







































_____________________________________________________________________________________________________ 
 
*Corresponding author: E-mail: celso@alergoimuno.med.br; 
 
Asian J. Immunol., vol. 7, no. 1, pp. 63-70, 2024 
 
 
 

Asian Journal of Immunology 
 
Volume 7, Issue 1, Page 63-70, 2024; Article no.AJI.115657 
 

 
 

 

 

Exploring the Role of Leukocyte 
Adherence Inhibition Test in Assessing 
Non-IgE Mediated Immunoreactivity to 

Benzoic Acid in Allergic Patients 
 

Celso Eduardo Olivier a*, Daiana Guedes Pinto a,  
Ana Paula Monezzi Teixeira a,  

Jhéssica Letícia Santos Santana a,  
Raquel Acácia Pereira Gonçalves Santos a,  

and Regiane Patussi Santos Lima b 
 

a Instituto Alergoimuno de Americana, Brazil. 
b Lavoisier’s laboratories, São Paulo, Brazil. 

 

Authors’ contributions  
 

This work was carried out in collaboration among all authors. All authors read and approved the final 
manuscript. 

 

Article Information 
 

DOI: https://doi.org/10.9734/aji/2024/v7i1130  
 

Open Peer Review History: 
This journal follows the Advanced Open Peer Review policy. Identity of the Reviewers, Editor(s) and additional Reviewers,  

peer review comments, different versions of the manuscript, comments of the editors, etc are available here: 
https://www.sdiarticle5.com/review-history/115657 

 
 
 

Received: 02/04/2024 
Accepted: 13/04/2024 
Published: 17/04/2024 

 
 

ABSTRACT 
 

Background: Several publications report benzoic acid as responsible for non–IgE-mediated allergic 
reactions. No standardized lab exam identifies these reactions besides in vivo provocation tests. 
Aim: To evaluate the potential of the Leukocyte Adherence Inhibition Test (LAIT) to discriminate 
non–IgE-mediated immunoreactivity against benzoic acid in patients with non–IgE-mediated allergic 
phenotypes. 

Original Research Article 

https://doi.org/10.9734/aji/2024/v7i1130


 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 63-70, 2024; Article no.AJI.115657 
 
 

 
64 

 

Study Design: We retrospectively examined the medical charts of 100 patients diagnosed with 
allergic rhinitis, allergic bronchitis, asthma, sinus headache, atopic dermatitis, and/or urticaria with 
clinical suspicion of non–IgE-mediated benzoic acid hypersensitivity who were investigated with ex 
vivo challenge test monitored by LAIT against benzoic acid. The cohort counted 30 males; the 
mean age was 41.9 years, the SD was 20.4 years, and the range was 2 to 90 years. 
Place and Duration of Study: Instituto Alergoimuno de Americana – São Paulo – Brazil –  
between January 2018 and March 2024. 
Methodology: The percentage of Leukocyte Adherence Inhibition (LAI) promoted by the ex vivo 
challenges against 1 mg/mL benzoic acid was distributed in ranges through a cascade distribution 
chart to outline the variability of the results. 
Results: The LAI ranged from 0% to 97%; the Mean was 41.1%; the Median was 40.5%; the 
Standard Deviation was 24.2%; the Mode was 0 and 59 (each appeared four times). The cascade 
distribution demonstrates a wide distribution of LAI results. This extensive distribution of LAI results 
suggests that some patients had mild, moderate, or severe non–IgE-mediated immunoreactivity 
against benzoic acid, while others did not present any immunoreactivity against it. 
Conclusion: Our preliminary results support that the LAIT performed with benzoic acid may 
discriminate diverse degrees of ex vivo immunoreactivity in patients suffering from diversified 
allergic phenotypes. It is worth carrying out more in-depth studies to evaluate the usefulness of 
LAIT in diagnosing non–IgE-mediated benzoic acid allergies. 
 

 
Keywords:  Allergy; asthma; atopic dermatitis; bronchitis; diagnosis; exposome-wide association 

study; hypersensitivity; leukocyte adherence inhibition test; non–ige-mediated 
immunoreactivity; precision medicine; rhinitis; sinus headache; urticaria. 

 

ABBREVIATIONS 
 
LAI  : Leukocyte Adherence Inhibition 
LAIT : Leukocyte Adherence Inhibition Test 

 

1. INTRODUCTION  
 

Benzoic acid is an organic compound formed by 
an aromatic ring and a carboxyl group [1].   
Benzoic acid is a parent and a metabolic 
common pathway of a large group of structurally 
related substances (aromatic salts, alcohols, 
aldehydes, esters, and acetals) legally regulated 
to be used as additives to industrialized foods, 
cosmetics, and medicaments [2]. Used as an 
antimicrobial, the legally acceptable daily intake 
uppermost limit for benzoic acid (or the 
benzyl/benzoic moiety) is 5.0 mg/kg body weight 
[3]. “There is evidence that benzyl benzoate is 
hydrolyzed to benzyl alcohol and benzoic acid; 
as well, benzyl alcohol and benzaldehyde suffer 
in vivo oxidation to benzoic acid” [4]. “In plants 
and animals, benzoic acid is produced 
endogenously through the phenylalanine–
tyrosine pathway” [5]. “Several of its derivatives 
occur naturally in foods, such as fruits (apple, 
avocado, blackberry, blueberry, cherry, 
cranberry, melon, papaya, plum, raspberry, 
strawberry, tomato), vegetables (artichokes, 
asparagus, beans, cabbage, corn, leek, 
mushroom, potatoes), meats (beef, chicken, 
pork, shellfish), cheeses, teas and wines” [6]. 

Food additives (among them benzoic acid) have 
long been described as sensitizer agents 
responsible for human allergic reactions [7]. A 
double-blinded provocation study done with 
benzoic acid produced objective reactions in 7% 
of patients with urticaria [8]. There is a report of a 
child who developed chronic cheilitis when 
ingesting daily benzoates-preserved 
industrialized foods [9]. There are reports of 
asthmatic patients who had a crisis of 
bronchospasm after the intake of benzoate-
containing antiasthmatic medicines [10]. Reports 
of cross-reactivity among benzoates, azo dyes, 
and aspirin in patients with urticaria are also 
common [11].  
 
Most provocation tests performed with benzoic 
acid do not elicit immediate reactions; instead, 
the reactions appear within 14 hours after the 
challenge [12]. The immunoreactivity elicited 
against benzoic acid is non–IgE-mediated and is 
not yet acknowledged explicitly among the 
recently classified hypersensitivity mechanisms 
[13]. The main knowledge about the mechanism 
of hypersensitivity against benzoate (and similar 
food additives) was brought by an ex vivo 
leukocyte challenge test determining the 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 63-70, 2024; Article no.AJI.115657 
 
 

 
65 

 

increasing sulfidoleukotriene production [14]. 
Since leukotrienes are known mediators of the 
leukocyte adherence inhibition phenomenon, we 
hypothesize that the employ of the Leukocyte 
Adherence Inhibition Test (LAIT) could help 
identify the endotype responsible for benzoic 
acid hypersensitivity [15-17]. 
 
“We routinely employ the LAIT in our facilities to 
evaluate non–IgE-mediated immunoreactivity 
against suspected allergens, previously engaging 
in exhaustive provocation tests” [18-24]. To 
evaluate the potential of the LAIT to discriminate 
non–IgE-mediated immunoreactivity against 
benzoic acid, we retrospectively compiled the 
electronic medical charts of patients with non–
IgE-mediated allergic rhinitis, allergic bronchitis, 
asthma, sinus headache, atopic dermatitis, 
and/or urticaria who were investigated with this 
procedure.  
 
The present study hypothesizes that the LAIT 
may differentiate diverse degrees of 
immunoreactivity against benzoic acid                        
among patients suffering from allergic 
phenotypes. 
 

2. MATERIALS AND METHODS  
 

2.1 Subjects 
 
After receiving Institutional Review Board 
approval from the Instituto Alergoimuno de 
Americana (Brazil; 03/2024), we proceeded with 
the electronic chart review of 8,500 outpatients 
who attended our facility from January 2018 to 
March 2024. A cohort of 100 outside patients had 
been submitted to an ex vivo allergen challenge 
test with benzoic acid 1mg/mL monitored with 
LAIT for presenting non–IgE-mediated allergic 
rhinitis, allergic bronchitis, asthma,                               
sinus headache, atopic dermatitis, and/or 
urticaria.  
 
This study did not include pregnant women, 
breastfeeding, and patients under biological 
and/or systemic anti-inflammatory therapy 
(corticoids, cyclosporin). “The cohort counted 30 
males; mean age 41.9 years; SD 20.4 years; 
range 2 to 90 years; median 43 years; modes = 
26; 28; 43; 43 and 53 (each appeared four 
times); geometric mean = 34.4 years.             
This procedure was offered to patients with 
clinical suspicion of benzoate hypersensitivity          
who demonstrated a non-reactive or                  
inconclusive skin test against benzoic                              
acid”  [25]. 

2.2 Ex vivo Investigation: Leukocyte 
Adherence Inhibition Test 

 
“We performed the LAIT as previously described” 
[26-34]. Shortly, each donor's fresh plasma was 
divided into two parts and used in paralleled ex 
vivo challenging tests with benzoic acid 1 mg/mL 
and the unchallenged plasma assay. We 
collected the plasma with high leukocyte content 
(buffy coat) from the heparinized tube after one 
hour of sedimentation at 37 °C. Then we 
distributed aliquots of 100 μL into Eppendorf 
tubes kept under agitation for 30 minutes (200 
rpm at 37 °C) with benzoic acid (10μL of a 
solution with 1mg/mL and pH 7.5) or without 
benzoic acid (when used as control). After 
incubation, the plasma was allocated into a 
standard Neubauer hemocytometer counting 
chamber with a plain, non-metallic glass surface 
and left to stand for 2 hours at 37 °C in the 
humidified atmosphere of the covered water bath 
to allow leukocytes to adhere to the glass. Next, 
we counted the leukocytes, removed the 
coverslip, and washed the chamber by 
immersion in a beaker with PBS at 37 °C. Then, 
we added a drop of PBS to the hemocytometer's 
chamber and allocated a clean coverslip over it. 
The remaining cells were counted in the same 
squares as previously examined. The percentage 
of Leukocyte Adherence (LA) of each assay was 
estimated as: (the number of leukocytes 
observed on the hemocytometry chamber after 
washing divided by the number of leukocytes 
observed on the hemocytometry chamber before 
washing) and multiplied by 100 (%). The 
Leukocyte Adherence Ratio (LAR) was estimated 
based on the ratio between the LA from the 
antigen-specific challenged plasma and the LA 
from the unchallenged control plasma: LAR = LA 
of the challenged sample divided by LA of 
unchallenged control plasma multiplied by 100 
(%). To further calculate the Leukocyte 
Adherence Inhibition (LAI), we subtracted the 
LAR from 100 (%). We employed the LAI results 
for the cascade distribution chart and the 
statistics calculations, both performed with the 
help of the Microsoft Excel® statistical package. 
 

3. RESULTS 
 

As a retrospective survey, there was no research 
protocol; therefore, we report the incidentally 
immune investigation as registered in the digital 
medical charts. The LAI ranged from 0% to 97%; 
the Mean was 41.1%; the Median was 40.5%; 
the Standard Deviation was 24.2%; the Mode 
was 0 and 59 (each appeared four times). 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 63-70, 2024; Article no.AJI.115657 
 
 

 
66 

 

The cascade distribution demonstrates a wide 
range of distribution of LAI results (Fig.1). Four 
patients ignored the presence of the allergen on 
the plasma and presented no inhibition of 
leukocyte adherence (LAI = 0%) after contact 
with benzoic acid (4% of the tests). Some 
patients showed low or moderate 
immunoreactivity during the ex vivo challenge 
test. In contrast, others displayed strong 
immunoreactivity, which could possibly reflect the 
participation of benzoic acid in a theoretical non–
IgE-mediated hypersensitivity condition to be 
further corroborated by in vivo provocation tests. 
 

4. DISCUSSION 
 
The non-IgE-mediated hypersensitivities are 
characterized by a challenging technical 
diagnosis due to the lack of standardized 
immunoassays [35]. Diagnosing these conditions 
among allergic patients is based on individual 
laborious medical work accomplishing 
anamnesis, cutaneous tests, and challenge in 
vivo provocation tests performed after meticulous 
exclusion diets [36].  
 
“As an increaser of the sulfidoleukotriene 
production, hypersensitivity to benzoic acid may 
be, in a certain way, similar (and potentially an 
enhancer) of the hypersensitivity produced by 
non-steroidal anti-inflammatory drugs (NSAIDs). 

The principal pharmacological action of NSAIDs 
is the inhibition of the cyclooxygenase enzymes, 
which catalyze the synthesis of prostaglandins 
and thromboxanes” [37]. “Cyclooxygenase 
enzymes catalyze the conversion of arachidonic 
acid released from the cellular membrane by 
cytosolic phospholipases activated by 
nociceptive mechanisms” [38]. “The 
cyclooxygenases and the lipoxygenases oxidize 
the arachidonic acid liberated into the cytosol. 
The cyclooxygenases pathway generates pro-
inflammatory autacoids such as prostaglandins 
and thromboxanes. The lipoxygenase pathway 
generates leukotrienes” [39]. The pharmacologic 
inhibition of the cyclooxygenases increases the 
lipoxygenases' activity, increasing the 
leukotrienes' production. Any substance that 
(pharmacologically or immunologically) increases 
the production of leukotrienes affects the 
autacoid balance, adversely producing allergic 
symptoms.  
 

The LAIT theoretically explores every immune 

pathway as an ex vivo challenge test with a 

viable leukocyte buffy coat, allowing the 

interaction of all immune-circulating participants 

[40]. However, as an observant of the final 

phenomenon, the LAIT did not indicate which 

pathways were involved in inhibiting the 

adherence (or increasing the production

 

 
 
Fig. 1. Cascade distribution chart of the range groups of Leukocyte Adherence Inhibition (LAI) 
results (x-axis %) of ex vivo benzoic acid challenges monitored by the Leukocyte Adherence 

Inhibition Test (LAIT), according to the respective number of outcomes over 100 tests/subjects 
(y-axis) 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 63-70, 2024; Article no.AJI.115657 
 
 

 
67 

 

of leukotrienes), whether pharmacological or 
immunological [41-44]. As proposed by the 
exposome-wide association study, the LAIT also 
configures itself as an exposome measurement, 
qualifying itself as an immune marker of the 
contact and the response to a specific antigen 
instead of being associated with a specific 
phenotype [45]. 
 
This preliminary retrospective survey 
demonstrated an extensive range of results from 
the ex vivo challenge test with benzoic acid 
monitored by LAIT  in a cohort of patients with 
various allergic phenotypes. We routinely employ 
the LAIT as a complementary triage test to select 
worthwhile antigens to proceed with more 
laborious in vivo provocation tests when the 
specific IgE is undetectable. None of our patients 
presented an exclusive reaction to benzoic acid. 
Every patient was simultaneously tested with 
protein allergens (mites, fungi, food allergens), 
demonstrating positive results for some of them 
[46]. Our results may suggest that allergic 
patients may impair their symptoms by a 
pharmacological or an immune additional action 
of benzoic acid over the hypersensitivity 
response. 
 

5. CONCLUSIONS 
 

Our preliminary results show that the LAIT may 
differentiate diverse degrees of ex vivo 
immunoreactivity against benzoic acid in patients 
clinically diagnosed with non–IgE-mediated 
allergies. The propaedeutic meaning of these 
results, however, must be established. More 
studies with prospective larger double-blind 
cohorts need to evaluate the potential 
contribution of LAIT for the etiologic diagnosis of 
patients suspected of symptomatic 
hypersensitivity against benzoic acid and other 
similar food, cosmetic, and pharmaceutical 
additives.  
 

6. LIMITATIONS 
   

This study is a retrospective analysis of data 
collected over six years. There was no protocol 
research, and the subject's data were limited to 
the essentials available on our electronic sheets. 
The number of subjects is appropriate for a 
preliminary study; however, future studies must 
be more comprehensive. The lack of a research 
protocol implies the possibility of a bias produced 
by the point of view of the physician who 
indicated the exam (CEO) based on a clinical 
suspicion led purely by the anamnesis. The study 

lost many of these patients to follow-up, so 
assessing the relationship between LAIT 
outcome and the patient's subsequent clinical 
outcome was impossible. 

  

CONSENT  
 
As per international standards or university 
standards, patient(s) written consent has been 
collected and preserved by the author(s). 
 

ETHICAL APPROVAL 
 
As per international standards or university 
standards written ethical approval has been 
collected and preserved by the author(s). 

 
COMPETING INTERESTS 
 
Authors have declared that they have no known 
competing financial interests OR non-financial 
interests OR personal relationships that could 
have appeared to influence the work reported in 
this paper. 
 

REFERENCES 
 

1. Nair B. Final report on the safety 
assessment of Benzyl Alcohol, Benzoic 
Acid, and Sodium Benzoate. Int J Toxicol. 
2001:20(suppl 3):23-50. 

2. Adams TB, Cohen SM, Doull J, Feron VJ, 
Goodman JI,  Marnett LJ. Et al. The FEMA 
GRAS assessment of benzyl derivatives 
used as flavor ingredients. Food Chem 
Toxicol. 2005;43(8):1207-40. 

3. FAO-WHO Expert Committee. 
Toxicological evaluation of certain food 
additives with a review of general 
principles and of specifications. 
Seventeenth report of the joint FAO-WHO 
Expert Committee on Food Additives. 
World Health Organization technical report 
series. 1974;539:1-40. 

4. FAO-WHO Expert Committee. Evaluation 
of certain food additives. Twenty-third 
Report of the Joint FAO/WHO Expert 
Committee on Food Additives. World 
Health Organization technical report 
series. 1980;648:1-45. 

5. Jia W, Wang X, Shi L. Endogenous 
benzoic acid interferes with the signatures 
of amino acids and thiol compounds 
through perturbing N-methyltransferase, 
glutamate-cysteine ligase, and glutathione 
S-transferase activity in dairy products. 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 63-70, 2024; Article no.AJI.115657 
 
 

 
68 

 

Food Res Int (Ottawa, Ont.) 2022; 
161:111857. 

6. Boelens MH, Visscher CA, Willemsens LC, 
Maarse H. Volatile compounds in food: 
qualitative and quantitative data. 6. Ed. 
Zeist Food Analysis Institute. Zeist. 1989. 

7. Simon RA. Adverse reactions to drug 
additives. J Allergy Clin Immunol. 1984; 
74(4 Pt 2):623-30. 

8. Lahti A, Hannuksela M. Is benzoic acid 
really harmful in cases of atopy and 
urticaria? Lancet. 1981;2(8254):1055. 

9. Jacob SE, Hill H, Lucero H, Nedorost S. 
Benzoate Allergy in Children - From Foods 
to Personal Hygiene Products. Pediatr 
Dermatol. 2016;33(2):213-5. 

10. Balatsinou L, Di Gioacchino G, Sabatino 
G, Cavallucci E, Caruso R, Gabriele E. 
Asthma worsened by benzoate contained 
in some antiasthmatic drugs. Int J 
Immunopathol Pharmacol. 2004;17(2):225-
6. 

11. Ros AM, Juhlin L, Michaëlsson G. A 
follow-up study of patients with recurrent 
urticaria and hypersensitivity to aspirin, 
benzoates and azo dyes. Br J Dermatol. 
1976;95 (1):19-24. 

12. Michaëlsson, G.; Juhlin, L., Urticaria 
induced by preservatives and dye 
additives in food and drugs. Br J Dermatol. 
1973;88 (6), 525-32. 

13. Jutel M, Agache I, Zemelka-Wiacek M, 
Akdis M, Chivato, T,  del Giacco S, et al. 
Nomenclature of allergic diseases and 
hypersensitivity reactions: Adapted to 
modern needs: An EAACI position paper. 
Allergy. 2023;78(11):2851-2874. 

14. Worm M, Vieth W, Ehlers I, Sterry W, 
Zuberbier T. Increased leukotriene 
production by food additives in patients 
with atopic dermatitis and proven food 
intolerance. Clin Exp Allergy. 2001;31(2): 
265-273. 

15. Fink A, Bibi H, Eliraz A, Tabachnik E, 
Bentwich Z. Leukotrienes (LTC4, LTD4) 
confer glass non-adherence on leukocytes 
of asthmatic individuals. Dependency on 
cyclooxygenase products and calcium ion. 
Immunol Lett. 1985;10(6):319-23. 

16. Fink A, Bibi H, Eliraz A, Schlesinger M, 
Bentwich Z. Ketotifen, disodium 
cromoglycate, and verapamil inhibit 
leukotriene activity: determination by tube 
leukocyte adherence inhibition assay. Ann 
Allergy. 1986;57(2):103-6. 

17. Fink A, Shahin R, Eliraz A, Bibi H, 
Berkenstadt H, Levin S, et al. Interferon 

modulates the leukotriene C4-induced 
non-adherence properties of leukocytes: 
acquisition of an asthmatic phenotype. 
Immunol Lett. 1985;10(3-4):159-63. 

18. Kuratsuji T. Studies on leukocyte 
adherence inhibition test. Part II. Clinical 
applications of LAI test to detect delayed 
type hypersensitivity in infants and 
children. Keio J Med. 1981;30(2):65-9. 

19. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS, 
et al. Evaluating Non-IgE-mediated 
Allergens' Immunoreactivity in Patients 
with "Intrinsic" Persistent Rhinitis with Help 
of the Leukocyte Adherence Inhibition 
Test. Eur J Med Health Sci. 2023;5(1):17-
22. 

20. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS, 
et al. Evaluating Non-IgE-Mediated 
Allergens' Immunoreactivity in Patients 
Formerly Classified as "Intrinsic" 
Asthmatics with Help of the Leukocyte 
Adherence Inhibition Test. Eur J Clin Med. 
2023:4(2):1-7. 

21. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS, 
et al. Contribution of the Leukocyte 
Adherence Inhibition Test to the Diagnosis 
of Innate Non–IgE-mediated 
Immunoreactivity against Alternaria 
alternata. Asian J Immunol. 2023;6(1):243-
251. 

22. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS. 
Contribution of the Leukocyte Adherence 
Inhibition Test to the Diagnosis of Innate 
Non–IgE-mediated Immunoreactivity 
against Saccharomyces cerevisiae.                  
Asian J Immunol. 2023;6(1):185-                       
195. 

23. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS, 
et al. Contribution of the Leukocyte 
Adherence Inhibition Test to the Diagnosis 
of Non–IgE-mediated Immunoreactivity 
against Candida albicans in Patients with 
Atopic Dermatitis. Asian J Immunol. 
2023;6(1):268-276. 

24. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS, 
et al. Contribution of the Leukocyte 
Adherence Inhibition Test in Diagnosing 
Non–IgE-Mediated Immunoreactivity 
against Aspergillus fumigatus in Patients 
with Allergic Rhinitis and Asthma. Asian J 
Immunol. 2024;7(1):12-20. 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 63-70, 2024; Article no.AJI.115657 
 
 

 
69 

 

25. Olivier CE, Argentão DGP, Santos RAPG, 
Silva MD, Lima RPS, Zollner RL. Skin 
scrape test: an inexpensive and painless 
skin test for recognition of immediate 
hypersensitivity in children and adults. 
Open Allergy J. 2013;6:9-17. 

26. Olivier CE, Lima RPS, Pinto DG, Santos 
RAPG, Silva GKM, Lorena SLS, et al. In 
search of a tolerance-induction strategy for 
cow's milk allergies: significant reduction of 
beta-lactoglobulin allergenicity via 
transglutaminase/cysteine polymerization. 
Clinics. 2012;67(10):1171-1179. 

27. Olivier CE, Santos RAPG, Lima RPS, 
Argentão DGP, Silva GKM, Silva MD. A 
Novel Utility for an Old Method: The 
Leukocyte Adherence Inhibition Test Is an 
Easy Way to Detect the Immunoreactive 
Interference of the Collection Tube 
Anticoagulant on Cellular Immunoassays. 
J Cell Adhesion. 2014:1-6 Article ID 
860427 

Available:http://dx.doi.org/10.1155/2014/86
0427 

28. Olivier CE, Pinto DG, Lima RPS, Silva MD, 
Santos RAPG, Teixeira, et al. Assessment 
of Immunoreactivity against Therapeutic 
Options Employing the Leukocyte 
Adherence Inhibition Test as a Tool for 
Precision Medicine. Eur J Clin Med. 
2021;2(3):40-45. 

29. Olivier CE, Pinto DG, Santos RAPG, Lima 
RPS. Dextran's interference over the 
Leukocyte Adherence Inhibition Test. 
Academia Letter 2021, Article (number), 
3792. 

30. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS. 
Immunoreactivity against 
Dermatophagoides pteronyssinus 
Assessed by the Leukocyte Adherence 
Inhibition Test in Patients with Intrinsic 
Atopic Dermatitis and Correlated "Intrinsic" 
Non–IgE-mediated Allergic Conditions. Eur 
J Clin Med. 2021;2(6):45-50. 

31. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS, 
et al. Contribution of the Leukocyte 
Adherence Inhibition Test to the Evaluation 
of Cellular Immunoreactivity against Latex 
Extracts for Non—IgE-Mediated Latex-
Fruit-Pollen Syndrome in Allergic 
Candidates to Exclusion Diets and Allergic 
Desensitization. Eur J Clin Med. 
2022;3(1):11-17. 

32. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS, 

et al. Contribution of the Leukocyte 
Adherence Inhibition Test for the 
evaluation of immunoreactivity against 
gluten extracts in non—IgE-mediated / 
non-autoimmune Gluten-Related 
Disorders. Eur J Clin Med. 2022;3(2):1-7. 

33. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS, 
et al. Leukocyte Adherence Inhibition Test 
to the Assessment of Immunoreactivity 
Against Cow's Milk Proteins in Non—IgE-
Mediated Gastrointestinal Food Allergy. 
Eur J Clin Med. 2022;3(2):38-43. 

34. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS, 
et al. Contribution of the Leukocyte 
Adherence Inhibition Test to the Diagnosis 
of Immunoreactivity against Cobalt. Asian 
J Immunol. 2023;6(1):174-184. 

35. Venter C, Vieira MC, Fleischer D. 
Tolerance development in non-IgE 
mediated food allergies: lessons from 
Brazil. Jornal de Pediatria. 2023;2(30):1-4. 

36. Asero R. Sodium benzoate-induced 
pruritus. Allergy. 2006;61(10):1240-1. 

37. Smith WL, Garavito RM, DeWitt DL. 
Prostaglandin Endoperoxide H Synthases 
(Cyclooxygenases) 1 and 2. J Biol Chem. 
1996;271(52):33157-33160. 

38. Shimizu T. Lipid Mediators in Health and 
Disease: Enzymes and Receptors as 
Therapeutic Targets for the Regulation of 
Immunity and Inflammation. Annual Rev 
Pharmacol Toxicol. 2009;49(1):123-150. 

39. Liu M, Yokomizo T. The role of 
leukotrienes in allergic diseases. Allergol 
Int. 2015;64(1):17-26. 

40. Olivier CE, Lima RPS, Pinto DG, Santos 
RAPG. The Plasma Preincubation with 
Papain Before the Assay Suggests that a 
Gell and Coombs Type II Reaction is Been 
Demonstrated by the Leukocyte 
Adherence Inhibition Test. Biom J Sci 
Tech Res. 2021;36(3):28647-28655. 

41. Thomson DMP. Assessment of immune 
status by the leukocyte adherence 
inhibition test. Academic Press: New York. 
1982:380p. 

42. Tong AW, Burger DR, Finke P, Barney C, 
Vandenbark AA, Vetto RM. Assessment of 
the mechanism of the leukocyte adherence 
inhibition test. Cancer Res. 
1979;39(2Pt2):597-603. 

43. Fink A, Heller L, Eliraz A, Weisman Z, 
Miskin A, Schlezinger M, et al. Allergen-
specific leukocyte adherence inhibition 
(LAI) assay: sensitivity, specificity and 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 63-70, 2024; Article no.AJI.115657 
 
 

 
70 

 

mechanism. Immunol Lett. 1987;16(1):65-
70. 

44. Halliday WJ, Maluish A, Miller S. Blocking 
and unblocking of cell-mediated anti-tumor 
immunity in mice, as detected by the 
leucocyte adherence inhibition test. Cell 
Immunol. 1974;10(3):467-475. 

45. Chung MK, House JS, Akhtari FS, Makris 
KC, Langston MA Islam, et al. Decoding 

the exposome: data science 
methodologies and implications in 
exposome-wide association studies 
(ExWASs). Exposome. 2024;4(1):osae0       
01. 

46. World Medical Association Declaration of 
Helsinki: ethical principles for medical 
research involving human subjects. JAMA. 
2013;310(20):2191-4. 

_________________________________________________________________________________ 
© Copyright (2024): Author(s). The licensee is the journal publisher. This is an Open Access article distributed under the terms 
of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, 
distribution, and reproduction in any medium, provided the original work is properly cited. 

 
 

 

Peer-review history: 
The peer review history for this paper can be accessed here: 

https://www.sdiarticle5.com/review-history/115657 


