Publications
Selected Publications
2023

Maternal antibodies induced by a live attenuated vaccine protect neonatal mice from cytomegalovirus
Le-Trilling VTK, Jagnjić A, Brizić I, Eilbrecht M, Wohlgemuth K, Rožmanić C, Herdman A, Hoffmann K, Westendorf AM, Hengel H, Jonjić S, Trilling M. Maternal antibodies induced by a live attenuated vaccine protect neonatal mice from cytomegalovirus. NPJ Vaccines. 2023 Feb 3;8(1):8. doi: 10.1038/s41541-023-00602-4. PMID: 36737485; PMCID: PMC9898546
2022

Cell-intrinsic ceramides determine T cell function during melanoma progression
Matthias Hose, Anne Günther, Eyad Naser, Fabian Schumacher, Tina Schönberger, Julia Falkenstein, Athanasios Papadamakis, Burkhard Kleuser, Katrin Anne Becker, Erich Gulbins, Adriana Haimovitz-Friedman, Jan Buer, Astrid M Westendorf, Wiebke Hansen (2022) Cell-intrinsic ceramides determine T cell function during melanoma progression eLife 11:e83073 https://doi.org/10.7554/eLife.83073

CD47 restricts antiviral function of alveolar macrophages during influenza virus infection
Christina Wenzek, Philine Steinbach, Florian Wirsdörfer, Kathrin Sutter, Julia D. Boehme, Robert Geffers, Robert Klopfleisch, Dunja Bruder, Verena Jendrossek, Jan Buer, Astrid M. Westendorf, Torben Knuschke, CD47 restricts antiviral function of alveolar macrophages during influenza virus infection, iScience, Volume 25, Issue 12, 2022, 105540, ISSN 2589-0042, https://doi.org/10.1016/j.isci.2022.105540.

Acute cytomegalovirus infection modulates the intestinal microbiota and targets intestinal epithelial cells
Le-Trilling, V.T.K., Ebel, J.-F., Baier, F., Wohlgemuth, K., Pfeifer, K.R., Mookhoek, A., Krebs, P., Determann, M., Katschinski, B., Adamczyk, A., Lange, E., Klopfleisch, R., Lange, C.M., Sokolova, V., Trilling, M. and Westendorf, A.M. (2023), Acute cytomegalovirus infection modulates the intestinal microbiota and targets intestinal epithelial cells. Eur. J. Immunol., 53: 2249940. https://doi.org/10.1002/eji.202249940

Superior antiviral activity of IFNb in genital HSV-1 infection
Schmitz Y, Schwerdtfeger M, Westmeier J, Littwitz-Salomon E, Alt M, Brochhagen L, Krawczyk A and Sutter K (2022) Superior antiviral activity of IFNb in genital HSV-1 infection. Front. Cell. Infect. Microbiol. 12:949036. doi: 10.3389/fcimb.2022.949036

Adjusted COVID-19 booster schedules balance age-dependent differences in antibody titers benefitting risk populations
Müller L, Andrée M, Moskorz W, Drexler I, Hauka S, Ptok J, Walotka L, Grothmann R, Hillebrandt J, Ritchie A, Peter L, Walker A, Timm J, Adams O and Schaal H (2022), Adjusted COVID-19 booster schedules balance age-dependent differences in antibody titers benefitting risk populations. Front. Aging 3:1027885. doi: 10.3389/fragi.2022.1027885

TLR2-induced CD8+ T-cell deactivation shapes dendritic cell differentiation in the bone marrow during sepsis
Antoni A-C, Pylaeva E, Budeus B, Jablonska J, Klein-Hitpaß L, Dudda M and Flohé SB (2022) TLR2-induced CD8+ T-cell deactivation shapes dendritic cell differentiation in the bone marrow during sepsis. Front. Immunol. 13:945409. doi:10.3389/fimmu.2022.945409

Distinct type I Interferon Subtypes Differentially Stimulate T Cell Responses in HIV-1-Infected Individuals
Karakoese Z, Schwerdtfeger M, Karsten CB, Esser S, Dittmer U and Sutter K (2022) Distinct type I Interferon Subtypes Differentially Stimulate T Cell Responses in HIV-1-Infected Individuals. Front. Immunol. 13:936918. doi: 10.3389/fimmu.2022.936918

The SARS-CoV-2 accessory factor ORF7a downregulates MHC class I surface expression

The Mycotoxin Beauvericin Exhibits Immunostimulatory Effects on Dendritic Cells via Activating the TLR4 Signaling Pathway
Yang X, Ali S, Zhao M, Richter L, Schäfer V, Schliehe-Diecks J, Frank M, Qi J, Larsen P-K, Skerra J, Islam H, Wachtmeister T, Alter C, Huang A, Bhatia S, Köhrer K, Kirschning C, Weighardt H, Kalinke U, Kalscheuer R, Uhrberg M and Scheu S (2022) The Mycotoxin Beauvericin Exhibits Immunostimulatory Effects on Dendritic Cells via Activating the TLR4 Signaling Pathway. Front. Immunol. 13:856230. doi: 10.3389/fimmu.2022.856230

The interplay of thyroid hormones and the immune system - where we stand and why we need to know about it
Wenzek C, Boelen A, Westendorf AM, Engel DR, Moeller LC, Führer D. The interplay of thyroid hormones and the immune system - where we stand and why we need to know about it. Eur J Endocrinol. 2022 Feb 1:EJE-21-1171. doi: 10.1530/EJE-21-1171. Epub ahead of print. PMID: 35175936.

A Hybrid Soluble gp130/Spike-Nanobody Fusion Protein Simultaneously Blocks Interleukin-6 trans-Signaling and Cellular Infection with SARS-CoV-2
Julia Ettich, Julia Werner, Hendrik T. Weitz, Eva Mueller, Roland Schwarzer, Philipp A. Lang, Jürgen Scheller, Jens M. Moll, A Hybrid Soluble gp130/Spike-Nanobody Fusion Protein Simultaneously Blocks Interleukin-6 trans-Signaling and Cellular Infection with SARS-CoV-2, Journal Article, 2022, Journal of Virology, e01622-21, 96, 4, doi:10.1128/jvi.01622-21

Immunotherapy With Interferon α11, But Not Interferon Beta, Controls Persistent Retroviral Infection
Schwerdtfeger M, Dickow J, Schmitz Y, Francois S, Karakoese Z, Malyshkina A, Knuschke T, Dittmer U and Sutter K (2022) Immunotherapy With Interferon a11, But Not Interferon Beta, Controls Persistent Retroviral Infection. Front. Immunol. 12:809774. doi: 10.3389/fimmu.2021.809774
2021

Peripheral blood iNKT cell activation correlates with liver damage during acute hepatitis C
Senff, T., C. Menne, C. Cosmovici, L. L. Lewis-Ximenez, J. Aneja, R. Broering, A. Y. Kim, A. M. Westendorf, U. Dittmer, N. Scherbaum, G. M. Lauer and J. Timm. "Peripheral blood iNKT cell activation correlates with liver damage during acute hepatitis C." JCI Insight (2021) https://doi.org/10.1172/jci.insight.155432

Protein Signatures of NK Cell-Mediated Melanoma Killing Predict Response to Immunotherapies
Cappello S, Sung HM, Ickes C, Gibhardt CS, Vultur A, Bhat H, Hu Z, Brafford P, Denger A, Stejerean-Todoran I, Köhn RM, Lorenz V, Künzel N, Salinas G, Stanisz H, Legler T, Rehling P, Schön MP, Lang KS, Helms V, Herlyn M, Hoth M, Kummerow C, Bogeski I. Protein Signatures of NK Cell-Mediated Melanoma Killing Predict Response to Immunotherapies. Cancer Res. 2021 Nov 1;81(21):5540-5554. doi: 10.1158/0008-5472.CAN-21-0164. Epub 2021 Sep 13. PMID: 34518212; PMCID: PMC8727679.

Neuropilin-1 Is Expressed on Highly Activated CD4 + Effector T Cells and Dysfunctional CD4 + Conventional T Cells from Naive Mice
Abberger, H., R. Barthel, J. Bahr, J. Thiel, S. Luppus, J. Buer, A. M. Westendorf and W. Hansen. (2021) "Neuropilin-1 Is Expressed on Highly Activated CD4 + Effector T Cells and Dysfunctional CD4 + Conventional T Cells from Naive Mice." The Journal of Immunology doi:10.4049/jimmunol.2100222

Nedd8-Activating Enzyme Is a Druggable Host Dependency Factor of Human and Mouse Cytomegalovirus
Flores-Martínez YA, Le-Trilling VTK, Trilling M. Nedd8-Activating Enzyme Is a Druggable Host Dependency Factor of Human and Mouse Cytomegalovirus. Viruses. 2021; 13(8):1610. https://doi.org/10.3390/v13081610

IL-33 Drives Expansion of Type 2 Innate Lymphoid Cells and Regulatory T Cells and Protects Mice From Severe, Acute Colitis
Ngo Thi Phuong N, Palmieri V, Adamczyk A, Klopfleisch R, Langhorst J, Hansen W, Westendorf AM and Pastille E (2021) IL-33 Drives Expansion of Type 2 Innate Lymphoid Cells and Regulatory T Cells and Protects Mice From Severe, Acute Colitis. Front. Immunol. 12:669787. doi: 10.3389/fimmu.2021.669787

Inhibition of TLR4 Signaling Impedes Tumor Growth in Colitis-Associated Colon Cancer
Pastille E, Faßnacht T, Adamczyk A, Ngo Thi Phuong N, Buer J and Westendorf AM (2021) Inhibition of TLR4 Signaling Impedes Tumor Growth in Colitis-Associated Colon Cancer. Front. Immunol. 12:669747. doi: 10.3389/fimmu.2021.669747

Deep Transfer Learning Approach for Automatic Recognition of Drug Toxicity and Inhibition of SARS-CoV-2
Werner, J.; Kronberg, R.M.; Stachura, P.; Ostermann, P.N.; Müller, L.; Schaal, H.; Bhatia, S.; Kather, J.N.; Borkhardt, A.; Pandyra, A.A.; Lang, K.S.; Lang, P.A. Deep Transfer Learning Approach for Automatic Recognition of Drug Toxicity and Inhibition of SARS-CoV-2. Viruses 2021, 13, 610. https://doi.org/10.3390/v13040610

Interleukin-33 signaling exacerbates experimental infectious colitis by enhancing gut permeability and inhibiting protective Th17 immunity
Palmieri, V., Ebel, JF., Ngo Thi Phuong, N. et al. Interleukin-33 signaling exacerbates experimental infectious colitis by enhancing gut permeability and inhibiting protective Th17 immunity. Mucosal Immunol 14, 923–936 (2021). https://doi.org/10.1038/s41385-021-00386-7

A Combination of Anti-PD-L1 Treatment and Therapeutic Vaccination Facilitates Improved Retroviral Clearance via Reactivation of Highly Exhausted T Cells

Hepatitis B virus particles activate B cells through the TLR2–MyD88–mTOR axis
Li Q, Wang J, Islam H, Kirschning C, Lu H, Hoffmann D, Dittmer U, Lu M. Hepatitis B virus particles activate B cells through the TLR2-MyD88-mTOR axis. Cell Death Dis. 2021 Jan 4;12(1):34. doi: 10.1038/s41419-020-03284-1. PMID: 33414473; PMCID: PMC7791069.
2020

Efficient Induction of Cytotoxic T Cells by Viral Vector Vaccination Requires STING-Dependent DC Functions
Barnowski C, Ciupka G, Tao R, Jin L, Busch DH, Tao S and Drexler I (2020) Efficient Induction of Cytotoxic T Cells by Viral Vector Vaccination Requires STING-Dependent DC Functions. Front. Immunol. 11:1458. doi: 10.3389/fimmu.2020.01458

Interplay between Cellular Autophagy and Hepatitis B Virus Replication: A Systematic Review
Lin Y, Zhao Z, Huang A, Lu M. Interplay between Cellular Autophagy and Hepatitis B Virus Replication: A Systematic Review. Cells. 2020; 9(9):2101. https://doi.org/10.3390/cells9092101

A Broad-Spectrum Antiviral Peptide Blocks Infection of Viruses by Binding to Phosphatidylserine in the Viral Envelope
Luteijn RD, Praest P, Thiele F, Sadasivam SM, Singethan K, Drijfhout JW, Bach C, de Boer SM, Lebbink RJ, Tao S, Helfer M, Bach NC, Protzer U, Costa AI, Killian JA, Drexler I, Wiertz EJHJ. A Broad-Spectrum Antiviral Peptide Blocks Infection of Viruses by Binding to Phosphatidylserine in the Viral Envelope. Cells. 2020; 9(9):1989. https://doi.org/10.3390/cells9091989

Efficient induction of cytotoxic T cells
by viral vector vaccination requires
STING-dependent DC functions
Barnowski C, Ciupka G, Tao R, Jin L, Busch DH, Tao S, and Drexler I. Efficient Induction of Cytotoxic T Cells by Viral Vector Vaccination Requires STING-Dependent DC Functions. Front. Immunol. 16 July 2020. doi.org/10.3389/fimmu.2020.01458.

An Inverse Relationship Between c-Kit/CD117 and mTOR Confers NK Cell Dysregulation Late After Severe Injury
Bösken B, Hepner-Schefczyk M, Vonderhagen S, Dudda M and Flohé SB (2020) An Inverse Relationship Between c-Kit/CD117 and mTOR Confers NK Cell Dysregulation Late After Severe Injury. Front. Immunol. 11:1200. doi: 10.3389/fimmu.2020.01200

CEACAM1 regulates CD8 + T Cell immunity and protects from severe pathology during Citrobacter rodentium induced colitis
Zöller J, Ebel J-F, Khairnar V, Schmitt V, Klopfleisch R, Meiners J, Seiffart V, Hansen W, Buer J, Singer BB, Lang KS, Westendorf AM. CEACAM1 regulates CD8 + T cell immunity and protects from severe pathology during Citrobacter rodentium induced colitis. Gut Microbes, 2020 Jun 10;1-16. doi: 10.1080/19490976.2020.1775464.

Mouse Cytomegalovirus M34 encodes
a non-essential, nuclear, early-late
expressed protein required for efficient
viral replication
Eilbrecht M, Le-Trilling VTK, Trilling M. Mouse Cytomegalovirus M34 encodes a non-essential, nuclear, early-late expressed protein required for efficient viral replication. Front. Cell. Infect. Microbiol. 05 May 2020;10:171. doi: 10.3389/fcimb.2020.00171.

O-GlcNAc transferase promotes influenza A virus–induced cytokine storm by targeting interferon regulatory factor–5
Wang Q, Fang P, He R, Li M, Yu H, Zhou L, Yi Y, Wang F, Rong Y, Zhang Y, Chen A, Peng N, Lin Y, Lu M, Zhu Y, Peng G, Rao L, Liu S. O GlcNAc transferase promotes influenza A virus-induced cytokine storm by targeting interferon regulatory factor-5. Sci Adv. 2020 Apr 15;6(16):eaaz708

Immunotherapeutic blockade of CD47
inhibitory signaling enhances innate & adaptive immune responses to viral infection
Cham LB, Torrez Dulgeroff LB, Caspi Tal M, Adomati T, Li F, Bhat H, Huang A, Lang PA, Moreno ME, Rivera JM, Galkina SA, Kosikova G, Stoddart CA, McCune JM, Myers LM, Weissman IL, Lang KS, Hasenkrug KJ. Immunotherapeutic blockade of CD47 inhibitory signaling [...]. Cell Rep. 2020 Apr 14;31(2):107494. doi: 10.1016.

Combination immunotherapy with anti-PD-L1 antibody and depletion of regulatory T cells during acute viral infections results in improved virus control but lethal immunopathology
David P, Drabczyk-Pluta M, Pastille E, Knuschke T, Werner T, [...], Dittmer U, Zelinskyy G. Combination immunotherapy with anti-PD-L1 antibody [...]. PLoS Pathog. 2020 Mar 30;16(3):e1008340. doi: 10.1371/journal.ppat.1008340.

Current gaps in sepsis immunology: new opportunities for translational research
Ignacio Rubio, Marcin F Osuchowski, Manu Shankar-Hari, Tomasz Skirecki, Martin Sebastian Winkler, Gunnar Lachmann, Paul La Rosée, Guillaume Monneret, Fabienne Venet, Michael Bauer, Frank M Brunkhorst, Matthijs Kox, Jean-Marc Cavaillon, Florian Uhle, Markus A Weigand, Stefanie B Flohé, W Joost Wiersinga, Marta Martin-Fernandez, Raquel Almansa, Ignacio Martin-Loeches, Antoni Torres, Evangelos J Giamarellos-Bourboulis, Massimo Girardis, Andrea Cossarizza, Mihai G Netea, Tom van der Poll, André Scherag, Christian Meisel, Joerg C Schefold, Jesús F Bermejo-Martín, Current gaps in sepsis immunology: new opportunities for translational research, The Lancet Infectious Diseases, Volume 19, Issue 12, 2019, Pages e422-e436, ISSN 1473-3099, https://doi.org/10.1016/S1473-3099(19)30567-5.

Targeting Autophagy in Innate Immune Cells: Angel or Demon During Infection and Vaccination?
Tao S and Drexler I (2020) Targeting Autophagy in Innate Immune Cells: Angel or Demon During Infection and Vaccination? Front. Immunol. 11:460. doi: 10.3389/fimmu.2020.00460

Dead cells induce innate anergy via Mertk after acute viral infection
Adomati T, Cham LB, Hamdan TA, Bhat H, Duhan V, Li F, Ali M, Lang E, Huang A, Naser E, Khairnar V, Friedrich SK, Lang J, Friebus-Kardash J, Bergerhausen M, Schiller M, Machlah YM, Lang F, Häussinger D, Ferencik S, Hardt C, Lang PA, Lang KS. Dead cells induce innate anergy via Mertk [...]. Cell Rep. 2020 Mar 17;30(11):3671-3681.e5. doi: 10.1016/j.celrep.2020.02.101.

Effective Activation of Human Antigen-Presenting Cells and Cytotoxic CD8+ T Cells by a Calcium Phosphate-Based Nanoparticle Vaccine Delivery System
Scheffel F, Knuschke T, Otto L, Kollenda S, Sokolova V, Cosmovici C, Buer J, Timm J, Epple M, Westendorf AM. Effective Activation of Human Antigen-Presenting Cells and Cytotoxic CD8+ T Cells by a Calcium Phosphate-Based Nanoparticle Vaccine Delivery System. Vaccines. 2020; 8(1):110. https://doi.org/10.3390/vaccines8010110

The human cytomegalovirus pUL145
isoforms act as viral DDB1-cullin-associated
factors to instruct host protein degradation
to impede innate immunity
Le-Trilling VTK, Becker T, Nachshon A, Stern-Ginossar N, Schöler L, Voigt S, Hengel H, Trilling M. The human CMV pUL145 isoforms act as viral DDB1-cullin-associated factors to instruct host protein degradation to impede innate immunity. Cell Rep. 2020 Feb 18;30(7):2248-2260.e5.

Map3k14 as a regulator of innate and
adaptive immune response during acute
viral infection
Hamdan TA, Bhat H, Cham LB, Adomati T, Lang J, Li F, Murtaza A, Hardt C, Lang PA, Duhan V, Lang KS. Map3k14 as a regulator of innate and adaptive immune response during acute viral infection. Pathogens. 2020 Feb 4;9(2). pii: E96. doi: 10.3390/pathogens9020096.
2019

Major Vault Protein Promotes Hepatocellular Carcinoma Through Targeting Interferon Regulatory Factor 2 and Decreasing p53 Activity

Depletion of Foxp3+ regulatory T cells is
accompanied by an increase in the relative
abundance of Firmicutes in the murine
gut microbiome

Diverse immunomodulatory effects of
individual IFNα subtypes on virus-specific CD8+ T cell responses

Toll-like receptor 7 activation enhances
CD8+ T cell effector functions by promoting
cellular glycolysis

Tamoxifen protects from
vesicular stomatitis virus infection

Glucosamine promotes hepatitis B virus replication through its dual effects in suppressing autophagic degradation and inhibiting MTORC1 signaling

Intestinal acid sphingomyelinase protects
from severe pathogen-driven colitis

The IL-33/ST2 pathway shapes the
regulatory T cell phenotype to promote
intestinal cancer

T cell-specific overexpression of
acid sphingomyelinase results in elevated
T cell activation and reduced parasitemia during Plasmodium yoelii infection

Friend retrovirus studies reveal complex interactions between intrinsic, innate and adaptive immunity

Expansion of stem-cell-like CD4+ memory
T cells during acute HIV-1 infection is linked
to rapid disease progression

A tumor-peptide based nanoparticle vaccine elicits efficient tumor growth control in
anti-tumor immunotherapy

Glycosylation of HIV Env impacts IgG
subtype responses to vaccination
2018

A mass spectrometry-based profiling of
interactomes of viral DDB1- and cullin
ubiquitin ligase-binding proteins reveals
NF-κB inhibitory activity of the HIV-2-encoded Vpx

A systematic electron microscopic
study on the uptake of barium sulphate
nano-, submicro-, microparticles by
bone marrow-derived phagocytosing cells

CEACAM1 promotes CD8+ T cell
responses and improves control
of a chronic viral infection

Pre-activation of toll-like receptor 2
enhances CD8+ T-cell responses and
accelerates Hepatitis B virus clearance
in the mouse models

STAT2-dependent immune responses ensure host survival despite the presence of a
potent viral antagonist

Impact of the TAP-like transporter in antigen presentation and phagosome maturation

Hepatitis B virus is degraded by autophagosome-lysosome fusion mediated by Rab7 and related components

Mutagenic repair of double-stranded DNA breaks in vaccinia virus genomes requires cellular DNA ligase IV activity in the cytosol

Cancer, drugs, and bugs -
Bacteriotherapy on the rise?

Imaging of cytotoxic antiviral immunity
while considering the 3R principle of
animal research

IL-1β as mucosal vaccine adjuvant:
the specific induction of tissue-resident
memory T cells improves the heterosubtypic immunity against influenza A viruses
2017

Sphingosine 1-phosphate- and C-C
chemokine receptor 2-dependent activation
of CD4+ plasmacytoid dendritic cells
in the bone marrow contributes to signs
of sepsis-induced immunosuppression

Conventional CD11chigh dendritic cells are important for T cell priming during the initial phase of Plasmodium yoelii infection,
but are dispensable at later time points

Broad recruitment of mGBP family members to Chlamydia trachomatis inclusions

Intestinal helminth infection drives
carcinogenesis in colitis-associated
colon cancer

Deciphering of the human interferon-regulated proteome by mass spectrometry-based quantitative analysis reveals extent and dynamics of protein induction
and repression

Local delivery of siRNA-loaded calcium
phosphate nanoparticles abates
pulmonary inflammation

SGLT1 deficiency turns Listeria infection into a lethal disease in mice

Acquired IFNg resistance impairs anti-tumor immunity and gives rise to T-cell-resistant melanoma lesions

Interference of retroviral envelope with vaccine-induced CD8+ T cell responses is
relieved by co-administration of
cytokine-encoding vectors

Differential expression of GPR15 on T cells during ulcerative colitis

Hypoxia enhances immunosuppression by inhibiting CD4+ effector T cell function
and promoting Treg activity

Different antiviral effects of IFNα and IFNβ in an HBV mouse model

DC-derived IL-10 modulates pro-inflammatory cytokine production
and promotes induction of CD4+IL-10+
regulatory T cells during Plasmodium yoelii infection
2016

Antigen-dependent cross-competition shapes the local repertoire of resident
memory CD8+ T cells in the skin

Opposing development of cytotoxic
and follicular helper CD4 T cells
controlled by the TCF-1-Bcl6 nexus

Targeted antibody-mediated depletion of CD19 CAR-T cells permanently reverses
B cell aplasia

Mucosal expression of DEC-205 targeted
allergen alleviates an asthmatic phenotype in mice

Brain-resident memory T cells represent
an autonomous cytotoxic barrier
to viral infection

Bile acids act as soluble host restriction
factors limiting Cytomegalovirus replication
in hepatocytes

Interferon alpha subtype-specific
suppression of HIV-1 infection in vivo

Combination of nanoparticle-based
therapeutic vaccination and transient
ablation of regulatory T cells enhances
anti-viral immunity during chronic
retroviral infection

Broad and potent antiviral activity
of the NAE inhibitor MLN4924
2015

The transcription and translation landscapes
during human cytomegalovirus infection
reveal novel host-pathogen interactions

Human TLR8 senses UR/URR motifs
in bacterial and mitochondrial RNA

Virus-induced type I Interferon deteriorates control of systemic Pseudomonas Aeruginosa infection

IL10-deficiency in CD4+ T cells exacerbates the IFNγ and IL17 response during bacteria induced Colitis

Modified vaccinia virus Ankara-infected
dendritic cells present CD4+ T-cell epitopes by endogenous major histocompatibility complex class II presentation pathways
