Journal club

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Journal club

• Dai Jianmin
• 2014-5-9

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Background
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????? -2010?,30???,308???
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?????????
????Williams hematology
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ABO??????????

• ABO?? 9q34.1-9q34.2
• 7????,?????DNA 18kb
• ???17 ,28bp 691bp??
• ??????exon 6 ? exon 7

Exon1-5?? ?????????????? Exon6-7??
???????(?????)

• A or B subgroups (with only few exceptions)
  result from a variety of nucleotide changes in
  exon 7 that cause alterations in the catalytic
  domain of the glycosyltransferase.

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• A?B????????????????????????
• A??????N-???????
• B??????D-????

O??????????
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A?B?O????????

• ABO???????A101????
• ?A1????????????????????????????????

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?????????????
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???????????????
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ABO??

• A??????A1?A2
• ??A??A3?Ax?Aend?Am?Ay??Ael
• B?????A????B3?Bx?Bm?Bel?

ABO subgroups the detection rate in Shanghai is
approximately 0.015.
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Samples
n 2.1 million Chinese blood donors,2003 to
2011 322 ABO subgroup individuals 29
family members 120 healthy random Chinese
donors included Aint, AintB, A3, A3B, Ax, AxB,
AmB, Ael,AelB, AB3,B3, A3B3, Bx, AxBx, ABx, ABm,
Bm, Bel, B(A), A2B, and A2Bx.
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Methods I

• Serology
• monoclonal anti-A, anti-B, anti-AB, anti-H
• polyclonal anti-A, anti-B, anti-AB
• an ABO red blood cell (RBC) kit
• Dolichos bijorus for anti-A1

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??A???????
Phenotype Red cells tested With anti-sera Red cells tested With anti-sera Red cells tested With anti-sera Red cells tested With anti-sera Red cells tested With anti-sera Serum tested with Known red cells Serum tested with Known red cells Serum tested with Known red cells Serum tested with Known red cells Salivaof secretors
Anti-A -B -AB -A1 -H A1 A2 B O
A1 V _ V V _ _ _ V _ A and H
Aint _ V _ _ V _ A and H
A2 _ V _ Some-times. _ V _ A and H
A3 mf _ mf _ Occa-sionally. _ V _ A and H
Am -/w _ -/w _ V _ _ V _ A and H
Ax -/W _ / _ V Fre-quently _ V _ H
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???A???????
??? A3/ Ax/ Aend ????(Mf)A60 A3
??? A3/ Ax/ Aend 10 ????????mf?? Aend
??? A3/ Ax/ Aend ???-AB???? Ax
??? ?????-A Am/ Ay / Ael ???????-A Am
??? ?????-A Am/ Ay / Ael ???????????A??
??? ?????-A Am/ Ay / Ael ????????-A Ay
??? ?????-A Am/ Ay / Ael ?????????A??
??? ?????-A Am/ Ay / Ael ?????-A Ael
??? ?????-A Am/ Ay / Ael ???????H???A??
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Methods II

• Polymerase chain reaction
• ABO gene sequences Exons 6 to 7
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  Exons 1 to 5 and their splice sites
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  the promoter region
• DNA sequencing of subgroup PCR products
• gel-purified PCR products containing the
  mutation sites
• the pMD18-T vector
• Screening for subgroup-specific mutations by
  sequencing
• Nomenclature of mutations and ABO alleles

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Methods III

• Analysis of ABO promoter activity
• pGL2-Basic vector
• K562 cells(Turbofect)
• renilla luciferase vector phRL-TK(cotransfecte
  d as
• an internal control)
• dual luciferases assay system firefly and
  renilla
• luciferase activities

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Results I

• Identification of 62 rare ABO alleles
• 29 novel subgroup alleles were newly linked
  to different kinds of ABO variations
• Promoter variant phenotype and genotype analysis

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more than half occurred in conserved or invariant
sites
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Results II

• Identification of 62 rare ABO alleles
• Promoter variant phenotype and genotype analysis
• the first evidence that promoter abnormality
  is involved in the formation of weak ABO
  phenotypes
• deletion at Positions -35 to -18 in their
  promoter region
• Ael and Bel phenotypesthe first naturally
  occurring ABO alleles with premature terminal
  codons in the 5'-region

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decreased by more than 50
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Results III

• Identification of 62 rare ABO alleles
• Promoter variant phenotype and genotype analysis
• Ael and Bel phenotypesthe first naturally
  occurring ABO alleles with premature terminal
  codons in the 5'-region
• the ABO mutations 52CgtT and 7GgtT, R18X and
  E3X in GTs

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The end ??!