Correlation Analysis of Mutation Severity and BTK-expression by Clinical Manifestations in Patients with X-linked Agammaglobulinemia

Document Type: Original Article

Authors

1 Research Center for Immunodeficiencies, Pediatrics Center of Excellence, Children's Medical Center, Tehran University of Medical Sciences, Tehran, Iran

2 Cellular and Molecular Research Center, Birjand University of Medical Sciences, Birjand, Iran.

Abstract

Backgrounds/Objectives: X-linked agammaglobulinemia (XLA) is a primary immunodeficiency disorder caused by mutations in the Bruton tyrosine kinase (BTK) gene. It is characterized by severely reduced numbers of peripheral B cells and a significant deficiency in all serum immunoglobulins. In the present study, the impact of mutation severity on the clinical and immunological characteristics of XLA patients was evaluated.
Methods: Mutation analysis was performed in 19 XLA patients by PCR assay to identify variations in theBTK gene. Subsequently, the western blotting technique was applied for measuring BTKexpression and function. A genotype-phenotype correlation was investigated regarding the impact of mutation severity on clinical and immunological parameters.
Results: Mutation detection in theBTK gene revealed missense mutations in 9 patients, nonsense mutations in 3 cases, splicing site defects in 5 patients, and small in-frame deletions in 2 patients; 31% of patients displayed normal BTK expression. A significant correlation was found between types of BTK mutation and BTK expression.
Discussion: Generally, genotype-phenotype correlation studies on XLA disease seem to be very controversial. The results of the correlation analysis in the present study could indicate that evolution of the disorder is not completely similar in all cases, even with the same mutation.

Keywords


1. Ochs HD, Smith CI. X-linked agammaglobulinemia. A clinical and molecular analysis. Medicine. 1996 Nov;75(6):287-99. 2. Yazdani R, Abolhassani H, Asgardoon M, Shaghaghi M, Modaresi M, Azizi G, et al. Infectious and Noninfectious Pulmonary Complications in Patients With Primary Immunodeficiency Disorders. J Investig Allergol Clin Immunol. 2017;27(4):213-24. 3. Conley ME, Howard V. Clinical findings leading to the diagnosis of X-linked agammaglobulinemia. The Journal of pediatrics. 2002 Oct;141(4):566-71. 4. Yu L, Mohamed AJ, Vargas L, Berglof A, Finn G, Lu KP, et al. Regulation of Bruton tyrosine kinase by the peptidylprolyl isomerase Pin1. J Biol Chem. 2006;281(26):18201-7. 5. Sideras P, Smith CI. Molecular and cellular aspects of X-linked agammaglobulinemia. Advances in immunology. 1995;59:135-223. 6. Kwan SP, Kunkel L, Bruns G, Wedgwood RJ, Latt S, Rosen FS. Mapping of the X-linked agammaglobulinemia locus by use of restriction fragment-length polymorphism. The Journal of clinical investigation. 1986 Feb;77(2):649-52. 7. Abolhassani H, Vitali M, Lougaris V, Giliani S, Parvaneh N, Parvaneh L, et al. Cohort of Iranian Patients with Congenital Agammaglobulinemia: Mutation Analysis and Novel Gene Defects. Expert review of clinical immunology. 2016;12(4):479-86. 8. Lopez-Granados E, Perez de Diego R, Ferreira Cerdan A, Fontan Casariego G, Garcia Rodriguez MC. A genotype-phenotype correlation study in a group of 54 patients with X-linked agammaglobulinemia. The Journal of allergy and clinical immunology. 2005 Sep;116(3):690-7. 9. Gaspar HB, Lester T, Levinsky RJ, Kinnon C. Bruton's tyrosine kinase expression and activity in X-linked agammaglobulinaemia (XLA): the use of protein analysis as a diagnostic indicator of XLA. Clinical and experimental immunology. 1998 Feb;111(2):334-8. 10. Futatani T, Miyawaki T, Tsukada S, Hashimoto S, Kunikata T, Arai S, et al. Deficient expression of Bruton's tyrosine kinase in monocytes from Xlinked agammaglobulinemia as evaluated by a flow cytometric analysis and its clinical application to carrier detection. Blood. 1998 Jan 15;91(2):595- 602. 11. Conley ME, Broides A, Hernandez-Trujillo V, Howard V, Kanegane H, Miyawaki T, et al. Genetic analysis of patients with defects in early B-cell development. Immunological reviews. 2005 Feb;203:216-34. 12. Gaspar HB, Bradley LA, Katz F, Lovering RC, Roifman CM, Morgan G, et al. Mutation analysis in Bruton's tyrosine kinase, the X-linked agammaglobulinaemia gene, including identification of an insertional hotspot. Human molecular genetics. 1995 Apr;4(4):755-7. 13. Jin H, Webster AD, Vihinen M, Sideras P, Vorechovsky I, Hammarstrom L, et al. Identification of Btk mutations in 20 unrelated patients with X-linked agammaglobulinaemia (XLA). Human molecular genetics. 1995 Apr;4(4):693-700. 14. Holinski-Feder E, Weiss M, Brandau O, Jedele KB, Nore B, Backesjo CM, et al. Mutation screening of the BTK gene in 56 families with Xlinked agammaglobulinemia (XLA): 47 unique mutations without correlation to clinical course. Pediatrics. 1998 Feb;101(2):276-84. 15. Kobayashi S, Iwata T, Saito M, Iwasaki R, Matsumoto H, Naritaka S, et al. Mutations of the Btk gene in 12 unrelated families with X-linked agammaglobulinemia in Japan. Human genetics. 1996 Apr;97(4):424-30. 16. Broides A, Yang W, Conley ME. Genotype/phenotype correlations in X-linked agammaglobulinemia. Clinical immunology (Orlando, Fla). 2006 Feb-Mar;118(2-3):195-200. 17. Aghamohammadi A, Fiorini M, Moin M, Parvaneh N, Teimourian S, Yeganeh M, et al. Clinical, immunological and molecular characteristics of 37 Iranian patients with X-linked agammaglobulinemia. International archives of allergy and immunology. 2006;141(4):408-14. 18. Lee PP, Chen TX, Jiang LP, Chan KW, Yang W, Lee BW, et al. Clinical characteristics and genotype-phenotype correlation in 62 patients with X-linked agammaglobulinemia. Journal of clinical immunology. 2010 Jan;30(1):121-31. 19. Vorechovsky I, Vihinen M, de Saint Basile G, Honsova S, Hammarstrom L, Muller S, et al. DNAbased mutation analysis of Bruton's tyrosine kinase gene in patients with X-linked agammaglobulinaemia. Human molecular genetics. 1995 Jan;4(1):51-8. 20. Teimourian S, Nasseri S, Pouladi N, Yeganeh M, Aghamohammadi A. Genotype-phenotype correlation in Bruton's tyrosine kinase deficiency. Journal of pediatric hematology/oncology. 2008 Sep;30(9):679-83. 21. Jefferies CA, Doyle S, Brunner C, Dunne A, Brint E, Wietek C, et al. Bruton's tyrosine kinase is a Toll/interleukin-1 receptor domain-binding protein that participates in nuclear factor kappaB activation by Toll-like receptor 4. J Biol Chem. 2003 Jul 11;278(28):26258-64. 22. Aghamohammadi A, Parvaneh N, Kanegana H, Moin M, Amirzargar AA, Farhoudi A, et al. Screening of the Bruton Tyrosine Kinase (BTK) Gene Mutations in 13 Iranian Patients with Presumed X-Linked Agammaglobulinemia. Iranian journal of allergy, asthma, and immunology. 2004 Dec;3(4):175-9. 23. Nasseri S, Sorouri R, Pourpak Z, Yeganeh M, Aghamohammadi A, Fiorini M, et al. Molecular characterization of Bruton's tyrosine kinase deficiency in 12 Iranian patients with presumed Xlinked agammaglobulinemia. Journal of investigational allergology & clinical immunology. 2011;21(7):572-4. 24. Kanegane H, Futatani T, Wang Y, Nomura K, Shinozaki K, Matsukura H, et al. Clinical and mutational characteristics of X-linked agammaglobulinemia and its carrier identified by flow cytometric assessment combined with genetic analysis. The Journal of allergy and clinical immunology. 2001 Dec;108(6):1012-20. 25. Rawlings DJ, Scharenberg AM, Park H, Wahl MI, Lin S, Kato RM, et al. Activation of BTK by a phosphorylation mechanism initiated by SRC family kinases. Science. 1996 Feb 9;271(5250):822- 5.