Abstract
Many investigations of the replication and pathogenesis of human T-cell leukemia virus type 1 (HTLV-1) employ chronically infected cell lines, cell lines stabilized from primary adult T-cell leukemia cells, and noninfected T-cell lines. The validity of data obtained from such studies depends on the unambiguous identification of each cell line, which can be performed by short-tandem-repeat (STR) profiling (DNA fingerprinting). While kit-based profiling represents the standard method for cell line authentication, not all labs have ready access to the required capillary electrophoresis equipment, and the costs of such tests can become substantial, especially if the cell lines are to be tested frequently. We analyzed DNA from a panel of HTLV-1-infected cell lines and noninfected T-cell lines using a commercial STR kit and then analyzed the same DNA for individual STR markers followed by nondenaturing polyacrylamide gel electrophoresis. This simplified method should facilitate routine confirmation of cell line identity in diverse laboratory settings.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
American Type Culture Collection Standards Development Organization Workgroup ASN (2010) Cell line misidentification: the beginning of the end. Nat Rev Cancer 10:441–448
Capes-Davis A, Theodosopoulos G, Atkin I, Drexler HG, Kohara A, MacLeod RA, Masters JR, Nakamura Y, Reid YA, Reddel RR, Freshney RI (2010) Check your cultures! A list of cross-contaminated or misidentified cell lines. Int J Cancer 127:1–8
Masters JR, Thomson JA, Daly-Burns B, Reid YA, Dirks WG, Packer P, Toji LH, Ohno T, Tanabe H, Arlett CF, Kelland LR, Harrison M, Virmani A, Ward TH, Ayres KL, Debenham PG (2001) Short tandem repeat profiling provides an international reference standard for human cell lines. Proc Natl Acad Sci U S A 98:8012–8017
Butler JM (2006) Genetics and genomics of core short tandem repeat loci used in human identity testing. J Forensic Sci 51:253–265
Popovic M, Lange-Wantzin G, Sarin PS, Mann D, Gallo RC (1983) Transformation of human umbilical cord blood T cells by human T-cell leukemia/lymphoma virus. Proc Natl Acad Sci U S A 80:5402–5406
Miyoshi I, Kubonishi I, Yoshimoto S, Akagi T, Ohtsuki Y, Shiraishi Y, Nagata K, Hinuma Y (1981) Type C virus particles in a cord T-cell line derived by co-cultivating normal human cord leukocytes and human leukaemic T cells. Nature 294:770–771
Salahuddin SZ, Markham PD, Wong-Staal F, Franchini G, Kalyanaraman VS, Gallo RC (1983) Restricted expression of human T-cell leukemia-lymphoma virus (HTLV) in transformed human umbilical cord blood lymphocytes. Virology 129:51–64
Poiesz BJ, Ruscetti FW, Gazdar AF, Bunn PA, Minna JD, Gallo RC (1980) Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma. Proc Natl Acad Sci U S A 77:7415–7419
Schneider U, Schwenk HU, Bornkamm G (1977) Characterization of EBV-genome negative “null” and “T” cell lines derived from children with acute lymphoblastic leukemia and leukemic transformed non-Hodgkin lymphoma. Int J Cancer 19:621–626
Ensenberger MG, Thompson J, Hill B, Homick K, Kearney V, Mayntz-Press KA, Mazur P, McGuckian A, Myers J, Raley K, Raley SG, Rothove R, Wilson J, Wieczorek D, Fulmer PM, Storts DR, Krenke BE (2010) Developmental validation of the PowerPlex 16 HS System: an improved 16-locus fluorescent STR multiplex. Forensic Sci Int Genet 4:257–264
Krenke BE, Tereba A, Anderson SJ, Buel E, Culhane S, Finis CJ, Tomsey CS, Zachetti JM, Masibay A, Rabbach DR, Amiott EA, Sprecher CJ (2002) Validation of a 16-locus fluorescent multiplex system. J Forensic Sci 47:773–785
Norman MR, Thompson EB (1977) Characterization of a glucocorticoid-sensitive human lymphoid cell line. Cancer Res 37:3785–3791
Azari S, Ahmadi N, Tehrani MJ, Shokri F (2007) Profiling and authentication of human cell lines using short tandem repeat (STR) loci: Report from the National Cell Bank of Iran. Biologicals 35:195–202
Sajantila A, Lukka M (1993) Improved separation of PCR amplified VNTR alleles by a vertical polyacrylamide gel electrophoresis. Int J Leg Med 105:355–359
Umetsu K, Yuasa I, Hashiyada M, Adachi N, Watanabe G, Haneda T, Yamazaki K (2016) The art of traditional native PAGE: The APLP 48-ID assay for human identification. Leg Med (Tokyo) 19:28–31
Kozulic B (1995) Models of gel electrophoresis. Anal Biochem 231:1–12
Huang NE, Schumm J, Budowle B (1995) Chinese population data on three tetrameric short tandem repeat loci-HUMTHO1, TPOX, and CSF1PO-derived using multiplex PCR and manual typing. Forensic Sci Int 71:131–136
Urquhart A, Kimpton CP, Downes TJ, Gill P (1994) Variation in short tandem repeat sequences—a survey of twelve microsatellite loci for use as forensic identification markers. Int J Leg Med 107:13–20
Barber MD, Parkin BH (1996) Sequence analysis and allelic designation of the two short tandem repeat loci D18S51 and D8S1179. Int J Leg Med 109:62–65
Mannucci A, Sullivan KM, Ivanov PL, Gill P (1994) Forensic application of a rapid and quantitative DNA sex test by amplification of the X-Y homologous gene amelogenin. Int J Leg Med 106:190–193
Acknowledgments
We thank Luca Persano for the Jurkat and CCRF-CEM cell lines. This work was supported by an investigator grant from the Associazione per la Ricerca sul Cancro (AIRC, awarded to V.C.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Raimondi, V., Minuzzo, S., Ciminale, V., D’Agostino, D.M. (2017). STR Profiling of HTLV-1-Infected Cell Lines. In: Casoli, C. (eds) Human T-Lymphotropic Viruses. Methods in Molecular Biology, vol 1582. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6872-5_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6872-5_11
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6870-1
Online ISBN: 978-1-4939-6872-5
eBook Packages: Springer Protocols