Human CD8+ herpes simplex virus-specific cytotoxic T-lymphocyte clones recognize diverse virion protein antigens

Abstract

The role of the HLA class I-restricted, CD8+, herpes simplex virus (HSV)-specific cytotoxic T lymphocytes (CTL) in the control of human HSV infections is controversial because previous reports suggest that a substantial portion of the antigen-specific lytic response is mediated by CD4+ cells. To address this question directly, we isolated HSV-specific CD8+ CTL clones from a patient with recurrent genital herpes. These CTL were cloned by coculturing responder peripheral blood mononuclear cells (PBMC) with phytohemagglutininstimulated PBMC that had been infected with live HSV-2 and then irradiated prior to the addition of responder cells. After 1 week, CTL were cloned by limiting dilution using phytohemagglutinin stimulation and allogeneic feeder PBMC. Seven clones were isolated; all seven clones were CD8+ CD4-CD3+ DRbeght, six lysed only HSV-2-infected targets, and one lysed both HSV-1-and HSV-2-infected targets. Antigen presentation was restricted by two to three different HLA class I loci. To determine the antigens recognized by these HSV-specific CTL, target cells were infected with HSV in the presence of acyclovir, 5,6-dichloro-1-f-D-ribofuranosylbenzimidazole, or cycloheximide in a series of drug block/release protocols to limit the repertoire of viral gene expression to select transcriptional classes. Five of the clones exhibited a different pattern of cytotoxicity, suggesting that each recognized a distinct HSV antigen.

CD8+ immunodominance among Epstein-Barr virus lytic cycle antigens directly reflects the efficiency of antigen presentation in lytically infected cells

“…It is interesting to contrast this situation with that seen for other human herpesviruses. The human CD8 ϩ T cell response to HSV, an ␣-herpesvirus predominantly replicating in skin and mucosal epithelium, is still quite poorly characterized; however, the evidence to date, at least for HSV type 2, suggests that virus structural proteins of the late lytic cycle constitute some of the major targets (9,10). CMV, a ␤-herpesvirus with a wider cell tropism, is more closely studied and its immunodominant antigens include late structural proteins-in particular the pp65 tegument component-as well as some IE and E proteins (11)(12)(13)(14).…”

“…As a result, their existence in herpesvirus systems is likely to emerge more clearly from studies in man, rather than in mouse models where work tends to focus on a small number of inbred strains (6)(7)(8). Studies to date on human CD8 ϩ T cell responses to herpes simplex virus, an ␣ -herpesvirus, suggest that they are preferentially directed against late structural proteins of the virus (9,10). Likewise with cytomegalovirus, a ␤ -herpesvirus, the pp65 viral tegument protein expressed late in lytic cycle is well established as an immunodominant antigen (11,12)-although not to the exclusion of some more recently discovered IE and E target proteins (13,14).…”

Cytotoxic T lymphocyte response to a wild type hepatitis B virus epitope in patients chronically infected by variant viruses carrying substitutions within the epitope.

“…In contrast to these TCR transgenic mice, the CTL response during human viral infections is usually polyclonal and multispecific which virtually precludes the emergence of escape mutants (21)(22)(23)(24)(25)(26). Nonetheless, HIV genomes containing mutations within an HLA-B8-restricted HIV gag CTL epitope have been shown to emerge in an HIVseropositive donor (27), and a dominant EBV genotype that contains an inactivating substitution within an immunodominant HLA-A11-restricted CTL epitope has been reported in a high HLA-All frequency human population (28).…”

The impact of interspecies recombination on human herpes simplex virus evolution and host immune recognition

“…To obtain bulk HSV-specific CD4 T-cells, HSV-1-reactive CD4 T-cells were enriched from PBMC from a Seattle HSV-1 seropositive, HSV-2 seronegative donor with no clinical history of herpes infection, as published (Jing et al, 2012). Epstein-Barr virus-transformed lymphocyte continuous lines (LCL) from persons of defined HLA type were cultured as described (Tigges et al, 1992) for use as antigen-presenting cells (APC). For functional readouts of CD8 T-cells and cloned CD4 T-cells, LCL were infected overnight with HSV and then co-cultured for an additional 24 hours with T-cells (50,000 LCL and 50,000 T-cells/well in 200 μL in triplicate).…”