Novel interferon-γ enzyme-linked immunoSpot assay using activated cells for identifying hypersensitivity-inducing drug culprits

Kenichi Kato; Asami Kawase; Hiroaki Azukizawa; Takaaki Hanafusa; Yukinobu Nakagawa; Hiroyuki Murota; Shimon Sakaguchi; Hideo Asada; Ichiro Katayama

doi:10.1016/j.jdermsci.2017.03.007

Novel interferon-γ enzyme-linked immunoSpot assay using activated cells for identifying hypersensitivity-inducing drug culprits

J Dermatol Sci. 2017 Jun;86(3):222-229. doi: 10.1016/j.jdermsci.2017.03.007. Epub 2017 Mar 14.

Authors

Kenichi Kato¹, Asami Kawase², Hiroaki Azukizawa³, Takaaki Hanafusa⁴, Yukinobu Nakagawa⁵, Hiroyuki Murota⁶, Shimon Sakaguchi⁷, Hideo Asada⁸, Ichiro Katayama⁹

Affiliations

¹ Department of Dermatology, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: bykenbykenbyken@yahoo.co.jp.
² Department of Dermatology, Nara Medical University, 840, Shijo-cho, Kashihara, Nara 634-8521, Japan. Electronic address: 41600401a@naramed-u.ac.jp.
³ Department of Dermatology, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan; Department of Dermatology, Nara Medical University, 840, Shijo-cho, Kashihara, Nara 634-8521, Japan. Electronic address: azukizaw@naramed-u.ac.jp.
⁴ Department of Dermatology, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan; Department of Dermatology, Tokyo Medical and Dental University, Yushima 1-5-45, Bunkyo-ku, Tokyo 113-8519, Japan. Electronic address: takafussa0710@gmail.com.
⁵ Department of Dermatology, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: nakagawa@derma.med.osaka-u.ac.jp.
⁶ Department of Dermatology, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: h-murota@derma.med.osaka-u.ac.jp.
⁷ Department of Experimental Immunology, Immunology Frontier Research Center, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan; Department of Experimental Pathology, Institute for Frontier Medical Sciences, Kyoto University, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan. Electronic address: shimon@ifrec.osaka-u.ac.jp.
⁸ Department of Dermatology, Nara Medical University, 840, Shijo-cho, Kashihara, Nara 634-8521, Japan. Electronic address: asadah@naramed-u.ac.jp.
⁹ Department of Dermatology, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: katayama@derma.med.osaka-u.ac.jp.

PMID: 28330775
DOI: 10.1016/j.jdermsci.2017.03.007

Abstract

Background: The drug-induced lymphocyte stimulation test (DLST), also referred to as lymphocyte transformation test (LTT), is used to identify the culprit drug in cases of cutaneous adverse drug reactions (cADR). Although DLST is a widely used in vitro test, its sensitivity and specificity are unsatisfactory. Recent reports suggest that the detection of drug-induced interferon (IFN)-γ production using enzyme-linked immunoSpot (ELISpot) assay (conventional IFN-γ ELISpot) is useful for identifying culprit drugs in cADR cases.

Objective: The aim of this study was to establish a novel method for identifying culprit drugs in patients with cADR by efficiently detecting drug-specific IFN-γ production using activated cells.

Methods: Sixteen patients with cADR, including drug-induced hypersensitivity syndrome, erythema multiforme-like eruption, maculopapular exanthema, Stevens-Johnson syndrome, and toxic epidermal necrolysis, caused by clinically convincing culprit drugs were enrolled in this study. In some cases, the blood samples were obtained at two or three different time points. Peripheral blood mononuclear cells (PBMCs) from total 20 samples were analyzed using both the DLST and drug-induced conventional IFN-γ ELISpot. In addition, drug-induced IFN-γ ELISpot was performed using PBMCs, which were stimulated with anti-cluster of differentiation (CD)-3/CD28 antibody-coated microbeads and interleukin (IL)-2 for 7 days before exposure to the culprit drugs (modified IFN-γ ELISpot).

Results: Among the culprit drugs tested in each patient, the modified IFN-γ ELISpot was positive in 17 samples (13 patients) while DLST and conventional IFN-γ ELISpot were positive in eight and four samples (six and three patients), respectively.

Conclusion: The modified IFN-γ ELISpot using activated PBMCs was more sensitive than the conventional IFN-γ ELISpot was for detecting drug-induced IFN-γ production, which could be a useful in vitro tool for identifying culprit drugs in cADR cases.

Keywords: Cutaneous adverse drug reactions; Drug hypersensitivity; ELISpot; IFN-γ; LTT.

Publication types

Comparative Study

MeSH terms

Adult
Aged
Aged, 80 and over
Biomarkers / blood
Cells, Cultured
Drug Eruptions / blood
Drug Eruptions / diagnosis*
Drug Eruptions / immunology
Enzyme-Linked Immunospot Assay
Female
Humans
Interferon-gamma / blood*
Interferon-gamma / immunology
Interferon-gamma Release Tests*
Lymphocyte Activation / drug effects*
Male
Middle Aged
Predictive Value of Tests
Reproducibility of Results
T-Lymphocytes / drug effects*
T-Lymphocytes / immunology
T-Lymphocytes / metabolism
Young Adult

Substances

Biomarkers
IFNG protein, human
Interferon-gamma