In the past grant period, we have shown that HSV-2 specific CD8+ T cells reside and persist at the dermal-epidermal junction (DEJ) contiguous to the sensory nerve termini innervating the basal keratinocytes of the genital skin. Using laser capture microdissection we have isolated single CD8+ DEJ T cells in situ and interrogating these cells with transcriptional arrays and quantitative RT PCR assays. Our data indicates these cells express a wide variety of cytolytic and antiviral cytokines, as well as a unique homodimeric ?? chain signature. We have also developed rigorous and specific sampling methods to precisely pinpoint the sites of HSV-2 reactivation in the genitalia in time and space. Our preliminary data have shown wide differences in the number of CD8+ DEJ T cells in areas of genital skin. We now propose to apply these and other advanced methodologies to understand the immune surveillance mechanism of HSV-2 specific CD8+ T cells. Specifically, in Aim 1 we will probe the relationship between immunosurveillance and duration, severity, and frequency of HSV-2 reactivation. We hypothesize that the number of CD8+ T cells and/or their levels of expression of cytolytic and antiviral cytokines/chemokines will be inversely correlated with frequency of HSV-2 reactivation. Areas in which DEJ CD8+ T cells are in low numbers or express low functional activity will have a greater likelihood of reactivation than areas which are populated with high CD8+ T cells at the DEJ. We will also explore the relationships between clonal diversity and reactivation.
Aim 2 is directed at defining host-viral interactions among HIV+ persons. We hypothesize that HIV+ persons will exhibit altered quantitative and functional differences in immune surveillance and that these differences will be reflected in poorer containment of HSV-2. The comparisons between HIV+/HSV-2+ and HIV-/HSV-2+ persons will provide further evidence of the importance of CD8+ T cells at the DEJ in immune surveillance and corroborate the mechanistic functions define in S.A. 1.

Public Health Relevance

This proposal aims to understand the mechanism by which host immune cells, specifically resident mucosal CD8+ T cells, contain herpes simplex virus-2 (HSV-2) reactivation at the source of their release from neuronal termini into basal keratinocytes of the genital skin/mucosa. Using laser capture microdissection and transcriptional arrays, we have shown that HSV-2 specific CD8+ T cells persist at the dermal-epidermal junction and that these cells express several cytolytic and antiviral cytokines. We now aim to further study the immune surveillance function of these cells and how they contain viral reactivation, a key concept that will lay the groundwork for better design and development of HSV-2 vaccines.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI042528-15
Application #
8262112
Study Section
AIDS-associated Opportunistic Infections and Cancer Study Section (AOIC)
Program Officer
David, Hagit S
Project Start
1997-12-01
Project End
2017-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
15
Fiscal Year
2012
Total Cost
$453,463
Indirect Cost
$103,786
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
078200995
City
Seattle
State
WA
Country
United States
Zip Code
98109
Milman, Neta; Zhu, Jia; Johnston, Christine et al. (2016) In Situ Detection of Regulatory T Cells in Human Genital Herpes Simplex Virus Type 2 (HSV-2) Reactivation and Their Influence on Spontaneous HSV-2 Reactivation. J Infect Dis 214:23-31
Tjernlund, Annelie; Burgener, Adam; Lindvall, Jessica M et al. (2016) In Situ Staining and Laser Capture Microdissection of Lymph Node Residing SIV Gag-Specific CD8+ T cells--A Tool to Interrogate a Functional Immune Response Ex Vivo. PLoS One 11:e0149907
Posavad, C M; Zhao, L; Mueller, D E et al. (2015) Persistence of mucosal T-cell responses to herpes simplex virus type 2 in the female genital tract. Mucosal Immunol 8:115-26
Diem, Kurt; Magaret, Amalia; Klock, Alexis et al. (2015) Image analysis for accurately counting CD4+ and CD8+ T cells in human tissue. J Virol Methods 222:117-21
Johnston, Christine; Zhu, Jia; Jing, Lichen et al. (2014) Virologic and immunologic evidence of multifocal genital herpes simplex virus 2 infection. J Virol 88:4921-31
Schiffer, Joshua T; Swan, David; Al Sallaq, Ramzi et al. (2013) Rapid localized spread and immunologic containment define Herpes simplex virus-2 reactivation in the human genital tract. Elife 2:e00288
Zhu, Jia; Peng, Tao; Johnston, Christine et al. (2013) Immune surveillance by CD8ýýýý+ skin-resident T cells in human herpes virus infection. Nature 497:494-7
Schiffer, Joshua T; Corey, Lawrence (2013) Rapid host immune response and viral dynamics in herpes simplex virus-2 infection. Nat Med 19:280-90
Schiffer, Joshua T; Swan, David A; Corey, Lawrence et al. (2013) Rapid viral expansion and short drug half-life explain the incomplete effectiveness of current herpes simplex virus 2-directed antiviral agents. Antimicrob Agents Chemother 57:5820-9
Jing, Lichen; Haas, Jürgen; Chong, Tiana M et al. (2012) Cross-presentation and genome-wide screening reveal candidate T cells antigens for a herpes simplex virus type 1 vaccine. J Clin Invest 122:654-73

Showing the most recent 10 out of 26 publications