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Our Science

Research topics and highlighted literature.

For nearly three decades, the Vignali Laboratory has advanced our understanding of immune regulation through pioneering work on inhibitory receptors, regulatory T cells, TCR–CD3 signaling, systems immunology, and therapeutic innovation.

Research topic

Inhibitory Receptors: LAG3, PD1 & NRP1

Our lab has been instrumental in studying the inhibitory receptor LAG3 for the last ~25 years. Our lab and collaborators were the first to show that LAG3 regulates Treg activity and function, and co-operates with PD1 to regulate T cell exhaustion following chronic viral infection. Importantly, our lab led a team that was the first to demonstrate synergistic co-operation between LAG3 and PD1 in limiting anti-tumor immunity. This landmark study demonstrated that combined LAG3 and PD1 blockade substantially improved tumor control and survival compared with either treatment alone; readers can explore the highlighted publication for details. This study, combined with IP licensed to BMS, served as a direct impetus for a clinical trial with anti-LAG3 (Relatlimab) with or without anti-PD1 (Nivolumab) that has recently progressed to a Phase II/III trial (RELATIVITY-047) in treatment-naive patients with metastatic melanoma and recently met its primary endpoint of progression-free survival. There are now over 10 LAG3-targeting therapeutics in clinical trials, including prospective registrational trials. More recently, we have expanded our interest in inhibitory receptors to PD1 and NRP1.

Highlighted publications

  1. Huang C-T, Workman CJ, Flies D, Pan X, Marson AL, Zhou G, Hipkiss EL, Ravi S, Kowalski J, Lavitsky HI, Powell JD, Pardoll DM, Drake CG, Vignali DAA (2004). Role of LAG-3 in regulatory T cells. Immunity 21:503-13 [PMID: 15485628].
  2. Woo S-R*, Turnis ME*, Goldberg MV*, Bankoti J, Selby M, Nirschl CJ, Bettini ML, Vogel P, Liu C-L, Tangsombatvisit S, Grosso JF, Netto G, Smeltzer MP, Chaux A, Utz PJ, Workman CJ, Pardoll DM, Korman AJ, Drake CG, Vignali DAA (2012). Immune inhibitory molecules LAG-3 and PD-1 synergistically regulate T cell function to promote tumoral immune escape. Cancer Research 72:917-927 [PMCID: 3288154].
  3. Zhang Q, Chikina M, Szymczak-Workman AL, Horne W, Kolls JK, Vignali KM, Normolle D, Bettini M, Workman CJ, Vignali DAA (2017). LAG-3 limits regulatory T cell proliferation and function in autoimmune diabetes. Science Immunology 2:eaah4569 [PMCID: 5609824].
  4. Liu C, Somasundaram A, Manne S, Gocher AM, Szymczak-Workman AL, Vignali KM, Scott EN, Normolle DP, Wherry EJ, Lipson EJ, Ferris RL, Bruno TC, Workman CJ, Vignali DAA (2020). Neuropilin-1 is a T cell memory checkpoint limiting long-term anti-tumor immunity. Nature Immunology, 21:1010-1021 [PMCID: 7442600].
  5. Andrews LP, Somasundaram A, Moskovitz JM, Szymczak-Workman AL, Liu C, Cillo AR, Lin H, Normolle DP, Moynihan KD, Taniuchi I, Irvine DJ, Kirkwood JM, Lipson EJ, Ferris RL, Bruno TC, Workman CJ, Vignali DAA (2020). Resistance to PD1 blockade in the absence of metalloprotease-mediated LAG3 shedding. Science Immunology. 5:eabc2728 [PMCID: 32680952].
  6. Guy C, Mitrea DM, Chou P-C, Temirov J, Vignali KM, Liu X, Zhang H, Kriwacki R, Bruchez M, Watkins S, Workman CJ†, Vignali DAA† (2022). LAG3 associates with TCR-CD3 complexes and suppresses signaling by driving co-receptor-Lck dissociation. Nature Immunology 23:757-767 [PMCID: 9106921].
  7. Grebinoski S*, Zhang Q*, Cillo AR, Manne S, Burnazzi EA, Tabib T, Cardello C, Lian C, Murphy GF, Lafyatis R, Wherry EJ, Das J, Workman CJ, Vignali DAA (2022). Autoreactive CD8+ T cells are restrained by a divergent exhaustion program. Nature Immunology 23:868-877 [PMCID: 9179227].
  8. Andrews LP*, Butler SC*, Cui J, Cillo AR, Cardello C, Liu C, Brunazzi EA, Baessler A, Xie B, Kunning SR, Ngiow SF, Huang YJ, Manne S, Sharpe AH, Delgoffe GM, Wherry EJ, Kirkwood JM, Bruno TC, Workman CJ, Vignali DAA (2024). LAG3 and PD1 synergize on CD8+ T cells to drive T cell exhaustion and hinder autocrine IFNγ-dependent anti-tumor immunity. Cell 187:4355-4372 [PMCID: 11323044].
  9. Cillo AR†, Cardello C, Shan F, Karapetyan L, Kunning SR, Sander C, Rush E, Li A, Karunamurthy A, Massa RC, Rohatgi A, Workman C, Kirkwood JM†, Bruno TC†, Vignali DAA† (2024). Relatlimab plus nivolumab rewires dysfunctional CD8+ T cells by coupling cytotoxic and exhaustion gene modules to promote antitumor immunity. Cell 187:4373-4388 [PMCID: 11346583].

Research topic

Treg Suppression

A major focus of our lab for over 15 years has been identifying and dissecting the pathways that mediate and control Treg function, with particular emphasis on pathways preferentially used in tumors that may offer targets for immunotherapy. Our lab has made two major discoveries in this area. In 2007, we were the first to describe interleukin-35 (IL-35), an inhibitory cytokine produced by Tregs. We subsequently showed that IL-35 can drive the generation of an induced regulatory T cell population called iTr35, which appears particularly prevalent in the tumor microenvironment. We were also the first to identify the IL-35 receptor and uncover its unique signaling pathway. More recently, our work has shown that neutralization or Treg-restricted genetic deletion of IL-35 limits tumor growth and reduces inhibitory receptor expression on tumor-infiltrating lymphocytes. We further found that distinct IL-35- and IL-10-producing Treg populations cooperate within the tumor microenvironment to drive multiple inhibitory receptors, including PD1, LAG3, TIM3, and TIGIT, on effector T cells through a BLIMP1-dependent program; readers can explore the highlighted publications for details.

Highlighted publications

  1. Collison LW, Workman CJ, Kuo TK, Boyd K, Wang Y, Vignali K, Cross R, Sehy D, Blumberg RS, Vignali DAA (2007). The inhibitory cytokine IL-35 contributes to regulatory T cell function. Nature 450: 566-569 [PMID: 18033300].
  2. Collison LW, Chaturvedi V, Henderson AL, Giacomin PR, Guy C, Bankoti J, Finkelstein D, Forbes K, Workman CJ, Brown SA, Rehg JE, Jones ML, Ni H-T, Artis D, Turk MJ, Vignali DAA (2010). Interleukin-35-mediated induction of a potent regulatory T cell population. Nature Immunology 11:1093-1101 [PMCID: 3008395].
  3. Collison LW*, Delgoffe GM*, Guy C, Vignali KM, Chaturvedi V, Fairweather D, Satoskar AR, Garcia KC, Hunter CA, Drake CG, Murray PJ, Vignali DAA (2012). The composition and signaling of the IL-35 receptor are unconventional. Nature Immunology 13:290-299 [PMCID: 3529151].
  4. Turnis ME*, Sawant DV*, Szymczak-Workman A, Andrews LP, Delgoffe GM, Yano H., Beres AJ, Vogel P, Workman CJ, Vignali DAA (2016). Interleukin-35 limits anti-tumor immunity. Immunity. 44:316-29 [PMCID: 4758699].
  5. Sawant DV*, Yano H*, Chikina M, Zhang Q, Liao M, Liu C, Sun Z, Sun T, Tabib T, Pennathur A, Luketich JD, Lafyatis R, Chen W, Poholek A, Bruno TC, Workman CJ, Vignali DAA (2019). Adaptive plasticity of IL10+ and IL35+ regulatory T cells cooperatively promote intratumoral T cell exhaustion. Nature Immunology. 20:724-735 [PMCID: 6531353].

Research topic

Treg Function & Stability

A second major area of our work concerns the pathways that control Treg stability and function. Our lab was the first to identify a neuropilin-1 (NRP1):semaphorin-4A (SEMA4A) pathway that maintains intratumoral Treg stability, function, and survival while remaining dispensable for the maintenance of peripheral tolerance. Our studies showed that disrupting this pathway can markedly restrict tumor growth without producing the autoimmune or inflammatory consequences associated with broad Treg loss. We have also shown that targeting NRP1 or SEMA4A can limit tumor growth, supporting further preclinical investigation and validation; readers can explore the highlighted publications for details.

Highlighted publications

  1. Collison LW, Pillai MR, Chaturvedi V, Vignali DAA (2009). Regulatory T cell suppression is potentiated by target T cells in a cell contact, IL-35- and IL-10-dependent manner. J. Immunol. 182:6121-6128 [PMID: 19414764; PMCID: 2698997].
  2. Delgoffe GM*, Woo S-R*, Turnis ME, Gravano DM, Guy C, Overacre AE, Bettini ML, Vogel P, Finkelstein D, Bonnevier J, Workman CJ, Vignali DAA (2013). Stability and function of regulatory T cells is maintained by a neuropilin-1:semaphorin-4a axis. Nature 501:252-256 [PMCID: 3867145].
  3. Overacre-Delgoffe AE, Chikina M, Dadey RE, Yano H, Brunazzi EA, Shayan G, Horne W, Moskovitz JM, Kolls JK, Sander C, Shuai Y, Normolle DP, Kirkwood J, Ferris RL, Delgoffe GM, Bruno TC, Workman CJ, Vignali DAA (2017). Interferon- drives Treg fragility to promote anti-tumor immunity. Cell 169:1130-1141 [PMCID: 5509332].
  4. Liu C, Chikina M, Deshpande R, Menk AV, Wang T, Tabib T, Brunazzi EA, Vignali KM, Sun M, Stolz DB, Lafyatis RA, Chen W, Delgoffe GM, Workman CJ, Wendell SG, Vignali DAA (2019). Treg cells promote the SREBP1-dependent metabolic fitness of M2-like intratumoral macrophages via CD8+ T cells. Immunity 51:381-397 [PMCID: 6703933].
  5. Chuckran CA, Cillo AR, Moskovitz J, Overacre-Delgoffe A, Somasundaram AS, Shan F, Magnon GC, Kunning SR, Abecassis I, Zureikat AH, Luketich J, Pennathur A, Sembrat J, Rojas M, Merrick DT, Taylor SE, Orr B, Modugno F, Bukanovich R, Schoen RE, Kim S, Duvvuri U, Zeh H, Edwards R, Kirkwood JM, Coffman L, Ferris RL, Bruno TC, Vignali DAA (2021). Prevalence of intratumoral regulatory T cells expressing neuropilin-1 is associated with poorer outcomes in patients with cancer. Science Translational Medicine 13:eabf8495 [PMCID: 9022491].
  6. Shan F, Cillo AR, Cardello C, Yuan D, Kunning SR, Cui J, Lampenfeld C, Williams AM, McDonough AP, Pennathur A, Luketich JD, Kirkwood JM, Ferris RL, Bruno TC, Workman CJ, Benos PV, Vignali DAA (2023). Integrated BATF transcriptional network regulates suppressive intratumoral regulatory T cells. Science Immunology 8: eadf6717 [PMCID: 11045170].
  7. Gocher-Demske AM, Cui J, Szymczak-Workman AL, Vignali KM, Latini JN, Pieklo GP, Kimball, JC, Avery L, Cipolla WL, Huckenstein BR, Hedden L, Meisel M, Alcorn JF, Kane LP, Workman CJ, Vignali DAA (2023). Interferon gamma-induction of TH1-like regulatory T cells controls anti-viral responses. Nature Immunology, 24: 841-854 [PMID: 36928412].

Research topic

TCR–CD3 Signaling

Our lab has maintained a long-standing interest in TCR–CD3 complex signaling and function for approximately 25 years. This work has examined cell trafficking, regulation of cell-surface expression, mechanisms of signal transduction, and the contributions of distinct CD3 signaling motifs, including ITAMs, to different T cell functions. We continue to investigate how this essential and highly complex receptor system coordinates T cell development and immune responses; readers can explore the highlighted publications for details.

Highlighted publications

  1. Liu HY, Rhodes M, Wiest D, Vignali DAA (2000). On the dynamics of TCR:CD3 complex cell surface expression and downmodulation. Immunity 13: 665-675 [PMID: 11114379].
  2. Holst J, Wang H, Durick-Eder K, Workman CJ, Boyd K, Baquet Z, Singh H, Forbes K, Chruscinski A, Smeyne R, van Oers NSC, Utz PJ, Vignali DAA (2008). Scalable signaling mediated by T cell antigen receptor-CD3 ITAMs ensures effective negative selection and prevents autoimmunity. Nature Immunology 9:658-666 [PMID: 18469818].
  3. Wang H, Holst J, Woo S-R, Guy C, Bettini ML, Wang Y, Shafer A, Naramura M, Mingueneau M, Dragone LL, Hayes SM, Malissen B, Band H, Vignali DAA (2010). Tonic ubiquitylation controls T cell receptor:CD3 complex expression during T cell development. EMBOJ 29: 1285-1298 [PMCID: 2857457].
  4. Guy C, Vignali KM, Temirov J, Bettini ML, Overacre AE, Smeltzer M, Zhang H, Huppa JB, Tsai Y-H, Lobry C, Xie J, Dempsey PJ, Crawford HC, Aifantis I, Davis MM, Vignali DAA (2013). Distinct TCR signaling pathways drive proliferation and cytokine production in T cells. Nature Immunology 14:262-270 [PMCID: 3577985].

Research topic

Systems Immunology

Our lab has incorporated transcriptional analysis into its research for over a decade. We continue to expand our systems immunology program through advanced single-cell transcriptomic approaches and computational tools, including methods developed within our collaborative research efforts. These approaches allow us to define immune-cell states, interactions, and disease-associated programs across cancer and other immune contexts; readers can explore the highlighted publications for details.

Highlighted publications

  1. Cillo AR, Kürten C, Tabib T, Qi Z, Onkar S, Wang T, Liu L, Duvvuri U, Kim S, Soose RJ, Oesterreich S, Chen W, Lafyatis R, Bruno TC†, Ferris RL†, Vignali DAA† (2020). Immune landscape of viral- and carcinogen-driven head and neck cancer. Immunity 52:183-199 [PMID: 31924475].
  2. Ruffin AT*, Cillo AR*, Tabib T, Liu A, Onkar S, Kunning S, Lampenfeld C, Abecassis I, Qi Z, Soose R, Duvvuri U, Kim S, Oesterrich S, Lafyatis R, Ferris RL, Vignali DAA†, Bruno TC† (2021). Distinct B cell signatures and tertiary lymphoid structures are driven by two etiologies in head and neck cancer. Nature Communications 12:3349 [PMID: 34099645].
  3. Onkar SS, Cui J, Zou J, Cardello C, Cillo AR, Uddin MR, Sagan A, Joy M, Osmanbeyoglu HU, Pogue-Geile K, McAuliffe P, Lucas PC, Tsang GC, Lee AV, Bruno TC, Oesterreich S*, Vignali DAA* (2023). Immune landscape in invasive ductal and lobular breast cancer reveals a divergent macrophage-driven microenvironment. Nature Cancer 4(4):516-534 [PMID: 36927792].

Research topic

Innovation

Our lab was among the first to develop bead-based multiplexed cytokine assays using Luminex technology, an approach now widely used by cancer immunologists and incorporated into commercial platforms. We also helped popularize 2A peptide-based self-cleaving multicistronic expression systems and were the first to apply them in viral vectors and genetically modified mice. These advances supported the development of TCR retrogenic mice and helped broaden the use of 2A-linked multicistronic vectors in TCR- and CAR-based adoptive T cell approaches for cancer immunotherapy; readers can explore the highlighted publications for details.

Highlighted publications

  1. Carson RT & Vignali DAA (1999). Simultaneous quantitation of fifteen cytokines using a multiplexed flow cytometric assay. J. Immunol. Methods 227:41-52 [PMID: 10485253].
  2. Szymczak AL, Workman CJ, Wang Y, Vignali KM, Dilioglou S., Vanin E, Vignali, DAA (2004). Correction of multi-gene deficiency in vivo using a single 'self-cleaving' 2A peptide-based retroviral vector. Nature Biotechnology 22:589-594 [PMID: 15064769].
  3. Holst J, Vignali KM, Burton AR, Vignali DAA (2006). Rapid analysis of T cell selection and function in vivo using T cell receptor retrogenic mice. Nature Methods 3:191-197 [PMID: 16489336].
  4. Holst J, Szymczak-Workman AL, Vignali, KM, Burton AR, Workman CJ, Vignali, DAA (2006). Generation of T cell receptor retrogenic mice. Nature Protocols 1:406-417 (Updated: Bettini et al (2013). Nature Protocols 8:1837-1840 [PMCID: 3832243]).
  5. Andrews LA, Vignali KM, Szymczak-Workman AL, Burton AR, Brunazzi EA, Ngiow SF, Harusato A, Sharpe AH, Wherry EJ, Taniuchi I, Workman CJ, Vignali DAA (2021). A Cre-driven allele-conditioning line to interrogate CD4+ conventional T cells. Immunity. 54:2209-2217 [PMID: 34551314].