Role of nanoscale antigen organization on B-cell activation probed using DNA origami
Source abstract
Abstract
Vaccine efficacy can be increased by arraying immunogens in multivalent form on virus-like nanoparticles to enhance B-cell activation. However, the effects of antigen copy number, spacing and affinity, as well as the dimensionality and rigidity of scaffold presentation on B-cell activation remain poorly understood. Here, we display the clinical vaccine immunogen eOD-GT8, an engineered outer domain of the HIV-1 glycoprotein-120, on DNA origami nanoparticles to systematically interrogate the impact of these nanoscale parameters on B-cell activation in vitro. We find that B-cell signalling is maximized by as few as five antigens maximally spaced on the surface of a 40-nm viral-like nanoparticle. Increasing antigen spacing up to ~25-30 nm monotonically increases B-cell receptor activation. Moreover, scaffold rigidity is essential for robust B-cell triggering. These results reveal molecular vaccine design principles that may be used to drive functional B-cell responses.
Source: europepmc · Quality: candidate unverified · Source record ↗
Research context
OpenAlex topics: Advanced biosensing and bioanalysis techniques · RNA Interference and Gene Delivery · Immunotherapy and Immune Responses
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
References (49)
Show source-provided references
1. Bachmann, M. F. & Jennings, G. T. Vaccine delivery: a matter of size, geometry, kinetics and molecular patterns. Nat. Rev. Immunol. 10, 787–796 (2010).10.1038/nri2868 ↗
2. Puffer, E. B., Pontrello, J. K., Hollenbeck, J. J., Kink, J. A. & Kiessling, L. L. Activating B cell signaling with defined multivalent ligands. ACS Chem. Biol. 2, 252–262 (2007).10.1021/cb600489g ↗
3. Minguet, S., Dopfer, E.-P. & Schamel, W. W. A. Low-valency, but not monovalent, antigens trigger the B-cell antigen receptor (BCR). Int. Immunol. 22, 205–212 (2010).10.1093/intimm/dxp129 ↗
4. Batista, F. D., Iber, D. & Neuberger, M. S. B cells acquire antigen from target cells after synapse formation. Nature 411, 489–494 (2001).10.1038/35078099 ↗
5. Dintzis, H. M., Dintzis, R. Z. & Vogelstein, B. Molecular determinants of immunogenicity: the immunon model of immune response. Proc. Natl Acad. Sci. USA 73, 3671–3675 (1976).10.1073/pnas.73.10.3671 ↗
6. Brouwer, P. J. M. et al. Enhancing and shaping the immunogenicity of native-like HIV-1 envelope trimers with a two-component protein nanoparticle. Nat. Commun. 10, 4272 (2019).10.1038/s41467-019-12080-1 ↗
7. Steichen, J. M. et al. HIV vaccine design to target germline precursors of glycan-dependent broadly neutralizing antibodies. Immunity 45, 483–496 (2016).10.1016/j.immuni.2016.08.016 ↗
8. Abbott, R. K. et al. Precursor frequency and affinity determine B cell competitive fitness in germinal centers, tested with germline-targeting HIV vaccine immunogens. Immunity 48, 133–146.e6 (2018).10.1016/j.immuni.2017.11.023 ↗
9. Stamatatos, L., Pancera, M. & McGuire, A. T. Germline-targeting immunogens. Immunol. Rev. 275, 203–216 (2017).10.1111/imr.12483 ↗
10. Steichen, J. M. et al. A generalized HIV vaccine design strategy for priming of broadly neutralizing antibody responses. Science 366, eaax4380 (2019).10.1126/science.aax4380 ↗
11. Saunders, K. O. et al. Targeted selection of HIV-specific antibody mutations by engineering B cell maturation. Science 366, eaay7199 (2019).10.1126/science.aay7199 ↗
12. Jardine, J. G. et al. Priming a broadly neutralizing antibody response to HIV-1 using a germline-targeting immunogen. Science 349, 156–161 (2015).10.1126/science.aac5894 ↗
13. Sok, D. et al. Priming HIV-1 broadly neutralizing antibody precursors in human Ig loci transgenic mice. Science 353, 1557–1560 (2016).10.1126/science.aah3945 ↗
14. Kim, Y.-M. et al. Monovalent ligation of the B cell receptor induces receptor activation but fails to promote antigen presentation. Proc. Natl Acad. Sci. USA 103, 3327–3332 (2006).10.1073/pnas.0511315103 ↗
15. Bennett, N. R., Zwick, D. B., Courtney, A. H. & Kiessling, L. L. Multivalent antigens for promoting B and T cell activation. ACS Chem. Biol. 10, 1817–1824 (2015).10.1021/acschembio.5b00239 ↗
16. Bachmann, M. F. et al. The influence of antigen organization on B cell responsiveness. Science 262, 1448–1451 (1993).10.1126/science.8248784 ↗
17. Rothemund, P. W. K. Folding DNA to create nanoscale shapes and patterns. Nature 440, 297–302 (2006).10.1038/nature04586 ↗
18. Veneziano, R. et al. Designer nanoscale DNA assemblies programmed from the top down. Science 352, 1534–1534 (2016).10.1126/science.aaf4388 ↗
19. Wamhoff, E.-C. et al. Programming structured DNA assemblies to probe biophysical processes. Annu. Rev. Biophys. 48, 395–419 (2019).10.1146/annurev-biophys-052118-115259 ↗
20. Jardine, J. G. et al. HIV-1 broadly neutralizing antibody precursor B cells revealed by germline-targeting immunogen. Science 351, 1458–1463 (2016).10.1126/science.aad9195 ↗
21. Jardine, J. et al. Rational HIV immunogen design to target specific germline B cell receptors. Science 340, 711–716 (2013).10.1126/science.1234150 ↗
22. Havenar-Daughton, C. et al. The human naive B cell repertoire contains distinct subclasses for a germline-targeting HIV-1 vaccine immunogen. Sci. Transl. Med. 10, eaat0381 (2018).10.1126/scitranslmed.aat0381 ↗
23. Schwarz, F. P., Robinson, S. & Butler, J. M. Thermodynamic comparison of PNA/DNA and DNA/DNA hybridization reactions at ambient temperature. Nucleic Acids Res. 27, 4792–4800 (1999).10.1093/nar/27.24.4792 ↗
24. Scharenberg, A. M., Humphries, L. A. & Rawlings, D. J. Calcium signalling and cell-fate choice in B cells. Nat. Rev. Immunol. 7, 778–789 (2007).10.1038/nri2172 ↗
25. Sil, D., Lee, J. B., Luo, D., Holowka, D. & Baird, B. Trivalent ligands with rigid DNA spacers reveal structural requirements for IgE receptor signaling in RBL mast cells. ACS Chem. Biol. 2, 674–684 (2007).10.1021/cb7001472 ↗
26. Cochran, J. R., Cameron, T. O., Stone, J. D., Lubetsky, J. B. & Stern, L. J. Receptor proximity, not intermolecular orientation, is critical for triggering T-cell activation. J. Biol. Chem. 276, 28068–28074 (2001).10.1074/jbc.M103280200 ↗
27. Yang, J. & Reth, M. Oligomeric organization of the B-cell antigen receptor on resting cells. Nature 467, 465–469 (2010).10.1038/nature09357 ↗
28. Liu, W., Wang, H. & Xu, C. Antigen receptor nanoclusters: small units with big functions. Trends Immunol. 37, 680–689 (2016).10.1016/j.it.2016.07.007 ↗
29. Gold, M. R. & Reth, M. G. Antigen receptor function in the context of the nanoscale organization of the B cell membrane. Annu. Rev. Immunol. 37, 97–123 (2019).10.1146/annurev-immunol-042718-041704 ↗
30. Tolar, P. & Pierce, S. K. in Immunological Synapse (eds Saito, T. & Batista, F. D.) Vol. 340, 155–169 (Springer, 2010).10.1007/978-3-642-03858-7_8 ↗
31. Shaw, A. et al. Binding to nanopatterned antigens is dominated by the spatial tolerance of antibodies. Nat. Nanotechnol. 14, 184–190 (2019).10.1038/s41565-018-0336-3 ↗
32. Kouskoff, V. et al. Antigens varying in affinity for the B cell receptor induce differential B lymphocyte responses. J. Exp. Med. 188, 1453–1464 (1998).10.1084/jem.188.8.1453 ↗
33. Harwood, N. E. & Batista, F. D. Early events in B cell activation. Annu. Rev. Immunol. 28, 185–210 (2010).10.1146/annurev-immunol-030409-101216 ↗
34. Stone, M. B., Shelby, S. A., Núñez, M. F., Wisser, K. & Veatch, S. L. Protein sorting by lipid phase-like domains supports emergent signaling function in B lymphocyte plasma membranes. eLife 6, e19891 (2017).10.7554/eLife.19891 ↗
35. Tolar, P. Cytoskeletal control of B cell responses to antigens. Nat. Rev. Immunol. 17, 621–634 (2017).10.1038/nri.2017.67 ↗
36. Mukherjee, S. et al. Monovalent and multivalent ligation of the B cell receptor exhibit differential dependence upon Syk and Src family kinases. Sci. Signal. 6, ra1 (2013).10.1126/scisignal.2003220 ↗
37. Kläsener, K., Maity, P. C., Hobeika, E., Yang, J. & Reth, M. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. eLife 3, e02069 (2014).10.7554/eLife.02069 ↗
38. Tolar, P., Hanna, J., Krueger, P. D. & Pierce, S. K. The constant region of the membrane immunoglobulin mediates B cell-receptor clustering and signaling in response to membrane antigens. Immunity 30, 44–55 (2009).10.1016/j.immuni.2008.11.007 ↗
39. Lee, J., Sengupta, P., Brzostowski, J., Lippincott-Schwartz, J. & Pierce, S. K. The nanoscale spatial organization of B-cell receptors on immunoglobulin M- and G-expressing human B-cells. Mol. Biol. Cell 28, 511–523 (2017).10.1091/mbc.e16-06-0452 ↗
40. Liu, W., Meckel, T., Tolar, P., Sohn, H. W. & Pierce, S. K. Antigen affinity discrimination is an intrinsic function of the B cell receptor. J. Exp. Med. 207, 1095–1111 (2010).10.1084/jem.20092123 ↗
41. Zhu, P. et al. Distribution and three-dimensional structure of AIDS virus envelope spikes. Nature 441, 847–852 (2006).
42. Marcandalli, J. et al. Induction of potent neutralizing antibody responses by a designed protein nanoparticle vaccine for respiratory syncytial virus. Cell 176, 1420–1431.e17 (2019).10.1016/j.cell.2019.01.046 ↗
43. Weaver, G. C. et al. In vitro reconstitution of B cell receptor–antigen interactions to evaluate potential vaccine candidates. Nat. Protoc. 11, 193–213 (2016).10.1038/nprot.2016.009 ↗
44. Villar, R. F. et al. Reconstituted B cell receptor signaling reveals carbohydrate-dependent mode of activation. Sci. Rep. 6, 36298 (2016).10.1038/srep36298 ↗
45. Veneziano, R. et al. In vitro synthesis of gene-length single-stranded DNA. Sci. Rep. 8, 6548 (2018).10.1038/s41598-018-24677-5 ↗
46. Shepherd, T. R., Du, R. R., Huang, H., Wamhoff, E.-C. & Bathe, M. Bioproduction of pure, kilobase-scale single-stranded DNA. Sci. Rep. 9, 6121 (2019).10.1038/s41598-019-42665-1 ↗
47. Havenar-Daughton, C. et al. Rapid germinal center and antibody responses in non-human primates after a single nanoparticle vaccine immunization. Cell Rep. 29, 1756–1766.e8 (2019).10.1016/j.celrep.2019.10.008 ↗
48. Marras, A. E., Zhou, L., Su, H.-J. & Castro, C. E. Programmable motion of DNA origami mechanisms. Proc. Natl Acad. Sci. USA 112, 713–718 (2015).10.1073/pnas.1408869112 ↗
49. Schneider, C. A., Rasband, W. S. & Eliceiri, K. W. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9, 671–675 (2012).10.1038/nmeth.2089 ↗