HMGB1 and cancer: Indeed, induction of necrosis by freeze-thawing of otherwise untreated cancer cells failed to yield an immunogenic preparation; and freeze-thawing actually destroyed the immunogenic properties of anthracycline-treated cells.4 Other hallmarks of ICD, namely the exodus of HMGB1 from the nuclei of dead cells and the induction of a type-1 interferon response occur in a largely caspase-independent manner.9, 23