Interrogation of individual intratumoral B lymphocytes from lung cancer patients for molecular target discovery.

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2016-02

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Abstract

Intratumoral B lymphocytes are an integral part of the lung tumor microenvironment. Interrogation of the antibodies they express may improve our understanding of the host response to cancer and could be useful in elucidating novel molecular targets. We used two strategies to explore the repertoire of intratumoral B cell antibodies. First, we cloned VH and VL genes from single intratumoral B lymphocytes isolated from one lung tumor, expressed the genes as recombinant mAbs, and used the mAbs to identify the cognate tumor antigens. The Igs derived from intratumoral B cells demonstrated class switching, with a mean VH mutation frequency of 4%. Although there was no evidence for clonal expansion, these data are consistent with antigen-driven somatic hypermutation. Individual recombinant antibodies were polyreactive, although one clone demonstrated preferential immunoreactivity with tropomyosin 4 (TPM4). We found that higher levels of TPM4 antibodies were more common in cancer patients, but measurement of TPM4 antibody levels was not a sensitive test for detecting cancer. Second, in an effort to focus our recombinant antibody expression efforts on those B cells that displayed evidence of clonal expansion driven by antigen stimulation, we performed deep sequencing of the Ig genes of B cells collected from seven different tumors. Deep sequencing demonstrated somatic hypermutation but no dominant clones. These strategies may be useful for the study of B cell antibody expression, although identification of a dominant clone and unique therapeutic targets may require extensive investigation.

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Intratumoral B lymphocytes, Non-small cell lung cancer, Target discovery, Tropomyosin 4, Tumor-infiltrating lymphocytes, Aged, Amino Acid Sequence, Antibodies, Antigens, Neoplasm, Antineoplastic Agents, B-Lymphocyte Subsets, Case-Control Studies, Clonal Evolution, Cloning, Molecular, Female, Gene Expression, Humans, Lung Neoplasms, Lymphocytes, Tumor-Infiltrating, Male, Middle Aged, Molecular Sequence Data, Molecular Targeted Therapy, Neoplasm Staging, Sequence Alignment, Sequence Analysis, DNA

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https://hdl.handle.net/10161/11569

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10.1007/s00262-015-1787-0

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Campa, Michael J, M Anthony Moody, Ruijun Zhang, Hua-Xin Liao, Elizabeth B Gottlin and Edward F Patz (2016). Interrogation of individual intratumoral B lymphocytes from lung cancer patients for molecular target discovery. Cancer Immunol Immunother, 65(2). pp. 171–180. 10.1007/s00262-015-1787-0 Retrieved from https://hdl.handle.net/10161/11569.

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Moody

Michael Anthony Moody

Professor of Pediatrics

Tony Moody, MD is a Professor in the Department of Pediatrics, Division of Infectious Diseases and Professor in the Department of Integrative Immunobiology at Duke University Medical Center. Research in the Moody lab is focused on understanding the B cell responses during infection, vaccination, and disease. The lab has become a resource for human phenotyping, flow characterization, staining and analysis at the Duke Human Vaccine Institute (DHVI). The Moody lab is currently funded to study influenza, syphilis, HIV-1, and emerging infectious diseases.

Dr. Moody is the director of the Duke CIVICs Vaccine Center (DCVC) at (DHVI) and co-director of the Centers for Research of Emerging Infectious Disease Coordinating Center (CREID-CC). Dr. Moody is mPI of a U01 program to develop a syphilis vaccine; this program is a collaboration with mPI Dr. Justin Radolf at the University of Connecticut. Dr. Moody is also the director of the DHVI Accessioning Unit, a biorepository that provides support for work occurring at DHVI and with its many collaborators around the world by providing processing, shipping, and inventory support for a wide array of projects.

Dr. Moody and his team are involved in many networks studying vaccine response including the Collaborative Influenza Vaccine Innovation Centers (CIVICs) and the COVID-19 Prevention Network (CoVPN).

Patz

Edward F. Patz

James and Alice Chen Distinguished Professor of Radiology

There are numerous ongoing clinical studies primarily focused on the early detection of cancer.

The basic science investigations in our laboratory concentration on three fundamental translational areas,

1) Development of molecular imaging probes - We have used several different approaches to develop novel imaging probes that characterize and phenotype tumors.

2) Discovery of novel lung cancer biomarkers - We explored the use of proteomics, autoantibodies, and genomics to discover blood and tissue biomarkers for early cancer detection and phenotyping of cancer.

3) Host response to cancer - We study the native immune response to tumors as this may provide cues to relevant diagnostic and therapeutic targets. Most recently we have focused on intratumoral lymphocytes and their specific tumor antigens.

 

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