A recombinant rhesus monkey rhadinovirus deleted of glycoprotein L establishes persistent infection of rhesus macaques and elicits conventional T cell responses

Alexander S. Hahn, Georg F. Bischof, Anna K. Großkopf, Young C. Shin, Aline Domingues, Lucas Gonzalez-Nieto, Eva G. Rakasz, David I. Watkins, Armin Ensser, Mauricio A. Martins, Ronald C. Desrosiers

Research output: Contribution to journalArticle

Abstract

A replication-competent, recombinant strain of rhesus monkey rhadinovirus (RRV) expressing the Gag protein of SIVmac239 was constructed in the context of a glycoprotein L (gL) deletion mutation. Deletion of gL detargets the virus from Eph family receptors. The ability of this gL-minus Gag recombinant RRV to infect, persist, and elicit immune responses was evaluated after intravenous inoculation of two Mamu-A*01 RRV-naive rhesus monkeys. Both monkeys responded with an anti-RRV antibody response, and quantitation of RRV DNA in peripheral blood mononuclear cells (PBMC) by real-time PCR revealed levels similar to those in monkeys infected with recombinant gL RRV. Comparison of RRV DNA levels in sorted CD3 versus CD20 versus CD14 PBMC subpopulations indicated infection of the CD20 subpopulation by the gL-minus RRV. This contrasts with results obtained with transformed B cell lines in vitro, in which deletion of gL resulted in markedly reduced infectivity. Over a period of 20 weeks, Gag-specific CD8 T cell responses were documented by major histocompatibility complex class I (MHC-I) tetramer staining. Vaccine-induced CD8 T cell responses, which were predominantly directed against the Mamu-A*01-restricted Gag181-189CM9 epitope, could be inhibited by blockade of MHC-I presentation. Our results indicate that gL and the interaction with Eph family receptors are dispensable for the colonization of the B cell compartment following high-dose infection by the intravenous route, which suggests the existence of alternative receptors. Further, gL-minus RRV elicits cellular immune responses that are predominantly canonical in nature. IMPORTANCE Kaposi’s sarcoma-associated herpesvirus (KSHV) is associated with a substantial disease burden in sub-Saharan Africa, often in the context of human immunodeficiency virus (HIV) infection. The related rhesus monkey rhadinovirus (RRV) has shown potential as a vector to immunize monkeys with antigens from simian immunodeficiency virus (SIV), the macaque model for HIV. KSHV and RRV engage cellular receptors from the Eph family via the viral gH/gL glycoprotein complex. We have now generated a recombinant RRV that expresses the SIV Gag antigen and does not express gL. This recombinant RRV was infectious by the intravenous route, established persistent infection in the B cell compartment, and elicited strong immune responses to the SIV Gag antigen. These results argue against a role for gL and Eph family receptors in B cell infection by RRV in vivo and have implications for the development of a live-attenuated KSHV vaccine or vaccine vector.

Original languageEnglish (US)
Article numbere01093-19
JournalJournal of virology
Volume94
Issue number2
DOIs
StatePublished - Jan 1 2020

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Rhadinovirus
Macaca mulatta
glycoproteins
Glycoproteins
T-lymphocytes
T-Lymphocytes
Infection
infection
Eph Family Receptors
Human herpesvirus 8
Simian immunodeficiency virus
gag Gene Products
Human Herpesvirus 8
Simian Immunodeficiency Virus
B-lymphocytes
B-Lymphocytes
receptors
Haplorhini
monkeys
major histocompatibility complex

Keywords

  • Glycoproteins
  • Live vector vaccines
  • Rhadinovirus
  • Simian immunodeficiency virus

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

A recombinant rhesus monkey rhadinovirus deleted of glycoprotein L establishes persistent infection of rhesus macaques and elicits conventional T cell responses. / Hahn, Alexander S.; Bischof, Georg F.; Großkopf, Anna K.; Shin, Young C.; Domingues, Aline; Gonzalez-Nieto, Lucas; Rakasz, Eva G.; Watkins, David I.; Ensser, Armin; Martins, Mauricio A.; Desrosiers, Ronald C.

In: Journal of virology, Vol. 94, No. 2, e01093-19, 01.01.2020.

Research output: Contribution to journalArticle

Hahn, Alexander S. ; Bischof, Georg F. ; Großkopf, Anna K. ; Shin, Young C. ; Domingues, Aline ; Gonzalez-Nieto, Lucas ; Rakasz, Eva G. ; Watkins, David I. ; Ensser, Armin ; Martins, Mauricio A. ; Desrosiers, Ronald C. / A recombinant rhesus monkey rhadinovirus deleted of glycoprotein L establishes persistent infection of rhesus macaques and elicits conventional T cell responses. In: Journal of virology. 2020 ; Vol. 94, No. 2.
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abstract = "A replication-competent, recombinant strain of rhesus monkey rhadinovirus (RRV) expressing the Gag protein of SIVmac239 was constructed in the context of a glycoprotein L (gL) deletion mutation. Deletion of gL detargets the virus from Eph family receptors. The ability of this gL-minus Gag recombinant RRV to infect, persist, and elicit immune responses was evaluated after intravenous inoculation of two Mamu-A*01 RRV-naive rhesus monkeys. Both monkeys responded with an anti-RRV antibody response, and quantitation of RRV DNA in peripheral blood mononuclear cells (PBMC) by real-time PCR revealed levels similar to those in monkeys infected with recombinant gL RRV. Comparison of RRV DNA levels in sorted CD3 versus CD20 versus CD14 PBMC subpopulations indicated infection of the CD20 subpopulation by the gL-minus RRV. This contrasts with results obtained with transformed B cell lines in vitro, in which deletion of gL resulted in markedly reduced infectivity. Over a period of 20 weeks, Gag-specific CD8 T cell responses were documented by major histocompatibility complex class I (MHC-I) tetramer staining. Vaccine-induced CD8 T cell responses, which were predominantly directed against the Mamu-A*01-restricted Gag181-189CM9 epitope, could be inhibited by blockade of MHC-I presentation. Our results indicate that gL and the interaction with Eph family receptors are dispensable for the colonization of the B cell compartment following high-dose infection by the intravenous route, which suggests the existence of alternative receptors. Further, gL-minus RRV elicits cellular immune responses that are predominantly canonical in nature. IMPORTANCE Kaposi’s sarcoma-associated herpesvirus (KSHV) is associated with a substantial disease burden in sub-Saharan Africa, often in the context of human immunodeficiency virus (HIV) infection. The related rhesus monkey rhadinovirus (RRV) has shown potential as a vector to immunize monkeys with antigens from simian immunodeficiency virus (SIV), the macaque model for HIV. KSHV and RRV engage cellular receptors from the Eph family via the viral gH/gL glycoprotein complex. We have now generated a recombinant RRV that expresses the SIV Gag antigen and does not express gL. This recombinant RRV was infectious by the intravenous route, established persistent infection in the B cell compartment, and elicited strong immune responses to the SIV Gag antigen. These results argue against a role for gL and Eph family receptors in B cell infection by RRV in vivo and have implications for the development of a live-attenuated KSHV vaccine or vaccine vector.",
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T1 - A recombinant rhesus monkey rhadinovirus deleted of glycoprotein L establishes persistent infection of rhesus macaques and elicits conventional T cell responses

AU - Hahn, Alexander S.

AU - Bischof, Georg F.

AU - Großkopf, Anna K.

AU - Shin, Young C.

AU - Domingues, Aline

AU - Gonzalez-Nieto, Lucas

AU - Rakasz, Eva G.

AU - Watkins, David I.

AU - Ensser, Armin

AU - Martins, Mauricio A.

AU - Desrosiers, Ronald C.

PY - 2020/1/1

Y1 - 2020/1/1

N2 - A replication-competent, recombinant strain of rhesus monkey rhadinovirus (RRV) expressing the Gag protein of SIVmac239 was constructed in the context of a glycoprotein L (gL) deletion mutation. Deletion of gL detargets the virus from Eph family receptors. The ability of this gL-minus Gag recombinant RRV to infect, persist, and elicit immune responses was evaluated after intravenous inoculation of two Mamu-A*01 RRV-naive rhesus monkeys. Both monkeys responded with an anti-RRV antibody response, and quantitation of RRV DNA in peripheral blood mononuclear cells (PBMC) by real-time PCR revealed levels similar to those in monkeys infected with recombinant gL RRV. Comparison of RRV DNA levels in sorted CD3 versus CD20 versus CD14 PBMC subpopulations indicated infection of the CD20 subpopulation by the gL-minus RRV. This contrasts with results obtained with transformed B cell lines in vitro, in which deletion of gL resulted in markedly reduced infectivity. Over a period of 20 weeks, Gag-specific CD8 T cell responses were documented by major histocompatibility complex class I (MHC-I) tetramer staining. Vaccine-induced CD8 T cell responses, which were predominantly directed against the Mamu-A*01-restricted Gag181-189CM9 epitope, could be inhibited by blockade of MHC-I presentation. Our results indicate that gL and the interaction with Eph family receptors are dispensable for the colonization of the B cell compartment following high-dose infection by the intravenous route, which suggests the existence of alternative receptors. Further, gL-minus RRV elicits cellular immune responses that are predominantly canonical in nature. IMPORTANCE Kaposi’s sarcoma-associated herpesvirus (KSHV) is associated with a substantial disease burden in sub-Saharan Africa, often in the context of human immunodeficiency virus (HIV) infection. The related rhesus monkey rhadinovirus (RRV) has shown potential as a vector to immunize monkeys with antigens from simian immunodeficiency virus (SIV), the macaque model for HIV. KSHV and RRV engage cellular receptors from the Eph family via the viral gH/gL glycoprotein complex. We have now generated a recombinant RRV that expresses the SIV Gag antigen and does not express gL. This recombinant RRV was infectious by the intravenous route, established persistent infection in the B cell compartment, and elicited strong immune responses to the SIV Gag antigen. These results argue against a role for gL and Eph family receptors in B cell infection by RRV in vivo and have implications for the development of a live-attenuated KSHV vaccine or vaccine vector.

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KW - Glycoproteins

KW - Live vector vaccines

KW - Rhadinovirus

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