T Cells Specific for the Triggering Virus Infiltrate the Eye in Patients with Herpes Simplex Virus-Mediated Acute Retinal Necrosis

Abstract

Acute retinal necrosis (ARN) is a rare, potentially blinding retinal disease resulting from ocular infections with herpes simplex virus (HSV) or varicella-zoster virus (VZV). To determine the antigen specificity and functional characteristics of ocular infiltrating T cells in ARN, T cells were isolated and expanded nonspecifically from intraocular fluid (IOF) samples from 2 patients with HSV-1- and 3 with VZV-mediated ARN. HSV-specific T cell reactivity could be detected only in the IOF-derived T cell lines (TCLs) of the 2 patients with HSV-mediated ARN. These TCLs consisted of both HSV type-common and type-specific CD4+ and CD8+ T cell clones (TCCs) with differential T cell receptor usage. Irrespective of their phenotype, the TCCs were cytolytic and secreted interferon-gamma, tumor necrosis factor-alpha, interleukin-4, and interleukin-5. In both patients, the antigen specificity of a substantial number of HSV-1-specific TCCs could be mapped to approximately 0.67-0.73 HSV-1 map units. The data presented suggest the contribution of T cells, specific for the triggering virus, to the pathogenesis of ARN.

Selective retention of herpes simplex virus-specific T cells in latently infected human trigeminal ganglia

“…TG-derived TCLs were generated by phytohemagglutinin (PHA) stimulation of one-fifth of the TG single-cell suspension in the presence of ␥-irradiated (3,000 rad) allogeneic PBMCs in RPMI medium 1640 supplemented with 10% human pooled serum, antibiotics, and recombinant human IL-2 (50 units/ml) essentially as described in ref. 46. The limited number of T cells necessitated two rounds of PHA stimulation to obtain sufficient T cells for cryopreservation and subsequent functional assays.…”

“…46 and 47. An rVV poly 186 without insert was used as control (46). The antigen-specificity of TCLs was determined in an IFN-␥ ELISPOT assay, as described in ref.…”

Functional Characterization of Ocular-Derived Human Alphaherpesvirus Cross-Reactive CD4 T Cells

“…The IOF-derived HSV-1-specific CD4 TCC 44 was previously obtained from the affected eye of an HSV-1 ARN patient (patient 1 in Ref. 21). Autologous B cell lines (BLCL) were obtained by transformation of peripheral blood B cells with EBV as described previously (19,21).…”

“…21). Autologous B cell lines (BLCL) were obtained by transformation of peripheral blood B cells with EBV as described previously (19,21). TCC were cultured in RPMI 1640 medium (Lonza) supplemented with 10% heat-inactivated human pooled serum and antibiotics (hereafter referred to as T cell medium [TCM]) (18,21).…”

“…Studies on the experimental HSV-1 ARN mouse model have demonstrated the pathogenic role of ocular infiltrating T cells (11,16). The intraocular virus-specific T cell response in aHHV uveitis patients, mediated by cytotoxic CD4 and CD8 Th0/Th1-like T cells, is preferentially directed to the causative herpesvirus (17)(18)(19)(20)(21). Herpesvirus particles and Ags have been detected in the neural retina and retinal pigment epithelial (RPE) cell layer of patients with HSV and VZV posterior uveitis (22)(23)(24).…”