广东地区CD36Ⅱ型缺失献血者CD36基因突变类型及其对CD36单核细胞表达的影响分析
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摘要
目的了解广东地区CD36Ⅱ型缺失人群CD36基因突变类型及其对CD36单核细胞表达量的影响。方法选择本中心2012年6月—2016年6月的广东地区献血者2 300名,采用流式细胞术筛查出CD36Ⅱ型缺失表达者,并借助DNA测序仪对CD36编码序列做测序分析以弄清该人群CD36基因突变类型;运用Graph Pad Prism v5. 0统计分析CD36基因突变对CD36Ⅱ型缺失者单核细胞表达的影响。结果本组广东地区献血者CD36Ⅱ型缺失者比例为0. 96%(22/2 300),CD36Ⅱ型缺失者的CD36编码基因的突变率为27. 27%(6/22),突变基因类型分别为A1163T、1228—1239del ATTGTGCCTATT、198—205del GATCTTTG、C1156T、C268T和C380T。22名CD36Ⅱ型缺失者单核细胞的CD36表达量(平均荧光强度,MFI):6名存在单链基因突变的CD36Ⅱ型缺失者[Mt(+/-)组]与16名无基因突变的CD36Ⅱ型缺失者[Mt(-/-)组]及8名正常CD36表达献血者(WT组)分别为58. 10±7. 57 vs 139. 90±16. 33 vs 141. 80±15. 59(P<0. 01)。结论广东地区CD36Ⅱ型缺失献血者中仅部分人存在CD36编码基因的单链基因突变,但突变可影响CD36在其单核细胞的表达量。
Objective To obtain the mutation types of CD36 gene among CD36 type Ⅱ deletion blood donors in Guangdong region,and analyze the mutation effect on CD36 expression in monocytes. Methods 2300 blood donors of our center recorded during June 2012 to June 2016 in Guangdong region were selected. The expression of CD36 in platelets were measured by flow cytometry. The coding region of CD36 were sequenced by 3130 XL DNA sequencer to obtain their mutation types of CD36 gene. The effect of CD36 mutations on expression of CD36 in monocytes among type Ⅱ deletion blood donors was analyzed using Graph Pad Prism v5. 0. Results The frequency of CD36 type Ⅱ deletion in Guangdong region was about0. 96%( 2/2 300),and the mutation rate of CD36-encoding gene among CD36 type Ⅱ deficient individuals was 27. 27%. 6 CD36 type Ⅱ deletion mutation types were identified,including A1163 T、1228—1239 delATTGTGCCTATT、C1156 T、198—205 del GATCTTTG、C268 T and C380 T. The expression level of CD36 in monocytes( mean fluorescence intensity,MFI) among the 22 CD36 type Ⅱ deletion individuals: the level of CD36 expression in the Mt( +/-) group carrying mutations in one chain was significant lower than the Mt(-/-) group lacking CD36 mutations in both chains( 58. 10± 7. 57 vs 139. 90±16. 33,P<0. 01),even lower than the WT group which have normal CD36 expression both in platelets and monocytes and without CD36 gene mutations( 58. 10±7. 57 vs 141. 80± 15. 59,P< 0. 01). Conclusion The CD36 mutation in one chain has been only found in some of CD36 type Ⅱ deletion individuals in Guangdong region; however,it can affect the level of CD36 expression in monocytes.
Objective To obtain the mutation types of CD36 gene among CD36 type Ⅱ deletion blood donors in Guangdong region,and analyze the mutation effect on CD36 expression in monocytes. Methods 2300 blood donors of our center recorded during June 2012 to June 2016 in Guangdong region were selected. The expression of CD36 in platelets were measured by flow cytometry. The coding region of CD36 were sequenced by 3130 XL DNA sequencer to obtain their mutation types of CD36 gene. The effect of CD36 mutations on expression of CD36 in monocytes among type Ⅱ deletion blood donors was analyzed using Graph Pad Prism v5. 0. Results The frequency of CD36 type Ⅱ deletion in Guangdong region was about0. 96%( 2/2 300),and the mutation rate of CD36-encoding gene among CD36 type Ⅱ deficient individuals was 27. 27%. 6 CD36 type Ⅱ deletion mutation types were identified,including A1163 T、1228—1239 delATTGTGCCTATT、C1156 T、198—205 del GATCTTTG、C268 T and C380 T. The expression level of CD36 in monocytes( mean fluorescence intensity,MFI) among the 22 CD36 type Ⅱ deletion individuals: the level of CD36 expression in the Mt( +/-) group carrying mutations in one chain was significant lower than the Mt(-/-) group lacking CD36 mutations in both chains( 58. 10± 7. 57 vs 139. 90±16. 33,P<0. 01),even lower than the WT group which have normal CD36 expression both in platelets and monocytes and without CD36 gene mutations( 58. 10±7. 57 vs 141. 80± 15. 59,P< 0. 01). Conclusion The CD36 mutation in one chain has been only found in some of CD36 type Ⅱ deletion individuals in Guangdong region; however,it can affect the level of CD36 expression in monocytes.
引文
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[7] Li R,Qiao Z,ling B,et al. Incidence and molecular basis of CD36 deficiency in Shanghai population. Transfusion,2015,55(3):666-673.
[8] Wu G,Zhou Y,Li L,et al. Platelet immunology in China:research and clinical applications. Transfus Med Rev,2017,31(2):118-125.
[9] Bierling P,Godeau B,Fromont P,et al. Posttransfusion purpuralike syndrome associated with CD36(Naka)isoimmunization.Transfusion,1995,35(9):777-782.
[10] Curtis BR,Ali S,Glazier AM,et al. Isoimmunization against CD36(glycoproteinⅣ):description of four cases of neonatal isoimmune thrombocytopenia and brief review of the literature. Transfusion,2002,42(9):1173-1179.
[11] Taketani T,Ito K,Mishima S,et al. Neonatal isoimmune thrombocytopenia caused by type I CD36 deficiency having novel splicing isoforms of the CD36 gene. Eur J Haematol,2008,81(1):70-74.
[12] Fujino H,Ohta K,Taniue J,et al. Primary refractoriness to platelet transfusion caused by Nak(a)antibody alone. Vox Sang,2001,81(1):42-44.
[13] Nakajima F,Nishimura M,Hashimoto S,et al. Role of anti-Nak(a)antibody,monocytes and platelets in the development of transfusion-related acute lung injury. Vox Sang,2008,95(4):318-323.
[14] Yanai H,Chiba H,Morimoto M,et al. Human CD36 deficiency is associated with elevation in low-density lipoprotein-cholesterol. Am J Med Genet,2000,93(4):299-304.
[15] Chiba H,Yanai H,Fujiwara H,et al.CD36 deficiency and insulin resistance. Lancet,2001,358(9277):243-244.
[16] Okamoto F,Tanaka T,Sohmiya K,et al. CD36 abnormality and impaired myocardial long-chain fatty acid uptake in patients with hypertrophic cardiomyopathy. Jpn Circ J,1998,62(7):499-504.
[17] Xu X,liu Y,Hong X,et al. Variants of CD36 gene and their association with CD36 protein expression in platelets. Blood Transfusion,2014,12(4):557-564.
[18] Imai M,Tanaka T,Kintaka T,et al. Genomic heterogeneity of typeⅡCD36 deficiency. Clin Chim Acta,2002,321(1-2):97-106.
[19] Kashiwagi H,Tomiyama Y,Kosugi S et al. Family studies of typeⅡCD36 deficient subjects:linkage of a CD36 allele to a plateletspecific mRNA expression defect(s)causing typeⅡCD36 deficiency. Thromb Haemost,1995,74(2):758-763.
[2] Yamamoto N,Akamatsu N,Sakuraba H,et al. Platelet glycoproteinⅣ(CD36)deficiency is associated with the absence(typeⅠ)or the presence(typeⅡ)of glycoproteinⅣon monocytes.Blood,1994,83(2):392-397.
[3] Yanai H,Chiba H,Morimoto M,et al. Human CD36 deficiency is associated with elevation in low-density lipoprotein-cholesterol. Am J Med Genet,2000,93(4):299-304.
[4] Yanai H,Chiba H,Fujiwara H,et al. Phenotype-genotype correlation in CD36 deficiency typesⅠandⅡ. Thromb Haemost,2000,84(3):436-441.
[5] Masuda Y,Tamura S,Matsuno K,et al. Diverse CD36 expression among Japanese population:defective CD36 mutations cause platelet and monocyte CD36 reductions in not only deficient but also normal phenotype subjects. Thromb Res,2015,135(5):951-957.
[6] Xu X,Ye X,Xia W,et al. Studies on CD36 deficiency in South China:two cases demonstrating the clinical impact of anti-CD36antibodies. Thromb Haemost,2013,110(6):1199-1206.
[7] Li R,Qiao Z,ling B,et al. Incidence and molecular basis of CD36 deficiency in Shanghai population. Transfusion,2015,55(3):666-673.
[8] Wu G,Zhou Y,Li L,et al. Platelet immunology in China:research and clinical applications. Transfus Med Rev,2017,31(2):118-125.
[9] Bierling P,Godeau B,Fromont P,et al. Posttransfusion purpuralike syndrome associated with CD36(Naka)isoimmunization.Transfusion,1995,35(9):777-782.
[10] Curtis BR,Ali S,Glazier AM,et al. Isoimmunization against CD36(glycoproteinⅣ):description of four cases of neonatal isoimmune thrombocytopenia and brief review of the literature. Transfusion,2002,42(9):1173-1179.
[11] Taketani T,Ito K,Mishima S,et al. Neonatal isoimmune thrombocytopenia caused by type I CD36 deficiency having novel splicing isoforms of the CD36 gene. Eur J Haematol,2008,81(1):70-74.
[12] Fujino H,Ohta K,Taniue J,et al. Primary refractoriness to platelet transfusion caused by Nak(a)antibody alone. Vox Sang,2001,81(1):42-44.
[13] Nakajima F,Nishimura M,Hashimoto S,et al. Role of anti-Nak(a)antibody,monocytes and platelets in the development of transfusion-related acute lung injury. Vox Sang,2008,95(4):318-323.
[14] Yanai H,Chiba H,Morimoto M,et al. Human CD36 deficiency is associated with elevation in low-density lipoprotein-cholesterol. Am J Med Genet,2000,93(4):299-304.
[15] Chiba H,Yanai H,Fujiwara H,et al.CD36 deficiency and insulin resistance. Lancet,2001,358(9277):243-244.
[16] Okamoto F,Tanaka T,Sohmiya K,et al. CD36 abnormality and impaired myocardial long-chain fatty acid uptake in patients with hypertrophic cardiomyopathy. Jpn Circ J,1998,62(7):499-504.
[17] Xu X,liu Y,Hong X,et al. Variants of CD36 gene and their association with CD36 protein expression in platelets. Blood Transfusion,2014,12(4):557-564.
[18] Imai M,Tanaka T,Kintaka T,et al. Genomic heterogeneity of typeⅡCD36 deficiency. Clin Chim Acta,2002,321(1-2):97-106.
[19] Kashiwagi H,Tomiyama Y,Kosugi S et al. Family studies of typeⅡCD36 deficient subjects:linkage of a CD36 allele to a plateletspecific mRNA expression defect(s)causing typeⅡCD36 deficiency. Thromb Haemost,1995,74(2):758-763.