







































 

_____________________________________________________________________________________________________ 
 
*Corresponding author: E-mail: celso@alergoimuno.med.br; 
 
Cite as: Olivier, C. E., Pinto, D. G., Teixeira, A. P. M., Santana, J. L. S., Santos, R. A. P. G., Lima, R. P. S., & Monteiro, E. S. 
(2024). Endotyping Non IgE Mediated Immunoreactivity to Dermatophagoides farinae: Implications for Allergic Patients: A 
Retrospective Study. Asian Journal of Immunology, 7(1), 90–99. Retrieved from 
https://journalaji.com/index.php/AJI/article/view/134 

 

Asian Journal of Immunology 
 
Volume 7, Issue 1, Page 90-99, 2024; Article no.AJI.117841 
 

 
 

 

 

Endotyping Non IgE Mediated 
Immunoreactivity to 

Dermatophagoides farinae:  
Implications for Allergic Patients: A 

Retrospective Study 
 

Celso Eduardo Olivier a*, Daiana Guedes Pinto a,  

Ana Paula Monezzi Teixeira a,  

Jhéssica Letícia Santos Santana a, 

Raquel Acácia Pereira Gonçalves Santos a, 

Regiane Patussi Santos Lima b  

and Everton Salgado Monteiro c 
 

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

c Department of Allergy and Immunopathology, Faculty of Medicine, São Paulo University, 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/v7i1134  

 
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/117841 

 
 
 

Received: 19/05/2024 
Accepted: 28/05/2004 
Published: 31/05/2024 

 

Original Research Article 

https://doi.org/10.9734/aji/2024/v7i1134
https://www.sdiarticle5.com/review-history/117841


 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 90-99, 2024; Article no.AJI.117841 
 
 

 
91 

 

ABSTRACT 
 

Background: Several publications report Dermatophagoides farinae allergens as responsible for 
several types of non–IgE-mediated allergic reactions. There is no standardized lab exam to 
endotype (quantify the participation of these mechanisms) in the context of allergic disease 
pathophysiology. 
Aims: To evaluate the potential of the Leukocyte Adherence Inhibition Test (LAIT) and the Tube 
Titration of Precipitins (TTP) on serum to discriminate non–IgE-mediated immunoreactivity against 
D. farinae in patients with non–IgE-mediated allergic phenotype with clinically suspected 
hypersensitivity to their allergens. 
Study Design: We retrospectively examined the medical charts of 134 allergic patients 
investigated with LAIT and 100 allergic patients investigated with TTP against an extract of D. 
farinae.  
Place and Duration of Study: Instituto Alergoimuno de Americana – São Paulo – Brazil –  
between January 2018 and May 2024. 
Methodology: The registered results of the semi-quantitative serum TTP against D. farinae extract 
were distributed in ranges through a cascade distribution chart to outline the variability of the 
results. The registered results of the percentage of Leukocyte Adherence Inhibition (LAI) promoted 
by the ex vivo challenges with D. farinae extract were distributed in ranges through a cascade 
distribution chart to outline the variability of results. The statistical characteristics of these cohorts 
were calculated. 
Results: The TTP showed a wide distribution range, with four negative and most positive results 
concentrating on the higher dilutions. The mean was estimated at 1:242; the median was 1:128; the 
standard deviation was estimated at 1:210; the mode was 1:512 (appeared 36 times). The LAI 
ranged from 0% to 100%. The mean was 52%; the median was 55.5%; the standard deviation was 
23.6%; the mode was 63% (appeared seven times). The cascade distribution demonstrates a wide 
range of LAI results.   
Conclusion: Our preliminary results support that the semi-quantitative TTP and the LAIT have the 
potential to endotype and clinically discriminate diverse degrees of cellular and humoral non–IgE-
mediated immunoreactivity against D. farinae in allergic patients. 
 

 
Keywords: Allergy; asthma; bronchitis; Dermatophagoides farinae; diagnosis; hypersensitivity; 

leukocyte adherence inhibition test; non–IgE-mediated immunoreactivity; precipitins; 
rhinitis. 

 

ABBREVIATIONS 
 
LAI : Leukocyte Adherence Inhibition 
LAIT : Leukocyte Adherence Inhibition Test 
TTP : Tube Titration of Precipitins 
 

1. INTRODUCTION  
 

Storing food and sharing mattresses with mites 
exposed humanity to commensalism, which was 
only noticed about a hundred years ago [1]. 
Storage mites have been reported as causes of 
allergic diseases since 1924 when Willem Storm 
van Leeuwen described several cases of asthma 
in patients exposed to mite-infested wheat and 
oats [2]. Storage mites are eight-legged 
members of the Arachnid class and a significant 
cause of allergic diseases through atopic 
sensitization to their multiple allergens [3]. 
Dermatophagoides farinae is a storage mite 
belonging to the phylum Arthropoda, subphylum 

Chelicerata, class Arachnida; subclass Acari; 
superorder Acariformes; order Sarcoptiformes; 
suborder Psoroptidia; family Pyroglyphidae; 
subfamily Dermatophagoidinae [4]. The allergic 
reactions produced by the ingestion of storage 
mites are frequently reported as "oral mite 
anaphylaxis" or "pancake syndrome" [5]. Several 
IgE-mediated and Non–IgE-mediated 
hypersensitivity mechanisms were associated 
with these conditions [6]. 
 
Besides a typical storage mite, D. farinae is also 
a house dust mite found in mattresses and 
pillows, which provides food, moisture, and a 
thermal source [7]. Besides corporal allergens, 
several allergens are identified in the feces, 
eggs, and excretions of D. farinae, which 
accumulate in the environment and are 
recognized as significant immunogens [8]. The 
Allergen Nomenclature Sub-Committee from the 
World Health Organization and the International 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 90-99, 2024; Article no.AJI.117841 
 
 

 
92 

 

Union of Immunological Societies (WHO/IUIS) 
had already classified dozens of allergens 
belonging to D. farinae [9]. Most D. farinae 
allergens were identified by proteomics, using 
IgE-based techniques such as enzyme-linked 
immunosorbent assay inhibition tests, 
immunoblots, basophil activation tests, and skin 
prick tests [10]. Besides the IgE-mediated 
allergenicity, the immunoreactivity of the D. 
farinae allergens on the innate immune system 
and its contribution to the production of 
symptoms in allergic patients have also been 
studied [11]. Several IgG epitopes of D. farinae 
allergens were already identified by peptide 
microarray immunoassay [12]. Specific D. farinae 
allergens, such as the Der f 38, stimulate Innate 
immunoreactivity by binding to Toll-like receptors 
(such as TLR4), acting as allergy inducers [13]. 
D. farinae also stimulates the transcription of 
long noncoding RNAs (non-protein-coding RNA 
that exert transcriptional and post-transcriptional 
regulation on messenger RNAs and microRNAs), 
causing cell dysfunction and dysregulation of 
circulating CD8+ T cells of patients with asthma 
[14]. Some D. farinae allergens, besides acting 
as IgE ligands, also stimulate Innate immune 
cells, such as the α-Tubulin (Der f 33), which 
increases the expression of interleukin-4 and 
upregulates CD80 and TNF-α levels in dendritic 
cells [15]. Exosomes from D. farinae induce 
immunogenic inflammation by stimulating 
epithelial cells and macrophages to release 
inflammatory-related cytokines such as 
interleukin-33, thymic stromal lymphopoietin, 
TNF-alpha, and IL-6 [16]. D. farinae induces 
innate inflammation via Interleukin-33 via 
Receptor-interacting protein kinase signaling 
[17]. 
 
Besides their allergens, mites also harbor a 
microbiome that produces toxins and a 
diversified set of allergens that can trigger innate 
and adaptive allergic responses [18]. 
Environmental allergens such as mites and 
microorganisms may produce innate immune 
dysregulation, resulting in epithelial damage, 
inadequate adaptive response, and a persistent 
inflammatory state [19]. The mite's microbiome is 
rich in pathogen-associated molecular patterns 
(PAMPs), which are also involved in mast cell 
activation and non–IgE-mediated allergic 
responses [20]. The microbial Damage-
Associated Molecular Patterns (DAMPs) promote 
type IVc hypersensitivity reactions through Th17 
cell cytokines and Group 3 Innate Lymphoid 
Cells, leading to the liberation of Neutrophil 
Extracellular Traps (NETs), stimulating the innate 

immune response and inflammation [21,22]. 
DAMPs trigger the type VII Hypersensitivity 
reaction through Pattern Recognition              
Receptors (PRRs) [23]. Non–IgE-mediated 
hypersensitivity to mite bacterial microbiome is 
documented in patients with acne rosacea 
[24,25]. 
 
To endotype non–IgE-mediated immunoreactivity 
against suspected allergens, we routinely employ 
the semi-quantitative Tube Titration of Precipitins 
(TTP) and the Leukocyte Adherence Inhibition 
Test (LAIT), an ex vivo challenge immunoassay 
made with viable leukocytes already reported to 
demonstrate allergen-specific immunoreactivity 
against D. farinae in allergic patients [26]. To 
evaluate the potential of these procedures to 
discriminate non–IgE-mediated immunoreactivity 
against D. farinae, we retrospectively compiled 
the electronic medical charts of patients with 
non–IgE-mediated allergic rhinitis, allergic 
bronchitis, allergic conjunctivitis, and/or atopic 
dermatitis who were investigated with them. 
Patients diagnosed with these allergic conditions 
were eligible for this investigation after 
demonstrating non-reactive or inconclusive skin 
tests against D. farinae extract, a normal range 
total IgE, and undetectable specific IgE for D. 
farinae.  
 

2. MATERIALS AND METHODS  
 

2.1 Subjects 
 
After receiving Institutional Review Board 
approval from the Instituto Alergoimuno de 
Americana (Brazil; 04/2024), we proceeded with 
the electronic chart review of 8,800 allergic 
patients who attended our outpatient facility from 
January 2018 to May 2024.  
 
A cohort of 134 patients had been submitted to 
an ex vivo allergen challenge test with D. farinae 
extract monitored with LAIT for presenting non–
IgE-mediated allergic conditions. The LAIT 
cohort counted 44 males; mean age 42.9 years; 
SD 17.8 years; range 17 to 89 years; modes = 
19 and 25 (each appeared seven times); 
geometric mean = 39.3 years.  
 

A cohort of 100 patients had been submitted to 
TTP with D. farinae extract for presenting non–
IgE-mediated allergic conditions. The TTP cohort 
counted 35 males; mean age 40.8 years; SD 
20.5 years; range 3 to 86 years; modes = 94 
(appeared five times); geometric mean = 33 
years.  



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 90-99, 2024; Article no.AJI.117841 
 
 

 
93 

 

“These procedures were offered to patients with 
allergic rhinitis, allergic bronchitis, allergic 
conjunctivitis, atopic dermatitis, and/or urticaria 
with a normal range total IgE, undetectable 
specific IgE against D. farinae (investigated 
through ImmunoCAP®), and a non-reagent or 
inconclusive investigation performed with allergic 
skin tests done with the D. farinae extract” [27]. 
 

2.2 Antigen Preparation 
 

“The D. farinae extracts were obtained from 
frozen cultures. The contents of the bottles 
containing mites (adults, nymphs, larvae, feces, 
and eggs) and culture medium were weighed 
and left at a rate of 10 ml of PBS buffer per gram 
of material (10%) by gentle magnetic stirring 
(1,000 rpm) for four hours at 4ºC. The material 
was centrifuged at 5,000 rpm for 30 minutes. The 
supernatant was kept apart. The sediment was 
resuspended in the same conditions as the 
previous step and stirred at four °C for 24 hours. 
The centrifugation was repeated, and the 
supernatants were mixed into an Erlenmeyer 
flask. The solution was filtered through a double 
paper filter and later through a 0.2 μm pore-size 
filter with the help of a vacuum flask and a 
vacuum pump. The extract was dialyzed with the 
aid of the Thermo Fisher Scientific SnakeSkin™ 
88244 Dialysis Tubing in distilled water (1:50 
ratio) with three water changes for 24 hours to 
eliminate molecules of low molecular mass (< 
5,000 Da). After dialysis, the extract was 
centrifuged at 10,000 rpm for 30 minutes at 4ºC 
and frozen at -40 ºC. The protein concentration 
was estimated spectrophotometrically and diluted 
to 1 mg/mL in saline (NaCl 0.9%) to perform the 
allergic skin tests, LAIT, and TTP” [28].  
 

2.3 Ex vivo Investigation: Leukocyte 
Adherence Inhibition Test (LAIT) 

 

2.3.1 Procedure for allergen Ex vivo 
challenging  

 

LAIT was performed as previously described [29-
39]. Shortly, each donor's fresh plasma was 
divided into two parts and used in paralleled ex 
vivo challenging tests with D, farinae extract, 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 antigen extract (10μL of a solution with 
1mg/mL and pH 7.5) or with the antigen dilution 
solution (when used as control).  

2.3.2 Procedure for adherence assay  
 
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.  
 
2.3.3 Procedure for calculation  
 
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. 

 
2.4 In vitro Investigation: Tube Titration 

of Precipitins (TTP) 
 
As previously reported, the semi-quantitative 
tube titration of precipitins (TTP) against D. 
farinae extract was performed in a transparent 
vitreous tube [40]. Shortly, the patient's blood 
was collected in a clot-activator collecting tube. 
After separation, the serum was centrifugated at 
2,000 rpm for 10 minutes. The allergen extracts 
were allocated in sets of eleven glass tubes at 
progressive duplicated serum dilutions. The 
progressive dilutions were combined with the 15 
μL of the antigen (1 mg/mL) with 250 μL of the 
patient's serum, progressively diluted into 
physiological saline solution (NaCl 0,9%) in the 
dilution ratios of 1:1; 1:2; 1:4; 1:8; 1:16; 1:32; 
1:64; 1:128; 1:256; and 1:512. One tube was a 
blank control done with the water and serum to 
observe occasional spontaneous precipitation 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 90-99, 2024; Article no.AJI.117841 
 
 

 
94 

 

(Sia Test) to detect circulating immune 
complexes [41]. After 24 hours, one of us 
examined the tubes, and the titers (the highest 
dilution factor yielding a positive reading) were 
recorded [42]. 
 

3. RESULTS 
 
As a retrospective survey, there was no research 
protocol; therefore, we report the incidental 
immune investigation as registered in the digital 
medical charts. Therefore, we could not establish 
a cross-comparison between positive and 
negative controls to validate the results. 
 
The TTP showed a wide distribution range, with 
four negative results and most positive results 
concentrated on the higher dilutions (Fig. 1). The 
mean was estimated at 1:242; the median was 

1:128; the standard deviation was estimated at 
1:210; the mode was 1:512 (appeared 36 times). 
All Sia tests were negative.  
 
The LAI ranged from 0% to 100%. The mean 
was 52%; the median was 55.5%; the standard 
deviation was 23.6%; the mode was 63% 
(appeared seven times). The cascade 
distribution demonstrates a wide range of 
distribution of LAI results (Fig. 2). Six patients 
ignored the presence of the allergen on the 
plasma and presented no inhibition of leukocyte 
adherence (LAI = 0%) after contact with D. 
farinae extract (4.5% of the tests). Some patients 
showed low or moderate immunoreactivity during 
the ex vivo challenge test, while others displayed 
strong immunoreactivity, which could reflect the 
participation of D. farinae allergens in a non–IgE-
mediated hypersensitivity condition. 

 

 
 

Fig. 1. Cascade distribution chart of the tube titration of precipitins (x-axis %) resulting from 
the D. farinae extract against the serum of a cohort of 100 tests/subjects (y-axis) 

 

 
 

Fig. 2. Cascade distribution chart of the range groups of Leukocyte Adherence Inhibition (LAI) 
results (x-axis %) of ex vivo D. farinae extract challenges monitored by the Leukocyte 

Adherence Inhibition Test (LAIT), according to the respective number of outcomes over a 
cohort with 134 tests/subjects (y-axis) 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 90-99, 2024; Article no.AJI.117841 
 
 

 
95 

 

4. DISCUSSION 
 

Diagnosing hypersensitivity reactions is essential 
in increasing patients' awareness of their 
sensitivities and commitment to avoiding 
allergens. The development of several lab 
methods to detect specific IgE has raised 
concerns about the eviction of house dust mite 
allergens, mainly by chemical treatment and 
encasement methods for pillows and bed 
mattresses [43]. However, non-IgE-mediated 
food-storage mite hypersensitivity may be a 
challenging diagnosis in the field of food allergies 
[44].  
 

Significant mite mortality and reduction of Der f 1 
fecal allergen were observed in carpets and 
mattresses after the treatment with hard surface 
steam cleaners [45]. Some studies demonstrated 
that negative ions produced by ionizers kill dust 
mites and can be used to reduce mite 
populations on surfaces such as floors and 
clothes [46]. Sublingual immunotherapy drops 
are a highly effective treatment for D. farinae 
allergy by increasing the subsets of T immune 
cells, specifically Th17 cells and CD4+CD25+ 
regulatory T cells (Treg cells), in peripheral blood 
[47]. 
 

Before the discovery and dissemination of lab 
methods to detect allergen-specific IgE, 
endotyping of allergic diseases was performed 
with the help of allergic skin tests, Complement 
fixation assays, and the research of precipitins 
[48]. However, diagnosing non-IgE-mediated 
hypersensitivity is difficult in medical practice 
since clinical analysis diagnostic laboratories do 
not offer specific tests to support this diagnosis, 
which is only possible in academic research 
institutions. 
 

An ex vivo challenge test with a viable leukocyte 
buffy coat, the LAIT allows the living interaction 
of all immune-circulating participants with the 
tested allergen, such as the innate and adaptive 
immune cells, cytokines, alarmins, and 
antibodies [49]. However, as an observant of the 
final phenomenon, the LAIT does not indicate 
which pathways were involved [26,50-52].  
 

The TTP classically proves that patients produce 
circulating antibodies against the specific 
allergen added to the serum in sufficient 
quantities to produce immune complexes large 
enough to precipitate visibly at the bottom of the 
tubes. TTP is the most basic laboratory exam 
upon which Immunology was established as a 
science [53,54].  

This preliminary retrospective survey 
demonstrated extensive results from the ex vivo 
challenge test monitored by LAIT and the TTP 
with D. farinae in two heterogeneous cohorts of 
allergic patients. These exams mostly 
demonstrate that IgE-mediated hypersensitivity is 
not responsible for the allergic diseases in these 
patients. These allergic diseases should instead 
be viewed as a conjoint of intricate immune 
hypersensitivity mechanisms that act conjointly to 
produce damage. Endotyping these mechanisms 
helps to clarify the individual's immune response 
to better prescribe the management and the 
treatment in personalized medical care.  
 
We routinely employ the LAIT and the research 
of tube precipitins as a complementary triage test 
to select worthwhile antigens to proceed with 
more laborious in vivo provocation tests when 
the specific IgE is undetectable. Our results 
suggest that most allergic patients present some 
immunoreactivity against D. farinae allergens, 
while some do not. However, the complete 
significance of these results is not yet fully 
established. As an isolated piece of information, 
the LAIT immunoreactivity or the presence of 
precipitins do not prove that the tested antigens 
explain the allergic symptoms exactly. Indeed, 
the clinical diagnosis must be accomplished by 
the in vivo challenges, the degree of colonization 
of the patient's environment, and the benefits of 
a change of ambient and an occasional 
desensitization treatment [55-57]. More studies 
with prospective larger double-blind cohorts need 
to evaluate the potential contribution of these 
methods to diagnosing patients suspected of D. 
farinae non–IgE-mediated hypersensitivity.  
 

5. CONCLUSION 
 
Our preliminary results support that the semi-
quantitative TTP and the LAIT have the potential 
to endotype and clinically discriminate diverse 
degrees of cellular and humoral non–IgE-
mediated immunoreactivity against D. farinae in 
allergic patients. This methodology can provide a 
socioeconomic impact since the technology to 
perform TIAL and TTP is inexpensive and can be 
performed in a single room attached to the 
facilities with minimum laboratory equipment.  
 
However, the propaedeutic meaning of these 
results must be better established, as well as the 
effects of possible interferents [58]. More studies 
focused on the quality-by-design approach, 
including prospective larger double-blind cohorts, 
need to evaluate the potential contribution of 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 90-99, 2024; Article no.AJI.117841 
 
 

 
96 

 

LAIT and TTP for endotyping immunoreactivity of 
patients with allergic phenotypes suspected of 
presenting D. farinae non-IgE-mediated 
hypersensitivity [59].  
 

CONSENT 
 
As a retrospective survey of results recorded 
incognito, consent was given collectively by the 
institution's ethics committee following the 
principles of the Declaration of Helsinki [60]. 
 

ETHICAL APPROVALS 
 
The authors have collected and preserved 
written ethical approval per international 
standards. 
 

ACKNOWLEDGEMENTS 
 

The Instituto Alergoimuno de Americana funded 
this work. We want to thank our new laboratory 
technician, Alessandra Vieira de Oliveira, for 
participating in the exams. 
 

COMPETING INTERESTS 
 

Authors have declared that no competing 
interests exist. 
 

REFERENCES 
 
1. Vackova T, Pekar S, Klimov PB, Hubert J. 

Sharing a bed with mites: Preferences of 
the house dust mite Dermatophagoides 
farinae in a temperature gradient. Exp Appl 
Acarol. 2021;84(4):755-767. 

2. Spieksma FT, Dieges PH. The history of 
the finding of the house dust mite. J Allergy 
Clin Immunol. 2004;113(3):573-6. 

3. Miller JD. The role of dust mites in allergy. 
Clin Rev Allergy Immunol. 2019;57(3):312-
329. 

4. Schoch CL et al. NCBI Taxonomy: A 
comprehensive update on curation, 
resources and tools. Database (Oxford); 
2020: baaa062.  
PubMed: 32761142  
PMC: PMC7408187.  
Available:https://130.14.29.110/Taxonomy/
Browser/wwwtax.cgi  
(accessed 02/11/2023) 

5. Sánchez-Borges M, Suárez-Chacon R, 
Capriles-Hulett A, Caballero-Fonseca F,  
Iraola V, Fernández-Caldas E. Pancake 
syndrome (oral mite anaphylaxis). World 
Allergy Organ J. 2009:2(5):91-96. 

6. Sánchez-Borges M, Fernandez-Caldas E. 
Hidden allergens and oral mite 
anaphylaxis: The pancake syndrome 
revisited. Curr Opin Allergy Clin Immunol. 
2015;15(4):337-43. 

7. Vackova T, Pekar S, Klimov PB, Hubert J. 
Population growth and respiration in the 
dust mite Dermatophagoides farinae under 
different temperature and humidity 
regimes. Exp Appl Acarol.  2023;89(2): 
157-169. 

8. Erban, T.; Hubert, J., Two-dimensional gel 
proteomic analysis of Dermatophagoides 
farinae feces. Exp Appl Acarol. 2015; 
65(1):73-87. 

9. World Allergen Nomenclature Sub-
Committee from the World Health 
Organization and the International Union of 
Immunological Societies (WHO/IUIS)   
Available:https://allergen.org/search.php?a
llergensource=Dermatophagoides+farinae
&searchsource=Search. 

10. An S, Chen L, Long C, Liu X,  Xu X, Lu X, 
et al. Dermatophagoides farinae allergens 
diversity identification by proteomics. Mol 
Cell Proteomics. 2013:12(7):1818-28. 

11. Cao H, Liu Z. Clinical significance of dust 
mite allergens. Mol Biol Rep. 
2020;47(8):6239-6246. 

12. Teng F, Han F, Zhu X, Yu L, Gai D, Xu C, 
et al. Identification of continuous 
immunoglobulin G epitopes of 
Dermatophagoides farinae allergens by 
peptide microarray immunoassay. IUBMB 
Life. 2020:72(9):1976-1985. 

13. Kim G, Hong M, Kashif A, Hong Y, Park 
BS, Mun JY, et al. Der f 38 Is a Novel 
TLR4-Binding Allergen Related to Allergy 
Pathogenesis from Dermatophagoides 
farinae. Int J Mol Sci. 2021;22(16). 

14. Wang L, Zhou Y, Cui Y. lncRNA profiling in 
NCI-H292 cells after stimulation with 
Dermatophagoides farinae extracts. Int J 
Immunopathol Pharmacol. 2018;32: 
394632017750997. 

15. Wang H, Lin J, Liu X, Liang Z, Yang P, 
Ran P, et al. Identification of α-tubulin, Der 
f 33, as a novel allergen from 
Dermatophagoides farinae. Immunobiol. 
2016;221(8):911-7. 

16. Yang T, Xu Z, Yu J, Liu J, Wang W, Hong 
S. Exosomes Derived from 
Dermatophagoides farinae Induce Allergic 
Airway Inflammation. Microbiol Spectrum. 
2023;11(4):e0505422. 

17. Jin M, Bang JS, Ha DL, Kim JY, Park KD, 
Lee WJ, et al. Dermatophagoides farinae 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 90-99, 2024; Article no.AJI.117841 
 
 

 
97 

 

Extract Induces Interleukin 33-Mediated 
Atopic Skin Inflammation via Activation of 
RIP1. Int J Mol Sci. 2023;24(6):5228. 

18. Lee J, Kim JY, Yi MH, Hwang Y, Lee IY, 
Nam SH, et al. Comparative microbiome 
analysis of Dermatophagoides farinae, 
Dermatophagoides pteronyssinus, and 
Tyrophagus putrescentiae. J Allergy Clin 
Immunol. 2019;143(4):1620-1623. 

19. Kuo IH, Yoshida T, Benedetto AD, Beck 
LA. The cutaneous innate immune 
response in patients with atopic dermatitis. 
J Allergy Clin Immunol. 2013;131(2):266-
78. 

20. Krystel-Whittemore M, Dileepan KN, Wood 
JG. Mast Cell: A Multi-Functional Master 
Cell. Front Immunol. 2016;6(6):620. 

21. Keir HR, Chalmers JD. Neutrophil 
extracellular traps in chronic lung disease: 
Implications for pathogenesis and therapy. 
Eur Resp Rev. 2022;31(163):1-19. 

22. Croxatto D, Micheletti A, Montaldo E, 
Orecchia P, Loiacono F, Canegallo, F, et 
al. Group 3 innate lymphoid cells regulate 
neutrophil migration and function in human 
decidua. Mucosal Immunol. 2016:9(6): 
1372-1383. 

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

24. Lacey N, Delaney S, Kavanagh K, Powell 
FC. Mite-related bacterial antigens 
stimulate inflammatory cells in rosacea. Br 
J Dermatol. 2007;157(3):474-81. 

25. Roihu T, Kariniemi AL. Demodex mites in 
acne rosacea. J Cutan Pathol. 1998; 
25(10):550-2. 

26. Fink A, Heller L, Eliraz A, Weisman Z. 
Miskin A. Schlezinger, et al. Allergen-
specific leukocyte adherence inhibition 
(LAI) assay: Sensitivity, specificity and 
mechanism. Immunol Lett. 1987;16(1):65-
70. 

27. 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. 

28. Bradford MM. A rapid and sensitive 
method for the quantitation of microgram 
quantities of protein utilizing the principle of 
protein-dye binding. Anal Biochem. 
1976:72:248-54. 

29. 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. 

30. 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 

31. 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. 

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

33. 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. 

34. 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. 

35. 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. 

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



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 90-99, 2024; Article no.AJI.117841 
 
 

 
98 

 

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. 

37. 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. 

38. 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. 

39. 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. 

40. Olivier CE, Pinto DG, Teixeira APM, 
Santana JLS, Santos RAPGS, Lima RPS. 
Intrinsic atopic dermatitis: titration of 
precipitins in the screening of food 
allergens for prescription of elimination 
diets and desensitization strategies. Eur J 
Clin Med. 2021;2(6):1-9. 

41. Keystone E, Pruzanski W. 
Immunochemical and physical studies of 
the Sia test. Am J Med Sci. 1976;271(2): 
151-7. 

42. Williams CA, Chase MW. Chapter 13 - 
Precipitation reactions. In: Reactions of 
antibodies with soluble antigens. Academic 
Press: 1971;3:1-102. 

43. Marks GB, Tovey ER, Green W, Shearer 
M, Salome CM, Woolcock AJ. House dust 
mite allergen avoidance: A randomized 
controlled trial of surface chemical 
treatment and encasement of bedding. Clin 
Exp Allergy. 1994;24(11):1078-83. 

44. 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. 

45. Glass EV, Needham GR. Eliminating 
Dermatophagoides farinae spp. (Acari: 
Pyroglyphidae) and their allergens through 
high temperature treatment of textiles. J 
Med Entomol. 2004;41(3):529-532. 

46. Abidin SZ, Ming HT. Effect of a commercial 
air ionizer on dust mites 
Dermatophagoides pteronyssinus and 
Dermatophagoides farinae (Acari: 
Pyroglyphidae) in the laboratory. Asian 
Pacific J Trop Biomed. 2012:2(2):156-8. 

47. Tian M, Wang Y, Lu Y, Jiang YH, Zhao 
DY. Effects of sublingual immunotherapy 
for Dermatophagoides farinae on Th17 
cells and CD4(+) CD25(+) regulatory T 
cells in peripheral blood of children with 
allergic asthma. Int Forum Allergy Rhinol. 
2014:4(5):371-5. 

48. Walker IC. Study XII: Complement Fixation 
and Precipitin Reactions with the serum of 
Bronchial Asthmatics who are sensitive to 
the proteins of wheat, horse dandruff, cat 
hair, and bacteria, using these proteins as 
antigens, and the cutaneous reaction as an 
index of sensitization. J Med Res. 1917; 
36(2):243-66. 

49. 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. 

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

51. 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(2): 
597-603. 

52. 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. 

53. Gell PGH, Harington CR, Rivers RP. The 
antigenic function of simple chemical 
compounds; production of precipitins in 
rabbits. British Journal of Experimental 
Pathology. 1946, 27 (5), 267-286. 

54. Wells HG. Studies on the chemistry of 
anaphylaxis (III). Experiments with isolated 
proteins, especially those of the hen's egg. 
J Infec Dis. 1911;9:147-171. 

55. Adly MN, Mandour MF, Abd Elaziz MF, 
Saafan ME. Efficacy of allergen specific 
immunotherapy for treatment of allergic 
rhinitis: A systematic review and meta-
analysis. Journal of Advances in Medicine 



 
 
 
 

Olivier et al.; Asian J. Immunol., vol. 7, no. 1, pp. 90-99, 2024; Article no.AJI.117841 
 
 

 
99 

 

and Medical Research. 2021;33(22):200–
213.  
Available:https://doi.org/10.9734/jammr/20
21/v33i2231172  

56. JW G, NA. Plasma Kinins- A 
pharmacological perspective. Journal of 
Pharmaceutical Research International. 
2022;34(57):1–15.  
DOI: 10.9734/jpri/2022/v34i577254 

57. Romagnani S. Cytokines and 
chemoattractants in allergic inflammation. 
Molecular Immunology. 2002 May 1;38(12-
13):881-5. 

58. Anouar S. Hazim R. Brahim A. 
Interferences in Immunological Assays: 
Causes, Detection, and Prevention. Asian 
J Immunol. 2024;7(1):71-78. 

59. Chiarentin L, Gonçalves C, Augusto C, 
Miranda M, Cardoso C, Vitorino C. Drilling 
into "Quality by Design" Approach for 
Analytical Methods. Crit Rev Anal Chem. 
2023;4:1-42.  

60. 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/117841 

https://doi.org/10.9734/jammr/2021/v33i2231172
https://doi.org/10.9734/jammr/2021/v33i2231172
https://www.sdiarticle5.com/review-history/117841

