Module 11: Transplant Immunology Obj. 3 SA Flashcards Preview

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Flashcards in Module 11: Transplant Immunology Obj. 3 SA Deck (10)
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1

Tissue typing involves testing for(3)

HLA antigens of donor and recipient
HLA Ab present in recipient serum

2

Tissue crossmatch

predicts compatibility of the transplant

3

HLA Ag detection techniques (2)

Serological (Old technique), may then require Mixed lymphocyte culture
Molecular technique

4

Serological technique for HLA Ag detection

Commercial antisera containing known ab
Incubate patient or donor lymphs with known antisera
Add complement
Cells which bind HLA Ab will be damaged due to complement binding
Damaged cells take up dye (then quantitated)

Lymphs must be from LIVE donor

5

Mixed lymphocyte culture

test for Class II MHC Ag

Culture 2 pts cells together
Cell proliferation is detected by uptake of radioactive thymidine (From DNA synthesis)

Low MLC = better success for transplant

6

Molecular techniques fro HLA Ag detection

Sequencing DNA by PCR

Basic steps:
1. Obtain DNA from nuclei of a cell
2. Double stranded DNA is denatured by heat
3. Oligonucleotide primer sequences are then chosen to identify the region of interest
-if primer chosen is close to region of interest, only that part of the DNA will replicate when DNA polymerase enzyme and deoxyribonucleotide triphosphates are added
-if a primer is used that is specific to a certain sequence, the product will only be produced if the patient has that particular sequence

7

HLA Ab detection techniques (2)

ELISA
Flow Cytometry

8

What happens if a patient has Ab against certain HLA's

Ab will attach to corresponding Ag on the transplanted tissue and lead to tissue destruction

9

ELISA

Pure HLA Ag bound to well wall
Binding of Ab from pt is detected by second Anti-IgG Ab labeled with alkaline phosphatase
Addition of enzyme substrate for detection

10

Flow cytometry

2nd Ab is a fluorescent dye-labelled anti human Ab which binds to the pt Ab attached to the reagent T and B lymphs