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Monoculture-derived T lymphocytes specific for multiple viruses expand and produce clinically relevant effects in immunocompromised individuals

Abstract

Immunocompromised individuals are at high risk for life-threatening diseases, especially those caused by cytomegalovirus (CMV), Epstein-Barr virus (EBV) and adenovirus. Conventional therapeutics are primarily active only against CMV, and resistance is frequent. Adoptive transfer of polyclonal cytotoxic T lymphocytes (CTLs) specific for CMV or EBV seems promising, but it is unclear whether this strategy can be extended to adenovirus, which comprises many serotypes. In addition, the preparation of a specific CTL line for each virus in every eligible individual would be impractical. Here we describe genetic modification of antigen-presenting cell lines to facilitate the production of CD4+ and CD8+ T lymphocytes specific for CMV, EBV and several serotypes of adenovirus from a single cell culture. When administered to immunocompromised individuals, the single T lymphocyte line expands into multiple discrete virus-specific populations that supply clinically measurable antiviral activity. Monoculture-derived multispecific CTL infusion could provide a safe and efficient means to restore virus-specific immunity in the immunocompromised host.

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Figure 1: Immunophenotype and multivirus specificity of CTL lines generated for clinical use.
Figure 2: Expansion of CMV-specific CTLs in vivo.
Figure 3: In vivo expansion and clinical benefits of EBV-specific T cells.
Figure 4: Effects of adenovirus-specific CTLs in vivo.

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Acknowledgements

We thank V. Torrano for research coordination; J. Gilbert for editorial assistance; T. Gotsolva and F. Jimenez for technical assistance; and T. Lopez and staff in the Good Manufacturing Practice facilities for assisting in CTL preparation and quality assurance. This work was supported in part by a Production Assistance for Cellular Therapies (contract NO1HB37163–C001) from the National Heart, Lung, and Blood Institute (to C.M.R and A.P.G), grants from the National Gene Vector Laboratories (NIH-NCRR U42 RR16578) and the General Clinical Research Center at Baylor College of Medicine (RR00188), a Doris Duke Distinguished Clinical Scientist award (to H.E.H.), the Amy Strelzer Manasevit Scholar Award (to C.M.B.), and an award from the Dana Foundation (to C.M.R. and A.M.L).

Author information

Authors and Affiliations

Authors

Contributions

This study was developed and designed by C.M.R., H.E.H., M.K.B. and C.M.B. The principal investigator on the clinical trial was C.M.B. A.M.L., U.S. and C.M.R. developed the new method for generating the tri-virus specific CTLs. G.D.M., A.M.L. and C.M.B. performed the PCR and immune reconstitution studies. R.A.K., G.C. and K.S.L. cared for the transplant subjects enrolled in this study. H.W. provided statistical support. A.P.G. and M.H.H. supervised C.T.L. preparation and quality assurance. C.-C.C. and J.J.M. provided advice and support for CMV PCR studies and immune monitoring. C.M.R., H.E.H., M.K.B., A.M.L. and C.M.B. contributed to the writing of the paper.

Corresponding author

Correspondence to Catherine M Bollard.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Effects of adenovirus-specific CTL in vivo. (PDF 229 kb)

Supplementary Fig. 2

Activity of polyclonal CTL line for a representative patient, sorted for both CD4+ and CD8+ T cells against CMV. (PDF 143 kb)

Supplementary Table 1

Virus-specific activity of CTL lines. (PDF 58 kb)

Supplementary Table 2

Epitopes recognized in each virus by multivirus-specific CTL compared to monospecific CTL. (PDF 58 kb)

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Leen, A., Myers, G., Sili, U. et al. Monoculture-derived T lymphocytes specific for multiple viruses expand and produce clinically relevant effects in immunocompromised individuals. Nat Med 12, 1160–1166 (2006). https://doi.org/10.1038/nm1475

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