Generation of hepatitis C virus-specific cytotoxic T lymphocytes from healthy individuals with peptide-pulsed dendritic cells.
OBJECTIVE
In hepatitis C virus (HCV) infection, cytotoxic T lymphocytes (CTL) are involved in liver inflammation and contribute to the reduction of viral load. Antibodies for HCV-CTL precursor frequencies (CTLpf) are relatively low in chronic hepatitis C, and this may be related to the poor CTL response in vivo. The aim of this study was to assess the efficacy of dendritic cells (DC) as antigen-presenting cells in CTL generation from low CTLpf.


METHODS
To confirm the rationale of using DC to prime naive T cells, five HCV-uninfected individuals were enrolled in the study. We obtained DC by maturation from peripheral progenitors under stimulation with granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-4 and IL-1alpha. Autologous T cells were cultured with DC or concanavalin-A-induced blasts loaded with four HCV-derived peptides bearing human leukocyte antigen (HLA)-A*0201 or -A24 motifs for 28 days under IL-7 and IL-2 stimulation. The lytic activity against peptide-pulsed targets was assessed by using a [51Cr]-releasing assay.


RESULTS
The DC strongly expressed HLA class I, II, B7-1 and B7-2, but not phenotypic markers of T-, B-, natural killer (NK)-cells or monocytes. The CD8-positive, HLA-class I-restricted and HCV peptide-specific CTL were generated with DC from HLA-A antigen-matched subjects, whereas no CTL activity was detected with concavalin (Con-A) blasts. We were thus able to generate HCV specific CTL from naive precursors with peptide-pulsed DC.


CONCLUSIONS
This DC-based system can be used to generate CTL of desired antigen specificity, even from a source with low CTLpf.
Ito
A
A
Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita, Japan.
Kanto
T
T
Kuzushita
N
N
Tatsumi
T
T
Sugimoto
Y
Y
Miyagi
T
T
Takehara
T
T
Katayama
K
K
Mochizuki
K
K
Hiramatsu
N
N
Kasahara
A
A
Yoshiya
I
I
Sasaki
Y
Y
Hori
M
M
Hayashi
N
N
D000328:Adult; D000938:Antigen-Presenting Cells / Q000276:immunology; D002455:Cell Division; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology / Q000502:physiology*; D000939:Epitopes; D006680:HLA Antigens / Q000032:analysis; D015234:HLA-A Antigens / Q000032:analysis; D015789:HLA-A2 Antigen; D016174:Hepacivirus / Q000276:immunology*; D006801:Humans; D008297:Male; D010455:Peptides / Q000494:pharmacology*; D010641:Phenotype; D012016:Reference Values; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology* / Q000276:immunology*; D014764:Viral Proteins / Q000494:pharmacology
D000939:Epitopes; D006680:HLA Antigens; D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D010455:Peptides; D014764:Viral Proteins