Evaluation of the immunogenicity of the synthetic α-melanocyte-stimulating hormone (α-MSH) analogue afamelanotide ([Nle4-D-Phe7]-α-MSH, Scenesse®) in erythropoietic protoporphyria patients by ELISA detecting both anti-afamelanotide and anti-α-MSH antibodies.

UNLABELLED: Afamelanotide is an α-melanocyte-stimulating hormone (α-MSH) agonist with proven efficacy in photodermatoses such as erythropoietic protoporphyria (EPP). This peptide drug, repeatedly administered over prolonged time, may induce anti-drug antibodies (ADA). Here, we describe a new ELISA method developed to monitor the occurrence of ADA against afamelanotide as well as against α-MSH. Covalent binding instead of absorption of antigen onto the microtitre wells prevented antigen leakage and enabled extensive washings followed by lower background. The cut-off between antibody-negative and -positive sera was determined. Inhibition of the antigen-antibody reaction by excess soluble antigen tested for specificity. The sensitivity of the ELISA was 608 and 1,390 ng/ml of specific ADA against afamelanotide and α-MSH, respectively. This ELISA method enabled us to investigate the occurrence of ADA during long-term administration of afamelanotide. No immunoreactivity was found in 23 of the 26 EPP patients exposed to the drug for up to 6 years. Pre-existing immunoreactivity against afamelanotide as well as α-MSH was found in 3 patients, whose titres did not change during afamelanotide administration. CONCLUSION: The new ELISA is suitable to determine ADA against afamelanotide and α-MSH. Afamelanotide did not elicit ADA during long-term administration in patients with EPP.

Fat marrow embolism during intramedullary bone endoscopy: an experimental study in sheep.

We present a study designed to investigate whether the intramedullary bone endoscopy (IBE) procedure within the cavity of an intact long bone will create embolic loads on the lungs similar to that of other orthopedic procedures (e.g., stem implantation in total hip arthroplasty [THA]). In a sheep model, 10 animals underwent the IBE procedure with complete perioperative anesthesiology monitoring. The lungs were harvested postoperatively and examined for fat embolisms. One animal showed evidence of intraoperative fat embolism with temporary increases in mean pulmonary arterial pressure (MPAD) and the mean CO(2)-gradient. The histological examination in this animal revealed fat embolism with a 2% surface area of the investigated fields covered with fat vacuoles. All peri- and postoperative data on the other nine animals were normal. Our findings indicate that, as with other intramedullary manipulation in intact long bones, there is a potential risk for systemic fat excavation during IBE. However, the embolic load is much lower than the rates reported for other orthopedic interventions.