Measurement of Varicella‐Zoster Virus (VZV)–Specific Cell‐Mediated Immunity: Comparison between VZV Skin Test and Interferon‐γ Enzyme‐Linked Immunospot Assay

Abstract

Cell-mediated immunity (CMI) is critical for the prevention and control of varicella-zoster virus (VZV)-related disease. To assess CMI to VZV, a varicella skin test and interferon-gamma enzyme-linked immunospot (ELISPOT) assay were both performed in healthy volunteers, and the results were compared. A total of 151 subjects were examined: 16 aged 20-29 years, 26 aged 30-39 years, 18 aged 40-49 years, 73 aged 50-59 years, and 18 aged 60-69 years. All were seropositive by a glycoprotein antigen-based enzyme-linked immunosorbent assay (gpELISA). Skin test reactivity was significantly correlated with the ELISPOT count, and both decreased with increasing age, indicating an age-dependent decline in CMI to VZV. In contrast, the antibody titer obtained by the gpELISA did not correlate with skin test reactivity. The results suggest that the skin test and ELISPOT assay are both reliable for assessing CMI to VZV and can easily be applied to screen individuals susceptible to the development of herpes zoster.

Development of an IFN-γ ELISpot Assay to Assess Varicella-Zoster Virus-specific Cell-mediated Immunity Following Umbilical Cord Blood Transplantation

“…monocytes, dendritic cells) from the PBMC pool, leading to reduced levels of presentation of VZV antigens to responder CD8 + T lymphocytes. The use of irradiated VZV is unlikely to have contributed to the skewing of the response towards CD4 dominance, as similar results were obtained by other groups using different techniques and antigen sources 14,21,22 . A low magnification micrograph of representative ELISpot wells is presented in Figure 4.…”

“…In our case, dilutions of live attenuated VZV vaccine were preferred over cell culture derived VZV in order to insure uniformity in antigen source and preparation and to limit culture and manipulation of live wildtype VZV strains in a clinical setting where nosocomial transmission is a serious concern 10 . In addition, γ irradiation was preferred over UV inactivation 21 because of the comparative precision with which doses of γ radiation can be delivered and because, unlike UV, γ-rays efficiently penetrate sealed VZV containing vials. Irradiation of viral stocks led to consistently higher estimates of the frequency of VZV specific IFN-γ producing cells in PBMC samples obtained from a healthy donor at all dilutions of VZV antigen tested (Figure 1).…”

VZV T Cell-Mediated Immunity

“…Both effector and memory VZV-specific T cells have been identified (Asanuma et al 2000;Diaz et al 1989;Rotbart et al 1993;Patterson-Bartlett et al 2007;Sadaoka et al 2008). These cells recognize epitopes in immediate early IE62, IE63, ORF4, and ORF10 gene products and in late gB, gC, gE, gH, and gI surface glycoproteins (Arvin et al 1986b(Arvin et al , 1991(Arvin et al , 2002Bergen et al 1991;Diaz et al 1988;Giller et al 1989).…”

Reducing the burden of Herpes Zoster in Italy

“…10,11 Approximately 20% of subjects aged >50 y do not show a measurable specific CMI, human Vaccines & Immunotherapeutics Volume 11 Issue 1 notwithstanding persistence of specific antibodies and a positive history of varicella. 12,13 HZ is strictly related to a reduction in VZV-specific T lymphocytes; a Zoster episode reactivates specific T-cell response.…”