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Title: The effect of apoptotic cells on virus-specific immune responses detected using IFN-gamma ELISPOT
Authors: Lenders, Kevin
Ogunjimi, Benson
Beutels, Philippe
HENS, Niel 
Van Damme, Pierre
Berneman, Zwi N.
Van Tendeloo, Viggo F. I.
Smits, Evelien L. J. M.
Issue Date: 2010
Source: JOURNAL OF IMMUNOLOGICAL METHODS, 357(1-2). p. 51-54
Abstract: Interferon (IFN)-gamma ELISPOT can be used to monitor the magnitude of virus-specific cellular immune responses in vaccine trials. Often, IFN-gamma ELISPOT is performed with cryopreserved peripheral blood mononuclear cells (PBMC). However, it has not been well defined yet to what extent diminished cell viability of PBMC following cryopreseivation affects IFN-gamma responses in ELISPOT. Therefore, we assessed the influence of apoptotic cells on the number of spot-forming cells (SFC) in IFN-gamma ELISPOT using a gradient of UV-irradiated apoptotic PBMC and viral antigens derived from varicella zoster virus (VZV) and cytomegalovirus (CMV). No SFC were observed when UV-irradiated apoptotic cells were stimulated with VZV or CMV antigens. Moreover, presence of apoptotic cells among viable T cells hampered the detection of SFC following stimulation with VZV or CMV cell lysates, but not with CMVpp65 peptide pool. Statistical analysis showed that mainly late apoptotic cells, staining both Annexin V and 7-amino-actinomycin D (7-AAD), were associated with a decreased number of SFC. In conclusion, it is recommended to use highly viable thawed PBMC for the detection of virus-specific cellular immune responses by IFN-gamma ELISPOT, since the detection of CMV- and VZV-specific T cell responses stimulated by cell lysates was significantly impeded by the presence of apoptotic cells. (C) 2010 Elsevier B.V. All rights reserved.
Notes: [Lenders, Kevin; Berneman, Zwi N.; Van Tendeloo, Viggo F. I.; Smits, Evelien L. J. M.] Univ Antwerp, Lab Expt Hematol, Vaccine & Infect Dis Inst Vaxinfectio, Univ Antwerp Hosp, B-2650 Edegem, Belgium. [Ogunjimi, Benson; Beutels, Philippe; Hens, Niel; Van Damme, Pierre] Univ Antwerp, CHERMID, B-2610 Antwerp, Belgium. [Ogunjimi, Benson; Beutels, Philippe; Hens, Niel; Van Damme, Pierre] Univ Antwerp, CEV, Vaccine & Infect Dis Inst Vaxinfectio, B-2610 Antwerp, Belgium. [Hens, Niel] Hasselt Univ, Interuniv Inst Biostat & Stat Bioinformat, B-3590 Diepenbeek, Belgium.
Keywords: ELISPOT; Apoptosis; IFN-gamma; Vaccine trial; Cellular immune response
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ISSN: 0022-1759
e-ISSN: 1872-7905
DOI: 10.1016/j.jim.2010.03.001
ISI #: 000279361100008
Category: A1
Type: Journal Contribution
Validations: ecoom 2011
Appears in Collections:Research publications

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