Novel Strategy for Non-Invasive Sampling of Epidermal Cytokines Using a Skin Sampling Disc in Acute Skin Graft Versus Host Disease and Engraftment Syndrome

Novel Strategy for Non-invasive Sampling of Epidermal Cytokines Using a Skin Sampling Disc in Acute Skin Graft versus Host Disease and Engraftment syndrome.

Introduction

The local cytokine milieu of target organs is poorly described in acute graft versus host disease (aGVHD) or engraftment syndrome (ES).  A novel and noninvasive methodology using D-Squame (Cuderm Dallas, TX) skin sampling discs was utilized to collect the superficial layer of the stratum corneum in patients with acute skin GVHD or ES and cytokines were measured from the keratinocytes which adhered to the disc.

Methods

D-squame skin sampling discs were placed on the forearm/wrist of 13 pediatric patients (ages 0.25 years-14 years) for 2 minutes and removed. In patients with acute skin GVHD or ES, samples were obtained from the involved area of the forearm/wrist. This procedure was noted to be painless and safe even for young infants.  Superficial keratinocytes were sampled by the fully cured medical grade polyacrylate ester adhesive on the disc. Cytokines were extracted from the sampling discs using a buffer made of phosphate buffered saline and 0.25 molar saline and analyzed using a 38 plex human cytokine Milliplex MAP magnetic bead panel (EMD Millipore Corp, Billerica, MA). Patients were divided into 4 groups; acute skin GVHD (n=5), isolated engraftment syndrome (n=2), no acute GVHD or ES (n=2) and healthy pediatric controls (n=4).

               

Results

IL1a was elevated in patients with aGVHD as compared to healthy controls and patients who did not develop aGVHD or ES ( p= 0.02 and p= 0.04)  Similarly IL1a was elevated in ES as compared to patients who did not develop aGVHD/ES or to healthy pediatric controls ( p=0.05 and p=0.01).

Conclusions

These preliminary results demonstrate a novel methodology to measure inflammatory cytokines locally in acute skin GVHD and ES.  With further patients and analysis, new approaches to management may be indicated in the future.

Figure 1. Average values of IL1a measured from the epidermis in patients with ES, aGVHD, no ES or aGVHD and healthy pediatric controls.