Quantifying Antibody Responses Induced by Antigen-Agnostic Immunotherapies

Jacob P. van Vloten, Elaine M. Klafuric, Khalil Karimi, Douglas McFadden, James J. Petrik, Sarah K. Wootton, Byram W. Bridle

Research output: Contribution to journalArticle

Abstract

As the development and clinical application of cancer immunotherapies continue to expand, so does the need for novel methods to dissect their mechanisms of action. Antibodies are important effector molecules in cancer therapies due to their potential to bind directly to surface-expressed antigens and facilitate Fc receptor-mediated uptake of antigens by antigen-presenting cells. Quantifying antibodies that are specific for defined antigens is straightforward. However, we describe herein a preclinical method to evaluate tumor-associated and virus-specific antibody responses to antigen-agnostic immunotherapies. This method uses autologous tumor cells as reservoirs of bulk tumor antigens, which can be bound by antibodies from the serum or plasma of tumor-bearing mice. These antibodies can then be detected and quantified using isotype-specific secondary antibodies conjugated to a fluorochrome. Alternatively, virus-infected cells can be used as a source of viral antigens. This method will enable researchers to assess antibody responses following immunotherapies without requiring pre-defined antigens. Alternatively, total virus-specific antibody responses could be studied as an alternative to more limited virus-neutralizing antibody assays. Therefore, this method can facilitate studying the role of humoral responses in the context of immunotherapies, including those that rely on the use of viral vectors.

Original languageEnglish (US)
Pages (from-to)189-196
Number of pages8
JournalMolecular Therapy - Methods and Clinical Development
Volume14
DOIs
StatePublished - Sep 13 2019

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Immunotherapy
Antibody Formation
Antigens
Antibodies
Viruses
Neoplasms
Satellite Viruses
Oncogenic Viruses
Fc Receptors
Viral Antigens
Neoplasm Antigens
Antigen-Presenting Cells
Surface Antigens
Neutralizing Antibodies
Fluorescent Dyes
Research Personnel
Serum
Therapeutics

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Quantifying Antibody Responses Induced by Antigen-Agnostic Immunotherapies. / van Vloten, Jacob P.; Klafuric, Elaine M.; Karimi, Khalil; McFadden, Douglas; Petrik, James J.; Wootton, Sarah K.; Bridle, Byram W.

In: Molecular Therapy - Methods and Clinical Development, Vol. 14, 13.09.2019, p. 189-196.

Research output: Contribution to journalArticle

van Vloten, Jacob P. ; Klafuric, Elaine M. ; Karimi, Khalil ; McFadden, Douglas ; Petrik, James J. ; Wootton, Sarah K. ; Bridle, Byram W. / Quantifying Antibody Responses Induced by Antigen-Agnostic Immunotherapies. In: Molecular Therapy - Methods and Clinical Development. 2019 ; Vol. 14. pp. 189-196.
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