Abstract:
:A coupled luminescent method (CLM) based on glyceraldehyde-3-phosphate dehydrogenase released from injured target cells was used to evaluate the cytotoxicity of antigen-specific HLA class I-restricted CTLs. In contrast to established methods, CLM does not require the pretreatment of target cells with radioactive or toxic labeling substances. CTLs from healthy HLA-A2 positive donors were stimulated by autologous dendritic cells (DCs) pulsed with HLA-A2 restricted HCMV-pp65 nonamer peptides. HLA-A2 positive T2 cells or autologous monocytes pulsed with HCMV-pp65 nonamer peptide served as target cells. Lysis was detected only in HCMV-pp65-pulsed target cells incubated with CTLs from seropositive donors stimulated by HCMV-pp65-pulsed DCs. After 3 days, stimulation 38% of T2 cells and 17% of monocytes were lysed at an effector to target ratio of 8:1. In conclusion, CLM represents a highly sensitive, fast, material-saving and non-toxic/non-radioactive method for the measurement of antigen-specific CTL cytotoxic activity.
journal_name
Med Microbiol Immunoljournal_title
Medical microbiology and immunologyauthors
Ogbomo H,Geiler J,Leutz A,von Kietzell K,Michaelis M,Doerr HW,Cinatl J Jrdoi
10.1007/s00430-009-0126-5subject
Has Abstractpub_date
2009-11-01 00:00:00pages
257-62issue
4eissn
0300-8584issn
1432-1831journal_volume
198pub_type
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