. Cytokine and chemokine levels were measured 4 days after treatment with blinatumomab and 1 day prior to tocilizumab (toci) (red lines) and repeated 3 days later (2 days after tocilizumab) (blue lines), using Luminex bead array technology and kits purchased from Life Technologies (Invitrogen 30-plex; Carlsbad, CA). Assays were performed according to the manufacturer’s protocol with the 9-point standard curve generated using a 3-fold dilution series. Each sample was evaluated in duplicate at 1:3 dilution; calculated percentage (%) of coefficient of variation for the duplicate measures was in most cases less than 5% and always less than 15%. Data were acquired on a FlexMAP-3D and analyzed using XPonent 4.0 software and 5-parameter logistic regression analysis. Standard curve quantification ranges were determined by the 80% to 120% (observed/expected value) range. IL-2R, IL-6, IL-8, IL-10, monocyte chemoattractant protein (MCP)-1, macrophage-inflammatory protein (MIP)1B, and INF-γ were elevated, and IL-1B, IL-4, IL-5, IL-7, IL-12, IL-13, IL-17, tumor necrosis factor (TNF)-a, and granulocyte macrophage–colony-stimulating factor (GM-CSF) were normal. This cytokine prolife is identical to other published work investigating cytokines in children with HLH.7,8,14,15  IL-2 was normal in this patient. IL-2 can be normal or elevated in HLH; however, IL2R is universally elevated in HLH, as was found in this patient.7,8,14,15  The top half of the figure (above the black horizontal line) indicate cytokines expected to be elevated in HLH. The bottom half of the figure (below the black horizontal line) indicate cytokines expected to be normal in HLH. X-axis is cytokine level in pg/mL. Numbers beside red/blue line pair represent fold changes in cytokine level pre- and post-tocilizumab. A number of additional cytokines that have not been studied in the published HLH literature were also measured. A complete list of all cytokines tested with absolute values before and after tocilizumab treatment is included, which is described in supplemental Table 1.

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