1例NEMO-ID患儿临床与免疫学特征研究
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摘要
目的探讨1例无外胚层发育不良、反复感染伴淋巴结肿大患儿临床特征、IKBKG(NEMO)基因突变、NEMO蛋白表达及免疫学特征。方法对1例临床表现为反复发热、支气管肺炎及颈淋巴结肿大患儿行免疫学初筛、免疫相关基因高通量测序、蛋白印迹法检测NEMO蛋白表达及淋巴细胞增殖功能检测。结果基因分析示IKBKG基因第9号外显子拼接位点发生g.21819 G>A杂合突变,其母为该突变携带者。患儿NEMO蛋白表达量明显降低,B细胞增殖功能明显受损。结论在国内首次确诊1例IKBKG基因突变致NEMO-ID患儿,不具备外胚层发育不良的反复感染患儿应考虑本病可能。假基因可能干扰高通量测序结果,仔细分析序列原始数据对最终获得确诊结果很有帮助。
To investigate the clinical characteristics, gene mutation and immunological features of a 7 years old male patient who suffered from recurrent infection, lymph node enlargement without ectodermal dysplasia, the immunologic screening, high-throughput sequencing of immune-related genes, NEMO protein detection and lymphocyte proliferation analysis were conducted in a patient with recurrent fever, bronchopneumonia and cervical lymphadenopathy. Data showed the patient carried a heterozygous G>A substitution located in exon 9 splicing site(21819) resulting in false mRNA splicing, which led to significantly reduce of NEMO protein expression dramatically impair of B cell proliferation as compared with parents. Mother of the patient was identified as heterozygous carriers of the same mutation. In conclusion, a NEMO-ID patient with a splicing mutation of IKBKG gene has been firstly identified in China. For patients who have recurrent infections, even without the typical clinical symptoms such as ectodermal dysplasia, Nemo-ID should be considered and assessment for immune function,genetic sequencing should be done. Rigorous analysis of the sequence's raw data can be helpful for the final diagnosis due to pseudogenes may interfere with high-throughput sequencing.
To investigate the clinical characteristics, gene mutation and immunological features of a 7 years old male patient who suffered from recurrent infection, lymph node enlargement without ectodermal dysplasia, the immunologic screening, high-throughput sequencing of immune-related genes, NEMO protein detection and lymphocyte proliferation analysis were conducted in a patient with recurrent fever, bronchopneumonia and cervical lymphadenopathy. Data showed the patient carried a heterozygous G>A substitution located in exon 9 splicing site(21819) resulting in false mRNA splicing, which led to significantly reduce of NEMO protein expression dramatically impair of B cell proliferation as compared with parents. Mother of the patient was identified as heterozygous carriers of the same mutation. In conclusion, a NEMO-ID patient with a splicing mutation of IKBKG gene has been firstly identified in China. For patients who have recurrent infections, even without the typical clinical symptoms such as ectodermal dysplasia, Nemo-ID should be considered and assessment for immune function,genetic sequencing should be done. Rigorous analysis of the sequence's raw data can be helpful for the final diagnosis due to pseudogenes may interfere with high-throughput sequencing.
引文
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[4]陈乡,王慧君,周文浩.全外显子测序技术检测在原发性免疫缺陷病诊断中的应用[J].国际药学研究杂志,2017,44(2):194-203.
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[7]Frans G,Jutte TB,Moens L,et al.Functional evaluation of an IKBKG variant suspected to cause immunodeficiency without ectodermal dysplasia[J].J Clin Immunol,2017,37(8):801-810.
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[11]Niehues T,Reichenbach J,Neubert J,et al.Nuclear factorκB essential modulator-deficient child with immunodeficiency yet without anhidrotic ectodermal dysplasia[J].J Allergy Clin Immunol,2004,114(6):1456-1462.
[12]Haverkamp MH,Marciano BE,Frucht DM,et al.Correlating interleukin-12 stimulated interferon-γproduction and the absence of ectodermal dysplasia and anhidrosis(EDA)in patients with mutations in NF-κBessential modulator(NEMO)[J].J Clin Immunol,2014,34(4):436-443.
[13]Filipesantos O,Bustamante J,Haverkamp MH,et al.X-linked susceptibility to mycobacteria is caused by mutations in NEMO impairing CD40-dependent IL-12production[J].J Exp Med,2006,203(7):1745-1759.
[14]Ding Y,Zhou L,Xia Y,et al.Reference values for peripheral blood lymphocyte subsets of healthy children in China[J].J Allergy Clin Immunol,2018,142:970-973.
[15]Hsu AP,Zerbe CS,Foruraghi L,et al.IKBKG(NEMO)5’untranslated splice mutations lead to severe,chronic disseminated mycobacterial infections[J].Clin Infect Dis,2018,67:456-459.
[16]D?ffinger R,Smahi A,Bessia C,et al.X-linked anhidrotic ectodermal dysplasia with immunodeficiency is caused by impaired NF-kappaB signaling[J].Nat Genetics,2001,27(3):277-285.
[17]Vollmer J,Krieg AM.Immunotherapeutic applications of CpG oligodeoxynucleotide TLR9 agonists[J].Adv Drug Deliv Rev,2009,61(3):195-204.
[18]Kumagai Y,Takeuchi O,Akira S.TLR9 as a key receptor for the recognition of DNA[J].Adv Drug Deliv Rev,2008,60(7):795-804.
[2]Maubach G,Naumann M.NEMO links nuclear factor-κBto human diseases[J].Trends Mol Med,2017,23(12):1138.
[3]Hanson EP,Monaco Shawver L,Solt LA,et al.Hypomorphic NEMO mutation database and reconstitution system identifies phenotypic and immunologic diversity[J].J Allergy Clin Immunol,2008,122(6):1169.
[4]陈乡,王慧君,周文浩.全外显子测序技术检测在原发性免疫缺陷病诊断中的应用[J].国际药学研究杂志,2017,44(2):194-203.
[5]Fusco F,Paciolla M,Napolitano F,et al.Genomic architecture at the Incontinentia Pigmenti locus favours de novo pathological alleles through different mechanisms[J].Hum Mol Genet,2011,21(6):1260-1271.
[6]周丽娜,丁媛,李黎,等.全血淋巴细胞流式细胞术精细免疫分型方法的建立[J].免疫学杂志,2015,31(6):523-527.
[7]Frans G,Jutte TB,Moens L,et al.Functional evaluation of an IKBKG variant suspected to cause immunodeficiency without ectodermal dysplasia[J].J Clin Immunol,2017,37(8):801-810.
[8]Ku CL,Dupuisgirod S,Dittrich AM,et al.NEMO mutations in 2 unrelated boys with severe infections and conical teeth[J].Pediatrics,2005,115(5):e615.
[9]Keller MD,Petersen M,Ong P,et al.Hypohidrotic ectodermal dysplasia and immunodeficiency with coincident NEMO and EDA mutations[J].Front Immunol,2011,2:61.
[10]Orange JS,Levy O,Brodeur SR,et al.Human nuclear factorκB essential modulator mutation can result in immunodeficiency without ectodermal dysplasia[J].JAllergy Clin Immunol,2004,114(3):650-656.
[11]Niehues T,Reichenbach J,Neubert J,et al.Nuclear factorκB essential modulator-deficient child with immunodeficiency yet without anhidrotic ectodermal dysplasia[J].J Allergy Clin Immunol,2004,114(6):1456-1462.
[12]Haverkamp MH,Marciano BE,Frucht DM,et al.Correlating interleukin-12 stimulated interferon-γproduction and the absence of ectodermal dysplasia and anhidrosis(EDA)in patients with mutations in NF-κBessential modulator(NEMO)[J].J Clin Immunol,2014,34(4):436-443.
[13]Filipesantos O,Bustamante J,Haverkamp MH,et al.X-linked susceptibility to mycobacteria is caused by mutations in NEMO impairing CD40-dependent IL-12production[J].J Exp Med,2006,203(7):1745-1759.
[14]Ding Y,Zhou L,Xia Y,et al.Reference values for peripheral blood lymphocyte subsets of healthy children in China[J].J Allergy Clin Immunol,2018,142:970-973.
[15]Hsu AP,Zerbe CS,Foruraghi L,et al.IKBKG(NEMO)5’untranslated splice mutations lead to severe,chronic disseminated mycobacterial infections[J].Clin Infect Dis,2018,67:456-459.
[16]D?ffinger R,Smahi A,Bessia C,et al.X-linked anhidrotic ectodermal dysplasia with immunodeficiency is caused by impaired NF-kappaB signaling[J].Nat Genetics,2001,27(3):277-285.
[17]Vollmer J,Krieg AM.Immunotherapeutic applications of CpG oligodeoxynucleotide TLR9 agonists[J].Adv Drug Deliv Rev,2009,61(3):195-204.
[18]Kumagai Y,Takeuchi O,Akira S.TLR9 as a key receptor for the recognition of DNA[J].Adv Drug Deliv Rev,2008,60(7):795-804.