Supplementary Table S1, Figures S1 - S12 from Tumor-Derived α-Fetoprotein Directly Drives Human Natural Killer–Cell Activation and Subsequent Cell Death

Vujanovic, Lazar; Stahl, Elizabeth C.; Pardee, Angela D.; Geller, David A.; Tsung, Allan; Watkins, Simon C.; Gibson, Gregory A.; Storkus, Walter J.; Butterfield, Lisa H.

doi:10.1158/2326-6066.22538252.v1

Supplementary Table S1, Figures S1 - S12 from Tumor-Derived α-Fetoprotein Directly Drives Human Natural Killer–Cell Activation and Subsequent Cell Death

https://doi.org/10.1158/2326-6066.22538252

journal contribution

posted on 2023-04-03, 23:08 authored by Lazar Vujanovic, Elizabeth C. Stahl, Angela D. Pardee, David A. Geller, Allan Tsung, Simon C. Watkins, Gregory A. Gibson, Walter J. Storkus, Lisa H. Butterfield

<p>Supplementary Table S1. Donor median secretion levels (pg/ml). Supplementary Figure S1. Evaluation of AFP internalization mechanism. Supplementary Figure S2. nAFP and tAFP enhance IL-2-induced NK cell activation in a dosedependent manner. Supplementary Figure S3. Intracellular IL-6 and IL-1β staining of NK cells. Supplementary Figure S4. IL-1β and IL-6 production by CD3+ and CD14+ cell contaminants. Supplementary Figure S5. nAFP and tAFP stimulations induce a pro-inflammatory, IL-2-hyperresponsive activation profile in NK-cells. Supplementary Figure S6. Confirmation of protein observations by NanoString single molecule imaging. Supplementary Figure S7. nAFP and tAFP stimulations induce a pro-inflammatory, IL-2-hyperresponsive activation profile in NK-cells. Supplementary Figure S8. nAFP and tAFP stimulations enhance NK cell-mediated tumor killing. Supplementary Figure S9. FACS analysis strategy for evaluating internalized AFP by NK cells. Supplementary Figure S10. Gating strategy used to evaluate NK cell viability in long-term cultures. Supplementary Figure S11. tAFP inhibits proliferation and promotes the death of NK cells in extended cultures. Supplementary Figure S12. RP-HPLC Fractionation of tAFP and its cargo.</p>

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DOI - Is supplement to Tumor-Derived α-Fetoprotein Directly Drives Human Natural Killer–Cell Activation and Subsequent Cell Death

ARTICLE ABSTRACT

Hepatocellular carcinoma (HCC) patients with reduced natural killer (NK)–cell numbers and function have been shown to have a poor disease outcome. Mechanisms underlying NK-cell deficiency and dysfunction in HCC patients remain largely unresolved. α-Fetoprotein (AFP) is an oncofetal antigen produced by HCC. Previous studies demonstrated that tumor-derived AFP (tAFP) can indirectly impair NK-cell activity by suppressing dendritic cell function. However, a direct tAFP effect on NK cells remains unexplored. The purpose of this study was to examine the ability of cord blood-derived AFP (nAFP) and that of tAFP to directly modulate human NK-cell activity and longevity in vitro. Short-term exposure to tAFP and, especially, nAFP proteins induced a unique proinflammatory, IL2-hyperresponsive phenotype in NK cells as measured by IL1β, IL6, and TNF secretion, CD69 upregulation, and enhanced tumor cell killing. In contrast, extended coculture with tAFP, but not nAFP, negatively affected long-term NK-cell viability. NK-cell activation was directly mediated by the AFP protein itself, whereas their viability was affected by hydrophilic components within the low molecular mass cargo that copurified with tAFP. Identification of the distinct impact of circulating tAFP on NK-cell function and viability may be crucial to developing a strategy to ameliorate HCC patient NK-cell functional deficits. Cancer Immunol Res; 5(6); 493–502. ©2017 AACR.