In vivo administration of granulocyte colony-stimulating factor (G- CSF), granulocyte-macrophage CSF, interleukin-1 (IL-1), and IL-4, alone and in combination, after allogeneic murine hematopoietic stem cell transplantation

Abstract

BALB/c mice (H-2d) given 10 Gy total body irradiation (TBI) followed by 10(7) bone marrow (BM) and 10(6) spleen cells from C57BL/6 (H-2b) donor mice received recombinant cytokines intraperitoneally (IP) twice daily. The effect on neutrophil recovery rate, graft-v-host disease (GVHD), and survival was assessed. Four reagents were used: granulocyte-colony- stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF), interleukin-1 (IL-1) and IL-4, both alone and in combination. The most effective combination for increasing the circulating absolute neutrophil account (ANC) above the control value at day 7 posttransplant was the combination of G-CSF and IL-1 (mean ANC 2.4 +/- 1.6 x 10(9)/L as compared with control value of 0.07 +/- 0.05, P less than .02), followed by G-CSF alone (mean ANC 1.1 +/- 0.2, P less than .0001), the combination of GM-CSF plus IL-1 (mean ANC 0.8 +/- 0.3, P less than .002), and the combination of G-CSF plus GM-CSF (mean ANC 0.8 +/- 0.3, p less than .005). At day 10 posttransplant, the most effective combination in raising the ANC was the combination of G-CSF plus GM-CSF (mean ANC 7.5 +/- 2.3 as compared with control value of 3.5 +/- 1.1, P less than .01), followed by G-CSF alone (mean ANC 6.9 +/- 2.1, P less than .02). At the doses used, neither G-CSF nor GM-CSF had a deleterious effect on the incidence or severity of GVHD; indeed, GM- CSF was associated with improved survival. In contrast, IL-1 at doses greater than or equal to 100 ng twice daily caused marked early mortality, and there was a suggestion that IL-4 at doses of 500 ng twice daily resulted in increased late mortality, possibly owing to exacerbation of GVHD. This model appears to be of value for exploring the use of hematopoietic growth factors before they are used clinically in marrow allograft recipients.

The Immunopathophysiology of Acute Graft‐Versus‐Host‐Disease

“…A similar increase in mononuclear cell IL‐1 mRNA has been shown during clinical acute GVHD [99]. Indirect evidence for a role of IL‐1 in GVHD was obtained with administration of this cytokine to hosts in an allogeneic murine BMT model [100]. Mice receiving IL‐1 displayed a wasting syndrome and increased mortality that appeared to be an accelerated form of disease.…”

“…Because IL‐4 is more important than IL‐10 in directing precursor T cells toward a Th2 phenotype [124], the administration of IL‐4 to prevent GVHD may be more promising. Intraperitoneal injections of rIL‐4 led to an increase in the severity of GVHD and to enhanced late mortality in a fully allogeneic BMT model [100]. Recipients of IL‐4 showed the same histopathologic changes in skin and liver as mice receiving no cytokine.…”

Cytokine Dysregulation in Acute Graft-versus-Host Disease

“…Lesions in GVHD target tissues are clearly associated with high levels of TNF-o: and IL-l [87][88][89]. The systemic infusion of these inflammatory cytokines induces cell necrosis and cachexia [79,87] and TNF-o: appears to be partially responsible for the pronounced lymphocyte suppression associated with acute GVHD. The later effect is mediated, at !east in part, by excess nitric oxide secretion [90].…”

Validation of a serum‐free growth factor‐replenished in vitro culture system for hematopoietic progenitor cells in healthy donors and recipients of an allogeneic bone marrow graft

“…However, our study suggests that a combined use of IL-3 and GM-CSF might give support to initial reconstitution of hematopoiesis after BMT. Refinement of the in-~itro test conditions, also adding varying doses of various rH-GF's at different time intervals to the cultures, may be essential for a better insight into the optimal effects of rH-GF's (36).…”