|
|
 Previous Article | Table of Contents | Next Article 
Blood, Vol. 94 No. 9 (November 1), 1999:
pp. 3242-3250
Simultaneous Ex Vivo Expansion of Cytomegalovirus and Epstein-Barr
Virus-Specific Cytotoxic T Lymphocytes Using B-Lymphoblastoid Cell
Lines Expressing Cytomegalovirus pp65
Qi Sun,
Karen E. Pollok,
Robert L. Burton,
Li Jun Dai,
William Britt,
David J. Emanuel, and
Kenneth G. Lucas
From the Bone Marrow Transplantation Program, University of Alabama
at Birmingham, Birmingham, AL; the Herman B. Wells Center for Pediatric
Research, Riley Hospital for Children, Indianapolis, IN; the Department
of Pediatrics, The Children's Hospital of Alabama, Birmingham,
AL; and Schering-Plough Research Institute, Kenilworth,
NJ.
Cytomegalovirus (CMV) infection and Epstein-Barr virus (EBV)-induced
lymphoproliferative disease are serious complications associated with
allogeneic stem cell transplantation. Immunotherapy using ex vivo
expanded, virus-specific cytotoxic T lymphocytes (CTL) has been
explored and proven to be effective in therapeutic or prophylactic
regimens for CMV and EBV infections. To generate CTL specific for both
CMV and EBV, we engineered EBV-transformed B-lymphoblastoid cell lines
(BLCL) to express CMV pp65 for use as antigen-presenting cells (APC).
BLCL were transduced with a recombinant retrovirus encoding pp65, the
immunodominant CMV polypeptide. Western blot analysis and
immunocytochemistry confirmed the expression of pp65 in the transduced
cells. Peripheral blood mononuclear cells (PBMC) from healthy CMV
seropositive donors were stimulated with autologous pp65-expressing
BLCL weekly for 3 weeks. Chromium release assays showed that the
resulting CTL cultures possessed specific cytotoxicity against EBV and
CMV. Recombinant vaccinia viruses encoding individual CMV peptides were
used to demonstrate that this CMV-specific cytotoxicity was specific
for pp65. Assays on CD4- and CD8-depleted CTL fractions indicated that
CD8+ CTL mediated the pp65-specific cytotoxicity. These
CMV/EBV-specific CTL recognized CMV- and EBV-infected targets sharing
HLA class I antigens, but not HLA mismatched targets. Our results
demonstrate that BLCL can be used as APC to stimulate expansion of EBV-
and CMV-specific CTL simultaneously. These findings have potential implications for posttransplant CMV and EBV immunotherapy in recipients of allogeneic stem cell transplants.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
H. Zhang, K. M. Snyder, M. M. Suhoski, M. V. Maus, V. Kapoor, C. H. June, and C. L. Mackall
4-1BB Is Superior to CD28 Costimulation for Generating CD8+ Cytotoxic Lymphocytes for Adoptive Immunotherapy
J. Immunol.,
October 1, 2007;
179(7):
4910 - 4918.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Trivedi, R. Y. Williams, R. J. O'Reilly, and G. Koehne
Generation of CMV-specific T lymphocytes using protein-spanning pools of pp65-derived overlapping pentadecapeptides for adoptive immunotherapy
Blood,
April 1, 2005;
105(7):
2793 - 2801.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. A. Abate, S. Watanabe, and E. S. Mocarski
Major Human Cytomegalovirus Structural Protein pp65 (ppUL83) Prevents Interferon Response Factor 3 Activation in the Interferon Response
J. Virol.,
October 15, 2004;
78(20):
10995 - 11006.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Li Pira, L. Bottone, F. Ivaldi, R. Pelizzoli, F. Del Galdo, L. Lozzi, L. Bracci, A. Loregian, G. Palu, R. De Palma, et al.
Identification of new Th peptides from the cytomegalovirus protein pp65 to design a peptide library for generation of CD4 T cell lines for cellular immunoreconstitution
Int. Immunol.,
May 1, 2004;
16(5):
635 - 642.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. A. Papanicolaou, J.-B. Latouche, C. Tan, J. Dupont, J. Stiles, E. G. Pamer, and M. Sadelain
Rapid expansion of cytomegalovirus-specific cytotoxic T lymphocytes by artificial antigen-presenting cells expressing a single HLA allele
Blood,
October 1, 2003;
102(7):
2498 - 2505.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
V. V. Parekh, D. V. R. Prasad, P. P. Banerjee, B. N. Joshi, A. Kumar, and G. C. Mishra
B Cells Activated by Lipopolysaccharide, But Not By Anti-Ig and Anti-CD40 Antibody, Induce Anergy in CD8+ T Cells: Role of TGF-{beta}1
J. Immunol.,
June 15, 2003;
170(12):
5897 - 5911.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Kondo, M. S. Topp, H.-P. Kiem, Y. Obata, Y. Morishima, K. Kuzushima, M. Tanimoto, M. Harada, T. Takahashi, and Y. Akatsuka
Efficient Generation of Antigen-Specific Cytotoxic T Cells Using Retrovirally Transduced CD40-Activated B Cells
J. Immunol.,
August 15, 2002;
169(4):
2164 - 2171.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Moosmann, N. Khan, M. Cobbold, C. Zentz, H.-J. Delecluse, G. Hollweck, A. D. Hislop, N. W. Blake, D. Croom-Carter, B. Wollenberg, et al.
B cells immortalized by a mini-Epstein-Barr virus encoding a foreign antigen efficiently reactivate specific cytotoxic T cells
Blood,
August 13, 2002;
100(5):
1755 - 1764.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Sun, R. L. Burton, and K. G. Lucas
Cytokine production and cytolytic mechanism of CD4+ cytotoxic T lymphocytes in ex vivo expanded therapeutic Epstein-Barr virus-specific T-cell cultures
Blood,
May 1, 2002;
99(9):
3302 - 3309.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Vaz-Santiago, J. Lule, P. Rohrlich, C. Jacquier, N. Gibert, E. Le Roy, D. Betbeder, J.-L. Davignon, and C. Davrinche
Ex Vivo Stimulation and Expansion of both CD4+ and CD8+ T Cells from Peripheral Blood Mononuclear Cells of Human Cytomegalovirus-Seropositive Blood Donors by Using a Soluble Recombinant Chimeric Protein, IE1-pp65
J. Virol.,
September 1, 2001;
75(17):
7840 - 7847.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Szmania, A. Galloway, M. Bruorton, P. Musk, G. Aubert, A. Arthur, H. Pyle, N. Hensel, N. Ta, L. Lamb Jr, et al.
Isolation and expansion of cytomegalovirus-specific cytotoxic T lymphocytes to clinical scale from a single blood draw using dendritic cells and HLA-tetramers
Blood,
August 1, 2001;
98(3):
505 - 512.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Peggs, S. Verfuerth, and S. Mackinnon
Induction of cytomegalovirus (CMV)-specific T-cell responses using dendritic cells pulsed with CMV antigen: a novel culture system free of live CMV virions
Blood,
February 15, 2001;
97(4):
994 - 1000.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Sun, R. L. Burton, L.-J. Dai, W. J. Britt, and K. G. Lucas
B Lymphoblastoid Cell Lines as Efficient APC to Elicit CD8+ T Cell Responses Against a Cytomegalovirus Antigen
J. Immunol.,
October 1, 2000;
165(7):
4105 - 4111.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Retière, V. Prod'homme, B.-M. Imbert-Marcille, M. Bonneville, H. Vié, and M.-M. Hallet
Generation of Cytomegalovirus-Specific Human T-Lymphocyte Clones by Using Autologous B-Lymphoblastoid Cells with Stable Expression of pp65 or IE1 Proteins: a Tool To Study the Fine Specificity of the Antiviral Response
J. Virol.,
May 1, 2000;
74(9):
3948 - 3952.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
J. A. Zaia, J.G. P. Sissons, S. Riddell, D. J. Diamond, M.R. Wills, A.J. Carmichael, M.P. Weekes, M. Gandhi, C. La Rosa, M. Villacres, et al.
Status of Cytomegalovirus Prevention and Treatment in 2000
Hematology,
January 1, 2000;
2000(1):
339 - 355.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Brenner, C. Rossig, U. Sili, J. W. Young, and E. Goulmy
Transfusion Medicine: New Clinical Applications of Cellular Immunotherapy
Hematology,
January 1, 2000;
2000(1):
356 - 375.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
