@Article{ecn.2008.0132,
AUTHOR = {Georges Khalil, Marwan El-Sabban, Sara Al-Ghadban, Sandy Azzi, Sara Shamra, Simon Khalifé, Richard Maroun},
TITLE = {Cytokine expression profile of sensitized human T lymphocytes following in vitro stimulation with amoxicillin},
JOURNAL = {European Cytokine Network},
VOLUME = {19},
YEAR = {2008},
NUMBER = {3},
PAGES = {131--141},
URL = {http://www.techscience.com/ECN/v19n3/65932},
ISSN = {1952-4005},
ABSTRACT = {Since the withdrawal of penicillin determinants from the market, in addition to the hazard of
re-exposing the patient to the drug, skin testing for the diagnosis of penicillin allergy has become less accurate and
less standardized. The assay currently used, the lymphocyte transformation test (LTT), lacks sufficient sensitivity,
and requires the use of radioactive material. The objective of this study was to establish an accessible and reliable
method for the safe diagnosis of penicillin allergy. Peripheral blood mononuclear cells (PBMC) were isolated from
18 patients who were allergic to penicillin and 12 control subjects using the Ficoll-Hypaque method. The isolated,
sensitized cells were stimulated in vitro with amoxicillin (1 mg/mL). Stimulation with phytohemagglutinin A
(PHA) was used as the positive control. Transcriptional expression of specific cytokines (IL-2, -4, -5 and -13,
TGF-β, TNF-α and IFN-γ) was assessed by RT-PCR. IFN-γ expression was also evaluated by ELISPOT. Secreted
levels of IL-2, -5 and IFN-γ were measured by ELISA. All of these assays were performed two or five days,
post-stimulation. This study of the in vitro diagnosis of penicillin allergy by the measurement of cytokine
concentration in the supernatants of sensitized lymphocytes cultures involved the largest number of patients
to-date. The Δ values (difference in cytokine concentration in the supernatants before and after stimulation) were
compared between cases and controls using different statistical tests (Student’s t test and the Mann-Whitney rank
test). Of the various tests performed in this study, measurement of secreted cytokines using ELISA was the most
sensitive and specific (80% and 100% respectively). In vitro stimulation of human lymphocytes sensitized to
amoxicillin is a safe and useful test for the diagnosis of penicillin allergy if the ELISA is used to measure cytokine
expression. The advantages are that it can be performed by many laboratories since kits to determine cytokines
are widely available, and it can be done without the need for particularly specialized equipment.},
DOI = {10.1684/ecn.2008.0132}
}