播散性暗色丝孢霉病患者CARD9突变及相关免疫学研究
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摘要
暗色丝孢霉病是指由暗色真菌引起的皮肤、皮下组织乃至深部组织脏器的感染。本研究探讨1例由皮肤及皮下组织暗色丝孢霉病逐渐进展为播散性暗色丝孢霉病患者的遗传背景及其抗真菌免疫功能。收集患者10余年病情进展的临床资料及真菌检查结果,进行临床资料及真菌学研究;对外周血DNA采用Sanger测序,进行遗传学研究;提取患者及正常人外周血单个核细胞(peripheral blood mononuclear cell,PBMC),采用蛋白免疫印迹法、流式细胞术、化学发光法、酶联免疫吸附试验等进行免疫学研究。结果显示,患者胱天蛋白酶募集域蛋白9(caspase recruitment domain-containing protein 9,CARD9)基因存在新的复合杂合错义突变(p.R35Q和p.E81K),CARD9蛋白表达正常,外周血淋巴细胞(peripheral blood lymphocyte,PBL)Th1和Th17细胞比例均降低;PBMC抗疣状瓶霉感染存在天然及适应性免疫缺陷。本研究首次报道1例播散性暗色丝孢霉病患者存在CARD9基因新发错义突变,同时发现患者PBMC抗真菌的天然及适应性免疫存在缺陷,提示CARD9蛋白在暗色丝孢霉病发病中可能发挥重要作用。
Phaeohyphomycosis is a collection of superficial cutaneous,subcutaneous and(or)systemic infections caused by diverse dematiaceous fungi.The clinical records and clinical samples collected from a patient with a diagnosis of skin and subcutaneous phaeohyphomycosis progressed to disseminated phaeohyphomycosis were subjected to assays for potential genetic and immune defects that might associate with the infection.Two new compound heterozygous missense mutations on caspase recruitment domaincontaining protein 9gene(CARD9)(p.R35 Qand p.E81K)were detected.Compared with the healthy donors,the proportions of Th1 and Th17cells in the patient's peripheral blood lymphocytes(PBLs)were low and the patient's peripheral blood mononuclear cells(PBMCs)had impaired innate and adaptive immune responses against Phialophora verrucosa(P.verrucosa).The results indicated that CARD9 protein may play an important role in the pathogenesis of phaeohyphomycosis.
Phaeohyphomycosis is a collection of superficial cutaneous,subcutaneous and(or)systemic infections caused by diverse dematiaceous fungi.The clinical records and clinical samples collected from a patient with a diagnosis of skin and subcutaneous phaeohyphomycosis progressed to disseminated phaeohyphomycosis were subjected to assays for potential genetic and immune defects that might associate with the infection.Two new compound heterozygous missense mutations on caspase recruitment domaincontaining protein 9gene(CARD9)(p.R35 Qand p.E81K)were detected.Compared with the healthy donors,the proportions of Th1 and Th17cells in the patient's peripheral blood lymphocytes(PBLs)were low and the patient's peripheral blood mononuclear cells(PBMCs)had impaired innate and adaptive immune responses against Phialophora verrucosa(P.verrucosa).The results indicated that CARD9 protein may play an important role in the pathogenesis of phaeohyphomycosis.
引文
[1]Chowdhary A,Perfect J,de Hoog GS.Black molds and melanized yeasts pathogenic to humans[J].Cold Spring Harb Perspect Med,2014,5(8):a019570.
[2]Revankar SG,Sutton DA.Melanized fungi in human disease[J].Clin Microbiol Rev,2010,23(4):884-928.
[3]Engelhardt KR,Grimbacher B.Mendelian traits causing susceptibility to mucocutaneous fungal infections in human subjects[J].J Allergy Clin Immunol,2012,129(2):294-305.
[4]Wang X,Wang W,Lin Z,Wang X,Li T,Yu J,Liu W,Tong Z,Xu Y,Zhang J,Guan L,Dai L,Yang Y,Han W,Li R.CARD9 mutations linked to subcutaneous phaeohyphomycosis and TH17 cell deficiencies[J].JAllergy Clin Immunol,2014,133(3):905-908.
[5]Drewniak A,Gazendam RP,Tool AT,van Houdt M,Jansen MH,van Hamme JL,van Leeuwen EM,Roos D,Scalais E,de Beaufort C,Janssen H,van den Berg TK,Kuijpers TW.Invasive fungal infection and impaired neutrophil killing in human CARD9deficiency[J].Blood,2013,121(13):2385-2392.
[6]Gao LJ,Yu J,Wang DL,Li RY.Recalcitrant primary subcutaneous phaeohyphomycosis due to Phialophora verrucosa[J].Mycopathologia,2013,175(1-2):165-170.
[7]Li Y,Wan Z,Li R.In vitro activities of nine antifungal drugs and their combinations against Phialophora verrucosa[J].Antimicrob Agents Chemother,2014,58(9):5609-5612.
[8]Keyser A,Schmid FX,Linde HJ,Merk J,Birnbaum DE.Disseminated Cladophialophora bantiana infection in a heart transplant recipient[J].J Heart Lung Transplant,2002,21(4):503-505.
[9]叶枫,吴璐璐,苏丹虹,曾庆思,陈荣昌.肺部暗色丝孢霉病1例并文献复习[J].中国感染与化疗杂志,2014,14(3):229-234.
[10]常杏芝,李建国,李若瑜,包新华,万哲,秦炯.中枢神经系统暗色丝孢霉病1例及文献复习[J].实用儿科临床杂志,2006,21(10):619-621.
[11]Tong Z,Chen SC,Chen L,Dong B,Li R,Hu Z,Jiang P,Li D,Duan Y.Generalized subcutaneous phaeohyphomycosis caused by Phialophora verrucosa:report of a case and review of literature[J].Mycopathologia,2013,175(3-4):301-306.
[12]Lanternier F,Pathan S,Vincent QB,Liu L,Cypowyj S,Prando C,Migaud M,Taibi L,Ammar-Khodja A,Boudghene Stambouli O,Guellil B,Jacobs F,Goffard JC,Schepers K,del Marmol V,Boussofara L,Denguezli M,Larif M,Bachelez H,Michel L,Lefranc G,Hay R,Jouvion G,Chretien F,Fraitag S,Bougnoux ME,Boudia M,Abel L,Lortholary O,Casanova JL,Picard C,Grimbacher B,Puel A.Deep dermatophytosis and inherited CARD9deficiency[J].N Engl J Med,2013,369(18):1704-1714.
[13]Lanternier F,Barbati E,Meinzer U,Liu L,Pedergnana V,Migaud M,Héritier S,Chomton M,Frémond ML,Gonzales E,Galeotti C,Romana S,Jacquemin E,Angoulvant A,Bidault V,Canioni D,Lachenaud J,Mansouri D,Mahdaviani SA,Adimi P,Mansouri N,Jamshidi M,Bougnoux ME,Abel L,Lortholary O,Blanche S,Casanova JL,Picard C,Puel A.Inherited CARD9deficiency in 2unrelated patients with invasive Exophiala infection[J].J Infect Dis,2015,211(8):1241-1250.
[14]Lanternier F,Mahdaviani SA,Barbati E,Chaussade H,Koumar Y,Levy R,Denis B,Brunel AS,Martin S,Loop M,Peeters J,de Selys A,Vanclaire J,Vermylen C,Nassogne MC,Chatzis O,Liu L,Migaud M,Pedergnana V,Desoubeaux G,Jouvion G,Chretien F,Darazam IA,Schffer AA,Netea MG,De Bruycker JJ,Bernard L,Reynes J,Amazrine N,Abel L,Van der Linden D,Harrison T,Picard C,Lortholary O,Mansouri D,Casanova JL,Puel A.Inherited CARD9 deficiency in otherwise healthy children and adults with Candida speciesinduced meningoencephalitis,colitis,or both[J].J Allergy Clin Immunol,2015,135(6):1558-1568.
[15]Grumach AS,de Queiroz-Telles F,Migaud M,Lanternier F,Filho NR,Palma SM,Constantino-Silva RN,Casanova JL,Puel A.A homozygous CARD9mutation in a Brazilian patient with deep dermatophytosis[J].J Clin Immunol,2015,35(5):486-490.
[16]Alves de Medeiros AK,Lodewick E,Bogaert DJ,Haerynck F,Van Daele S,Lambrecht B,Bosma S,Vanderdonckt L,Lortholary O,Migaud M,Casanova JL,Puel A,Lanternier F,Lambert J,Brochez L,Dullaers M.Chronic and invasive fungal infections in a family with CARD9deficiency[J].J Clin Immunol,2016,36(3):204-209.
[17]Hsu YM,Zhang Y,You Y,Wang D,Li H,Duramad O,Qin XF,Dong C,Lin X.The adaptor protein CARD9is required for innate immune responses to intracellular pathogens[J].Nat Immunol,2007,8(2):198-205.
[18]Gross O,Gewies A,Finger K,Schfer M,Sparwasser T,Peschel C,F9rster I,Ruland J.Card9controls a non-TLRsignalling pathway for innate anti-fungal immunity[J].Nature,2006,442(7103):651-656.
[19]Liang P,Wang X,Wang R,Wan Z,Han W,Li R.CARD9deficiencies linked to impaired neutrophil functions against Phialophora verrucosa[J].Mycopathologia,2015,179(5-6):347-357.
[20]Wu W,Zhang R,Wang X,Song Y,Liu Z,Han W,Li R.Impairment of immune response against dematiaceous fungi in Card9knockout mice[J].Mycopathologia,2016,181(9-10):631-642.
[21]Gavino C,Cotter A,Lichtenstein D,Lejtenyi D,Fortin C,Legault C,Alirezaie N,Majewski J,Sheppard DC,Behr MA,Foulkes WD,Vinh DC.CARD9 deficiency and spontaneous central nervous system candidiasis:complete clinical remission with GM-CSF therapy[J].Clin Infect Dis,2014,59(1):81-84.
[22]Celmeli F,Oztoprak N,Turkkahraman D,Seyman D,Mutlu E,Frede N,K9ksoy S,Grimbacher B.Successful granulocyte colony-stimulating factor treatment of relapsing Candida albicans meningoencephalitis caused by CARD9deficiency[J].Pediatr Infect Dis J,2016,35(4):428-431.
[2]Revankar SG,Sutton DA.Melanized fungi in human disease[J].Clin Microbiol Rev,2010,23(4):884-928.
[3]Engelhardt KR,Grimbacher B.Mendelian traits causing susceptibility to mucocutaneous fungal infections in human subjects[J].J Allergy Clin Immunol,2012,129(2):294-305.
[4]Wang X,Wang W,Lin Z,Wang X,Li T,Yu J,Liu W,Tong Z,Xu Y,Zhang J,Guan L,Dai L,Yang Y,Han W,Li R.CARD9 mutations linked to subcutaneous phaeohyphomycosis and TH17 cell deficiencies[J].JAllergy Clin Immunol,2014,133(3):905-908.
[5]Drewniak A,Gazendam RP,Tool AT,van Houdt M,Jansen MH,van Hamme JL,van Leeuwen EM,Roos D,Scalais E,de Beaufort C,Janssen H,van den Berg TK,Kuijpers TW.Invasive fungal infection and impaired neutrophil killing in human CARD9deficiency[J].Blood,2013,121(13):2385-2392.
[6]Gao LJ,Yu J,Wang DL,Li RY.Recalcitrant primary subcutaneous phaeohyphomycosis due to Phialophora verrucosa[J].Mycopathologia,2013,175(1-2):165-170.
[7]Li Y,Wan Z,Li R.In vitro activities of nine antifungal drugs and their combinations against Phialophora verrucosa[J].Antimicrob Agents Chemother,2014,58(9):5609-5612.
[8]Keyser A,Schmid FX,Linde HJ,Merk J,Birnbaum DE.Disseminated Cladophialophora bantiana infection in a heart transplant recipient[J].J Heart Lung Transplant,2002,21(4):503-505.
[9]叶枫,吴璐璐,苏丹虹,曾庆思,陈荣昌.肺部暗色丝孢霉病1例并文献复习[J].中国感染与化疗杂志,2014,14(3):229-234.
[10]常杏芝,李建国,李若瑜,包新华,万哲,秦炯.中枢神经系统暗色丝孢霉病1例及文献复习[J].实用儿科临床杂志,2006,21(10):619-621.
[11]Tong Z,Chen SC,Chen L,Dong B,Li R,Hu Z,Jiang P,Li D,Duan Y.Generalized subcutaneous phaeohyphomycosis caused by Phialophora verrucosa:report of a case and review of literature[J].Mycopathologia,2013,175(3-4):301-306.
[12]Lanternier F,Pathan S,Vincent QB,Liu L,Cypowyj S,Prando C,Migaud M,Taibi L,Ammar-Khodja A,Boudghene Stambouli O,Guellil B,Jacobs F,Goffard JC,Schepers K,del Marmol V,Boussofara L,Denguezli M,Larif M,Bachelez H,Michel L,Lefranc G,Hay R,Jouvion G,Chretien F,Fraitag S,Bougnoux ME,Boudia M,Abel L,Lortholary O,Casanova JL,Picard C,Grimbacher B,Puel A.Deep dermatophytosis and inherited CARD9deficiency[J].N Engl J Med,2013,369(18):1704-1714.
[13]Lanternier F,Barbati E,Meinzer U,Liu L,Pedergnana V,Migaud M,Héritier S,Chomton M,Frémond ML,Gonzales E,Galeotti C,Romana S,Jacquemin E,Angoulvant A,Bidault V,Canioni D,Lachenaud J,Mansouri D,Mahdaviani SA,Adimi P,Mansouri N,Jamshidi M,Bougnoux ME,Abel L,Lortholary O,Blanche S,Casanova JL,Picard C,Puel A.Inherited CARD9deficiency in 2unrelated patients with invasive Exophiala infection[J].J Infect Dis,2015,211(8):1241-1250.
[14]Lanternier F,Mahdaviani SA,Barbati E,Chaussade H,Koumar Y,Levy R,Denis B,Brunel AS,Martin S,Loop M,Peeters J,de Selys A,Vanclaire J,Vermylen C,Nassogne MC,Chatzis O,Liu L,Migaud M,Pedergnana V,Desoubeaux G,Jouvion G,Chretien F,Darazam IA,Schffer AA,Netea MG,De Bruycker JJ,Bernard L,Reynes J,Amazrine N,Abel L,Van der Linden D,Harrison T,Picard C,Lortholary O,Mansouri D,Casanova JL,Puel A.Inherited CARD9 deficiency in otherwise healthy children and adults with Candida speciesinduced meningoencephalitis,colitis,or both[J].J Allergy Clin Immunol,2015,135(6):1558-1568.
[15]Grumach AS,de Queiroz-Telles F,Migaud M,Lanternier F,Filho NR,Palma SM,Constantino-Silva RN,Casanova JL,Puel A.A homozygous CARD9mutation in a Brazilian patient with deep dermatophytosis[J].J Clin Immunol,2015,35(5):486-490.
[16]Alves de Medeiros AK,Lodewick E,Bogaert DJ,Haerynck F,Van Daele S,Lambrecht B,Bosma S,Vanderdonckt L,Lortholary O,Migaud M,Casanova JL,Puel A,Lanternier F,Lambert J,Brochez L,Dullaers M.Chronic and invasive fungal infections in a family with CARD9deficiency[J].J Clin Immunol,2016,36(3):204-209.
[17]Hsu YM,Zhang Y,You Y,Wang D,Li H,Duramad O,Qin XF,Dong C,Lin X.The adaptor protein CARD9is required for innate immune responses to intracellular pathogens[J].Nat Immunol,2007,8(2):198-205.
[18]Gross O,Gewies A,Finger K,Schfer M,Sparwasser T,Peschel C,F9rster I,Ruland J.Card9controls a non-TLRsignalling pathway for innate anti-fungal immunity[J].Nature,2006,442(7103):651-656.
[19]Liang P,Wang X,Wang R,Wan Z,Han W,Li R.CARD9deficiencies linked to impaired neutrophil functions against Phialophora verrucosa[J].Mycopathologia,2015,179(5-6):347-357.
[20]Wu W,Zhang R,Wang X,Song Y,Liu Z,Han W,Li R.Impairment of immune response against dematiaceous fungi in Card9knockout mice[J].Mycopathologia,2016,181(9-10):631-642.
[21]Gavino C,Cotter A,Lichtenstein D,Lejtenyi D,Fortin C,Legault C,Alirezaie N,Majewski J,Sheppard DC,Behr MA,Foulkes WD,Vinh DC.CARD9 deficiency and spontaneous central nervous system candidiasis:complete clinical remission with GM-CSF therapy[J].Clin Infect Dis,2014,59(1):81-84.
[22]Celmeli F,Oztoprak N,Turkkahraman D,Seyman D,Mutlu E,Frede N,K9ksoy S,Grimbacher B.Successful granulocyte colony-stimulating factor treatment of relapsing Candida albicans meningoencephalitis caused by CARD9deficiency[J].Pediatr Infect Dis J,2016,35(4):428-431.