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Neutrophil elastase enhances antigen presentation by upregulating human leukocyte antigen class I expression on tumor cells

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Abstract

Neutrophil elastase (NE) is an innate immune cell-derived inflammatory mediator that we have shown increases the presentation of tumor-associated peptide antigens in breast cancer. In this study, we extend these observations to show that NE uptake has a broad effect on enhancing antigen presentation by breast cancer cells. We show that NE increases human leukocyte antigen (HLA) class I expression on the surface of breast cancer cells in a concentration and time-dependent manner. HLA class I upregulation requires internalization of enzymatically active NE. Western blots of NE-treated breast cancer cells confirm that the expression of total HLA class I as well as the antigen-processing machinery proteins TAP1, LMP2, and calnexin does not change following NE treatment. This suggests that NE does not increase the efficiency of antigen processing; rather, it mediates the upregulation of HLA class I by stabilizing and reducing membrane recycling of HLA class I molecules. Furthermore, the effects of NE extend beyond breast cancer since the uptake of NE by EBV–LCL increases the presentation of HLA class I-restricted viral peptides, as shown by their increased sensitivity to lysis by EBV-specific CD8+ T cells. Together, our results show that NE uptake increases the responsiveness of breast cancer cells to adaptive immunity by broad upregulation of membrane HLA class I and support the conclusion that the innate inflammatory mediator NE enhances tumor cell recognition and increases tumor sensitivity to the host adaptive immune response.

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Abbreviations

APM:

Antigen-processing machinery

CCNE:

Cyclin E

CPZ:

Chlorpromazine

CTLs:

Cytotoxic T lymphocytes

DC:

Dendritic cell

DMEM:

Dulbecco’s modified Eagle’s Medium

EBV:

Epstein–Barr virus

ER:

Endoplasmic reticulum

FBS:

Fetal bovine serum

HLA:

Human leukocyte antigen

IFN:

Interferon

LCLs:

Lymphoblastoid cell lines

LMW:

Low molecular weight

NE:

Neutrophil elastase

PBMCs:

Peripheral blood mononuclear cells

PE:

Phycoerythrin

PMSF:

Phenylmethanesulfonyl fluoride

TAA:

Tumor-associated antigen

TAMs:

Tumor-associated macrophages

TANs:

Tumor-associated neutrophils

TNF:

Tumor necrosis factor

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Acknowledgments

This work was supported by Grant R00CA133244 from the National Cancer Institute (NCI) (to Elizabeth A. Mittendorf), Leukemia SPORE CA100632 (to Jeffrey J. Molldrem), NCI P01 CA148600-01 (to Jeffrey J. Molldrem), Leukemia and Lymphoma Society Research Grant #6030-12 (to Jeffrey J. Molldrem), the T32 CA009599 training grant (to Akhil Chawla and Victor Gall) and the T32 CA009598 training grant (to Celine Kerros and Haley L. Peters) from the NCI. The Flow Cytometry and Cellular Imaging core were supported by Cancer Center Support Grant NCI #CA16672. STR DNA fingerprinting was done by the Cancer Center Support Grant funded Characterized Cell Line core, NCI #CA16672. Elizabeth A. Mittendorf is an R. Lee Clark Fellow of the University of Texas MD Anderson Cancer Center supported by the Jeanne F. Shelby Scholarship Fund.

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    Authors and Affiliations

    1. Department of Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

      Akhil Chawla, Anne V. Philips, Na Qiao, Victor Gall, Samantha Neal & Elizabeth A. Mittendorf

    2. Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0900, Houston, TX, 77030, USA

      Gheath Alatrash, Pariya Sukhumalchandra, Celine Kerros, Haley L. Peters, Karen Clise-Dwyer & Jeffrey J. Molldrem

    3. Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX, USA

      Iulia Diaconu

    4. Department of Breast Surgical Oncology, The University of Texas MD Anderson Cancer Center, 1400 Pressler Street, Unit 1434, Houston, TX, 77030, USA

      Elizabeth A. Mittendorf

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    Gheath Alatrash and Elizabeth A. Mittendorf have contributed equally to this work.

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    Chawla, A., Alatrash, G., Philips, A.V. et al. Neutrophil elastase enhances antigen presentation by upregulating human leukocyte antigen class I expression on tumor cells. Cancer Immunol Immunother 65, 741–751 (2016). https://doi.org/10.1007/s00262-016-1841-6

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