Simultaneous Detections of 27 Cytokines during Cerebral Wound Healing by Multiplexed Bead-Based Immunoassay for Wound Age Estimation

Abstract

Quantification of 27 cytokines following cerebral wounding was performed for wound age estimation. The cytokines evaluated included interleukin (IL)-1alpha, IL-1beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, IL-12 p40, IL-12 p70, IL-15, IL-17, IL-18, basic fibroblast growth factor (bFGF), granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), Interferon-gamma (IFN-gamma), keratinocyte derived cytokine (KC), leukemia inhibitory factor (LIF), macrophage-colony stimulating factor (M-CSF), monokine inducible by interferon gamma (MIG), macrophage inflammatory protein (MIP)-1 alpha, MIP 2, platelet-derived growth factor BB (PDGF BB), regulated upon activation, normal T-cell expressed, and secreted (Rantes), tumor necrosis factor-alpha (TNF-alpha), and vascular endothelial growth factor (VEGF). The proliferation of glial cells as well as the infiltration of inflammatory cells were also evaluated. Although astroglia proliferated from 72 hours post-injury, inflammatory cell dynamics were generally steady. Among cytokines analyzed in the present study, IL-1beta, IL-5, IL-6, IL-12 p40, G-CSF, IFN-gamma, KC, LIF, MIP2, and PDGF BB increased during the early phase of cerebral wound healing, and M-CSF increased during the middle phase, while IL-15, IL-18, and MIG increased during the late phase. In contrast, IL-1alpha, IL-10, IL-12 p70, and TNF-alpha were suppressed throughout the cerebral wound healing process. Based on our findings, quantitative cytokine analyses at the cerebral wound site may be a useful tool for wound age estimation. Further, this study suggests that multiplex data gained from the same sample using a single methodology demonstrates highly accurate cytokine interactions during the process of cerebral wound healing.

Screening of Biochemical and Molecular Mechanisms of Secondary Injury and Repair in the Brain after Experimental Blast-Induced Traumatic Brain Injury in Rats

“…No increase in MIP-1a was noted after a mouse stab wound, suggesting differences between blast and penetrating TBI. 43 Surprisingly, we noted an increase of IL-1a at 2 h, but not IL-1b. This corroborates our gene array data.…”

“…42 Previous work in central nervous system (CNS) injury has shown varied cytokine patterns, depending on the injury mechanism. Takamiya and colleagues 43 applied a 27-plex over 240 h after stab wound and noted elevations of proinflammatory cytokines (IL-1a, IL-1b, IL-6, IL-12p40, IL-12p70, IL-18, and TNF-a), anti-inflammatory cytokines (IL-5 and IL-10), and chemokines (G-CSF, GRO/KC, and MIP-2) at various time points. Buttram and colleagues 44 applied multiplex to CSF of TBI patients and noted increases of proinflammatory cytokines (IL-1b, IL-6, and IL-12p70), an anti-inflammatory cytokine (IL-10), and chemokines (IL-8 and MIP-1a).…”

Glioblastoma-synthesized G-CSF and GM-CSF contribute to growth and immunosuppression: Potential therapeutic benefit from dapsone, fenofibrate, and ribavirin

“…24,25 Post-traumatic hypoxia in patients with closed head trauma increases GM-CSF in the cerebrospinal fluid. 26 Whole body radiation that included brain induced increased synthesis of G-CSF that, although it contributes to BM recovery, 27 contributed to systemic immunosuppression (see section "Elevated G(M)-CSF is immunosuppressive in GB").…”

Vitality and wound-age estimation in forensic pathology: review and future prospects

“…For example, because brain damage is frequently involved in cases of violent death and is typically fatal, much research has focused on estimating wound age in cases involving brain damage [11][12][13]. Skeletal muscle and skin have also been the subjects of most of the recent experimental and investigative studies, respectively.…”