Comparison of Immune Responses Induced by Deferoxamine and Deferasirox

Yeong Suk Jung; E Young Bae; Nack Gyun Chung; Bin Cho; Hack Ki Kim; Chang Ki Min; Chi Wha Han; Ho Shik Kim; Dae Chul Jeong

doi:10.5045/kjh.2008.43.3.150

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Jung, Bae, Chung, Cho, Kim, Min, Han, Kim, and Jeong: Comparison of Immune Responses Induced by Deferoxamine and Deferasirox

Original Article

Korean J Hematol 2008;43(3):150-158.

Published online: 19 September 2008

DOI: https://doi.org/10.5045/kjh.2008.43.3.150

Comparison of Immune Responses Induced by Deferoxamine and Deferasirox

Yeong Suk Jung, M.D., E Young Bae, M.D., Nack Gyun Chung, M.D., Bin Cho, M.D., Hack Ki Kim, M.D., Chang Ki Min, M.D.¹, Chi Wha Han, M.D.¹, Ho Shik Kim, M.D.², Dae Chul Jeong, M.D.

Departments of Pediatrics, Seoul, Korea

¹Internal Medicine, Seoul, Korea

²Biochemistry, College of Medicine, The Catholic University of Korea, Seoul, Korea

Correspondence to：Dae Chul Jeong, M.D. Department of Pediatrics, College of Medicine, The Catholic University of Korea 665, Bupyeong 6-dong, Bupyeong-gu, Incheon 403-720, Korea Tel: ＋82-32-510-5687, Fax: ＋82-32-503-9724 E-mail: dcjeong@catholic.ac.kr

Received 27 May 2008 Revised 21 July 2008 Accepted 24 July 2008

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background:

The iron chelating agents (ICA) have various biological effects besides iron chelation. We investigated the immunomodulatory effects of Deferasirox (DFS) compared to Deferoxamine (DFO).

Methods:

Spleen cells (SP) were obtained from 5 week-old C57/BL6 (H-2^b). The cytotoxicity of ICAs was examined using the CCK8 method. For the cell proliferation assay, SP were cultured with irradiated in addition to 10, 50, 100μM of DFS or DFO and 200ng/mL of cyclosporin A (CSA). Cytokines and nitrite levels were evaluated from supernatants by ELISA.

Results:

The viability of ICA was reported to be over 100%. Both DFS and DFO inhibited cell proliferation in a manner comparable to CSA. Cell proliferation without iron was reduced at the concentration of 100μM of DFO. With iron treatment, the reduction of the stimulation index was dependent on DFO concentrations. DFS decreased the proliferation without reference to the concentrations. After stimulation of phytohemagglutinin, the nitrite concentrations increased with iron. With lipopolysaccharides, the nitrite levels were higher in DFO with iron than control, but similar in DFS regardless of iron treatment. The levels of interleukin-2 were not different. Interleukin-10 was more abundantly produced in 50μM of DFO compared to DFS. Transforming growth factor-β was higher in DFS than DFO at the low concentration, but opposite at the high concentration.

Conclusion:

These data suggested that both iron chelating agents possessed immune suppressive effects comparable to CSA. The immunosuppressive effect of DFS may be distinct from DFO. More experiments are required to determine the exact mechanism of the immunosuppressive effect of DFS.

Keywords: Deferoxamine, Deferasirox, Immunosuppressive effect

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Fig. 1

Cell viability according to concentrations of iron chelating agents (ICA) by CCK 8 method. (A) There is no difference in cell viability among various concentrations of ICA without ferric chloride (n=5). (B) The viability is significantly diminished at 20μM ferric chloride, but there are no difference in viability among ICA in spite of presence of iron (n=5) (∗: P＜0.05). Abbreviations: DFS, deferasirox; DFO, deferoxamine; FeCl₃, ferric chloride.

Fig. 2

Stimulation index (SI) of C57/BL6 spleen cells (SP) and irradiated BALB/c SP under 5μg/mL of phytohemagglutinin with 1：1 ratio according to the concentration of iron chelating agents (ICA) (n=7) without (A) and with (B) ferric chloride. There are significant differences between cyclosporin A (CSA) and ICA with or without ferric chloride (^†: P＜0.05). (A) The high level (100μM) of deferoxamine (DFO) exerts profound inhibition of cell proliferation than deferasirox (DFS) (∗: P＜0.05), although it is not different at 10 and 50μM of DFO. Deferasirox (DFS) showed no difference in SI among any concentrations. (B) The decrement of SI is dependent on concentration in DFO (^†: P＜0.01), but is not in DFS. There is a significant difference in SI between DFO and DFS at 10μM (∗: P＜0.01).

Fig. 3

The levels of nitrite in supernatants from cell proliferation assay with or without ferric chloride according to the concentration of iron chelating agents by non-specific mitogen (A; phytohemagglutinin (PHA), B; lipopolysaccharide (LPS)). All nitrite levels after stimulation are not different according to PHA or LPS. (A) There is lower nitrite in 10μM of Deferoxamine (DFO) and Deferasirox (DFS) among ICA without iron, and higher at 10μM of DFS than other ICA with iron (^†: P＜0.01). (B) The levels of nitrite are not different among ICA without iron, but higher only in DFO with iron (^‡: P＜0.01). ∗: P＜0.05 in each ICA concentration according to presence with iron. Abbreviations: DFS, deferasirox; DFO, deferoxamine.

Table 1.

The level of cytokines in supernatants from cell proliferation assay according to the concentration of iron chelating agents without ferric chloride (n=4∼6)

(pg/mL)	Control	CSA	10μM		50μM		100μM
(pg/mL)	Control	CSA	DFO	DFS	DFO	DFS	DFO	DFS
IL-2	49.7±7.2	17.9±10.4^†	41.4±16.3	46.9±7.3	36.9±14.3	40.8±12.0	55.7±9.9	25.1±16.5
IL-10	18.2±7.0	15.5±2.8	25.1±5.0	12.8±7.8	14.4±0.1∗	7.4±4.7	12.8±5.2	10.3±5.3
TGF-β	1,575.8±473.4	1,424.8±466.7	1,248.8±82.2	1,567.0±472.3∗	1,222.2±82.1 1	1,133.4±151.9	1,362.9±402.3∗	1,279.9±39.7

Abbreviations: CSA, cyclosporin A; DFO, deferrioxamine; DFS, deferasirox.

^∗ : P＜0.05 between DFO and DFS,

^† : P＜0.01 among experimental group. 9971267239

Table 2.

The level of cytokines in supernatants from cell proliferation assay according to the concentration of iron chelatingagents with ferric chloride (n=4∼6)

(pg/mL)	Control	CSA	10μM		50μM		100μM
(pg/mL)	Control	CSA	DFO	DFS	DFO	DFS	DFO	DFS
IL-2	42.4±10.6	29.8±16.8	30.5±17.3	41.7±12.4	62.6±23.8	33.41±15.3	49.3±32.3	47.9±23.3
IL-10	21.1±8.0	19.2±5.9	7.6±3.4	13.2±12.1	41.0±11.5	8.06±3.4	7.3±1.7	30.5∗±11.3
TGF-β	1,066.6±53.6	1,376.8±317.2	1,355.0±276.0	1,672.7±372.1^†	1,374.7±252.7 1	1,498.9±352.1	1 1,202.0±2.7	1,225.9±66.8

Abbreviations: CSA, cyclosporin A; DFO, deferrioxamine; DFS, deferasirox. ∗: P＜0.05 between DFO and DFS,

^† : P＜0.05 among experimental group.

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