KIR: B haplotipo įtaka haploidentinėms alokklt. Rita Čekauskienė VULSK, HOTC

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1 KIR: B haplotipo įtaka haploidentinėms alokklt Rita Čekauskienė VULSK, HOTC

2 KIR

3 NK ląstelės Sudaro apie 5 25% PK limfocitų Įgimto imuniteto ląstelės Veikia citotoksiškai be prieš tai įvykusios sensibilizacijos Atsako pikas kelios val dienos Sunaikina virusų pažeistas, vėžines ar kitaip pakitusias ląsteles, praradusias savo paviršinius ligandus (HLA) NK ląstelių receptoriams Subrendę ir funkciškai aktyvios NK CD56dim, CD16+, KIR+ ir/ar NKG2A+ Sekretuoja citokinus (IFN g, TNF a,gm CSF ), moduliuoja imuninį atsaką Citotoksiškai veikia: iš lizosomų išskirdamos perforinus ir granzimus Taip pat veikia per ADCC mechanizmą ar aktyvuotos citokinų NK atsistato vienos iš pirmųjų po KKLT

4 NK ląstelių receptoriai Inhibitoriniai Jie yra svarbiausi NK funkcijų reguliatoriai, žinomi jų ligandai, veikimas Pažindami ligandus jie blokuoja NK aktyvaciją t.y. neleidžia žudyti savų ląstelių KIR Ig tipo šeimos receptoriai; jų ligandai HLA A, B, C tam tikri alotipai NKG2A/CD 94 lectino tipo receptoriai jų ligandai HLA E Aktyvuojantys Jų reikšmė NK funkcijoms mažiau žinoma, ne visų ligandai nustatyti KIR aktyvuojantys NKp46 NKp30 NKp44 NKG2D KIR

5 Ląstelės, turinčios HLA ligandus inhibitoriniams receptoriams pažįstamos atitinkamų inhibuojančių KIR, NK ląstelė lieka neaktyvi MHC Class I NK - No Kill Normal Ląstelės, praradusios HLA ligandus inhibitoriniams receptoriams tampa NK taikiniais NK Kill Cancer Karre et al. Nature 1986

6 Inhibitorinių ir aktyvuojančių receptorių tarpusavio sąveika nulemia NK sprendimą sunaikinti ląstelę KIR MHC Class I KIR NK - Normal Disease NK Activating Receptor Activating Ligand No Kill Activating Ligand Kill Activating Receptor

7 KIR Killer Ig like Receptors KIR genų šeima turi 15 genų (KIR2DL1, KIR2DL2, KIR2DL3, KIR2DL4, KIR2DL5A, KIR2DL5B, KIR2DS1, KIR2DS2, KIR2DS3, KIR2DS4, KIR2DS5, KIR3DL1, KIR3DL2, KIR3DL3 and KIR3DS1) ir du pseudogenus (KIR2DP1 and KIR3DP1) kurie užkoduoti Kb Leukocitinių Receptorių Komplekso (LRC) srityje, esančioje 19 (19q13.4) chromosomoje KIR genų šeima polimorfiška 10 dažnų KIR haplotipų > 40 nustatyta

8 KIR struktūra ir funkcija Ekstraląsteliniai domenai: skaičius ir tipas (D0, D1, D2) Transmembraninė dalis Citoplazminė dalis: ilga (ITIM) inhibuojantys trumpa (ITAM) aktyvuojantys Lanier L Ann Rev Immunol 25: 225, 2005

9 KIR receptorių struktūra

10 Inhibitorinis KIR KIR2DL1 KIR2DL2/3 KIR3DL1 KIR Ligandas HLA-C group 2 (2,4,5,6) HLA-C group 1 (1,3,7,8) HLA-Bw4 (e.g. B27)

11 KIR haplotipai A tipas turi 4 pagrindinius inhibitorinius KIR ir vieną aktyvuojantį geną KIR2DS4 B tipas turi žymiai daugiau (iki 12) genų, vyrauja aktyvuonatys Šie haplotipai gali būti suskirstyti į telomerinius ir centromerinius motyvus (Cen A, Tel A, Cen B, and Tel B)

12 KIR haplotipai group A haplotypes are generally non variable in its gene organisation with all four framework genes present plus KIR2DL1, KIR2DL3, KIR3DL1, KIR2DS4 and KIR2DP1. Group B haplotypes show a lot more variation in the number and combination of KIR genes present and are characterized by the presence of one or more of KIR2DL2, KIR2DL5A/B, KIR2DS1, KIR2DS2, KIR2DS3, KIR2DS5 and KIR3DS1 genes (23, 24). Group B haplotypes possess a greater variability in the number of genes present. They possess from one to five activating KIR (i.e. KIR2DS1, KIR2DS2, KIR2DS3, KIR2DS5 and KIR3DS1) and can incorporate inhibitory KIR genes which are known to be absent in group A haplotypes (i.e. KIR2DL2 and KIR2DL5)

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15 KIR B content calculation

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17 Skirtingi NK ląstelių klonai ekspresuoja skirtingus KIR ir skirtingus jų kiekius Kiekvienas klonas turi ir pastoviai išlaiko bent po vieną inhibitorinį receptorių savam ligandui KIR ir HLA paveldimi skirtingai, todėl galimos įvairios jų kombinacijos, kurios turi reikšmės sergamumui autoimuninėmis, infekcinėmis ligomis, nėštumui, KKLT išeitims

18 KIR reikšmė alokklt Inhibitorinių KIR reikšmė Netapataus donoro KKLT ( missing self ) Haploidentinės KKLT Netapataus negiminingo donoro KKLT Virkštelinio kraujo KKLT Aktyvuojančių KIR reikšmė KIR haplotipai (B) Atskirų KIR svarba

19 missing self Apie 1/3 HLA I klasės netapačių donoro recipiento porų turės KIR ligando netapatumą Davies et al Blood 2002 Giebel et al Blood 2003

20

21 KIR reikšmė alokklt Farag et al. Blood 100:1935, 2002

22 KIR reikšmė alokklt

23 KIR ligand netapatumas turi naudos alogeninėms transplantacijoms: AML pacientai T deplecija Didelė KKL dozė Tiek giminingi haploidentiniai donorai, tiek negiminingi donorai, tiek virkštelinis kraujas Rugerri; Cooley;

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26 448 AML pacientai (dauguma didelės rizikos) UD KKLT, mieloabliacinis kondicionavimas. Be T deplecijos Donorų KIR haplotipo įtaka KKLT išeitims: A/A vs B/x haplotipai

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28 HLA matched HLA mismatched

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30 B/x haplotipo donorai pagerina AML pacientų išgyvenamumą po KKLT B/x haplotipo donorai 30% pagerina RFS (p=0.002) B/x haplotipo donorai daugiau cgvhd (RR 1.51, p=0.03) Siūlymas tipuoti donorų KIR, rinktis B/x haplotipų donorus

31 KIR B haplotipas labai polimorfiškas Tikslas ištirti, kurios haplotipų sritys turi didžiausią įtaką Ištirtas 1409 Tx porų (AML ir ALL, mieloabliac, be T deplecijos, UD) KIR genotipas With Cen B/B homozygous donors the cumulative incidence of relapse was 15.4% compared with 36.5% for Cen A/A donors (relative risk of relapse 0.34; 95% confidence interval ; P <.001). Overall, significantly reduced relapse was achieved with donors having 2 or more B gene content motifs (relative risk 0.64; 95% confidence interval ; P.003) for both HLA matched and mismatched transplants.

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34 Išvados Donorai su KIR B genotipu turinčiu daugiau KIR B genų bei ypač centromerinių B genų motyvų (CenB/CenB) sumažina ligos atkryčių bei prailgina OS ir DFS pacientams su AML Autorių siūlymas tipuoti keletos tapačių donorų KIR, rinktis donorus su palankiu KIR genotipu. Taip galima būtų parinkti donorą su palankiu KIR genotipu 67% pacientų (atsitiktinai 31%) Taip parenkant donorą 22% sumažėtų ligos atkryčių

35 614 NHL patients receiving unrelated donor (URD) T cell replete marrow or peripheral blood grafts NHL patients receiving 10/10 HLAematched URD grafts with KIR B/x donors experienced significantly lower relapse at 5 years (26%; 95% confidence interval [CI], 21% to 32% versus 37%; 95% CI, 27% to 46%; P ¼.05) compared with KIR A/A donors, resulting in improved 5 year progression free survival (PFS) (35%; 95% CI, 26% to 44% versus 22%; 95% CI, 11% to 35%; P ¼.007). In multivariate analysis, use of KIR B/x donors was associated with significantly reduced relapse risk (relative risk [RR],.63, P ¼.02) and improved PFS (RR,.71, P ¼.008).

36 86 patients with advanced hematologic malignancies who received nonmyeloablative, HLA-haploidentical HSCT with high-dose, post-transplantation cyclophosphamide (Cy).

37 Patients homozygous for the KIR A haplotype, which encodes only one activating KIR, had an improved OS (HR=0.30; CI: ; p=.004), EFS (HR=0.47; CI: ; p=.05) and NRM HR=0.13; CI: ; p=0.046) if their donor expressed at least one KIR B haplotype, which encodes several activating KIRs

38

39 Michaelis et al, 57 adults with hematological malignancies given T cell depleted (C3/C19) haploidentical HSCT (RIC) were found to have a reduced risk for relapse when transplanted from a KIR haplotype B donor There was no difference between donor KIR haplotypes in nonrelapse mortality (NRM, p=0.200) but had a significantly reduced incidence of relapse for patients with ahaplotype B donor (p=0.001).

40 85 vaikai, ALL Haplo donoras PBSC, ex vivo T deplecija

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42

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44 121 haplotx su T deplecija (dauguma AML) In the 69 transplant pairs with donor vs recipient NK cell alloreactivity, transplantation from donors with KIR2DS1 and/or KIR3DS1 was associated with reduced risk of non relapse mortality, largely infection related (KIR2DS1 present vs absent: hazard ratio [HR], 0.25; P 0.01; KIR3DS1 present vs absent: HR, 0.18; P 0.006), and better event free survival (KIR2DS1 present vs absent: HR, 0.31; P 0.011; KIR3DS1 present vs absent: HR, 0.30; P 0.008). Transplantation from donors with KIR2DS1 and/or KIR3DS1 was also associated with a 50% reduction in infection rate (P5.003).

45 + 106 pacientai, siblingai, MUD, be T depl.

46 Siūlymas Negiminingi donorai: Jei įmanoma (yra ištipuota), rinktis KIR B haplotipo donorus; ypač didelės rizikos ligoms, AML Haploidentinė transplantacija: Rinktis KIR B haplotipo donorus su didesniu KIR B score jei įmanoma, įvertinant ir kitus donoro pasirinkimo kriterijus

47 Gal aktyvūs donorai tikrai geresni?

48 KIR genotipavimo metodika A. Jakubauskas

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