| ?Objectives?To analyze the distribution and proportion of donor-specific aKIR genes and their immune reconstruct regularity profile.Then discuss the clinical significance of dynamically detecting the donor-specific aKIR genes'expression after unrelated allogeneic hematopoietic stem cell transplantation?allo-HSCT?.?Method?216 pairs of donors and recipients were performed KIR genotyping using sequence specific primers PCR?PCR-SSP?methods.Dynamically detected the mRNA expression of donor-specific KIRs for 86 cases of patients at different time points using real-time fluorescent quantitation PCR?RT-qPCR?methods after allo-HSCT,being defined as KIRs'copies per 10000 ABL copies in all nucleated cells.Analyzed the immune reconstruct regularity profile of donor-specific KIRs.And at the same time,combined with the clinical data,research the clinical significance of donor-specific KIR genes'expression after allo-HSCT.?Result?PART I Discussion about the distribution and the regularity of donor-specific activating KIR after unrelated allo-HSCT1)The frequency of donor-specific KIR genes was 53.7%?116/216?in the cohort,including 78.3%?112/143?in the KIR genes mismatched group and 5.5%?4/73?in matched group.And 99.1%?115/116?contained activating donor-specific KIR genes.2)Among 55 pairs of donors'KIR Bx genotype and patients'KIR AA genotype group,the most commonly observed genotypes were Bx1,Bx2,Bx3,Bx4,in which the donor-specific KIR genes were respectively KIR 3DS1,2DL5A,2DS5,2DS1;KIR 3DS1,2DL5A,2DS3,2DS1;KIR 2DS2,2DL2;KIR 2DS2,2DL2,3DS1,2DL5A,2DS5,2DS1.3)Among 44 pairs of donors'KIR AA genotype and patients'KIR AB genotype group,there was no donor-specific KIR for post transplant follow-up in addition to the detection of donor specific KIR 2DS4?FUL?,because of the common haplotype A.36.4%?16/44?recipients may have donor-specific KIR 2DS4?FUL?gene.4)In KIR mismatched group,the proportion of donor-specific aKIR genes were 2DS135.7%?51/143?,3DS1 32.9%?47/143?,2DS5 29.4%?42/143?,2DS4?FUL?25.9%?37/143?,2DS2 24.5%?35/143?,2DS3 21.7%?31/143?;The proportion of donor-specific inhibitory killer cell immunoglobulin-like receptor?iKIR?genes were 2DL2 25.2%?36/143?,3DL18.4%?12/143?.PART II The immune reconstruct regularity profile of donor-specific activating and inhibitory KIR after unrelated allo-HSCT1)The mRNA expression of donor-specific aKIR 2DS1,2DS2,2DS3,2DS4,2DS5,3DS1:On the day of transplantation,1M,2M and 3M after unrelated allo-HSCT,the mRNA expression of donor-specific 2DS1 were 103.65,104.26,109.49,140.63;The mRNA expression of donor-specific 2DS2 were 64.17,118.87,163.85,281.72;The m RNA expression of donor-specific 2DS3 were 9.65,19.76,30.16,23.25;The mRNA expression of donor-specific 2DS4 were 100.42,191.95,193.97,311.49;The mRNA expression of donor-specific 2DS5 were 107.11,111.45,195.77,179.84;The mRNA expression of donor-specific 3DS1 were 2896.24,3612.32,5136.43,6905.50.In the 2nd mouth after transplantation,KIR 2DS2,2DS3 mRNA expression was higher than the donors'on the day of transplantation,showed statistical difference?P=0.026,P=0.022?.And in the 3rd mouth,the donor-specific 2DS2,3DS1 mRNA expression was also showed statistical difference,compared with the donors'on the day of transplantation?P=0.032,P=0.039?.2)The mRNA expression of donor-specific iKIR 2DL2,3DL1:On the day of transplantation,1M,2M and 3M after unrelated allo-HSCT,the mRNA expression of2DL2 were 48.26,106.67,153.66,134.98;The mRNA expression of 3DL1 were 627.50,2031.52,1461.85,1436.07.2DL2,3DL1 mRNA expression was higher than the donors'on the day of transplantation in the 1stmouth after transplantation,showed statistical difference?P=0.027,P=0.035?.And in the next two mouths,the donor-specific 2DL2mRNA expression was also showed statistical difference,compared with the donors'on the day of transplantation?P=0.004,0.005?.3)Among donor-specific aKIR genes,the m RNA expression level of 2DS1,2DS2,2DS4 and 3DS1 at 3M and 2DS3,2DS5 at 2M after transplantation was higher than that of donor on the day of transplantation.And at the same time,the m RNA expression level of the donor-specific 2DL2/2DS2 genes respectively at 2M and 3M after transplantation exceeded the level of donor on the transplantation day.And the mRNA expression level of the donor-specific 3DL1/3DS1 genes respectively at 1M and 3M after transplantation exceeded the level of donor on the transplantation day.All donor-specific aKIR and iKIR genes achieved immune reconstitution after transplantation,but the reconstitution time of aKIR genes was later than that of the iKIR genes.PART III the clinical significance of dynamically detecting the donor-specific aKIR genes'expression after unrelated allo-HSCT1)Overall survival rate?OS?of the donor-recipient HLA-10/10 matched group was higher than the donor-recipient HLA?7/10 mismatched group,but showed no statistical difference?P=0.126?.Overall survival rate?OS?of donors for KIR-Bx genotype was higher than the donor for KIR-AA genotype,showed no statistical difference?P=0.592?.And in the HLA-10/10 matched group the aGVHD incidence rate of donors for KIR-AA genotype was higher than the donor for KIR-Bx genotype,both compared statistically significant?P=0.035?.2)In HLA-10/10 matched group,the mRNA expression of donor-specific KIR 2DS2,2DS3,2DS5 in the 2nd and 3rd mouths after transplantation the expression with aGVHD were higher than without aGVHD.The m RNA expression of donor-specific KIR 2DS4with aGVHD was higher than without aGVHD in the 3rd mouth.But all showed no statistical significance.?Conclusion?The donor-specific aKIR genes mainly existed in KIR mismatched group after unrelated allo-HSCT and there were higher specific aKIRs in donor KIR AA,Bx1,Bx2,Bx3,Bx4 genotypes with higher frequency.The donor-specific aKIR and iKIR genes exhibit different patterns of immune reconstitution after transplantation.Therefore,the dynamical detection of donor-specific KIR genes after transplantation can be used as a new predictor of clinical outcome. |
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