Abstract
Lymphocyte activation gene-3 (LAG3) is a transmembrane protein expressed on activated T cells and delivers inhibitory signals to render the T cells unable to effectively help B cells to produce antibodies to microbes and vaccines. Presumably, antagonizing LAG3 could enhance the antibody responses to vaccines, and LAG3 antagonists could facilitate vaccines to induce vigorous antibody responses. In this study, we designed a LAG3-interfering antisense oligonucleotide, designated as LIO-1. The LIO-1 is complementary to an identical region shared in human and mouse LAG3 mRNA. We demonstrated that LIO-1 induced the degradation of LAG3 mRNA in immune cells, decreased the LAG3 expression on CD4+ T cells, maintained the prolonged proliferation and promoted the activation of antigen-specific CD4+ T cells, and increased the production of IFN-γ, IL-2, and IL-6 in the antigen re-stimulated immune cells. In addition, we found that LIO-1 enhanced the antibody responses induced by ISA35-formulated recombinant antigen vaccine or ISA35-formulated inactivated influenza virus vaccines in mice. Thus, the LIO-1, a nucleic acid LAG3 antagonist, could facilitate vaccines to induce vigorous antibody responses and has the possibility to be used as a novel adjuvant.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andrews LP, Marciscano AE, Drake CG, Vignali DA (2017) LAG3 (CD223) as a cancer immunotherapy target. Immunol Rev 276(1):80–96. https://doi.org/10.1111/imr.12519
Brignone C, Grygar C, Marcu M, Perrin G, Triebel F (2007a) IMP321 (sLAG-3) safety and T cell response potentiation using an influenza vaccine as a model antigen: a single-blind phase I study. Vaccine 25(24):4641–4650. https://doi.org/10.1016/j.vaccine.2007.04.019
Brignone C, Grygar C, Marcu M, Perrin G, Triebel F (2007b) IMP321 (sLAG-3), an immunopotentiator for T cell responses against a HBsAg antigen in healthy adults: a single blind randomised controlled phase I study. J Immune Based Ther Vaccines 5(1):5. https://doi.org/10.1186/1476-8518-5-5
Castelli C, Triebel F, Rivoltini L, Camisaschi C (2014) Lymphocyte activation gene-3 (LAG-3, CD223) in plasmacytoid dendritic cells (pDCs): a molecular target for the restoration of active antitumor immunity. Oncoimmunology 3(11):e967146–e967146. https://doi.org/10.4161/21624011.2014.967146
Chan JH, Lim S, Wong WS (2006) Antisense oligonucleotides: from design to therapeutic application. Clin Exp Pharmacol Physiol 33(5-6):533–540. https://doi.org/10.1111/j.1440-1681.2006.04403.x
Cuevas VD, Anta L, Samaniego R, Orta-Zavalza E, Vladimir de la Rosa J, Baujat G, Domínguez-Soto Á, Sánchez-Mateos P, Escribese MM, Castrillo A, Cormier-Daire V, Vega MA, Corbí ÁL (2017) MAFB determines human macrophage anti-inflammatory polarization: relevance for the pathogenic mechanisms operating in multicentric carpotarsal osteolysis. J Immunol 198(5):2070–2081. https://doi.org/10.4049/jimmunol.1601667
Danelli L, Donnarumma T, Kassiotis G (2018) Correlates of follicular helper bias in the CD4 T Cell response to a retroviral antigen. Front Immunol 9:1260–1260. https://doi.org/10.3389/fimmu.2018.01260
Del Giudice G, Rappuoli R, Didierlaurent AM (2018) Correlates of adjuvanticity: a review on adjuvants in licensed vaccines. Semin Immunol 39:14–21. https://doi.org/10.1016/j.smim.2018.05.001
Ding Y, Chan CY, Lawrence CE (2004) Sfold web server for statistical folding and rational design of nucleic acids. Nucleic Acids Res 32(Web Server issue):W135-41. https://doi.org/10.1093/nar/gkh449
Elmir S, Triebel F (2000) A soluble lymphocyte activation gene-3 molecule used as a vaccine adjuvant elicits greater humoral and cellular immune responses to both particulate and soluble antigens. J Immunol 164(11):5583–5589. https://doi.org/10.4049/jimmunol.164.11.5583
Gemelli C, Montanari M, Tenedini E, Zanocco Marani T, Vignudelli T, Siena M, Zini R, Salati S, Tagliafico E, Manfredini R, Grande A, Ferrari S (2006) Virally mediated MafB transduction induces the monocyte commitment of human CD34+ hematopoietic stem/progenitor cells. Cell Death Differ 13:1686–1696. https://doi.org/10.1038/sj.cdd.4401860
Gemelli C, Zanocco Marani T, Bicciato S, Mazza EMC, Boraschi D, Salsi V, Zappavigna V, Parenti S, Selmi T, Tagliafico E, Ferrari S, Grande A (2014) MafB is a downstream target of the IL-10/STAT3 signaling pathway, involved in the regulation of macrophage de-activation. Biochim Biophys Acta 1843(5):955–964. https://doi.org/10.1016/j.bbamcr.2014.01.021
Grosso JF, Kelleher CC, Harris TJ, Maris CH, Hipkiss EL, De Marzo A, Anders R, Netto G, Getnet D, Bruno TC, Goldberg MV, Pardoll DM, Drake CG (2007) LAG-3 regulates CD8+ T cell accumulation and effector function in murine self- and tumor-tolerance systems. J Clin Invest 117(11):3383–3392. https://doi.org/10.1172/jci31184
Hannier S, Tournier M, Bismuth G, Triebel F (1998) CD3/TCR complex-associated lymphocyte activation gene-3 molecules inhibit CD3/TCR signaling. J Immunol 161(8):4058
Huang CT, Workman CJ, Flies D, Pan X, Marson AL, Zhou G, Hipkiss EL, Ravi S, Kowalski J, Levitsky HI, Powell JD, Pardoll DM, Drake CG, Vignali DA (2004) Role of LAG-3 in regulatory T cells. Immunity 21(4):503–513. https://doi.org/10.1016/j.immuni.2004.08.010
Huard B, Tournier M, Hercend T, Triebel F, Faure F (1994) Lymphocyte-activation gene 3/major histocompatibility complex class II interaction modulates the antigenic response of CD4+ T lymphocytes. Eur J Immunol 24(12):3216–3221. https://doi.org/10.1002/eji.1830241246
Huard B, Mastrangeli R, Prigent P, Bruniquel D, Donini S, El-Tayar N, Maigret B, Dreano M, Triebel F (1997) Characterization of the major histocompatibility complex class II binding site on LAG-3 protein. Proc Natl Acad Sci U S A 94(11):5744–5749. https://doi.org/10.1073/pnas.94.11.5744
Huard B, Tournier M, Triebel F (1998) LAG-3 does not define a specific mode of natural killing in human. Immunol Lett 61(2):109–112. https://doi.org/10.1016/S0165-2478(97)00170-3
Isakov N, Altman A (2002) Protein kinase Cθ in T cell activation. Annu Rev Immunol 20(1):761–794. https://doi.org/10.1146/annurev.immunol.20.100301.064807
Kim JH, Liepkalns J, Reber AJ, Lu X, Music N, Jacob J, Sambhara S (2016) Prior infection with influenza virus but not vaccination leaves a long-term immunological imprint that intensifies the protective efficacy of antigenically drifted vaccine strains. Vaccine 34(4):495–502. https://doi.org/10.1016/j.vaccine.2015.11.077
Kisielow M, Kisielow J, Capoferri-Sollami G, Karjalainen K (2005) Expression of lymphocyte activation gene 3 (LAG-3) on B cells is induced by T cells. Eur J Immunol 35(7):2081–2088. https://doi.org/10.1002/eji.200526090
Kouo T, Huang L, Pucsek AB, Cao M, Solt S, Armstrong T, Jaffee E (2015) Galectin-3 shapes antitumor immune responses by suppressing CD8<sup>+</sup> T cells via LAG-3 and inhibiting expansion of plasmacytoid dendritic cells. Cancer Immunol Res 3(4):412–423. https://doi.org/10.1158/2326-6066.cir-14-0150
Kretschmer-Kazemi Far R, Nedbal W, Sczakiel G (2001) Concepts to automate the theoretical design of effective antisense oligonucleotides. Bioinformatics 17(11):1058–1061. https://doi.org/10.1093/bioinformatics/17.11.1058
Li X, Yang L, Zhao P, Yao Y, Lu F, Tu L, Liu J, Li Z, Yu Y, Wang L (2017) Adjuvanticity of a CTLA-4 3' UTR complementary oligonucleotide for emulsion formulated recombinant subunit and inactivated vaccines. Vaccine 35(18):2379–2389. https://doi.org/10.1016/j.vaccine.2017.03.043
Liao W, Lin JX, Leonard WJ (2013) Interleukin-2 at the crossroads of effector responses, tolerance, and immunotherapy. Immunity 38(1):13–25. https://doi.org/10.1016/j.immuni.2013.01.004
Lima WF, De Hoyos CL, Liang XH, Crooke ST (2016) RNA cleavage products generated by antisense oligonucleotides and siRNAs are processed by the RNA surveillance machinery. Nucleic Acids Res 44(7):3351–3363. https://doi.org/10.1093/nar/gkw065
Lino AC, Dang VD, Lampropoulou V, Welle A, Joedicke J, Pohar J, Simon Q, Thalmensi J, Baures A, Flühler V, Sakwa I, Stervbo U, Ries S, Jouneau L, Boudinot P, Tsubata T, Adachi T, Hutloff A, Dörner T, Zimber-Strobl U, de Vos AF, Dahlke K, Loh G, Korniotis S, Goosmann C, Weill J-C, Reynaud C-A, Kaufmann SHE, Walter J, Fillatreau S (2018) LAG-3 Inhibitory receptor expression identifies immunosuppressive natural regulatory plasma cells. Immunity 49(1):120-133.e9. https://doi.org/10.1016/j.immuni.2018.06.007
Linterman MA, Pierson W, Lee SK, Kallies A, Kawamoto S, Rayner TF, Srivastava M, Divekar DP, Beaton L, Hogan JJ, Fagarasan S, Liston A, Smith KG, Vinuesa CG (2011) Foxp3+ follicular regulatory T cells control the germinal center response. Nat Med 17(8):975–982. https://doi.org/10.1038/nm.2425
Martin P, Del Hoyo GM, Anjuere F, Arias CF, Vargas HH, Fernandez LA, Parrillas V, Ardavin C (2002) Characterization of a new subpopulation of mouse CD8alpha+ B220+ dendritic cells endowed with type 1 interferon production capacity and tolerogenic potential. Blood 100(2):383–390. https://doi.org/10.1182/blood.V100.2.383
Martinez RJ, Evavold BD (2015) Lower affinity T cells are critical components and active participants of the immune response. Front Immunol 6:468–468. https://doi.org/10.3389/fimmu.2015.00468
Matsushima-Hibiya Y, Nishi S, Sakai M (1998) Rat Maf-related factors: the specificities of DNA binding and heterodimer formation. Biochem Biophys Res Commun 245(2):412–418. https://doi.org/10.1006/bbrc.1998.8447
Matsuzaki J, Gnjatic S, Mhawech-Fauceglia P, Beck A, Miller A, Tsuji T, Eppolito C, Qian F, Lele S, Shrikant P, Old LJ, Odunsi K (2010) Tumor-infiltrating NY-ESO-1–specific CD8+T cells are negatively regulated by LAG-3 and PD-1 in human ovarian cancer. Proc Natl Acad Sci U S A 107(17):7875–7880. https://doi.org/10.1073/pnas.1003345107
Matveeva OV, Tsodikov AD, Giddings M, Freier SM, Wyatt JR, Spiridonov AN, Shabalina SA, Gesteland RF, Atkins JF (2000) Identification of sequence motifs in oligonucleotides whose presence is correlated with antisense activity. Nucleic Acids Res 28(15):2862–2865. https://doi.org/10.1093/nar/28.15.2862
Miyazaki T, Dierich A, Benoist C, Mathis D (1996) Independent modes of natural killing distinguished in mice lacking Lag3. Science 272(5260):405–408. https://doi.org/10.1126/science.272.5260.405
Nascimbeni M, Perié L, Chorro L, Diocou S, Kreitmann L, Louis S, Garderet L, Fabiani B, Berger A, Schmitz J, Marie J-P, Molina TJ, Pacanowski J, Viard J-P, Oksenhendler E, Beq S, Abehsira-Amar O, Cheynier R, Hosmalin A (2009) Plasmacytoid dendritic cells accumulate in spleens from chronically HIV-infected patients but barely participate in interferon-α expression. Blood 113(24):6112–6119. https://doi.org/10.1182/blood-2008-07-170803
Naundorf S, Schröder M, Höflich C, Suman N, Volk H-D, Grütz G (2009) IL-10 interferes directly with TCR-induced IFN-γ but not IL-17 production in memory T cells. Eur J Immunol 39(4):1066–1077. https://doi.org/10.1002/eji.200838773
Nishida J, Li Y, Zhuang Y, Huang Z, Huang H (2013) IFN-gamma suppresses permissive chromatin remodeling in the regulatory region of the Il4 gene. Cytokine 62(1):91–95. https://doi.org/10.1016/j.cyto.2013.02.010
Okamura T, Sumitomo S, Morita K, Iwasaki Y, Inoue M, Nakachi S, Komai T, Shoda H, J-i M, Fujio K, Yamamoto K (2015) TGF-β3-expressing CD4(+)CD25(−)LAG3(+) regulatory T cells control humoral immune responses. Nat Commun 6:6329. https://doi.org/10.1038/ncomms7329
Poeck H, Wagner M, Battiany J, Rothenfusser S, Wellisch D, Hornung V, Jahrsdorfer B, Giese T, Endres S, Hartmann G (2004) Plasmacytoid dendritic cells, antigen, and CpG-C license human B cells for plasma cell differentiation and immunoglobulin production in the absence of T-cell help. Blood 103(8):3058–3064. https://doi.org/10.1182/blood-2003-08-2972
Richter K, Agnellini P, Oxenius A (2010) On the role of the inhibitory receptor LAG-3 in acute and chronic LCMV infection. Int Immunol 22(1):13–23. https://doi.org/10.1093/intimm/dxp107
Romano E, Michielin O, Voelter V, Laurent J, Bichat H, Stravodimou A, Romero P, Speiser DE, Triebel F, Leyvraz S, Harari A (2014) MART-1 peptide vaccination plus IMP321 (LAG-3Ig fusion protein) in patients receiving autologous PBMCs after lymphodepletion: results of a Phase I trial. J Transl Med 12:97. https://doi.org/10.1186/1479-5876-12-97
Schroeder HW Jr, Cavacini L (2010) Structure and function of immunoglobulins. J Allergy Clin Immunol 125(2 Suppl 2):S41–S52. https://doi.org/10.1016/j.jaci.2009.09.046
Shen P, Fillatreau S (2015) Antibody-independent functions of B cells: a focus on cytokines. Nat Rev Immunol 15:441–451. https://doi.org/10.1038/nri3857
Sierra-Filardi E, Nieto C, Domínguez-Soto Á, Barroso R, Sánchez-Mateos P, Puig-Kroger A, López-Bravo M, Joven J, Ardavín C, Rodríguez-Fernández JL, Sánchez-Torres C, Mellado M, Corbí ÁL (2014) CCL2 Shapes Macrophage Polarization by GM-CSF and M-CSF: Identification of CCL2/CCR2-Dependent Gene Expression Profile. J Immunol 192(8):3858–3867. https://doi.org/10.4049/jimmunol.1302821
Swiecki M, Colonna M (2015) The multifaceted biology of plasmacytoid dendritic cells. Nat Rev Immunol 15:471–485. https://doi.org/10.1038/nri3865
Triebel F, Jitsukawa S, Baixeras E, Roman-Roman S, Genevee C, Viegas-Pequignot E, Hercend T (1990) LAG-3, a novel lymphocyte activation gene closely related to CD4. J Exp Med 171(5):1393–1405. https://doi.org/10.1084/jem.171.5.1393
Vazquez MI, Catalan-Dibene J, Zlotnik A (2015) B cells responses and cytokine production are regulated by their immune microenvironment. Cytokine 74(2):318–326. https://doi.org/10.1016/j.cyto.2015.02.007
Workman CJ, Vignali DAA (2005) Negative regulation of T cell homeostasis by lymphocyte activation gene-3 (CD223). J Immunol 174(2):688–695. https://doi.org/10.4049/jimmunol.174.2.688
Workman CJ, Dugger KJ, Vignali DAA (2002) Cutting edge: molecular analysis of the negative regulatory function of lymphocyte activation gene-3. J Immunol 169(10):5392–5395. https://doi.org/10.4049/jimmunol.169.10.5392
Workman CJ, Cauley LS, Kim IJ, Blackman MA, Woodland DL, Vignali DAA (2004) Lymphocyte activation gene-3 (CD223) regulates the size of the expanding T cell population following antigen activation in vivo. J Immunol 172(9):5450–5455. https://doi.org/10.4049/jimmunol.172.9.5450
Workman CJ, Wang Y, El Kasmi KC, Pardoll DM, Murray PJ, Drake CG, Vignali DA (2009) LAG-3 regulates plasmacytoid dendritic cell homeostasis. J Immunol 182(4):1885–1891. https://doi.org/10.4049/jimmunol.0800185
Yao Q, Vuong V, Li M, Compans RW (2002) Intranasal immunization with SIV virus-like particles (VLPs) elicits systemic and mucosal immunity. Vaccine 20(19):2537–2545. https://doi.org/10.1016/S0264-410X(02)00160-3
Yu C, Li X, Liu J, Diao W, Zhang L, Xiao Y, Wei H, Yu Y, Yu Y, Wang L (2016) Replacing the decoy epitope of PCV2b capsid protein with a protective epitope enhances efficacy of PCV2b vaccine. Vaccine 34(50):6358–6366. https://doi.org/10.1016/j.vaccine.2016.10.044
Zhao Q, Elson CO (2018) Adaptive immune education by gut microbiota antigens. Immunology 154(1):28–37. https://doi.org/10.1111/imm.12896
Zuker M (2003) Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res 31(13):3406–3415. https://doi.org/10.1093/nar/gkg595
Funding
This study is financially supported by the National Nature Scientific Foundation of China (31670937).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
All animal experimental procedures were performed in accordance with guidelines of the Animal Ethical and Experimental Committee of Jilin University.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Li, Z., Song, Y., Cui, C. et al. A LAG3-interfering oligonucleotide acts as an adjuvant to enhance the antibody responses induced by recombinant protein vaccines and inactivated influenza virus vaccines. Appl Microbiol Biotechnol 103, 6543–6557 (2019). https://doi.org/10.1007/s00253-019-09919-4
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-019-09919-4