June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Ex vivo quantification and memory phenotyping of HSV-1-specific CD8+ T-cells in Herpes Simplex Keratitis
Author Affiliations & Notes
  • John Curnow
    Centre for Translational Inflammation Research, University of Birmingham, Birmingham, United Kingdom
    Academic Unit of Ophthalmology, University of Birmingham, Birmingham, United Kingdom
  • Lindsay Durant
    Centre for Translational Inflammation Research, University of Birmingham, Birmingham, United Kingdom
  • Sai Kolli
    Queen Elizabeth Hospital Birmingham, Birmingham, United Kingdom
  • Peter McDonnell
    Queen Elizabeth Hospital Birmingham, Birmingham, United Kingdom
  • Saaeha Rauz
    Centre for Translational Inflammation Research, University of Birmingham, Birmingham, United Kingdom
    Academic Unit of Ophthalmology, University of Birmingham, Birmingham, United Kingdom
  • Geraint P Williams
    Centre for Translational Inflammation Research, University of Birmingham, Birmingham, United Kingdom
    Academic Unit of Ophthalmology, University of Birmingham, Birmingham, United Kingdom
  • Footnotes
    Commercial Relationships John Curnow, None; Lindsay Durant, None; Sai Kolli, None; Peter McDonnell, None; Saaeha Rauz, None; Geraint Williams, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 1857. doi:
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      John Curnow, Lindsay Durant, Sai Kolli, Peter McDonnell, Saaeha Rauz, Geraint P Williams; Ex vivo quantification and memory phenotyping of HSV-1-specific CD8+ T-cells in Herpes Simplex Keratitis. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1857.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: Demonstration of human Herpes Simplex Virus (HSV) recognition by CD8+ T cells is challenging. They have been reported using MHC class I-peptide tetramers, but largely following expansion in vitro. We present a pilot study to quantify ex vivo the CD8+ T cell response to reported immunodominant HSV-1 peptides in recurrent herpes simplex keratitis (HSK) and healthy controls (HC).

Methods: 20 patients with HSK (median age 66 [range 35-90]) and 20 age-matched (58 [36-80]) HC were enrolled following ethical approval. Clinical phenotyping was undertaken together with peripheral blood/serum samples for HLA-A*0201 typing, HSV-1 sero-status and mononuclear cell isolation. CD8+ T cell memory status was demonstrated with antibodies specific for CD45RA and CCR7, and HSV-1 viral epitope recognition using MHC class I pentamers (HLA-A*02:01; gD53-61, gD70 -78, gD278 -286, gB183-191, gB441-449, gB561-569 and gB342-350). Samples were analysed by flow cytometry.

Results: The median duration of symptoms was 21 years [1-73] with 8/20 HSK patients requiring long-term oral Acyclovir prophylaxis (19 [12-36] months duration). The time since last relapse was significantly shorter in the prophylaxis group (p<0.05). There were no significant differences in the total CD8+ naïve and memory compartment frequencies between the HSK and HC groups. We were able to detect gD70 -78-specific memory CD8+ T cells only in HLA-A*02:01+ and HSV-1 seropositive individuals, with frequencies of 0.05% (0-1.74%) and 0.085% (0-6.39%) within the CD8+ cells, for HSK and HC respectively (p=0.82). The HSV-1-specific CD8+ expansions were predominantly of an effector memory (CD45RA-CCR7-) or effector memory RA+ (CD45RA+CCR7+) phenotype.

Conclusions: In this cross-sectional study of patients we have detected ex vivo HLA-A*02:01-restricted CD8+ T cells specific for gD70 -78 of HSV-1. These responses were only present in HLA-A*02:01+ HSV-1 seropositive individuals, and were found in both HC and those with HSK. These investigations suggest that further characterisation in a larger cohort is warranted, to establish any differences in epitope recognition during HSK.

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