Effects of rapamycin combined with low dose prednisone in patients with chronic immune thrombocytopenia

Jiaming Li, Zhaoyue Wang, Lan Dai, Lijuan Cao, Jian Su, Mingqing Zhu, Ziqiang Yu, Xia Bai, Changgeng Ruan, Jiaming Li, Zhaoyue Wang, Lan Dai, Lijuan Cao, Jian Su, Mingqing Zhu, Ziqiang Yu, Xia Bai, Changgeng Ruan

Abstract

We conducted this randomized trial to investigate the efficacy and safety of rapamycin treatment in adults with chronic immune thrombocytopenia (ITP). Eighty-eight patients were separated into the control (cyclosporine A plus prednisone) and experimental (rapamycin plus prednisone) groups. The CD4⁺CD25⁺CD127(low) regulatory T (Treg) cells level, Foxp3 mRNA expression, and the relevant cytokines levels were measured before and after treatment. The overall response (OR) was similar in both groups (experimental group versus control group: 58% versus 62%, P = 0.70). However, sustained response (SR) was more pronounced in the experimental group than in the control group (68% versus 39%, P < 0.05). Both groups showed similar incidence of adverse events (7% versus 11%, P = 0.51). As expected, the low pretreatment baseline level of Treg cells was seen in all patients (P < 0.001); however, the experimental group experienced a significant rise in Treg cell level, and there was a strong correlation between the levels of Treg cells and TGF-beta after the treatment. In addition, the upregulation maintained a stable level during the follow-up phase. Thus, rapamycin plus low dose prednisone could provide a new promising option for therapy of ITP.

Figures

Figure 1
Figure 1
Immunologic assessment before and after the rapamycin or cyclosporine treatment. (a) The Treg cells levels by the flow cytometry. Patients with CR and PR in the experimental group experienced a marked upregulation (CR group: 5.35 ± 1.49% versus 7.55 ± 1.74%, P = 0.013; PR group: 5.17 ± 1.65% versus 6.12 ± 1.43%, P = 0.015); (b) the expression of Foxp3 mRNA by quantitative RT-PCR assay. It was consistent with the increase in Treg cells level (CR group: 44 ± 19 versus 63 ± 13, P = 0.017; PR group: 44 ± 21 versus 58 ± 18, P = 0.005); (c) the plasma levels of TGF-β by the enzyme-linked immunosorbent assay. A significant increase in the level of TGF-β was observed after the rapamycin treatment (CR group: 2.38 ± 1.0 ng/mL versus 3.57 ± 1.1 ng/mL, P = 0.028; PR group: 2.04 ± 0.92 ng/mL versus 2.90 ± 0.8 ng/mL, P = 0.001).
Figure 2
Figure 2
The suppressive activity of Treg cells before and after the treatment in the experimental group; (a) the suppressive ability for CD4+ T cells; (b) the suppressive ability for CD8+ T cells. The pretreatment baseline levels of Treg cells suppression was significantly lower for CD4+ cells and CD8+ cells than those in the healthy controls (6.87 ± 1.24% versus 11.28 ± 1.60%, P < 0.001; 6.86 ± 0.95% versus 10.93 ± 2.04%, P < 0.005). Following the rapamycin treatment, the suppression for CD4+ cells (9.15 ± 0.64%, P = 0.007) and CD8+ cells (8.89 ± 0.73%, P = 0.001) was stronger than that before the treatment.
Figure 3
Figure 3
FCM analysis of suppressive Treg cells in vitro; (a) the suppression for CD4+ T cells by FCM analysis; (b) the suppression for CD8+ T cells by FCM analysis. The Treg cells were isolated from patient 5 (“A” means the Treg cells from the patient who was not given the rapamycin treatment; “B” means the Treg cells from the same patient who obtained CR), and their freshly purified autologous effector CD4+ T or CD8+ T cells were stimulated with anti-CD3 and anti-CD28. They were cocultured in culture medium. The cell division was assessed at day 5 after stimulation by FACS analysis of CFSE dilution. Suppression of responder cell proliferation is indicated as %.

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Source: PubMed

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