Asian Journal of Immunology

Contribution of the Leukocyte Adherence Inhibition Test to the Diagnosis of Non–IgE-mediated Immunoreactivity against Staphylococcus aureus in Patients with Intrinsic Atopic Dermatitis

Celso Eduardo Olivier *

Instituto Alergoimuno de Americana, Brazil.

Daiana Guedes Pinto

Instituto Alergoimuno de Americana, Brazil.

Ana Paula Monezzi Teixeira

Instituto Alergoimuno de Americana, Brazil.

Jhéssica Letícia Santos Santana

Instituto Alergoimuno de Americana, Brazil.

Raquel Acácia Pereira Gonçalves Santos

Instituto Alergoimuno de Americana, Brazil.

Regiane Patussi Santos Lima

Lavoisier’s Laboratories, São Paulo, Brazil.

*Author to whom correspondence should be addressed.

---

Abstract

Aims: The study aims to evaluate the potential of the Leukocyte Adherence Inhibition Test (LAIT) to discriminate non–IgE-mediated immunoreactivity against Staphylococcus aureus in patients with Intrinsic Atopic Dermatitis (IAD).

Study Design: We retrospectively examined the medical charts of 200 patients diagnosed with IAD who were investigated with an ex vivo challenge monitored by LAIT against an extract of S. aureus.

Place and Duration of Study: The study was conducted at Instituto Alergoimuno de Americana – São Paulo – Brazil – between January 2018 and October 2023.

Methodology: The percentage of Leukocyte Adherence Inhibition (LAI) promoted by the ex vivo challenges with S. aureus extract was distributed in ranges through a cascade distribution chart to outline the variability of the results.

Results: The LAI mean was 39.3%; SD 27.6%; ranging from 0% to 100%; mode = 0% (appeared 33 times). There was a wide range of distribution of LAI results, suggesting that some patients had immunoreactivity against the S. aureus allergens while others did not.

Conclusion: Our preliminary results support that the LAIT performed with S. aureus may differentiate diverse degrees of ex vivo immunoreactivity against their allergens in IAD patients.

Keywords: Allergy, Staphylococcus aureus, atopic dermatitis, diagnosis, hypersensitivity, leukocyte adherence inhibition test, non–IgE-mediated immunoreactivity

---

---

---

References

Mesjasz A, Zawadzka M, Chałubiński M, Trzeciak M. Is Atopic Dermatitis Only a Skin Disease? Int J Mol Sci. 2023;24(1):837.

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.

Roguedas-Contios AM, Misery L. What is Intrinsic Atopic Dermatitis? Clin Rev Allergy Immunol. 41(3):233.

Ashman RB, Papadimitriou JM, Ott AK, Warmington JR. Antigens and immune responses in Candida albicans infection. Immunol Cell Biol. 1990;68(1):1-13.

Shi B, Bangayan NJ, Curd E, Taylor PA, Gallo RL, Leung DYM, et al. The skin microbiome is different in pediatric versus adult atopic dermatitis. J Allergy Clin Immunol. 2016;138(4):1233-1236.

Otto M. Staphylococcus colonization of the skin and antimicrobial peptides. Exp Rev Dermatol. 2010;5(2):183-195.

Family Staphylococcaceae. List of Prokaryotic names with Standing in Nomenclature (LPSN). Retrieved October 22, 2023;

Available:https://lpsn.dsmz.de/family/staphylococcaceae.

Coico R. Gram Staining. Current Protocols in Microbiology, 00: A.3C.1-A.3C.2. Retrieved on October 22, 2023. Available:https://doi.org/10.1002/9780471729259.mca03cs00.

Lowy FD. Staphylococcus aureus Infections. N Eng J Med. 1998;339(8):520-532.

Frank DN, Feazel LM, Bessesen MT, Price CS, Janoff EN, Pace NR. The Human Nasal Microbiota and Staphylococcus aureus Carriage. PLoS ONE. 2010;5(5):e10598.

Li Z, Peres A, Damian A, Madrenas J. Immunomodulation and Disease Tolerance to Staphylococcus aureus. Pathogens. 2015;4(4):793-815.

Hanakawa Y, Schechter NM, Lin C, Garza L, Li H, Yamaguchi T, et al. Molecular mechanisms of blister formation in bullous impetigo and staphylococcal scalded skin syndrome. J. Clin. Invest. 2002;110(1):53-60.

Joubert O. Keller D. Pinck A, Monteil H, Prevost G. Sensitive and Specific Detection of Staphylococcal Epidermolysins A and B in Broth Cultures by Flow Cytometry-Assisted Multiplex Immunoassay. J Clin Microbiol. 2005:43(3):1076-1080.

Brezski RJ, Jordan RE. Cleavage of IgGs by proteases associated with invasive diseases: an evasion tactic against host immunity? MAbs. 2010;2(3):212-20.

Liu GY. Molecular pathogenesis of Staphylococcus aureus infection. Ped Res. 2009;65(5):71R-77R.

Von Eiff C, Becker K, Machka K, Stammer H, Peters G. Nasal carriage as a source of Staphylococcus aureus bacteremia. N Eng J Med. 2001;344(1):11-16.

Peschel A. How do bacteria resist human antimicrobial peptides? Trends Microbiol. 2002;10(4):179-186.

Lei J, Sun L, Huang S, Zhu C, Li P, He J, et al. The antimicrobial peptides and their potential clinical applications. Am J Transl Res. 2019;11(7):3919-3931.

Mahlapuu M, HÃ¥kansson J, Ringstad L, BjÃrn C. Antimicrobial Peptides: An Emerging Category of Therapeutic Agents. Front Cell Infect Microbiol. 2016;6(article 194).

Hancock RE, Chapple DS. Peptide antibiotics. Antimicrob Ag Chemother. 1999;43(6):1317-1323.

Young JDE, Peterson CGB, Venge P, Cohn ZA. Mechanism of membrane damage mediated by human eosinophil cationic protein. Nature. 1986;321(6070):613-616.

Thijs J, Krastev T, Weidinger S, Buckens CF, de Bruin-Weller M, Bruijnzeel-Koomen C, et al. Biomarkers for atopic dermatitis: a systematic review and meta-analysis. Curr Opin Allergy Clin Immunol. 2017:15(5):453-460.

Fournier B, Philpott DJ. Recognition of Staphylococcus aureus by the innate immune system. Clin Microbiol Rev. 2005;18(3):521-540.

Karauzum H, Datta SK. Adaptive Immunity Against Staphylococcus aureus. Curr Top Microbiol Immunol. 2017;409:419-439.

Brandt SL, Putnam NE, Cassat JE, Serezani CH. Innate Immunity to Staphylococcus aureus: Evolving Paradigms in Soft Tissue and Invasive Infections. J Immunol. 2018;200(12):3871-3880.

Richter M, Banerjee D, Sklar S. The antibody-independent cytotoxic activity of normal circulating human leucocytes. II. Failure to demonstrate effector cell-target cell interaction and target cell specificity of the circulating cytotoxic-enhancing factor. Immunology. 1981;44(1):109-118.

Gomes LC, Dikic I. Autophagy in Antimicrobial Immunity. Molecular Cell. 2014;54(2):224-233.

Maurer K, Reyes-Robles T, Alonzo F, Durbin J, Torres VJ, Cadwell, K. Autophagy Mediates Tolerance to Staphylococcus aureus Alpha-Toxin. Cell Host Microbe. 2015;17(4):429-440.

Pan Z, Dumas E, Lawrence C, Pate L, Longobardi S, Wang X, et al. Bacillus anthracis Edema Toxin Inhibits Efferocytosis in Human Macrophages and Alters Efferocytic Receptor Signaling. Int J Mol Sci. 2019;20(5):1167.

Fraser DA, Curiel L, Budin R, Pulanco MC. Complement protein C1q modulates lipoprotein metabolism in macrophage foam cells. J Immunol. 2017:198(Sup 1):208.10-208.10.

Ho MM, Manughian-Peter A, Spivia WR, Taylor A, Fraser DA. Macrophage molecular signaling and inflammatory responses during ingestion of atherogenic lipoproteins are modulated by complement protein C1q. Atherosclerosis. 2016;253:38-46.

Ali HA, El-Mahdy RH, Gaballah MA. Community-acquired methicillin-resistant Staphylococcus aureus colonization in atopic dermatitis patients in Mansoura, Egypt. Biomed Dermatol. 2019;3(1):2.

Kong HH, Oh J, Deming C, Conlan S, Grice EA, Beatson MA, et al. Temporal shifts in the skin microbiome associated with disease flares and treatment in children with atopic dermatitis. Gen Res. 2012;22(5):850-859.

Röcken M, Urban J, Shevach EM. Antigen-specific activation, tolerization, and reactivation of the interleukin 4 pathway in vivo. J Exp Med. 1994;179(6):1885-1893.

Mueller DL, Chiodetti L, Bacon PA, Schwartz RH. Clonal anergy blocks the response to IL-4, as well as the production of IL-2, in dual-producing T helper cell clones. J Immunol. 1991;147(12):4118-4125.

Wherry EJ. T cell exhaustion. Nat Immunol. 2011;12(6):492-499.

Hünefeld C, Mezger M, Röcken M, Röcken M. The Three Dimensions of Functional T-Cell Tolerance: From Research to Practice. J Invest Dermatol. 2012;132(3): 508-511.

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.

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.

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.

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.

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):234-241.

Powell AE, Sloss AM, Smith RN. Leukocyte-Adherence Inhibition: A Specific Assay of Cell-Mediated Immunity Dependent on Lymphokine-Mediated Collaboration between T Lymphocytes. J Immunol. 1978;120(6):1957-1966.

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.

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.

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. article ID 860427

Available:http://dx.doi.org/10.1155/2014/860427), 1-6.

Olivier CE, Pinto DG, Lima RPS, Silva MD, Santos RAPG, Teixeira APM, 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.

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

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.

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

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

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

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

Schlievert PM, Strandberg KL, Lin YC, Peterson ML, Leung DY. Secreted virulence factor comparison between methicillin-resistant and methicillin-sensitive Staphylococcus aureus, and its relevance to atopic dermatitis. J Allergy Clin Immunol. 2010;125(1):39-49.

Lin YT, Wang CT, Chiang BL. Role of bacterial pathogens in atopic dermatitis. Clin Rev Allergy Immunol. 2007;33(3):167-77.

Kim J, Kim BE, Ahn K, Leung DYM. Interactions Between Atopic Dermatitis and Staphylococcus aureus Infection: Clinical Implications. Allergy Asthma Immunol Res. 2019;11(5):593-603.

Brown AF, Leech JM, Rogers TR, McLoughlin RM. Staphylococcus aureus Colonization: Modulation of Host Immune Response and Impact on Human Vaccine Design. Front Immunol. 2014;4(article 507):1-20.

Geoghegan JA, Irvine AD, Foster TJ. Staphylococcus aureus and Atopic Dermatitis: A Complex and Evolving Relationship. Trends Microbiol. 2018;26(6):484-497.

Huang JT, Abrams M, Tlougan B, Rademaker A, Paller AS. Treatment of Staphylococcus aureus Colonization in Atopic Dermatitis Decreases Disease Severity. Pediatrics. 2009;123(5):e808-e814.

Halcón L, Milkus K. Staphylococcus aureus and wounds: A review of tea tree oil as a promising antimicrobial. Am J Infect Control. 2004;32(7):402-408.

Carson CF, Hammer KA, Riley TV. Melaleuca alternifolia (Tea Tree) Oil: a Review of Antimicrobial and Other Medicinal Properties. Clin Microbiol Rev. 2006;19(1):50-62.

Carson CF, Mee BJ, Riley TV. Mechanism of Action of Melaleuca alternifolia (Tea Tree) Oil on Staphylococcus aureus Determined by Time-Kill, Lysis, Leakage, and Salt Tolerance Assays and Electron Microscopy. Antimicrob Ag Chemother. 2002;46(6):1914-1920.

Bachert C, Humbert M, Hanania NA, Zhang N, Holgate S, Buhl R, et al. Staphylococcus aureus and its IgE-inducing enterotoxins in asthma: current knowledge. Eur Respir J. 2020;55(4):1901592.

Hauser C, Wuethrich B, Matter L, Wilhelm JA, Schopfer K. Immune Response to Staphylococcus aureus in Atopic Dermatitis. Dermatologica. 2009;170(3):114-120.

Leung DYM. Pathogenesis of atopic dermatitis. J Allergy Clin Immunol. 1999;104(3):S99-S108.

Rajan TV. The Gell-Coombs classification of hypersensitivity reactions: a re-interpretation. Trends Immunol. 2003;24(7):376-9.

Zheng C, Cao T, Ye C, Zou Y. Neutrophil recruitment by CD4 tissue-resident memory T cells induces chronic recurrent inflammation in atopic dermatitis. Clin Immunol. 2023;109805.

Jutel M, Agache I, Zemelka-Wiacek M, Akdis M, Chivato T, Del Giacco S, Gajdanowicz P, et al. Nomenclature of allergic diseases and hypersensitivity reactions: Adapted to modern needs: An EAACI position paper. Allergy; 2023.

Available:https://onlinelibrary.wiley.com/doi/epdf/10.1111/all.15889.

Shirakawa T, Kusaka Y, Fujimura N, Goto S, Morimoto K. The existence of specific antibodies to cobalt in hard metal asthma. Clin Exp Allergy. 1988;18(5):451-460.

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. Biomedic J Sci Tech Res. 2021;36(3):28647-28655.

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

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.

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

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.

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