KIR in Tr. Medular

7/28/2019 KIR in Tr. Medular

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KIR and Cytokine Gene Polymorphisms

as risk factors for Graft versus Host

Disease after Bone Marrow

Transplantation

Ileana Constantinescu*, Daniela Vasile, Daniela Nedelcu,

Adela Toader, Marilena Zaharia, Mihaela Sora, Anca Tica,

Mihai Magazin, Paul Nistor

Fundeni Centre for Immunogenetics and Virology



HLA alleles The considerable polymorphism of HLA is well-

known.

HLA polymorphism is reflected by allelicsubstitution of many amino acid residues in thepolypeptide chains, especially the externaldomains which contain the peptide binding site.

This affects the spectrum of antigenic peptidespresented by the different allelic types of HLAmolecules and the repertoire of responding T-cells.




The HLA genetics is complex

HLA polymorphism

Expression of HLA polymorphism –Typing

The transplanted graft represents a continuous source of HLA alleles that can induce a rejection response at anytime post-transplant.

HLA matching can have a dualistic effect on transplantoutcome: it reduces rejection but conversely, it maypromote other HLA-restricted mechanisms of allograftinjury.




GVHD

In GVH diseases donor-derived immuno-competent lymphocytesreact with HLA incompatible recipient cells and induce inflammatoryresponses in host tissues such as the skin and gastrointestinal tract.

GVH disease seems more likely in cases whereby the donor is wellmatched for the patient.

Direct and indirect HLA allorecognition mediate GVH reactions if immunocompetent donor cells recognize recipient incompatibilities.

During infection, microbial antigens are processed by APC andpresented via HLA molecules to T-cells that elicit cytotoxic and DTH-like inflammatory reactions in the allograft.




Cytokine gene polymorphism in human disease, N Haukim et al, Review –Cytokine gene polymorphism in human disease: on-line

databases, Supplement 2, Genes and Immunity (2002) 3, 313-330

Cytokine and

polymorphism

Disease Association

IFNg(CA)N intron 1 Graft-versus-host

disease

Yes (increased

severity)

TNFα (TNFSF2) –

308 (TNF2)

Acute graft-versus-

host diseases

Yes

IL-10 (IL10.G) Chronic graft-versus-host diseases Yes

IL-6-174C-G (NlaIII) Graft-versus-host-

disease

Yes (increased

severity)




Leucocytes complex receptor and KIR haplotype

(A. P. Williams, KIR and Their Role in Disease,Southampton, UK, 2006)



Methods

We have assessed eight pairs for allogeneic bone marrow transplantation (BMT).All related donors and recipients had a 100% HLA alleles match (HLA A, B C,DRB1, DQB1, DPB).

In addition all pairs were investigated for their cytokine gene polymorphisms andKIR haplotypes as long term prediction factors for graft versus host diseases

(GVHD) after BMT.

HLA typing was performed by high resolution sequence specific primers (SSP)method using Dynal (KIR Genotyping SSP Kit) high resolution SSP. HLA allelesambiguities were resolved by Sequencing Based Typing (SBT) Allele SEQR,ABBOTT.

Cytokine gene polymorphisms were performed by Dynal (Cytokine GenotypingSSP Kit) PCR SSP (IL1 alpha, IL1 beta, IL1R, IL1R alpha, IL2, IL4, IL4R alpha,IL6, IL10, IL12, gamma IFN, TGF beta and TNF alpha).

KIR genotyping for both donors and recipients were revealed using PCR SSP(Dynal).




Cytokine gene polymorphisms results in BMT patients

The following genes were identified: IL1 , IL1 , IL1R A, gIFN, TGF 1, IL4, IL6, IL10.

44% TNFα and IFNγ

56% TGFβ

RESULTSFundeni Centre for Immunogenetics and Virology



KI R genotypes resul ts in BMT patients

The following genes were identified :

2DL1, 2DL3, 2DL4, 2DS4*003-006, 3DL1, 3DL2, 3DL3, 3DP1*003.

2DL1, 2DL3, 2DL4, 2DS4*001/002, 2DS4*003-006, 3DL1, 3DL2, 3DL3, 3DP1*003.

KIR genotype: B, in 12 patients

A, in 4 patients.




Interpretation of resultsRecipient D.F. Donor D.C.

HLA HLA

A 02-24 A 02-24

B 18-18 B 18-18

C 07-07 C 07-07

DRB1 11-13 DRB1 11-13

DQB1 03-05 DQB1 03-05

KIR – B4 genotype KIR – B4 genotype

KIR –2DL1, 2DL2, 2DL4, 2DL5B, 2DS2,

2DS3, 2DS4004, 3DL2, 3DL3, 3DS1,

2DP1, 3DP1

KIR –2DL1, 2DL2, 2DL4, 2DL5B, 2DS2,

2DS3, 2DS4004, 3DL2, 3DL3, 3DS1,

2DP1, 3DP1




Recipient A.C. Donor C.R.

HLA HLA

A 03-24 A 03-24

B 18-44 B 18-44

C 05-12 C 05-12

DRB1 16-16 DRB1 16-16

DQB1 05-05 DQB1 05-05

DPB1 02-05 DPB1 02-05KIR – B4 genotype KIR – B4 genotype

KIR –2DL1, 2DL2, 2DL4, 2DL5B003-006,

2DS3, 2DS4, 2DL1, 3DL2, 3DL3, 2DP1,

3DP1003

KIR –2DL1, 2DL2, 2DL4, 2DL5B003-006,

2DS3, 2DS4, 2DL1, 3DL2, 3DL3, 2DP1,

3DP1003

Interpretation of results


F d i C f I i d Vi l



Interpretation of resultsRecipient F.C. Donor F.E.

HLA HLA

A 02-03 A 02-03

B 07-40 B 07-40

C 01-03 C 01-03

DRB1 04-10 DRB1 04-10

DQB1 03-07 DQB1 03-07

KIR – A genotype KIR – A genotype

KIR – 2DL1, 2DL3, 2DL4, 2DS1, 3DL1,

3DL2, 3D33, 2DP1, 3DP1003

KIR – 2DL1, 2DL3, 2DL4, 2DS4, 3DL1,

3DL2, 3DL3, 2DP1, 3DP1003


F d i C t f I ti d Vi l



Cytokine gene polymorphism, donor-recipientKIR genes match, are useful in prevention of

acute rejection episodes, maintaining, on a longterm the bone marrow allograft functionality.

Donor surface NK and T cells KIR genes

mismatch with the recipient could activatecellular immune response via MHC class I.

Preliminary conclusions


F d i C t f I ti d Vi l



Conclusions

Transplantation immunology is complex. Our Immunogenetic Centre provide expanded

immunological monitoring together with virological and

drug monitoring of BMT transplanted patients.

Our goal is to offer complete integrated monitoringdata and to fulfill EFI Standards.

We are open for collaborative work and research projects

within EFI.

Clinical and laboratory scientists should worktogether as a team in order to have a

complete overview of the transplanted

patients.


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KIR in Tr. Medular