Identification of tumor-reactive B cells and systemic IgG in breast cancer based on clonal frequency in the sentinel lymph node
- 578 Downloads
A better understanding of antitumor immune responses is the key to advancing the field of cancer immunotherapy. Endogenous immunity in cancer patients, such as circulating anticancer antibodies or tumor-reactive B cells, has been historically yet incompletely described. Here, we demonstrate that tumor-draining (sentinel) lymph node (SN) is a rich source for tumor-reactive B cells that give rise to systemic IgG anticancer antibodies circulating in the bloodstream of breast cancer patients. Using a synergistic combination of high-throughput B-cell sequencing and quantitative immunoproteomics, we describe the prospective identification of tumor-reactive SN B cells (based on clonal frequency) and also demonstrate an unequivocal link between affinity-matured expanded B-cell clones in the SN and antitumor IgG in the blood. This technology could facilitate the discovery of antitumor antibody therapeutics and conceivably identify novel tumor antigens. Lastly, these findings highlight the unique and specialized niche the SN can fill in the advancement of cancer immunotherapy.
KeywordsBreast cancer Antibody Sentinel Node Next-Generation Sequencing Repertoire Heavy–light (VH:VL) chain pairing
Liquid Chromatography–tandem Mass Spectrometry
Reverse Transcription Xenopolymerase
Surface Plasmon Resonance
Triple Negative Breast Cancer
Variable Heavy chain
Variable Light chain
The authors thank the patients for their willingness to participate, the UVM Larner College of Medicine Microscopy Imaging Center for tissue imaging; the Genome Sequencing and Analysis Facility at the University of Texas at Austin for performing Illumina next-generation sequencing; the clinical coordinators in the Vermont Cancer Center for consenting patients and collecting the tissues for this study; Andrew D. Ellington (UT Austin) for a generous gift of RTX polymerase; and members of our laboratories for critical reading of the manuscript.
Conception and design: JRM, SCP, GSS, NS, GG, GCI, DNK, development of methodology: JRM, SCP, WNV, GSS, JG, JWE. Acquisition of data: JRM, SCP, WNV, YS, SS, C-HL, CCK, YT, and SH. Acquired and managed patients: DNK and SH. Analysis and interpretation of data: JRM, SCP, WNV, GSS, YS, SS, C-HL, APH, GCI, DNK. Writing, review, and/or revision of the manuscript: JRM, SCP, WNV, GSS, SS, C-HL, APH, GG, GCI, DNK. Administrative, technical, or material support (i.e., reporting or organizing data, constructing databases): SCP, WNV, JG, JWE, YT, GG, DNK. Study Supervision: JRM, SCP, GG, GCI, DNK.
This study was supported by S.D. Ireland Cancer Research Foundation (David N. Krag), The Lake Champlain Cancer Research Foundation Grant (Stephanie C. Pero, Nikoletta Sidiropoulos), the Defense Threat Reduction Agency (DTRA) HDTRA1-12-C-0105 (George Georgiou), and the National Institutes of Health (NIH) 5R21AI119368 (Gregory C. Ippolito). The Zeiss 510 META laser scanning confocal microscope was supported by NIH Award Number 1S10RR019246 from the National Center for Research Resources.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
Ethical approval and ethical standards
This study was approved by the Institutional Review Board at the University of Vermont, CHRMS# M12-009, PI: David N. Krag.
Informed Consent was received from each of the patients involved in this study.
- 6.Krag DN, Anderson SJ, Julian TB et al (2007) Technical outcomes of sentinel-lymph-node resection and conventional axillary-lymph-node dissection in patients with clinically node-negative breast cancer: results from the NSABP B-32 randomised phase III trial. Lancet Oncol 8:881–888. https://doi.org/10.1016/S1470-2045(07)70278-4 CrossRefPubMedGoogle Scholar
- 16.Yuan J, Adamow M, Ginsberg BA et al (2011) Integrated NY-ESO-1 antibody and CD8+ T-cell responses correlate with clinical benefit in advanced melanoma patients treated with ipilimumab. Proc Natl Acad Sci U S A 108:16723–16728. https://doi.org/10.1073/pnas.1110814108 CrossRefPubMedPubMedCentralGoogle Scholar
- 18.Krag DN, Anderson SJ, Julian TB et al (2010) Sentinel-lymph-node resection compared with conventional axillary-lymph-node dissection in clinically node-negative patients with breast cancer: overall survival findings from the NSABP B-32 randomised phase 3 trial. Lancet Oncol 11:927–933. https://doi.org/10.1016/S1470-2045(10)70207-2 CrossRefPubMedPubMedCentralGoogle Scholar
- 24.Plotly Technologies Inc (2017) Collaborative data science. Plotly Technologies Inc. https://www.plot.ly. Accessed 24 Jul 2017