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The Profile of T Helper Subsets in Bone Marrow Microenvironment Is Distinct for Different Stages of Acute Myeloid Leukemia Patients and Chemotherapy Partly Ameliorates These Variations.

Tian T, Yu S, Liu L, Xue F, Yuan C, Wang M, Ji C, Ma D - PLoS ONE (2015)

Bottom Line: BM plasma Th-associated cytokines levels were determined by ELISA.Th22, Th17, Th1, Th2 cells, IL-22 and RORC expression were significantly decreased, while Treg cells, related cytokine IL-10 and transcription factor Foxp3 were markedly elevated in ND compared to CR patients or controls.Our findings suggest that these cells and cytokines may be implicated in AML pathogenesis and provided therapeutic insights.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Qilu Hospital, Shandong University, Jinan, China.

ABSTRACT

Background: T helper (Th) cells immune regulation is important for the pathogenesis of acute myeloid leukemia (AML). Recurrent Th abnormalities in AML peripheral blood were reported, while the comprehensive status of various Th subsets is rarely investigated in bone marrow (BM) microenvironment which is the origin of AML leukemic blast cells.

Methods: BM was extracted from 48 newly-diagnosed (ND), 34 complete-remission (CR), 19 relapsed-refractory AML patients and 15 controls. Slight iron deficiency anemia patients were used as controls. Th subsets frequencies were examined by flow cytometry. BM plasma Th-associated cytokines levels were determined by ELISA. The expression of key transcription factor was examined by RT-PCR.

Results: Th22, Th17, Th1, Th2 cells, IL-22 and RORC expression were significantly decreased, while Treg cells, related cytokine IL-10 and transcription factor Foxp3 were markedly elevated in ND compared to CR patients or controls. Meanwhile, the imbalanced Th1/Th2 and Th17/Treg ratio were observed in ND and relapsed-refractory patients. Negative correlation between Th1 or Th2 and peripheral WBC, between Th17/Treg or Th1/Th2 and leukemic blast existed in ND patients. Moreover, chemotherapy ameliorated these variations.

Conclusion: Th subsets in BM are distinct for different stages of AML and chemotherapy partly ameliorates the abnormality. Our findings suggest that these cells and cytokines may be implicated in AML pathogenesis and provided therapeutic insights.

No MeSH data available.


Related in: MedlinePlus

The percentage of Th22 cells in representative patients with ND, CR, relapsed-refractory AML patients or in controls.Heparinized bone marrow from all subjects was stimulated with phorbol myristate acetate, ionomycin, and monensin for 4 h or BM mononuclear cells were isolated by Ficoll-Hypaque gradient centrifugation and then stained with labeled antibodies, as described in the “Materials and methods” section. A. Lymphocytes were gated by flow cytometry. B. The percentage of CD4+IFN-γ- T cells in AML patients and controls. C, D, E, F. The percentage of Th22 (CD4+IFN-γ-IL-17-IL-22+) cells in AML patients and controls.
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pone.0131761.g001: The percentage of Th22 cells in representative patients with ND, CR, relapsed-refractory AML patients or in controls.Heparinized bone marrow from all subjects was stimulated with phorbol myristate acetate, ionomycin, and monensin for 4 h or BM mononuclear cells were isolated by Ficoll-Hypaque gradient centrifugation and then stained with labeled antibodies, as described in the “Materials and methods” section. A. Lymphocytes were gated by flow cytometry. B. The percentage of CD4+IFN-γ- T cells in AML patients and controls. C, D, E, F. The percentage of Th22 (CD4+IFN-γ-IL-17-IL-22+) cells in AML patients and controls.

Mentions: Th22 was defined as CD4+IFN-γ-IL-17-IL-22+ T cells to exclude Th1 or Th17 cells (Fig 1). Fig 1 is the representative Th22 figure. Th22 frequency in ND (median 1.03%; range, 0.14–7.02%) or relapsed-refractory AML patients (median, 1.02%; range, 0.33–1.66%) was significantly lower than that in controls (median, 2.03%; range, 0.70–19.08%, *P = 0.0263; *P = 0.0109; respectively). For the different AML stages, Th22 frequency in ND or relapsed-refractory AML patients was significantly lower than that in CR AML (median, 1.56%; range, 0.21–6.58%; *P = 0.0118; *P = 0.0067; respectively). No statistical decrease was shown in CR patients compared with controls (P>0.05) (Fig 2A). We also observed a significant decrease of BM plasma IL-22 level in ND patients (median, 77.63 pg/ml; range 71.73–83.53 pg/ml; *P = 0.0453) and CR patients (median, 68.28 pg/ml; range, 59.83–76.73 pg/ml; *P = 0.0340) compared with the level in controls (median, 100.78 pg/ml; range, 63.08–138.48 pg/ml). No significant difference of BM IL-22 level between the three AML groups (P >0.05) was observed (Fig 2B). No correlation was found between Th22 and IL-22 in BM of AML patients or controls. Meanwhile, BM Th1 and Th17 cells failed to show statistical correlations with IL-22 in our present research.


The Profile of T Helper Subsets in Bone Marrow Microenvironment Is Distinct for Different Stages of Acute Myeloid Leukemia Patients and Chemotherapy Partly Ameliorates These Variations.

Tian T, Yu S, Liu L, Xue F, Yuan C, Wang M, Ji C, Ma D - PLoS ONE (2015)

The percentage of Th22 cells in representative patients with ND, CR, relapsed-refractory AML patients or in controls.Heparinized bone marrow from all subjects was stimulated with phorbol myristate acetate, ionomycin, and monensin for 4 h or BM mononuclear cells were isolated by Ficoll-Hypaque gradient centrifugation and then stained with labeled antibodies, as described in the “Materials and methods” section. A. Lymphocytes were gated by flow cytometry. B. The percentage of CD4+IFN-γ- T cells in AML patients and controls. C, D, E, F. The percentage of Th22 (CD4+IFN-γ-IL-17-IL-22+) cells in AML patients and controls.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4489914&req=5

pone.0131761.g001: The percentage of Th22 cells in representative patients with ND, CR, relapsed-refractory AML patients or in controls.Heparinized bone marrow from all subjects was stimulated with phorbol myristate acetate, ionomycin, and monensin for 4 h or BM mononuclear cells were isolated by Ficoll-Hypaque gradient centrifugation and then stained with labeled antibodies, as described in the “Materials and methods” section. A. Lymphocytes were gated by flow cytometry. B. The percentage of CD4+IFN-γ- T cells in AML patients and controls. C, D, E, F. The percentage of Th22 (CD4+IFN-γ-IL-17-IL-22+) cells in AML patients and controls.
Mentions: Th22 was defined as CD4+IFN-γ-IL-17-IL-22+ T cells to exclude Th1 or Th17 cells (Fig 1). Fig 1 is the representative Th22 figure. Th22 frequency in ND (median 1.03%; range, 0.14–7.02%) or relapsed-refractory AML patients (median, 1.02%; range, 0.33–1.66%) was significantly lower than that in controls (median, 2.03%; range, 0.70–19.08%, *P = 0.0263; *P = 0.0109; respectively). For the different AML stages, Th22 frequency in ND or relapsed-refractory AML patients was significantly lower than that in CR AML (median, 1.56%; range, 0.21–6.58%; *P = 0.0118; *P = 0.0067; respectively). No statistical decrease was shown in CR patients compared with controls (P>0.05) (Fig 2A). We also observed a significant decrease of BM plasma IL-22 level in ND patients (median, 77.63 pg/ml; range 71.73–83.53 pg/ml; *P = 0.0453) and CR patients (median, 68.28 pg/ml; range, 59.83–76.73 pg/ml; *P = 0.0340) compared with the level in controls (median, 100.78 pg/ml; range, 63.08–138.48 pg/ml). No significant difference of BM IL-22 level between the three AML groups (P >0.05) was observed (Fig 2B). No correlation was found between Th22 and IL-22 in BM of AML patients or controls. Meanwhile, BM Th1 and Th17 cells failed to show statistical correlations with IL-22 in our present research.

Bottom Line: BM plasma Th-associated cytokines levels were determined by ELISA.Th22, Th17, Th1, Th2 cells, IL-22 and RORC expression were significantly decreased, while Treg cells, related cytokine IL-10 and transcription factor Foxp3 were markedly elevated in ND compared to CR patients or controls.Our findings suggest that these cells and cytokines may be implicated in AML pathogenesis and provided therapeutic insights.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Qilu Hospital, Shandong University, Jinan, China.

ABSTRACT

Background: T helper (Th) cells immune regulation is important for the pathogenesis of acute myeloid leukemia (AML). Recurrent Th abnormalities in AML peripheral blood were reported, while the comprehensive status of various Th subsets is rarely investigated in bone marrow (BM) microenvironment which is the origin of AML leukemic blast cells.

Methods: BM was extracted from 48 newly-diagnosed (ND), 34 complete-remission (CR), 19 relapsed-refractory AML patients and 15 controls. Slight iron deficiency anemia patients were used as controls. Th subsets frequencies were examined by flow cytometry. BM plasma Th-associated cytokines levels were determined by ELISA. The expression of key transcription factor was examined by RT-PCR.

Results: Th22, Th17, Th1, Th2 cells, IL-22 and RORC expression were significantly decreased, while Treg cells, related cytokine IL-10 and transcription factor Foxp3 were markedly elevated in ND compared to CR patients or controls. Meanwhile, the imbalanced Th1/Th2 and Th17/Treg ratio were observed in ND and relapsed-refractory patients. Negative correlation between Th1 or Th2 and peripheral WBC, between Th17/Treg or Th1/Th2 and leukemic blast existed in ND patients. Moreover, chemotherapy ameliorated these variations.

Conclusion: Th subsets in BM are distinct for different stages of AML and chemotherapy partly ameliorates the abnormality. Our findings suggest that these cells and cytokines may be implicated in AML pathogenesis and provided therapeutic insights.

No MeSH data available.


Related in: MedlinePlus