title: Development of a human whole blood assay for prediction of cytokine release similar to anti-CD28 superagonists using multiplex cytokine and hierarchical cluster analysis.
abstract: Anti-CD28 superagonist (SA) mediated cytokine release syndrome (CRS), an adverse event resulting in systemic release of cytokines, is an emergent issue in drug development. CRS is of potential concern for all monoclonal antibodies (mAbs) particularly those directed against cell surface targets on lymphocytes. Concern regarding patient safety requires development of novel methods to predict these adverse reactions. Due to the inability of animal studies to predict CRS, we have developed a whole blood in vitro screen to support First in Human studies and assess the potential for mAbs to cause anti-CD28 SA-like CRS. For this purpose we have immobilized marketed mAbs, whose potential for causing CRS and milder infusion reactions is known, on Protein A beads and used these beads to stimulate cytokine release. After culture, supernatants are harvested and frozen for later multiplex analysis of cytokines using Searchlight™ technology. We have employed hierarchicalluster analysis (HCA) to allow comparison of 12 different cytokine levels across numerous donors, treatments, and experiments. Results conclusively distinguish test mAb responses from an anti-CD28 superagonist mAb response. As part of a global analysis of preclinical data, the results of this assay can facilitate entry into First in Human clinical trials, help with selection of starting doses and may allow more rapid dose escalation using smaller cohorts.
authors.affiliation: Biologics Toxicology, Center of Excellence in Biotechnology, Centocor R&D Inc., Radnor, PA 19087, United States. mwalker2@its.jnj.com
authors.forename: Mindi R
authors.initials: MR
authors.lastname: Walker
authors.forename: Dorie A
authors.initials: DA
authors.lastname: Makropoulos
authors.forename: Ram
authors.initials: R
authors.lastname: Achuthanandam
authors.forename: Scott
authors.initials: S
authors.lastname: Van Arsdell
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Bugelski
mesh_terms: D000911:Antibodies, Monoclonal / Q000009:adverse effects*; D018106:CD28 Antigens / Q000276:immunology*; D016000:Cluster Analysis; D016207:Cytokines / Q000097:blood* / Q000276:immunology; D064420:Drug-Related Side Effects and Adverse Reactions / Q000097:blood* / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D007154:Immune System Diseases / Q000097:blood* / Q000139:chemically induced / Q000378:metabolism; D011237:Predictive Value of Tests; D011961:Receptors, Fc / Q000378:metabolism; D013577:Syndrome
chemical_list: D000911:Antibodies, Monoclonal; D018106:CD28 Antigens; D016207:Cytokines; D011961:Receptors, Fc