Epstein-Barr virus (EBV) is one of the common human herpesvirus types in the world. EBV is known to infect more than 95% of adults in the world. The virus mainly infects B lymphocytes and could immortalize and transform the cells into EBV-bearing lymphoblastoid cell lines (LCLs). Limited studies have been focused on characterizing the surface marker expression of the immortalized LCLs. This study demonstrates the generation of 15 LCLs from sixteen rheumatoid arthritis (RA) patients and a healthy volunteer using B95-8 marmoset-derived EBV. The success rate of LCL generation was 88.23%. All CD19+ LCLs expressed CD23 (16.94-58.9%) and CD27 (15.74-80.89%) on cell surface. Our data demonstrated two distinct categories of LCLs (fast- and slow-growing) (p<0.05) based on their doubling time. The slow-growing LCLs showed lower CD23 level (35.28%) compared to fast-growing LCLs (42.39%). In contrast, the slow-growing LCLs showed higher percentage in both CD27 alone and CD23+CD27+ in combination. Overall, these findings may suggest the correlations of cellular CD23 and CD27 expression with the proliferation rate of the generated LCLs. Increase expression of CD23 may play a role in EBV immortalization of B-cells and the growth and maintenance of the EBV-transformed LCLs while CD27 expression might have inhibitory effects on LCL proliferation. Further investigations are warranted to these speculations.

1. Stanfield, B. A., & Luftig, M. A. (2017). Recent advances in understanding Epstein–Barr virus. F1000Research, 6, Article 386. https://doi.org/10.12688/f1000research.10591.1

2. Hutt-Fletcher, L. M. (2017). The long and complicated relationship between Epstein–Barr virus and epithelial cells. Journal of Virology, 91(1), e01677-16. https://doi.org/10.1128/JVI.01677-16

3. Ko, Y.-H. (2015). EBV and human cancer. Experimental & Molecular Medicine, 47(1). https://doi.org/10.1038/emm.2014.109

4. Omi, N., Tokuda, Y., Ikeda, Y., Ueno, M., Mori, K., Sotozono, C., Kinoshita, S., Nakano, M., & Tashiro, K. (2017). Efficient and reliable establishment of lymphoblastoid cell lines by Epstein–Barr virus transformation from a limited amount of peripheral blood. Scientific Reports, 7, Article 43833. https://doi.org/10.1038/srep43833

5. Hussain, T., & Mulherkar, R. (2012). Lymphoblastoid cell lines: A continuous in vitro source of cells to study carcinogen sensitivity and DNA repair. International Journal of Molecular and Cellular Medicine, 1(2), 75–87.

6. Draube, A., & von Bergwelt-Baildon, M. (2008). Immortalized B cells: A never-ending source of antigen-presenting cells for vaccines? Expert Review of Vaccines, 7(4), 411–415. https://doi.org/10.1586/14760584.7.4.411

7. Sadreddini, S., Jadidi-Niaragh, F., Younesi, V., Pourlak, T., Afkham, A., Shokri, F., & Yousefi, M. (2016). Evaluation of EBV transformation of human memory B-cells isolated by FACS and MACS techniques. Journal of Immunotoxicology, 13(4), 490–497. https://doi.org/10.3109/1547691X.2015.1132288

8. Yap, H., Tee, S., Wong, M., Chow, S., Peh, S., & Teow, S. (2018). Pathogenic role of immune cells in rheumatoid arthritis: Implications in clinical treatment and biomarker development. Cells, 7(10), Article 161. https://doi.org/10.3390/cells7100161

9. Marston, B., Palanichamy, A., & Anolik, J. H. (2010). B cells in the pathogenesis and treatment of rheumatoid arthritis. Current Opinion in Rheumatology, 22(3), 307–315. https://doi.org/10.1097/BOR.0b013e3283369cb8

10. De Miguel, S., Galocha, B., Jover, J. A., Bañares, A., Hernández-García, C., García-Asenjo, J. A., & Fernández-Gutiérrez, B. (2001). Mechanisms of CD23 hyperexpression on B cells from patients with rheumatoid arthritis. The Journal of Rheumatology, 28(6), 1222–1228.

11. Zhang, X., Burch, E., Cai, L., So, E., Hubbard, F., Matteson, E. L., & Strome, S. E. (2013). CD40 mediates downregulation of CD32B on specific memory B cell populations in rheumatoid arthritis. The Journal of Immunology, 190(12), 6015–6022. https://doi.org/10.4049/jimmunol.1203366

12. Hu, F., Zhang, W., Shi, L., et al. (2018). Impaired CD27+IgD+ B cells with altered gene signature in rheumatoid arthritis. Frontiers in Immunology, 9. https://doi.org/10.3389/fimmu.2018.00626

13. Anderson, J., Caplan, L., Yazdany, J., et al. (2012). Rheumatoid arthritis disease activity measures: American College of Rheumatology recommendations for use in clinical practice. Arthritis Care & Research, 64(5), 640–647. https://doi.org/10.1002/acr.21649

14. Hui-Yuen, J., McAllister, S., Koganti, S., Hill, E., & Bhaduri-Mcintosh, S. (2011). Establishment of Epstein–Barr virus growth-transformed lymphoblastoid cell lines. Journal of Visualized Experiments, 57, e3321. https://doi.org/10.3791/3321

15. Roth, V. (2006). Doubling Time Calculator. http://www.doubling-time.com/compute.php

16. Wang, F., Gregory, C. D., Rowe, M., Rickinson, A. B., Birkenbach, M., Kikutani, H., Kishimoto, T., & Kieff, E. (1987). Epstein–Barr virus nuclear antigen 2 specifically induces expression of the B-cell activation antigen CD23. Proceedings of the National Academy of Sciences of the United States of America, 84(10), 3452–3456. https://doi.org/10.1073/pnas.84.10.3452

17. Agematsu, K., Hokibara, S., Nagumo, H., & Komiyama, A. (2000). CD27: A memory B-cell marker. Immunology Today, 21(5), 204–206. https://doi.org/10.1016/S0167-5699(00)01605-9

18. Rambert, J., Mamani-Matsuda, M., Moynet, D., Dubus, P., Desplat, V., Kauss, T., Dehais, J., Schaeverbeke, T., Ezzedine, K., Malvy, D., Vincendeau, P., Mossalayi, M. D., & Hartl, D. (2009). Molecular blocking of CD23 supports its role in the pathogenesis of arthritis. PLoS ONE, 4(3), e4834. https://doi.org/10.1371/journal.pone.0004834

19. Azim, T., Allday, M. J., & Crawford, D. H. (1990). Immortalization of Epstein–Barr virus-infected CD23-negative B lymphocytes by the addition of B cell growth factor. Journal of General Virology, 71(3), 665–671. https://doi.org/10.1099/0022-1317-71-3-665

20. Megyola, C., Ye, J., & Bhaduri-Mcintosh, S. (2011). Identification of a sub-population of B cells that proliferates after infection with Epstein–Barr virus. Virology Journal, 8, Article 84. https://doi.org/10.1186/1743-422X-8-84