Turk J Immunol. 2020; 8(3): 120-128 | DOI: 10.25002/tji.2020.1347

Cytokine Secretion and Proliferative Capacity of CD4+ T Cells in Delayed Type Hypersensitivity Reactions due to Ciprofloxacin

Belkýs Ertek¹, Semra Demir, Umut Can Küçüksezer, Leyla Pur Özyiðit, Aslý Gelincik, Suna Büyüköztürk, Günnur Deniz, Esin Aktaþ Çetin
¹Istanbul University, Aziz Sancar Institute of Experimental Medicine (Aziz Sancar DETAE), Department of Immunology, Istanbul, Turkey
²Istanbul University, Istanbul Faculty of Medicine, Department of Internal Medicine, Division of Allergy, Istanbul, Turkey
³University Hospitals of Leicester, Department of Allergy and Immunology, Leicester, UK

INTRODUCTION: Introduction: Ciprofloxacin (CPFX), a frequently prescribed quinolone, may induce cutaneous adverse drug reactions. Delayed type hypersensitivity reactions (DTHR) are often difficult to deal with, therefore, in vitro testing for DTHR is the long-anticipated method for their management. This study aimed to evaluate potential value of lymphocyte transformation test (LTT) and intracellular cytokine secretion of drug stimulated CD4+ T cells in patients with DTHR against ciprofloxacin.
Material and Methods: Patients experienced DTHR with CPFX (n=8) and healthy subjects (n=10) were enrolled. CPFX skin prick, patch and intradermal tests were performed. LTT by flow cytometry aimed to determine CPFXspecific CD4+ T cell proliferation. Intracellular IL-4, IL-10, IL-2 & IFN-γ levels were analysed by flow cytometry in CPFX-specific CD4+ T cells. Cytokine contents of cell culture supernatants were evaluated by ELISA.
Results: In patients with DTHR, 5 and 10 μg/mL CPFX induced significant CD4+ T cell proliferation (p=0.014 and p=0.05, respectively). IL-2 (p=0.02, p=0.001 and p=0.001, respectively) and IL-4 (p=0.001) secreting CD4+ T cell percentages were increased, while IFN-γ+ (p=0.001, p=0.011 and p=0.012, respectively) and IL-10+ (p=0.001, p=0.001 and p=0.002, respectively) CD4+ T cells were decreased. The cell culture supernatants revealed downregulated IL-10 (p<0.000, p=0.004, p=0.001 and p=0.0001, respectively) and upregulated IL-4 levels (p=0.003, p=0.013 and p=0.0001, respectively) in patients, regardless of CPFX stimulation. Intradermal test was positive in only one patient while all patch tests remained negative.
Conclusion: Our findings suggest that the increase of IL-2 and IL-4-secreting CD4+ T cells together with the decrease of IL-10 and IFN-γ-secreting CD4+ T cells is related to DTHR seen in patients with delayed-type CPFX allergy. Intracellular cytokine measurement, together with LTT could ease the management of CPFX hypersensitivity when in vivo tests are non-available, remain inconclusive or negative.
METHODS:
RESULTS:
DISCUSSION AND CONCLUSION:

Keywords: Alerji, CD4+ T hücresi, siprofloksasin, aþýrý duyarlýlýk, hücre içi sitokin, lenfosit transformasyonu

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Siprofloksasine Baðlý Geç Tip Aþýrý Duyarlýlýk Reaksiyonlarýnda CD4+ T Hücrelerinin Sitokin Salýnýmlarý ve Proliferatif Kapasiteleri

Belkýs Ertek¹, Semra Demir, Umut Can Küçüksezer, Leyla Pur Özyiðit, Aslý Gelincik, Suna Büyüköztürk, Günnur Deniz, Esin Aktaþ Çetin
¹Istanbul University, Aziz Sancar Institute of Experimental Medicine (Aziz Sancar DETAE), Department of Immunology, Istanbul, Turkey
²Istanbul University, Istanbul Faculty of Medicine, Department of Internal Medicine, Division of Allergy, Istanbul, Turkey
³University Hospitals of Leicester, Department of Allergy and Immunology, Leicester, UK

GÝRÝÞ ve AMAÇ: Giriþ: Siprofloksasin (SPFX), en yaygýn kullanýlan kinolon grubu antibiyotiklerden olup, kutanöz ters ilaç reaksiyonlarý oluþturabilme kapasitesine sahiptir. Gecikmiþ tip aþýrý duyarlýlýk reaksiyonlarýnda (ADR) kesin taný konulmasý zor olmakla beraber in vitro taný testlerinin geç tip ilaç ADR’de kullanýmý önemlidir. Çalýþmamýzda SPFX’e baðlý geç tip ADR geçiren hastalarda, ilaçlar ile uyarýlmýþ CD4+ T hücrelerinde, hücre içi sitokin üretimleri ve lenfosit transformasyon (LTT) yanýtlarý araþtýrýlmýþtýr.
Gereç ve Yöntemler: Çalýþmaya SPFX ile geç tip hipersensitivite reaksiyonu (n=8) geçiren hastalar ve saðlýklý kontroller (n=10) dâhil edilmiþ, intradermal testler ve yama testleri uygulanmýþtýr. LTT metodu ile SPFX’e özgül CD4+ T hücre proliferasyonu belirlenmiþtir. SPFX’e özgül CD4+ T hücrelerinde hücre içi IL-4, IL-10, IL-2 ve IFN-γ düzeyleri akan hücre ölçer kullanýmý ile deðerlendirilmiþtir. Kültür üst sývýlarýndaki sitokin düzeyleri ELISA aracýlýðý ile ölçülmüþtür.
Bulgular: Geç tip ADR geçiren hastalarda 5 ve 10 μg/ml SPFX uyarýmý CD4+ T hücre proliferasyonunu arttýrmýþtýr (p=0.014 ve p=0.05, sýrasýyla). IL-2 (p=0.02, p=0.001 ve p=0.001, sýrasýyla) ve IL-4 (p=0.001) salgýlayan CD4+ T hücre yüzdesi artmasýna raðmen IFN-γ+ (p=0.001, p=0.011 ve p=0.012, sýrasýyla) ve IL-10+ (p=0.001, p=0.001 ve p=0.002, sýrasýyla) CD4+ T hücre oranlarý azalmýþtýr. Hücre kültür üst sývýlarýnda SPX uyarýmýndan baðýmsýz olarak, hastalarda IL-10 düzeyi (p<0.000, p=0.004, p=0.001 ve p=0.0001, sýrasýyla) azalýrken, IL-4 düzeyi (p=0.003, p=0.013 ve p=0.0001, sýrasýyla) artmýþtýr. Ýntradermal test uygulanan hastalarýn sadece birinde pozitif yanýt saptanýr iken, yama testlerinde pozitif sonuç elde edilmemiþtir.
Sonuç: Bulgularýmýz, IL-2 ve IL-4 üreten CD4+ T hücrelerindeki artýþla birlikte IL-10 ve IFN-γ üreten CD4+ T hücrelerindeki azalmanýn, gecikmiþ tipte CPFX alerjisine sahip hastalarda DTHR ile iliþkili olduðunu düþündürmektedir. LT ile birlikte hücre içi sitokin ölçümleri, in vivo testlerin negatif kaldýklarý, karar verdirici olamadýklarý ve hatta uygulanamadýklarý durumlarda, gecikmiþ tipte CPFX hipersensitivitesinin deðerlendirilmesinde fayda saðlayabilir.
YÖNTEM ve GEREÇLER:
BULGULAR:
TARTIÞMA ve SONUÇ:

Anahtar Kelimeler: Allergy, CD4+ T cells, ciprofloxacin, hypersensitivity, intracellular cytokine, LTT

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