Randomized comparison of single dose of recombinant human IL-12 versus placebo for restoration of hematopoiesis and improved survival in rhesus monkeys exposed to lethal radiation

Zoya Gluzman-Poltorak, Sarita R Mendonca, Vladimir Vainstein, Hue Kha, Lena A Basile, Zoya Gluzman-Poltorak, Sarita R Mendonca, Vladimir Vainstein, Hue Kha, Lena A Basile

Abstract

Background: The hematopoietic syndrome of the acute radiation syndrome (HSARS) is a life-threatening condition in humans exposed to total body irradiation (TBI); no drugs are approved for treating this condition. Recombinant human interleukin-12 (rHuIL-12) is being developed for HSARS mitigation under the FDA Animal Rule, where efficacy is proven in an appropriate animal model and safety is demonstrated in humans.

Methods: In this blinded study, rhesus monkeys (9 animals/sex/dose group) were randomized to receive a single subcutaneous injection of placebo (group 1) or rHuIL-12 at doses of 50, 100, 250, or 500 ng/kg (groups 2-5, respectively), without antibiotics, fluids or blood transfusions, 24-25 hours after TBI (700 cGy).

Results: Survival rates at Day 60 were 11%, 33%, 39%, 39%, and 50% for groups 1-5, respectively (log rank p < 0.05 for each dose vs. control). rHuIL-12 also significantly reduced the incidences of severe neutropenia, severe thrombocytopenia, and sepsis (positive hemoculture). Additionally, bone marrow regeneration following TBI was significantly greater in monkeys treated with rHuIL-12 than in controls.

Conclusions: Data from this study demonstrate that a single injection of rHuIL-12 delivered one day after TBI can significantly increase survival and reduce radiation-induced hematopoietic toxicity and infections. These data significantly advance development of rHuIL-12 toward approval under the Animal Rule as an effective stand-alone medical countermeasure against the lethal effects of radiation exposure.

Figures

Figure 1
Figure 1
Survival of rhesus monkeys following exposure to TBI and treatment 24 hours after TBI with either vehicle or rHuIL-12. Kaplan-Meier plots of survival over the study period are shown for each treatment group. Each dose group comprised 18 animals. Log rank p-values were 0.0305 0.0344, 0.0404, and 0.0265, respectively for the 50 ng/kg, 100 ng/kg, 250 ng/kg and 500 ng/kg dose groups vs. the vehicle-treated control group.
Figure 2
Figure 2
Blood counts over time in rhesus monkeys exposed to lethal TBI and treated 24 hours after TBI with either vehicle or rHuIL-12 (Average ± SEM). A) platelets; B) mean platelet volume; C) neutrophils; D) lymphocytes; E) reticulocytes; F) red blood cells. Normal ranges are as follows: platelets, 252 to 612 × 109/L; mean platelet volume, 6.3 to 9.4 × 109/L; neutrophils, 1.21 to 10.29 × 109/L; lymphocytes, 1.85 to 8.71 × 109/L; reticulocytes, 29.9 to 103.9 × 109/L; red blood cells, 4.95 to 6.94 × 1012/L.
Figure 3
Figure 3
Identification of Bone Marrow Regeneration Islands. (A) Histopathological identification of regenerating bone marrow. Clusters of cells appearing in otherwise ablated bone marrow were scored as one regenerating island. Left panel, ablated bone marrow; middle panel, regenerating bone marrow; right panel, non-irradiated bone marrow. (Olympus BX41 compound microscope; Infinity Analyze software v5.0, magnification: 10×). (B) Quantification of number of islands of regeneration for individual treatment groups (left panel, p < 0.01 for 500 ng/kg group vs. control) and the combined rHUIL-12-treated groups vs. vehicle-treated control (right panel, p < 0.05). (C) Quantification of area of regeneration for individual treatment groups (left panel, p < 0.05 for 50 and 500 ng/kg groups vs. control) and the combined rHuIL-12-treated groups vs. vehicle-treated control (right panel, p < 0.05). (D) Quantification of megakaryocytes for individual treatment groups (left panel) and the combined rHuIL-12-treated groups vs. vehicle-treated control (right panel).

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