Abstract
Here, retrotransposon-like 1 (RTL1) is introduced as a marker for circulating and tissue neutrophils, tissue macrophages, and tumor-associated macrophages (TAM) and neutrophils (TAN). Anti-RTL1 polyclonal and monoclonal antibodies were produced, and their reactivity was examined by Western blotting (WB), ELISA, and immunostaining of human normal and cancer tissues. The reactivity of the anti-RTL1 antibodies with peripheral blood leukocytes and a panel of hematopoietic cell lines was examined. The generated antibodies specifically detected RTL1 in the WB of the placenta and U937 cells. The polyclonal antibody showed excellent reactivity with tissue-resident macrophages, Hofbauer cells, alveolar and splenic macrophages, Kupffer cells, and inflammatory cells in the tonsil, appendix, and gallbladder. In vitro GM-CSF-differentiated macrophages also showed a high level of intracellular RTL1 expression. TAM and TAN also showed excellent reactivity with this antibody. Almost all circulating granulocytes but not lymphocytes or monocytes expressed RTL1 at their surface. Serial sections of the appendix stained with CD15 and RTL1 and placenta stained with CD68 and RTL1 showed a considerable overlap in RTL1 expression in CD15+ granulocytes and CD68+ macrophages. A small percentage of myelomonocytic cell lines was positive for surface RTL1, while promyelocytic, monocytic, megaloblastic, and lymphoblastic cell lines were negative. Endothelial cells of normal and cancer tissues highly expressed RTL1. RTL1 could be considered a new marker for different normal tissue macrophages, TAM, circulating and tissue neutrophils, and TAN.
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Data supporting these findings are available on request from the corresponding author.
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Acknowledgements
The results of this research work were patented in 2019 at Iran Patent and Intellectual Property Office under number: 139850140003006981. The authors thank Dr. Bozorgmehr for his contribution to flow cytometry data analysis.
Funding
Research reported in this publication was supported by Elite Researcher Grant Committee under award number [971138] from the National Institute for Medical Research Development (NIMAD), Tehran, Iran, and in part by the Iran University of Medical Sciences (IUMS) (Grant No: 95–03-138–29530), Tehran, Iran.
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SM produced monoclonal antibody, performed relevant experiments, and assisted in data collection and analysis and preparation of the first draft of the manuscript. ARM was actively involved in the production and characterization of polyclonal antibody, involved in data collection and analysis, and wrote the first draft of the manuscript. SS, SV, and AHB are actively involved in antibody characterization and IHC experiments. HS prepared all tissues used in this study and interpreted IHC results. FSH.designed and supervised the additional experiments requested for manuscript revision. AHZ and RG contributed to the study conception and design, supervised the study, and revised the manuscript. All authors approved the final version of the manuscript.
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Mortezagholi, S., Mahmoudi, AR., Shojaeian, S. et al. Discovery of a novel marker for human granulocytes and tissue macrophages: RTL1 revisited. Cell Tissue Res 394, 177–188 (2023). https://doi.org/10.1007/s00441-023-03817-y
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DOI: https://doi.org/10.1007/s00441-023-03817-y