Development of an additional method for X-linked lymphoproliferative syndrome type 2 diagnosis based on the assessment of muramyldipeptide-stimulated expression of tumor necrosis factor by monocytes

• Elvira Ya. Musaeva
• Elena A. Kulakovskaya
• Viktoria A. Vedmedskaya
• Julia A. Rodina
• Dmitry Е. Pershin
• Anna Yu. Shcherbina
• Michail M. Maschan

Abstract

X-linked lymphoproliferative syndrome type 2 (XLP2) is a rare primary immunodeficiency, characterized by high incidence of severe complications such as hemophagocytic lymphohistiocytosis and inflammatory bowel disease. The disease is caused by pathogenic variants in the XIAP (X-linked inhibitor of apoptosis) gene encoding XIAP protein that is involved in NOD2 signaling. Development of new methods for an early diagnosis of XLP2 is a matter of current interest.

The aim of the study was to evaluate tumor necrosis factor (TNF) expression in monocytes of XLP2 patients in response to stimulation by NOD2 ligand muramyldipeptide (MDP).

Material and methods. The study included 4 XLP2 patients, 3 mothers with hemizygous pathogenic variants in the XIAP gene, as well as 21 healthy donors over the age of 18. Fraction of mononuclear cells was isolated from peripheral blood samples, NOD2 receptor stimulation was carried out by incubation of cells in the presence of MDP. Lipopolysaccharide was used as a positive control, and a complete culture medium was used as a negative control. TNF expression in monocytes was evaluated by flow cytometry.

Results. A threshold value for evaluating the response to MDP stimulation was calculated. All samples of healthy donors showed significant increase in the proportion of TNF-producing monocytes in response to stimulation. Mothers with hemizygous pathogenic variants in the XIAP gene demonstrated response that was comparable to that of healthy donors. All patients included in the study, despite the presence of residual XIAP protein expression, showed no response to MDP stimulation. Additionally, the possibility of using cryopreserved mononuclear cells as a material for the assay was demonstrated on samples of 5 healthy donors and one patient.

Conclusion. Diagnostic method described in this study in combination with the assessment of XIAP protein expression allows for an early diagnosis of XLP2.

Keywords:X-linked lymphoproliferative syndrome type 2; inflammatory bowel disease; hemophagocytic lymphohistiocytosis; flow cytometry; NOD2; muramyl dipeptide

References

1. Mudde A.C.A., Booth C., Marsh R.A. Evolution of Our Understanding of XIAP Deficiency. Front Pediatr. 2021; 9: 660520. DOI: https://doi.org/10.3389/fped.2021.660520

2. Speckmann C., Lehmberg K., Albert M.H., Damgaard R.B., Fritsch M., Gyrd-Hansen M., Rensing-Ehl A., Vraetz T., Grimbacher B., Salzer U., Fuchs I., Ufheil H., Belohradsky B.H., Hassan A., Cale C.M., Elawad M., Strahm B., Schibli S., Lauten M., Kohl M., Meerpohl J.J., Rodeck B., Kolb R., Eberl W., Soerensen J., von Bernuth H., Lorenz M., Schwarz K., Zur Stadt U., Ehl S. X-linked inhibitor of apoptosis (XIAP) deficiency: the spectrum of presenting manifestations beyond hemophagocytic lymphohistiocytosis. Clin Immunol. 2013; 149 (1): 133–41. DOI: https://doi.org/10.1016/j.clim.2013.07.004

3. Basiaga M.L., Weiss P.F., Behrens E.M. BIRC4 Mutation: An Important Rare Cause of Uveitis. J Clin Rheumatol. 2015; 21 (8): 444–7. DOI: https://doi.org/10.1097/RHU.0000000000000327

4. Russian register of patients with primary immunodeficiency states analytical report. URL: https://naepid.ru/registr-pid/statistical-reports/ (date of access 14.12.2023)

5. Yang L., Booth C., Speckmann C., Seidel M.G., Worth A.J.J., Kindle G., Lankester A.C., Grimbacher B., ESID Clinical and Registry Working Parties, Gennery A.R., Seppanen M.R.J., Morris E.C., Burns S.O. Phenotype, genotype, treatment, and survival outcomes in patients with X-linked inhibitor of apoptosis deficiency. J Allergy Clin Immunol. 2022; 150 (2): 456–66. DOI: https://doi.org/10.1016/j.jaci.2021.10.037

6. Laberko A.L., Starichkova Yu.V., Khismatullina R.D., Persiantseva M.I., Maschan M.A., Balashov D.N., Rumyantsev A.G. Optimization of hematopoietic stem cell transplantation planning using medical information system in children with primary immunodeficiencies. Immunologiya. 2021; 42 (1): 49–59. DOI: https://doi.org/10.33029/0206-4952-2021-42-1-49-59 (in Russian)

7. Tsuma Y., Imamura T., Ichise E., Sakamoto K., Ouchi K., Osone S., Ishida H., Wada T., Hosoi H. Successful treatment of idiopathic colitis related to XIAP deficiency with allo-HSCT using reduced-intensity conditioning. Pediatr Transplant. 2015; 19 (1): E25–8. DOI: https://doi.org/10.1111/petr.12405

8. Chinn I.K., Orange J.S. A 2020 update on the use of genetic testing for patients with primary immunodeficiency. Expert Rev Clin Immunol. 2020; 16 (9): 897–909. DOI: https://doi.org/10.1080/1744666X.2020.1814145

9. Pershin D.Е., Vedmedskaya V.А., Fadeeva M.S., Vladimirov I.S., Kulakovskaya E.A., Roppelt A.A., Kieva A.M., Raykina E.V., Rodina Yu.A., Maschan M.A., Shcherbina A.Yu. Verification of X-linked lymphoproliferative syndrome type 1 and 2 using a flow cytometry method. Pediatric Hematology/Oncology and Immunopathology. 2020; 19 (4): 108–18. DOI: https://doi.org/10.24287/1726-1708-2020-19-4-108-118 (in Russian)

10. Damgaard R.B., Nachbur U., Yabal M., Wong W.W., Fiil B.K., Kastirr M., Rieser E., Rickard J.A., Bankovacki A., Peschel C., Ruland J., Bekker-Jensen S., Mailand N., Kaufmann T., Strasser A., Walczak H., Silke J., Jost P.J., Gyrd-Hansen M. The ubiquitin ligase XIAP recruits LUBAC for NOD2 signaling in inflammation and innate immunity. Mol Cell. 2012; 46 (6): 746–58. DOI: https://doi.org/10.1016/j.molcel.2012.04.014

11. Negroni A., Pierdomenico M., Cucchiara S., Stronati L. NOD2 and inflammation: current insights. J Inflamm Res. 2018; 11: 49–60. DOI: https://doi.org/10.2147/JIR.S137606

12. Lebedeva E.S., Bagaev A.V., Garaeva A.Y., Chulkina M.M., Pichugin A.V., Ataullakhanov R.I. The cooperative interaction of TLR4-, TLR9- and NOD2-signaling pathways in mouse macrophages. Immunologiya. 2018; 39 (1): 4–11. DOI: http://dx.doi.org/10.18821/0206-4952-2018-39-1-4-11 (in Russian)

13. Dagil Yu.A., Pashenkov M.V. A comparison of three types of cell-based test systems for the assessment of biological activity of NOD2 receptor agonists. Immunologiya. 2020; 41 (4): 304–11. DOI: https://doi.org/10.33029/0206-4952-2020-41-4-304-311 (in Russian)

14. Topal Y., Gyrd-Hansen M. RIPK2 NODs to XIAP and IBD. Semin Cell Dev Biol. 2021; 109: 144–50. DOI: https://doi.org/10.1016/j.semcdb.2020.07.001

15. Nielsen O.H., LaCasse E.C. How genetic testing can lead to targeted management of XIAP deficiency-related inflammatory bowel disease. Genet Med. 2017; 19 (2): 133–43. DOI: https://doi.org/10.1038/gim.2016.82

16. Quaranta M., Wilson R., Gonçalves Serra E., Pandey S., Schwerd T., Gilmour K., Klenerman P., Powrie F., Keshav S., Travis S.P.L., Anderson C.A., Uhlig H.H. Consequences of Identifying XIAP Deficiency in an Adult Patient With Inflammatory Bowel Disease. Gastroenterology. 2018; 155 (1): 231–4. DOI: https://doi.org/10.1053/j.gastro.2018.03.069

17. Roppelt A.A., Yukhacheva D.V., Myakova N.V., Smirnova N.V., Skvortsova Yu.V., Varlamova T.V., Raikina E.V., Abramov D.S., Ulanova N.B., Gabrusskaya T.V., Shcherbina A.Yu. X-Linked lymphoproliferative syndrome types 1 and 2. Vopr. gematol./onkol. immunopatol. pediatr. (Pediatric Haematology/Oncology and Immunopathology). 2016; 15 (1): 17–26. DOI: https://doi.org/10.20953/1726-1708-2016-1-17-26 (in Russian)

18. Pirozhkov S.V., Litvitskiy P.F. Inflammasomal diseases. Immunologiya. 2018; 39 (2-3): 158–65. DOI: http://dx.doi.org/10.18821/0206-4952-2018-39-2-3-158-165 (in Russian)

19. Ammann S., Elling R., Gyrd-Hansen M., Dückers G., Bredius R., Burns S.O., Edgar J.D., Worth A., Brandau H., Warnatz K., Zur Stadt U., Hasselblatt P., Schwarz K., Ehl S., Speckmann C. A new functional assay for the diagnosis of X-linked inhibitor of apoptosis (XIAP) deficiency. Clin Exp Immunol. 2014; 176 (3): 394–400. DOI: https://doi.org/10.1111/cei.12306

20. Tang J., Zhou X., Wang L., Hu G., Zheng B., Wang C., Lu Y., Jin Y., Guo H., Liu Z. Eosinophilic colitis in a boy with a novel XIAP mutation: a case report. BMC Pediatr. 2020; 20 (1): 171. DOI: https://doi.org/10.1186/s12887-020-02075-z

21. Chang I., Park S., Lee H.J., Kim I., Park S., Ahn M.K., Lee J., Kang M., Baek I.J., Sung Y.H., Pack C.G., Kang H.J., Lee K., Im H.J., Seo E.J., Kim K.M., Yang S.K., Song K., Oh S.H. Interpretation of XIAP Variants of Uncertain Significance in Paediatric Patients with Refractory Crohn’s Disease. J Crohns Colitis. 2021; 15 (8): 1291–304. DOI: https://doi.org/10.1093/ecco-jcc/jjab013

22. Ammann S., Fuchs S., Martin-Martin L., Castro C.N., Spielberger B., Klemann C., Elling R., Heeg M., Speckmann C., Hainmann I., Kaiser-Labusch P., Horneff G., Thalhammer J., Bredius R.G., Stadt U.Z., Lehmberg K., Fuchs I., von Spee-Mayer C., Henneke P., Ehl S. Functional flow cytometry of monocytes for routine diagnosis of innate primary immunodeficiencies. J Allergy Clin Immunol. 2020; 145 (1): 434–7.e4. DOI: https://doi.org/10.1016/j.jaci.2019.09.002

23. Schwerd T., Pandey S., Yang H.T., Bagola K., Jameson E., Jung J., Lachmann R.H., Shah N., Patel S.Y., Booth C., Runz H., Düker G., Bettels R., Rohrbach M., Kugathasan S., Chapel H., Keshav S., Elkadri A., Platt N., Muise A.M., Koletzko S., Xavier R.J., Marquardt T., Powrie F., Wraith J.E., Gyrd-Hansen M., Platt F.M., Uhlig H.H. Impaired antibacterial autophagy links granulomatous intestinal inflammation in Niemann-Pick disease type C1 and XIAP deficiency with NOD2 variants in Crohn’s disease. Gut. 2017; 66 (6): 1060–73. DOI: https://doi.org/10.1136/gutjnl-2015-310382

All articles in our journal are distributed under the Creative Commons Attribution 4.0 International License (CC BY 4.0 license)