SIGNIFICANT REDUCTION OF REGULATORY INNATE LYMPHOID CELLS (ILCREGS, CD45+LIN-CD127+IL-10+) AND CD45+LIN-CD127+IL-10- CELLS IN PATIENTS WITH ACUTE MYELOID LEUKEMIA DETECTED BY FLOW CYTOMETRY
Author(s): ,
Jifeng Yu
Affiliations:
Department of Hematology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou,China
,
Chen He
Affiliations:
Department of Hematology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou,China
,
Yingmei Li
Affiliations:
Department of Hematology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou,China
,
Qiutang Zhang
Affiliations:
Department of Hematology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou,China
Zhongxing Jiang
Affiliations:
Department of Hematology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou,China
EHA Library. Yu J. Jun 14, 2019; 266024; PF234
Jifeng Yu
Jifeng Yu
Contributions
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Abstract

Abstract: PF234

Type: Poster Presentation

Presentation during EHA24: On Friday, June 14, 2019 from 17:30 - 19:00

Location: Poster area

Background
A new regulatory subpopulation of ILCs, ILCregs has been identified in mouse and human intestines. ILCregs share characteristics with both innate lymphoid cells and regulatory cells. The significance of CD45+Lin-CD127+IL-10+ ILCregs in patients with AML remains unclear.  Also, the CD45+Lin-CD127+IL-10- subset population has not been classified and its significance remains unknown.

Aims
To explored the difference of CD45+Lin-CD127+IL-10+ ILCregs population and the CD45+Lin-CD127+IL-10- subset population by flow cytometry between the normal donors and patients with AML.

Methods
Flow cytometry assay: Bone marrow(BM) cells from 12 normal donors and 42 patients newly diagnosed with AML were collected and cultured in media containing Bredfeldin A for 4 hr at 37°C. Cells were harvested and stained with surface markers CD45-FITC, Lin-APC,CD127-PE-Cy7 for 15 minutes. Red blood cells were lysed with lysing buffer. Cells were then fixed and permeablized by Intracellular Fixation & Permeablization buffer set (eBioscience) after surface marker staining, followed by application of anti-IL-10-PE antibody (JES5-16E3) staining intracellular IL-10 before analyzed through flow cytometry (FACS Aria III, BD). Whole cell populations with SSC vs FSC were selected and gated on the CD45+Lin- population with further gating of CD127+IL-10+ subset and CD127+IL-10- subset. At least a total of 1 million cells were collected for analysis. ILCregs cells were defined as CD45+Lin-CD127+IL-10+ population.

Results
Using the 4 colors monoclonal antibody combination, we were able to detect the ILCregs defined as CD45+Lin-CD127+IL-10+ in the BM from both normal donor and AML patients. The frequency of ILCregs in normal donors were 0.9958±1.0730% and 2.2941±2.3533% in whole BM cells from all events gating and CD45+Lin- cell population gating, respectively. In addition, the frequency of ILCregs in AML patients were 0.2434±0.5344% and 0.8924±1.3791% in whole BM cells from all events gating and CD45+Lin- cell population gating, respectively. In comparison with the normal donors, the frequency of ILCregs cells in AML patients were significantly decreased for both  whole BM cells gating and CD45+Lin- cell population gating (both P<0.01).  The frequency of CD45+Lin-CD127+IL-10- subset in normal donors were 4.0869±6.7701% and 8.2445±11.1016% in whole BM cells from all events gating and CD45+Lin- cell population gating, respectively. In addition, the frequency of CD45+Lin-CD127+IL-10- cell frequency in AML patients were 0.2769±0.2526% and 1.2053±1.7382% in whole BM cells from all events gating and CD45+Lin- cell population gating, respectively.  In comparison with the normal donors, the frequency of CD45+Lin-CD127+IL-10- cells in AML patients were also significantly decreased for both  whole BM cells gating and CD45+Lin- cell population gating (both P<0.01).

Conclusion
By using the combination of surface and IL-10 for intracellular staining analyzed by flow cytometry, we were able to detect ILCregs in  BM from both normal donors and patients with AML. The frequency of the ILCregs and CD45+Lin-CD127+IL-10- subset populations were both significantly decreased in the patients with AML comparing to normal donors. Further studies need to be done to explore the significance of ILCregs and CD45+Lin-CD127+IL-10- subset populations in AML patients.

Session topic: 3. Acute myeloid leukemia - Biology & Translational Research

Keyword(s): Acute myeloid leukemia, Flow cytometry, Regulatory T cell

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