Analysis of the basophil activation test to quinolones using two

different activation markers

Ariza A1, Fernandez TD1, Rodriguez MJ1, Montañez MI1, Guerrero MA2, Blanca M2, Mayorga C1,2, Torres MJ2.
1
Research Laboratory-Allergy Unit. IBIMA-Regional University Hospital of Málaga, Spain; 2Allergy Service. IBIMA-Regional University Hospital of Málaga, Spain.

BACKGROUND RESULTS
Percentage of activation using CD203c or CD63 markers. CD203c and CD63 expression by clinical entity in
Quinolones are well-tolerated antibiotics Clinical characteristics moxifloxacin allergic patients.
increasingly prescribed (1). Hypersensitivity SEX AGE DRUG REACTION
CONTROLS

reactions as immediate urticaria and P1 F 54 MOXIFLOXACIN ANAPHILACTIC SHOCK

80
25

anaphylaxis, have been reported (1, 2). The P2 F 66 MOXIFLOXACIN ANAPHILACTIC SHOCK 60

diagnosis is complex since in vivo tests such as

% c e lls
20
P3 F 56 MOXIFLOXACIN ANAPHILACTIC SHOCK 40
skin test are not valuable and regarding in
P4 F 79 MOXIFLOXACIN ANAPHILACTIC SHOCK
vitro tests, only basophil activation test (BAT) 20 15

% cells
P5 F 50 MOXIFLOXACIN ANAPHILACTIC SHOCK
has shown to be useful (2,3). Recent studies 0
P6 F 68 MOXIFLOXACIN ANAPHYLAXIS
have shown contradictory results regarding Negative
Control
Moxifloxacin
2 mg/mL
Moxifloxacin
0.2 mg/mL
Ciprofloxacin
2 mg/mL
Ciprofloxacin
0.2 mg/mL
10
P7 F 48 MOXIFLOXACIN ANAPHYLAXIS
the use of different markers to assess basophil MOXIFLOXACIN ALLERGIC PATIENTS
P8 M 71 MOXIFLOXACIN ANAPHYLAXIS
activation that may be influenced by the drug 80 5

included in the test (2-4). P9 F 51 MOXIFLOXACIN ANAPHYLAXIS

60
P10 M 80 MOXIFLOXACIN URTICARIA 0

% c e lls
Anaphylactic Anaphylaxis Urticaria
P11 F 77 MOXIFLOXACIN URTICARIA 40
Shock
OBJECTIVES P12 M 74 CIPROFLOXACIN ANAPHYLAXIS
20
P13 M 75 CIPROFLOXACIN ANAPHYLAXIS Analysing the expression of both markers in basophils from
To evaluate the use of two different activations P14 F 60 CIPROFLOXACIN ANAPHYLAXIS
0
Negative
Control
Moxifloxacin
2 mg/mL
Moxifloxacin
0.2 mg/mL
Ciprofloxacin
2 mg/mL
Ciprofloxacin
0.2 mg/mL
moxifloxacin allergic patients stimulated with the culprit drug, we
markers, CD63 and CD203c, in the evaluation of P15 F 79 CIPROFLOXACIN URTICARIA found a higher expression of CD203c in patients suffering
patients with immediate allergic reactions to
CIPROFLOXACIN ALLERGIC PATIENTS
anaphylactic shock, whereas in patients with anaphylaxis was
P16 F 59 CIPROFLOXACIN URTICARIA 80
CD63 the marker that showed a higher up-regulation.
different quinolones, moxifloxacin and P17 F 57 CIPROFLOXACIN URTICARIA 0.0053

ciprofloxacin. 60

% c e lls
40 Sensitivity and specificity of the test using different
activation markers.
MATERIALS & METHODS 20
SPECIFICITY
Patient allergic to MOXIFLOXACIN CIPROFLOXACIN
0 Marker CD63 CD203c
Patients and controls: We studied 17 patients with Flow cytometry strategy Negative
Control
Moxifloxacin
2 mg/mL
Moxifloxacin
0.2 mg/mL
Ciprofloxacin
2 mg/mL
Ciprofloxacin
0.2 mg/mL
Marker CD63 CD203c CD63 CD203c

9.1% 36.4% Moxifloxacin 77.8% 94.4%

Drug used in the

immediate allergic reactions to quinolones (6 to Moxifloxacin 0% (0) 0% (0)
CD63 CD203c (1) (4)
ciprofloxacin and 11 to moxifloxacin) diagnosed by the

test
clinical history and drug provocation test when mild and Although ciprofloxacin and moxifloxacin are able to induced both Ciprofloxacin 88,9% 94.4%
36.4% 9.1% 83.3%
moderate reactions were reported. Eighteen subjects with Ciprofloxacin 0% (0)
CD63 and CD203c, we found a predominance in the expression (4) (1) (5)
tolerance to quinolones were also included as controls.
depending on the drug included in the test. Thus ciprofloxacin was
Basophil activation test: BAT was performed as described able to mainly increase the expression of CD63 whereas moxifloxacin The best results were observed using the culprit drug and CD203c as
(2) with moxifloxacin and ciprofloxacin at 2 different mainly increase the expression of CD203c. activation marker for moxifloxacin and CD63 for ciprofloxacin.
concentrations (2 mg/mL and 0.2 mg/mL) using CD193
(CCR3) for basophil selection and CD203c or CD63 as
Negative Result

activation markers. Results were expressed as percentage CONCLUSIONS REFERENCES

of activated cells. Positivity was calculated as the
percentage of cases with positive results (with stimulation
index (SI) > 3) to at least one of the concentrations used in • To diagnose quinolone allergy, BAT must be performed using the culprit 1. Blanca-López N, Ariza A, Doña I, Mayorga C, Montañez MI,
Garcia-Campos J,Gomez F, Rondón C, Blanca M, Torres MJ.. Clin
the test. SI was calculated as % drug activated basophils / drug and CD203c for moxifloxacin or CD63 for ciprofloxacin as activation Exp Allergy. 2013;43(5):560-7.
% spontaneous activated basophils ratio. marker. 2. Aranda A, Mayorga C, Ariza A, Dona I, Rosado A, Blanca-Lopez
N, Andreu ITorres MJ. Allergy. 2011; 66(2): 247-54.
Statistical analysis: Comparisons between different • The differential expression of both activation markers seems to be related
Positive Result

activation markers, drugs and cut off were performed by 3. Mayorga C, Andreu I, Aranda A, Doña I, Montañez MI, Blanca-
with the culprit drug and clinical entity. Lopez N, Ariza A,Nuin E, Blanca M, Miranda MA, Torres MJ. Int
Wilcoxon test. χ2 analysis was used for comparing the Arch Allergy Immunol.2013;160(4):377-82.
positivity. All reported p values < 0.05 were considered • Using this criteria and a cut-off of 3, we have obtained a better sensitivity
4. Ben Said B, Berard F, Bienvenu J, Nicolas JF, Rozieres A. Allergy.
statistically significant. for ciprofloxacin. 2010;65(4):535-6.