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Author Notes:

Ankur Singh, ankur.singh@gatech.edu

P.L.G. conducted all PEG-4MAL hydrogel studies, young and aged animal studies, bacterial antigen studies, and analyzed data with A.S. K.L performed comparisons between hydrogel chemistries and imaging. J.C. provided single-cell RNA sequencing on lymph node stromal cells and advice on germinal center biology. S.P and I.B. generated bacterial proteins, membranes, and western blots. A.S and P.G wrote the initial manuscript, revision, and all authors reviewed the manuscript and provided feedback. The concept was conceived by A.S. and funding was generated by A.S.

The authors acknowledge financial support from the National Institute of Allergy and Infectious Diseases of the US National Institutes of Health (5R01AI132738-03 awarded to A.S.), a US National Science Foundation CAREER award (DMR-1554275 awarded to A.S.), and the Innovative Molecular Analysis Technology program of the US National Cancer Institute (NIH R33-CA212968-01 awarded to A.S.). The authors acknowledge financial support from the Immunoengineering T32 training grant to S.P. (NIH and NIBIB, 1T32EB023860-01A1) and NIH 1S10RR025502 funding for the shared Zeiss LSM 710 Confocal Microscope, awarded to the Cornell University Biotechnology Resource Center. Opinions, interpretations, conclusions, and recommendations are those of the authors and are not necessarily endorsed by the funding agency.

The authors declare that they have no competing interests.

Subjects:

Keywords:

  • Science & Technology
  • Physical Sciences
  • Technology
  • Chemistry, Multidisciplinary
  • Chemistry, Physical
  • Nanoscience & Nanotechnology
  • Materials Science, Multidisciplinary
  • Physics, Applied
  • Physics, Condensed Matter
  • Chemistry
  • Science & Technology - Other Topics
  • Materials Science
  • Physics
  • aging
  • B cell
  • bacteria
  • EZH2
  • histone
  • hydrogel
  • IMMUNE ORGANOIDS
  • PEG HYDROGELS
  • DIFFERENTIATION
  • SELECTION
  • CULTURE
  • MATRIX
  • LIGHT
  • HYPERMUTATION
  • ORGANIZATION
  • EXPRESSION

Organoid Polymer Functionality and Mode of Klebsiella pneumoniae Membrane Antigen Presentation Regulates Ex Vivo Germinal Center Epigenetics in Young and Aged B Cells

Tools:

Journal Title:

ADVANCED FUNCTIONAL MATERIALS

Volume:

Volume 30, Number 48

Publisher:

Type of Work:

Article | Post-print: After Peer Review

Abstract:

Antibiotic-resistant bacteria are a major global health threat that continues to rise due to a lack of effective vaccines. Of concern are Klebsiella pneumoniae (K. pneumoniae) that fail to induce in vivo germinal center B cell responses, which facilitate antibody production to fight infection. Immunotherapies using antibodies targeting antibiotic-resistant bacteria are emerging as promising alternatives, however, they cannot be efficiently derived ex vivo, necessitating the need for immune technologies to develop therapeutics. Here, polyethylene glycol (PEG)-based immune organoids are developed to elucidate the effects of polymer end-point chemistry, integrin ligands, and mode of K. pneumoniae antigen presentation on germinal center-like B cell phenotype and epigenetics, to better define the lymph node microenvironment factors regulating ex vivo germinal center dynamics. Notably, PEG vinyl sulfone or acrylate fail to sustain primary immune cells, but functionalization with maleimide (PEG-4MAL) leads to B cell expansion and germinal center-like induction. RNA sequencing analysis of lymph node stromal and germinal center B cells shows niche associated heterogeneity of integrin-related genes. Incorporation of niche-mimicking peptides reveals that collagen-1 promotes germinal center-like dynamics and epigenetics. PEG-4MAL organoids elucidate the impact of K. pneumoniae outer membrane-embedded protein antigen versus soluble antigen presentation on germinal centers and preserve the response across young and aged mice.
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