EXAMPLE 7
PBMC isolation. Healthy individuals (negative for HIV, hepatitis B virus, and with or without any HSV infection history) were recruited at the University of California Irvine Institute for Clinical and Translational Science. Between 40 and 100 ml blood was drawn into yellow-top Vacutainer tubes (Becton Dickinson). The serum was isolated and stored at −80° C. for detection of antiHSV-1 and antiHSV-2 Abs, as previously described by Chentoufi, A. et al., Asymptomatic human CD4+ cytotoxic T-cell epitopes identified from herpes simplex virus glycoprotein B. J. Virol. 82: 11792-11802 (2008), the content of which is hereby incorporated by reference in its entirety. PBMCs were isolated by gradient centrifugation using leukocyte separation medium (Cellgro, Manassas, Va.). The cells were washed in PBS and resuspended in complete culture medium consisting of RPMI 1640 medium containing 10% FBS (Gemini Bio-Products, Woodland, Calif.) supplemented with 1× penicillin/L-glutamine/streptomycin, 1× sodium pyruvate, 1× nonessential amino acids, and 50 μM 2-ME (Life Technologies, Rockville, Md.). Aliquots of freshly isolated PBMCs were also cryopreserved in 90% FCS and 10% DMSO in liquid nitrogen.