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

doi:10.1002/cyto.a.24197

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 journal › Article › peer-review

2 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. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.

Original language	English (US)
Pages (from-to)	278-288
Number of pages	11
Journal	Cytometry Part A
Volume	99
Issue number	3
DOIs	https://doi.org/10.1002/cyto.a.24197
State	Published - 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.

Keywords

CD8 T-cell epitope
immunophenotyping
in situ tetramer staining
rhesus cytomegalovirus
Cytomegalovirus
Humans
Histocompatibility Antigens Class I
Cytomegalovirus Infections
Immunophenotyping
Macaca mulatta
Epitopes, T-Lymphocyte
CD8-Positive T-Lymphocytes
Animals
Simian Immunodeficiency Virus

PubMed: MeSH publication types

Journal Article
Research Support, N.I.H., Extramural

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1002/cyto.a.24197

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title = "Immunophenotyping of Rhesus CMV-Specific CD8 T-Cell Populations",

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. {\textcopyright} 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.",

keywords = "CD8 T-cell epitope, immunophenotyping, in situ tetramer staining, rhesus cytomegalovirus, Cytomegalovirus, Humans, Histocompatibility Antigens Class I, Cytomegalovirus Infections, Immunophenotyping, Macaca mulatta, Epitopes, T-Lymphocyte, CD8-Positive T-Lymphocytes, Animals, Simian Immunodeficiency Virus",

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

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: {\textcopyright} 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.",

year = "2021",

month = mar,

doi = "10.1002/cyto.a.24197",

language = "English (US)",

volume = "99",

pages = "278--288",

journal = "Cytometry Part A",

issn = "1552-4922",

publisher = "John Wiley and Sons Inc.",

number = "3",

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TY - JOUR

T1 - Immunophenotyping of Rhesus CMV-Specific CD8 T-Cell Populations

AU - Pomplun, Nicholas L.

AU - Vosler, Logan

AU - Weisgrau, Kim L.

AU - Furlott, Jessica

AU - Weiler, Andrea M.

AU - Abdelaal, Hadia M.

AU - Evans, David T.

AU - Watkins, David I.

AU - Matano, Tetsuro

AU - Skinner, Pamela J.

AU - Friedrich, Thomas C.

AU - Rakasz, Eva G.

N1 - 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.

PY - 2021/3

Y1 - 2021/3

N2 - 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. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.

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

KW - CD8 T-cell epitope

KW - immunophenotyping

KW - in situ tetramer staining

KW - rhesus cytomegalovirus

KW - Cytomegalovirus

KW - Humans

KW - Histocompatibility Antigens Class I

KW - Cytomegalovirus Infections

KW - Immunophenotyping

KW - Macaca mulatta

KW - Epitopes, T-Lymphocyte

KW - CD8-Positive T-Lymphocytes

KW - Animals

KW - Simian Immunodeficiency Virus

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UR - http://www.scopus.com/inward/citedby.url?scp=85089006258&partnerID=8YFLogxK

U2 - 10.1002/cyto.a.24197

DO - 10.1002/cyto.a.24197

M3 - Article

C2 - 32713108

AN - SCOPUS:85089006258

SN - 1552-4922

VL - 99

SP - 278

EP - 288

JO - Cytometry Part A

JF - Cytometry Part A

IS - 3

ER -

Immunophenotyping of Rhesus CMV-Specific CD8 T-Cell Populations

Abstract

Bibliographical note

Keywords

PubMed: MeSH publication types

UN SDGs

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