|Table of Contents|

Expression of KIR2DL4 in peripheral blood of patients with acute lymphoblastic leukemia and its effect on NK cell killing function

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2023 03
Page:
450-455
Research Field:
Publishing date:

Info

Title:
Expression of KIR2DL4 in peripheral blood of patients with acute lymphoblastic leukemia and its effect on NK cell killing function
Author(s):
Adina·Wutikuer; SHI Yuwei
Department of Hematology,the Second Affiliated Hospital of Xinjiang Medical University,Xinjiang Urumqi 830000，China.
Keywords:
acute lymphoblastic leukemia; killer cell Ig-like receptor 2DL4; NK cell; kill function
PACS:
R733
DOI:
10.3969/j.issn.1672-4992.2023.03.010
Abstract:
Objective:To investigate the expression of killer cell Ig-like receptor 2DL4（KIR2DL4) in peripheral blood of patients with acute lymphoblastic leukemia（ALL),and its effect on nature killer(NK) cells of the killing function.Methods:Mononuclear cells were isolated from the peripheral blood of ALL patients and healthy volunteers,and real-time fluorescence quantitative PCR was used to detect the difference in the expression of KIR2DL4 in mononuclear cells.NK cells were sorted from mononuclear cells.The proportion of CD3-CD56+ labeled NK cells was detected by flow cytometry,and the expression levels of KIR2DL4 on NK cells and human leukocyte antigen G(HLA-G) on Molt-4 cells were analyzed.The levels of interferon-gamma(IFN-γ) and tumor necrosis factor-α(TNF-α) in cell supernatants were measured by ELISA at different potency-to-target ratios（5∶1,10∶1,20∶1) using NK cells,NK cells after the action of KIR2DL4 blocking antibody,NK cells after the action of IgG1 antibody as effector cells,and Molt-4 cells as target cells,respectively.Lactate dehydrogenase(LDH) release assay was used to detect the killing effect of NK cells with different treatments on Molt-4 cells.At the effect-target ratio of 20∶1,the morphological changes of Molt-4 cells under each treatment were observed by laser confocal microscopy,and the degranulation of NK cells in different treatments was detected by flow cytometry.Results:The relative expression of KIR2DL4 mRNA in peripheral blood of ALL patients was significantly higher than that of healthy volunteers（P＜0.05).The purity of CD3-CD56+ labeled NK cells after sorting reached was more than 90%.Compared with the co-culture of NK cells and Molt-4 cells,blocking KIR2DL4 increased the levels of NK cell-secreted factors IFN-γ and TNF-α in the cell supernatant（P＜0.05),and the killing rate of NK cells to Molt-4 cells was increased（P＜0.05).The deformation and nuclear pyknosis of Molt-4 cells were more obvious,and the expression level of CD107α in NK cells was increased（P＜0.05).Conclusion:KIR2DL4 was highly expressed in the peripheral blood of ALL patients,and blocking KIR2DL4 could promote the secretion of IFN-γ,TNF-α and degranulation by NK cells,and improve the killing function of Molt-4 cells.

References:

[1]FREGONA V,BAYET M,GERBY B.Oncogene-induced reprogramming in acute lymphoblastic leukemia:towards targeted therapy of leukemia-initiating cells［J］.Cancers（Basel),2021,13(21):5511.
[2]INABA H,MULLIGHAN CG.Pediatric acute lymphoblastic leukemia［J］.Haematologica,2020,105(11):2524-2539.
[3]HAO Q,CAO M,ZHANG C,et al.Age-related differences of genetic susceptibility to patients with acute lymphoblastic leukemia［J］.Aging（Albany NY),2021,13(9):12456-12465.
[4]ALDOSS I,PILLAI R,YANG D,et al.Late and very late relapsed acute lymphoblastic leukemia:clinical and molecular features,and treatment outcomes［J］.Blood Cancer J,2021,11(7):125.
[5]RAJAGOPALAN S,LONG EO.KIR2DL4（CD158d):An activation receptor for HLA-G［J］.Front Immunol,2012,3:258.
[6]MORADI S,BERRY R,PYMM P,et al.The structure of the atypical killer cell immunoglobulin-like receptor,KIR2DL4［J］.J Biol Chem,2015,290(16):10460-10471.
[7]朱文怡,李生君,候倩倩.KIR2DL4基因多态性及促凋亡基因HtrA2在老年白血病病人中的表达及与预后的相关性研究［J］.实用老年医学,2021,35(6):620-623. ZHU WY,LI SJ，HOU QQ.Expression of KIR2DL4 gene polymorphism and pro-apoptotic gene HtrA2 in elderly leukemia patients and its correlation with their prognosis［J］.Practical Geriatrics,2021,35(6):620-623.
[8]DEANGELO DJ,JABBOUR E,ADVANI A.Recent advances in managing acute lymphoblastic leukemia［J］.Am Soc Clin Oncol Educ Book,2020,40:330-342.
[9]MYERS JA,MILLER JS.Exploring the NK cell platform for cancer immunotherapy［J］.Nat Rev Clin Oncol,2021,18(2):85-100.
[10]BECKER PS,SUCK G,NOWAKOWSKA P,et al.Selection and expansion of natural killer cells for NK cell-based immunotherapy［J］.Cancer Immunol Immunother,2016,65(4):477-484.
[11]LIU S,GALAT V,GALAT Y,et al.NK cell-based cancer immunotherapy:from basic biology to clinical development［J］.J Hematol Oncol,2021,14(1):7.
[12]DUAULT C,KUMAR A,TAGHI KHANI A,et al.Activated natural killer cells predict poor clinical prognosis in high-risk B-and T-cell acute lymphoblastic leukemia［J］.Blood,2021,138(16):1465-1480.
[13]MAKANGA DR,DA RIN DE LORENZO F,DAVID G,et al.Genetic and molecular basis of heterogeneous NK cell responses against acute leukemia［J］.Cancers（Basel),2020,12(7):1927.
[14]ALLISON M,MATHEWS J,GILLILAND T,et al.Natural killer cell-mediated immunotherapy for leukemia［J］.Cancers（Basel),2022,14(3):843.
[15]HE Y,BUNN PA,ZHOU C,et al.KIR 2D（L1,L3,L4,S4) and KIR 3DL1 protein expression in non-small cell lung cancer［J］.Oncotarget,2016,7(50):82104-82111.
[16]UESHIMA C,KATAOKA TR,HIRATA M,et al.The killer cell Ig-like receptor 2DL4 expression in human mast cells and its potential role in breast cancer invasion［J］.Cancer Immunol Res,2015,3(8):871-880.
[17]ATTIA JVD,DESSENS CE,VAN DE WATER R,et al.The molecular and functional characteristics of HLA-G and the interaction with its receptors:where to intervene for cancer immunotherapy［J］.Int J Mol Sci,2020,21(22):8678.
[18]KATAOKA TR,UESHIMA C,HIRATA M,et al.Killer immunoglobulin-like receptor 2DL4（CD158d) regulates human mast cells both positively and negatively:possible roles in pregnancy and cancer metastasis［J］.Int J Mol Sci,2020,21(3):954.
[19]PLATONOVA S,CHERFILS-VICINI J,DAMOTTE D,et al.Profound coordinated alterations of intratumoral NK cell phenotype and function in lung carcinoma［J］.Cancer Res,2011,71(16):5412-5422.
[20]KANG L,VOSKINARIAN-BERSE V,LAW E,et al.Characterization and ex vivo expansion of human placenta-derived natural killer cells for cancer immunotherapy［J］.Front Immunol,2013,4:101.
[21]BERRAONDO P,SANMAMED MF,OCHOA MC,et al.Cytokines in clinical cancer immunotherapy［J］.Br J Cancer,2019,120(1):6-15.

Memo

Memo:
新疆维吾尔自治区自然科学基金(编号：2019D01C315）
Last Update: 2022-12-30