Immune response after autologous hematopoietic stem cell transplantation in type 1 diabetes mellitus

Lei Ye, Li Li, Bing Wan, Minglan Yang, Jie Hong, Weiqiong Gu, Weiqing Wang, Guang Ning, Lei Ye, Li Li, Bing Wan, Minglan Yang, Jie Hong, Weiqiong Gu, Weiqing Wang, Guang Ning

Abstract

Background: This study explored the details of the immune response after autologous hematopoietic stem cell transplantation (AHSCT) treatment in type 1 diabetes mellitus.

Methods: Peripheral blood mononuclear cells (PBMCs) from 18 patients with type 1 diabetes mellitus were taken at baseline and 12 months after AHSCT or insulin-only therapy. The lymphocyte proliferation, mRNA expression and secretion of pro-inflammatory and anti-inflammatory cytokines belonging to T-helper type 1 (Th1), T-helper type 17 (Th17) and regulatory T (Treg) cells in PBMC culture supernatants were assessed.

Results: Compared with patients receiving insulin-only treatment, the patients receiving AHSCT treatment showed better residual C-peptide secretion, lower anti-GAD titers and less exogenous insulin dosages after 12 months of follow-up. AHSCT treatment was associated with significantly reduced Th1 and Th17 cell proportions as well as decreased IFN-γ, IL-2, IL-12p40 and IL-17A levels in the PBMC culture supernatants (all P < 0.05). Although there was no significant Treg cell expansion after AHSCT treatment, we observed increased IL-10, TGF-β and Foxp3 mRNA expression and increased TGF-β levels. However, we found no significant changes in the T-cell subpopulations after insulin treatment, except for higher IL-12p40 mRNA expression and a lower proportion of Treg cells.

Conclusions: AHSCT treatment was associated with decreased expansion and function of Th1 and Th17 cells, which may explain the better therapeutic effect of AHSCT compared with the traditional intensive insulin therapy.

Trial registration: Clinicaltrials.gov NCT00807651 . Registered 18 December 2008.

Keywords: Hematopoietic stem cell; Immune response; Regulartory T cell; Th1 cell; Th17 cell; Type 1 diabetes mellitus.

Figures

Fig. 1
Fig. 1
Flow cytometry analysis, cytokine proteins and mRNA measurements of Th1 cells from PBMCs in the AHSCT and Insulin-only groups before and after treatment (0 M and 12 M). a, b Representative flow cytometry plots of CD3+CD4+ (R2) and Th1 cells (R20). c Proportion of IFN-γ+ Th1 cells in PBMCs. d, e, f mRNA expression levels of IL-2, IL-12p40 and T-bet respectively. g, h, j Concentrations of IL-2, IL-12p40 and IFN-γ in the cell culture supernatants respectively. *P < 0.05, **P < 0.01, ***P < 0.001. AHSCT autologous hematopoietic stem cell transplantation, ns no significance, PBMC peripheral blood mononuclear cell
Fig. 2
Fig. 2
Flow cytometry analysis, cytokine proteins and mRNA measurements of Th17 cells from PBMCs in the AHSCT and Insulin-only groups before and after treatment (0 M and 12 M). a Representative flow cytometry plots of Th17 cells (R21). b Proportion of CD3+CD4+IL-17A+ Th17 cells in PBMCs. c, d mRNA expression levels of I IL-17A and ROR-rt respectively. e Concentration of IL-17A in the cell culture supernatants. *P < 0.05, **P < 0.01, ***P < 0.001. AHSCT autologous hematopoietic stem cell transplantation, ns no significance, PBMC peripheral blood mononuclear cell
Fig. 3
Fig. 3
Flow cytometry analysis, cytokine proteins and mRNA measurements of Treg cells from PBMCs in the AHSCT and Insulin-only groups before and after treatment (0 M and 12 M). a Representative flow cytometry plots of Treg cells (R3). b Proportion of CD4+CD25+CD127– Treg cells in PBMCs. c, d, e mRNA expression levels of IL-10, TGF-β and foxp3 respectively. f Concentration of TGF-β in the cell culture supernatants. *P < 0.05, **P < 0.01, ***P < 0.001. AHSCT autologous hematopoietic stem cell transplantation, ns no significance, PBMC peripheral blood mononuclear cell
Fig. 4
Fig. 4
PBMC proliferation levels detected with CCK-8 in the AHSCT and Insulin-only groups before and after treatment (0 M and 12 M). *P < 0.05, ***P < 0.001. AHSCT autologous hematopoietic stem cell transplantation, ns no significance

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