HLA ? ????

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HLA ? ????

• ????? ???

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HLA (Human Leukocyte Antigen)

• 1. ??? MHC
• (Major Histocompatibility Complex)
• 2. HLA? ??? ??? ?? class I, II, III?
• ?? ?.
• 1) class I HLA-A, B, C
• 2) class II HLA-DR, DQ, DP
• 3) class III C2C4, 21-hydroxylase, tumor
• necrosis factor ?
• 3. ?????? ??? ?? ( class I,II )

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Genomic Structure of MHC Region
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HLA ???

• 1. ??? ? 6? ???? ???? ??
• 2. ??
• haplotype (????) ?? ????
• ???? HLA ?? ??
• ?? ????? 50 ??
• ?? ??? ?? ??? 25

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• HLA? ??
• 1. ?? ??? ??
• HLA antigen? self recognition? ????. T
  cell dependent immune response ? ???? foreign
  substance ? ?? cell? ????? ????.
• ? class I Cytotoxic T cell? ????? ??? ??
  ? ????? ???? ???? antigen
  recognition ? ?????.
• ? class II Helper T cell? antigen
  receptor? ???? humoral ? cellular immune
  response? ????. ???? ??(?????)? ??? ??? ??? ?
  ?????.
• 2. ??? ?? ????
• 3. ?? ???

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HLA typing ? ??

• ???? ??? ?? ??? ??
• ???? ??? ???(HLA B27)
• ???? ???
• ???? ????, ????
• ?? HLA ?? ??

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HLA ?? ??

• (1) HLA??
• ? ???? ??
• ? ???? ?? ??????? ??
• (microlymphocytotoxicity test)
• HLA A, B, C lymphocytes or T lymphocytes
• HLA DR(DQ) B lymphocytes

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• ??
• ? HLA plate? ? well?? ?? HLA ??? ???? ???? ?? ??
  ???, ??? ???? lymphocytes? ??
• ? ??? ?? ???? ???? HLA ??? ??? ??? ?? ?? ??
  ????? ???
• ? Complement(rabbit plasma)? ???? complement
  fixation ? ?? cell lysis? ???
• ? Lysis? ??? ??? ???(eosin Y)? ???? ?? ???(????)
  ???? ??? ?? ??? lysis? ??? ???? ??(????). ?? ???
  ? ???? ??? ?

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?? ??? Microplate
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• ? ?????????
• (mixed lymphocyte test, MLC)
• T ???? HLA ??? ??? ?? ?? ???
• ???? ???? ??? ??? ??? ???
• ??? ??

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• ? ?????? ??( DNA typing )
• HLA ???? ????? ??? ????? PCR?
• ???? typing
• - Sequence specific primers (SSP)
• - Sequence specific oligonucleotide
• probes (SSOP)
• - Sequence-based typing (SBT)

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Hybridization Detection
Sequence specific oligonucleotide probes (SSOP)
reverse

• Prepare
• Hybridization, Wash and Citrate
• Solutions From Stocks Supplied

Hybridize At 50C For 30 Min.
Wash At 50C For 15 Min.

• Add
• PCR Product
• Hybridization Solution

• Aspirate Then Add
• Stringent Wash solution

• Aspirate Then Add
• SA-HRP

Incubate At Room Temperature For 15 Min.
Wash At Room Temperature For 5 Min, 2X

• Aspirate Then Add
• Wash Solution

• Aspirate Then Add
• Citrate, Incubate For 5 Min.
• Aspirate Then Add
• Substrates A and B,

• Incubate At Room Temperature For 10 Min.
• Aspirate Then Add
• Water, Wash For 5 Min, 3X

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• (2)HLA ??
• ? HLA ?? ??
• ??
• ??????? (hyperacuterejection) ??
• ??? ?? ??
• ???? ? HLA ?? ?? ??

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• ??
• ???? ???? ??? ??? ???? HLA??? ????
  ????(?????????)? ?? ?? well? ?? ?? ??? 10 15??
  ??? ? ???? ??.
• ???
• NIH, ????? ????? ?? ???, AHG(anti human
  globulin)?(?? kappa ??)? ? ??? ????? ??
• ?PRA (Panel Reactive Antibody)

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• ?? ???? ???????? ?? ??
• ?? ??
  ?? ?? ??
• ?? ???? ?? ???? ??
• ABO ???
  -
• HLA A,B -
  ??? ??? ???
• HLA-D/DR
  ???
• HLA crossmatch
  -
• ???? ???? ?? ?? ??
• ???? ???? ???? ??
• - ???? ?? ??? ?? ??

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?????