Expansion of EBV latent membrane protein 2a specific cytotoxic T cells for the adoptive immunotherapy of EBV latency type 2 malignancies: influence of recombinant IL12 and IL15.
BACKGROUND
EBV-associated malignancies with a Type II latency gene expression pattern, such as EBV-positive HD, or nasopharyngeal carcinoma, frequently express the EBV latency Ag LMP2a. Hence, they provide a potential target for adoptive immunotherapy using in vitro-generated LMP2a-specific cytotoxic T lymphocytes (CTL). In this study, LMP2a-specific CTL were specifically amplified and the influence of rIL12 and rIL15 on the culture outcome was tested.


METHODS
PBMC from donors were stimulated twice with autologous DC transduced with an adenovirus vector expressing LMP2a. This led to a significant expansion of LMP2a-tetramer-specific CTL, which were subsequently further expanded with autologous EBV-transformed B-lymphoblastoid cells (LCL). The addition of rIL12 and rIL15 to the standard IL2-containing culture medium enhanced the proliferation of LMP2a-specific CTL.


RESULTS
While rIL15 did not change the pattern of cytokines secreted by LMP2a-CTL, rIL12 enhanced the production of Th1/Tc1 cytokines, such as IFN-n, while suppressing the production of the Th2/Tc2 cytokine IL5.


CONCLUSIONS
Stimulation of CTL cultures with rIL12 or rIL15 will generate CTL more rapidly, facilitating the application of this approach for patients with these EBV-associated disorders.
Wagner
H-J
HJ
Department of Immunology, St Jude Children's Research Hospital, Memphis, TN, USA.
Sili
U
U
Gahn
B
B
Vigouroux
S
S
Huls
M H
MH
Xie
W
W
Vignali
D
D
Brenner
M K
MK
Heslop
H E
HE
Rooney
C M
CM
D000256:Adenoviridae / Q000235:genetics; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology; D020031:Epstein-Barr Virus Infections / Q000276:immunology* / Q000628:therapy; D005822:Genetic Vectors; D006801:Humans; D016219:Immunotherapy, Adoptive; D018664:Interleukin-12 / Q000235:genetics / Q000494:pharmacology*; D019409:Interleukin-15 / Q000235:genetics / Q000494:pharmacology*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation; D011994:Recombinant Proteins / Q000494:pharmacology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000637:transplantation; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*
D016207:Cytokines; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D019409:Interleukin-15; D011994:Recombinant Proteins; D014763:Viral Matrix Proteins; D018664:Interleukin-12