Identification of human plasma cells with a lamprey monoclonal antibody
Cuiling Yu, … , Max D. Cooper, Götz R.A. Ehrhardt
Cuiling Yu, … , Max D. Cooper, Götz R.A. Ehrhardt
Published March 17, 2016
Citation Information: JCI Insight. 2016;1(3):e84738. https://doi.org/10.1172/jci.insight.84738.
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Resource and Technical Advance Immunology

Identification of human plasma cells with a lamprey monoclonal antibody

Abstract

Ab-producing plasma cells (PCs) serve as key participants in countering pathogenic challenges as well as being contributors to autoimmune and malignant disorders. Thus far, only a limited number of PC–specific markers have been identified. The characterization of the unique variable lymphocyte receptor (VLR) Abs that are made by evolutionarily distant jawless vertebrates prompted us to investigate whether VLR Abs could detect novel PC antigens that have not been recognized by conventional Abs. Here, we describe a monoclonal lamprey Ab, VLRB MM3, that was raised against primary multiple myeloma cells. VLRB MM3 recognizes a unique epitope of the CD38 ectoenzyme that is present on plasmablasts and PCs from healthy individuals and on most, but not all, multiple myelomas. Binding by the VLRB MM3 Ab coincides with CD38 dimerization and NAD glycohydrolase activity. Our data demonstrate that the lamprey VLRB MM3 Ab is a unique reagent for the identification of plasmablasts and PCs, with potential applications in the diagnosis and therapeutic intervention of PC or autoimmune disorders.

Authors

Cuiling Yu, Yanling Liu, Justin Tze Ho Chan, Jiefei Tong, Zhihua Li, Mengyao Shi, Dariush Davani, Marion Parsons, Srijit Khan, Wei Zhan, Shuya Kyu, Eyal Grunebaum, Paolo Campisi, Evan J. Propst, David L. Jaye, Suzanne Trudel, Michael F. Moran, Mario Ostrowski, Brantley R. Herrin, F. Eun-Hyung Lee, Ignacio Sanz, Max D. Cooper, Götz R.A. Ehrhardt

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Figure 4

Recognition of CD38 by VLRB MM3.

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Recognition of CD38 by VLRB MM3. (A) Reactivity of panels of human cell ...
(A) Reactivity of panels of human cell lines (left) and primary human tonsillar lymphocyte populations (right) with VLRB MM3 (top row) and conventional anti-CD38 Abs (bottom row). Shown are mean the fluorescence intensities ± SD for cell lines (n = 3) and the median fluorescence intensities (n = 16) for primary cells normalized to negative control VLR4 or isotype control Abs, respectively. (B) Daudi B cells were transiently transfected with siRNA targeting CD38 or scrambled control siRNA and GFP expression plasmids and stained with VLRB MM3 and anti-CD38, respectively. In control experiments, cells were stained with VLRB EHT46 and anti-CD20, respectively. Shown are the mean fluorescent intensities normalized to the indicated controls ± SD (n = 5) gated on GFP-positive, propidium iodide–negative cell populations. *P < 0.05, by Student’s t test. (C) Ablation of VLRB MM3 reactivity by targeting CD38 in Daudi B cells using CRISPR/Cas9. Daudi B cells transfected with CRISPR/Cas9 constructs targeting CD38 or GFP as negative control were stained with VLRB MM3 and anti-CD38 Abs. Similar results were obtained with a second, independent CRISPR/Cas9 construct targeting CD38. BL, B lymphoid leukemia; DLBL, diffuse large B cell lymphoma; GC, germinal center; MFI, mean fluorescence intensity; Non-B/T, non–B/T cell; norm., normalized; VLRB, variable lymphocyte receptor B.