The dismal prognosis of pediatric and young adult patients with high-risk rhabdomyosarcoma (RMS) underscores the need for novel treatment options for this patient group. In previous studies, the tumor-associated surface antigen ERBB2 (HER2/neu) was identified as targetable in high-risk RMS. As a proof of concept, in this study, a novel treatment approach against RMS tumors using a genetically modified natural killer (NK)-92 cell line (NK-92/5.28.z) as an off-the-shelf ERBB2-chimeric antigen receptor (CAR)-engineered cell product was preclinically explored. In cytotoxicity assays, NK-92/5.28.z cells specifically recognized and efficiently eliminated RMS cell suspensions, tumor cell monolayers, and 3D tumor spheroids via the ERBB2-CAR even at effector-to-target ratios as low as 1:1. In contrast to unmodified parental NK-92 cells, which failed to lyse RMS cells, NK-92/5.28.z cells proliferated and became further activated through contact with ERBB2-positive tumor cells. Furthermore, high amounts of effector molecules, such as proinflammatory and antitumoral cytokines, were found in cocultures of NK-92/5.28.z cells with tumor cells. Taken together, our data suggest the enormous potential of this approach for improving the immunotherapy of treatment-resistant tumors, revealing the dual role of NK-92/5.28.z cells as CAR-targeted killers and modulators of endogenous adaptive immunity even in the inhibitory tumor microenvironment of high-risk RMS.

The behavior of an artificial immune complex was investigated in 15 rabbits. The immune complex was labeled with iodine-125 (125I). The advantage was that the immune complex could not be metabolized without being eliminated by the kidney. This artificial immune complex was injected in a dosage of 0.8 microgram, in 2 rabbits, 5.0 micrograms in 4 rabbits, 10 micrograms in 3 rabbits, 20 micrograms in 2 rabbits, and 40 micrograms in 4 rabbits on two consecutive days. One group (n = 9) was treated with plasmapheresis: the other (n = 6) was a control group that did not undergo plasmapheresis. A miniaturized plasmapheresis system eliminated the immune complex very effectively with an average exchange volume of 95.7 ml per treatment. The increasing concentration of the immune complex in the blood before the next plasmapheresis treatment was probably an expression of mobilization of the immune complex from different organs. The animals of the plasmapheresis group were in better condition than the control group.