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Tests performed on donors and recipients of kidney and BM transplants



How many?

3



List

Serological test


Cellular test


Molecular methods

Serological test


Cellular test


Molecular methods



Tests performed on ____ and _____ of kidney and BM transplants

Tests performed on donors and recipients of ____ and _____ transplants



List

BM


kidney



Serological, cellular test, molecular methods

Serological test



Principle

Use ab to identify



HLA antigen on patient cells


Antibodies in patient serum



What test it is?


how many examples there are?

Serological test



3

3 examples of serological test

ELISA


Microcytotoxicity test


Cross match test

Cellular test



Principle

Detect T cell response to HLA Ags on grafts



How many examples there are?

2

2 examples of cellular test

MLR


MLC



Full name

Mixed lymphocyte reaction


Mixed lymphocyte culture

Molecular test



Principles

Identify HLA alleles



Examples

PCR

Serological test



ELISA


_______________


Cross-match test

Microcytotoxicity test

Microcytotoxicity test



Aka

Terasaki method



test for?

HLA A, B, C, DR Ags

Microcytotoxicity test



Test for?

HLA A, B, C, DR ags



They are known as?

SD Ags

Microcytotoxicity test



Steps

1. Obtain a well that contains antiserum against different HLA Ags in each well.



2. Add peripheral blood to each well


For HLA A, B, C, add mononuclear cell


For HLA DR, add purified B cells



3. Add reagent complement (from rabbit serum)


Wells that have Ab to HLA ag will have cell lysis



4. Add viability stain


Dead cells: permeability is altered, colored


Live cells: colorless

Microcytotoxicity test



How cells are examined?

under phase contrast microscope

Microcytotoxicity test



Positive result for HLA antigen

50% of cells are pink

Cross match test



Application

Detect pre-formed Abs in recipient against donor's HLA abs



Purpose

prevent hyperacute rejection

Cross match test



procedure

similar to Microcytotoxicity test



1. Serum from recipient


+


Peripheral blood (mononuclear cells)from donor (not patients)


+


C




2. Incubate in microtiter plate



3. Add viability stain



Interpret the results?


+: bad donor


-: good donor

MLC



aka

Mixed lymphocyte reaction

MLC/MLR



What does it detect?

Differences between HLA-D Ags

MLC procedure

Lymphocytes from donor


is mixed with


Lymphocytes from recipient


in a microtiter plate



If HLA-D from donor is different from recipient,


recipient lymphocytes will proliferate



it is measured by 3-H, a radioactive dye that is incorporated into DNA

3-H: thymine

MLC



Limitations

Labor intensive



How long it takes?


Not applicable to?

6 months



Cadaver

Molecular methods



List

PCR


SOOP


Sequence




Function

PCR: Amplifies HLA gene


SOOP: identify HLA allele by binding to HLA gene


Sequence: HLA gene

Immunosuppressive agents



List

1. Anti-proliferative agents


2. Cortiosteroids


3. Calcineurine inhibitors


4. Anti-thymcyte globulin


5. Monoclonal antibodies

Immunosuppresive agents



1. ______________


2. Cortiosteroids


3. Calcineurine inhibitors


4. Anti-thymcyte globulin


5. Monoclonal antibodies

Anti-proliferative agents



Example: Mycophenolate Mofetil, Azathioprine, Sirolimus



Mechanism

Blocks DNA synthesis and therefore lymphocyte proliferation

immunosuppressive agents



1. Anti-proliferative agents


2. ______________


3. Calcineurine inhibitors


4. Anti-thymcyte globulin


5. Monoclonal antibodies

Corticosteroids



Example: Prednisone, Methylprednisolone



Mechanism

Block cytokine production by T and M cells



Antiinflammation

Imunosuppressive agents



1. Antiproliferative agents


2. Cortiosterioids


3. _______________


4. Anti-thromcyte globulin


5. Monoclonal antibodies

Calcineurine inhibitors



Example: Cyclosporin A, Tacrolimus



Mechanism

Decrease cytokine production

Immunosuppressive agents



1. Anti-proliferative agents


2. Cortiosteriods


3. Calcineurine ihibitors


4. ___________________


5. Monoclonal antibodies

Anti-thromcyte globulin



Mechanism

Destroy T cells

Immunosuppressive agents



1. Anti-proliferative agents


2. Cortiosteriods


3. Calcineurine inhibitors


4. Anti-thromcyte globulin


5. _________________

Monoclonal antibodies



Examples: to CD3, IL2 receptor, adhesion molecules, cytokines

Inhibits various steps of the immune response