Immunophenotyping of Rhesus CMV-Specific CD8 T-Cell Populations

Nicholas L. Pomplun, Logan Vosler, Kim L. Weisgrau, Jessica Furlott, Andrea M. Weiler, Hadia M. Abdelaal, David T. Evans, David I. Watkins, Tetsuro Matano, Pamela J. Skinner, Thomas C. Friedrich, Eva G. Rakasz

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

A vaccine to ameliorate cytomegalovirus (CMV)-related pathogenicity in transplantation patients is considered a top priority. A therapeutic vaccine must include components that elicit both neutralizing antibodies, and highly effective CD8 T-cell responses. The most important translational model of vaccine development is the captive-bred rhesus macaque (Macaca mulatta) of Indian origin. There is a dearth of information on rhesus cytomegalovirus (rhCMV)-specific CD8 T cells due to the absence of well-defined CD8 T-cell epitopes presented by classical MHC-I molecules. In the current study, we defined two CD8 T-cell epitopes restricted by high-frequency Mamu alleles: the Mamu-A1*002:01 restricted VY9 (VTTLGMALY aa291-299) epitope of protein IE-1, and the Mamu-A1*008:01 restricted NP8 (NPTDRPIP aa96-103) epitope of protein phosphoprotein 65-2. We developed tetramers and determined the level, phenotype, and functional capability of the two epitope-specific T-cell populations in circulation and various tissues. We demonstrated the value of these tetramers for in situ tetramer staining. Here, we first provided critical reagents and established a flow cytometric staining strategy to study rhCMV-specific T-cell responses in up to 40% of captive-bred rhesus macaques.

Original languageEnglish (US)
Pages (from-to)278-288
Number of pages11
JournalCytometry Part A
Volume99
Issue number3
DOIs
StatePublished - Mar 2021

Bibliographical note

Funding Information:
We wish to gratefully acknowledge the Scientific Protocol Implementation, the Veterinary Services, and Pathology units, and Genetics Services core laboratory of the Wisconsin National Primate Research Center. We would like to thank Dr. Roger Weisman for his helpful advice and Mr. Jason Weinfurter for his technical support. We acknowledge the NIH Tetramer Core Facility at Emory University for producing the tetramers. This study was supported by National Center for Research Resources grant number P51OD011106 for the Wisconsin National Primate Research Center.

Funding Information:
We wish to gratefully acknowledge the Scientific Protocol Implementation, the Veterinary Services, and Pathology units, and Genetics Services core laboratory of the Wisconsin National Primate Research Center. We would like to thank Dr. Roger Weisman for his helpful advice and Mr. Jason Weinfurter for his technical support. We acknowledge the NIH Tetramer Core Facility at Emory University for producing the tetramers. This study was supported by National Center for Research Resources grant number P51OD011106 for the Wisconsin National Primate Research Center.

Publisher Copyright:
© 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

Keywords

  • CD8 T-cell epitope
  • immunophenotyping
  • in situ tetramer staining
  • rhesus cytomegalovirus

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