Quantification of a cell-mediated immune response against varicella zoster virus by assessing responder CD4high memory cell proliferation in activated whole blood cultures

Abstract

BackgroundHerpes zoster (HZ) is caused by reactivation of a latent varicella zoster virus (VZV). The potential to develop HZ increases with age due to waning of memory cell-mediated immunity (CMI), mainly the CD4 response. Therefore, VZV-CD4-memory T cells (CD4-M) count in blood could serve as a barometer for HZ protection. However, direct quantification of these cells is known to be difficult because they are few in number in the blood. We thus developed a method to measure the proliferation level of CD4-M cells responding to VZV antigen in whole blood culture. MethodsBlood samples were collected from 32 children (2–15 years old) with or without a history of varicella infection, 18 young adults (28–45 years old), and 80 elderly (50–86 years old) with a history of varicella infection. The elderly group was vaccinated, and blood samples were taken 2 months and 1 year after VZV vaccination. Then, 1 mL of blood was mixed with VZV, diluted 1/10 in medium, and cultured. CD4-M cells were identified and measured by flow cytometry. ResultsThere was distinct proliferation of CD3+CD4highCD45RA−RO+ (CD4high-M) cells specific to VZV antigen at day 9. The majority of CD4high-M cells had the effector memory phenotype CCR7− and was granzyme B-positive. CD4high-M cells were detected in blood culture from varicella-immune but not varicella-non-immune children. Meanwhile, a higher level of CD4high-M proliferation was observed in young adults than in the elderly. The CD4high-M proliferation level was boosted 2 months after VZV vaccination and maintained for at least 1 year in the elderly. ConclusionQuantifying VZV responder CD4high -M cell proliferation is a convenient way to measure VZV CMI using small blood volumes. Our method can be applied to measure VZV vaccine-induced CMI in the elderly.Clinical study registry numbers: (www.clinicaltrials.jp) 173532 and 183985.