Variable Number of Tandem Repeats (VNTR) Disparity between Donor and Recipient has a Potential to Predict the Outcomes of HLA-identical Allogeneic Stem Cell Transplantation

Dong Hoon Kwack; Dong Hwan Kim; Shi Nae Kim; Byung Min Ahn; Joon Ho Moon; Yee Soo Chae; Jin Ho Baek; Jong Gwang Kim; Sang Kyun Sohn; Nan Young Lee; Jang Soo Suh; Kyu Bo Lee

doi:10.5045/kjh.2005.40.4.231

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Kwack, Kim, Kim, Ahn, Moon, Chae, Baek, Kim, Sohn, Lee, Suh, and Lee: Variable Number of Tandem Repeats (VNTR) Disparity between Donor and Recipient has a Potential to Predict the Outcomes of HLA-identical Allogeneic Stem Cell Transplantation

Original Article

Korean J Hematol 2005;40(4):231-241.

Published online: 21 December 2005

DOI: https://doi.org/10.5045/kjh.2005.40.4.231

Variable Number of Tandem Repeats (VNTR) Disparity between Donor and Recipient has a Potential to Predict the Outcomes of HLA-identical Allogeneic Stem Cell Transplantation

Dong Hoon Kwack, M.D.¹, Dong Hwan Kim, M.D.^1,², Shi Nae Kim, M.D.¹, Byung Min Ahn, M.D.¹, Joon Ho Moon, M.D.¹, Yee Soo Chae, M.D.¹, Jin Ho Baek, M.D.¹, Jong Gwang Kim, M.D.^1,², Sang Kyun Sohn, M.D.^1,², Nan Young Lee, M.D.², Jang Soo Suh, M.D.², Kyu Bo Lee, M.D.¹

¹Department of Hematology/Oncology, Kyungpook National University Hospital, Daegu, Korea

²Stem Cell Transplantation Center, Kyungpook National University Hospital, Daegu, Korea

Correspondence to: Sang Kyun Sohn, M.D., Ph.D. Department of Oncology/Hematology, Kyungpook National University Hospital 50 Samdeock 2-ga, Jung-gu, Daegu 700-721, Korea Tel: 82-53-420-5587, Fax: 82-53-426-2046 E-mail: sksohn@knu.ac.kr

Received 19 October 2005 Revised 15 November 2005 Accepted 30 October 2005

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

Detection of variable number of tandem repeats (VNTR) between recipient and donor has been adopted to monitor the degree of chimerism after allogeneic stem cell transplantation (SCT). In allogeneic SCT, besides MHC-disparity, the disparity of various polymorphous proteins encoded by several genes may play a critical role in the pathogenesis of graft-versus-host disease (GVHD). However, the biologic effect of VNTR disparity has been scarcely studied.

Methods

We analyzed 84 patients receiving SCT from HLA-identical sibling (n=68) or unrelated donors (n=16). Enrolled diseases included AML 48, ALL 8, CML 15, NHL 10, and high-risk MDS 3. The PCR was performed to amplify 3 VNTR regions (D1S80, D1S111, and D17S5).

Results

We observed strong correlation between the D1S80 disparity and transplant outcomes in terms of OS (P=0.0179) or non-relapse mortality (NRM) (P=0.0305), but not for D1S111 or D17S5 disparity. The D1S80-fully matched pair showed a better OS (72% vs 38%) and lower NRM (17% vs 50%) compared to partially matched or mismatched pairs. In multivariate analyses, D1S80-fully matched pair was found to be independent favorable prognostic factor for OS (P=0.03) or NRM (P=0.05). In addition, the D1S80 disparity was significantly associated with the myeloid engraftment speed (P=0.01) or the occurrence of gut chronic GVHD (P=0.05).

Conclusion

Our data suggest that disparities in D1S80-located on chromosome1-seemed to be associated with increased incidence of gut chronic GVHD and NRMs, thus suggesting the existence of unknown genes of minor histocompatibility antigens targeting gut or cytokine/cytokine receptor on chromosome

Keywords: Variable number of tandem repeats, Allogeneic stem cell transplantation, Graft-versus-host disease

REFERENCES

1). Kim TY, Park SH, Kwon EH, Kim KY, Suh JS, Sohn SK. The discrimination power and effectiveness of 3 kinds of LTR primers in the VNTR-PCR for evaluation of the engraftment of allogeneic peripheral blood stem cell transplantation. Korean J Clin Pathol. 2001; 21:527–33.

2). Kim TY, Park SH, Suh JS, Sohn SK. A case of DNA chimerism analysis as a marker of donor lymphocyte infusion. Korean J Hematol. 2001; 36:342–5.

3). Martinelli G, Trabetti E, Zaccaria A, et al. In vitro amplification of hypervariable DNA regions for the evaluation of chimerism after allogeneic BMT. Bone Marrow Transplant. 1993; 12:115–20.

4). van Leeuwen JE, van Tol MJ, Joosten AM, et al. Relationship between patterns of engraftment in peripheral blood and immune reconstitution after allogeneic bone marrow transplantation for (severe) combined immunodeficiency. Blood. 1994; 84:3936–47.

5). Sreenan JJ, Pettay JD, Tbakhi A, et al. The use of amplified variable number of tandem repeats (VNTR) in the detection of chimerism following bone marrow transplantation. A comparison with restriction fragment length polymorphism (RFLP) by Southern blotting. Am J Clin Pathol. 1997; 107:292–8.

6). Rothberg PG, Gamis AS, Baker D. Use of DNA polymorphisms to monitor engraftment after allogeneic bone marrow transplantation. Clin Lab Med. 1997; 17:109–18.

7). Thiede C. Diagnostic chimerism analysis after allogeneic stem cell transplantation: new methods and markers. Am J Pharmacogenomics. 2004; 4:177–87.

8). Falkenburg JH, van de Corput L, Marijt EW, Willemze R. Minor histocompatibility antigens in human stem cell transplantation. Exp Hematol. 2003; 31:743–51.

9). Chao NJ. Minors come of age: minor histocompatibility antigens and graft-versus-host disease. Biol Blood Marrow Transplant. 2004; 10:215–23.

10). Alcoceba M, Diez-Campelo M, Martin-Jimenez P, et al. Allogeneic transplantation with identical MHC: clinic-pronostic value of discrepances of microsa-tellite DNA regions between recipient and donor. Blood. 2004; 104:912a.

11). Stern M, Meyer-Monard S, Bucher C, et al. Prognostic value of discrepancies in micro-satellite DNA regions between donor and recipient in allogeneic stem cell transplantation. Bone Marrow Transplant. 2005; 35(Suppl 2):S44.

12). Sohn SK, Kim DH, Kim JG, et al. Transplantation outcome in allogeneic PBSCT patients according to a new chronic GVHD grading system, including extensive skin involvement, thrombocytopenia, and progressive-type onset. Bone Marrow Transplant. 2004; 34:63–8.

13). Kim DH, Kim JG, Sohn SK, et al. Clinical impact of early absolute lymphocyte count after allogeneic stem cell transplantation. Br J Haematol. 2004; 125:217–24.

14). Sohn SK, Kim JG, Chae YS, et al. Large-volume leukapheresis using femoral venous access for harvesting peripheral blood stem cells with the Fenwal CS 3000 Plus from normal healthy donors: predictors of CD34+ cell yield and collection efficiency. J Clin Apher. 2003; 18:10–5.

15). Sohn SK, Kim JG, Sung WJ, et al. Harvesting peripheral blood stem cells from healthy donors on 4th day of cytokine mobilization. J Clin Apher. 2003; 18:186–9.

16). Przepiorka D, Weisdorf D, Martin P, et al. 1994 Consensus Conference on Acute GVHD Grading. Bone Marrow Transplant. 1995; 15:825–8.

17). Shulman HM, Sullivan KM, Weiden PL, et al. Chronic graft-versus-host syndrome in man. A longterm clinicopathologic study of 20 Seattle patients. Am J Med. 1980; 69:204–17.

18). Roopenian D, Choi EY, Brown A. The immuno-genomics of minor histocompatibility antigens. Immunol Rev. 2002; 190:86–94.

19). Fuhlbrigge RC, Kieffer JD, Armerding D, Kupper TS. Cutaneous lymphocyte antigen is a specialized form of PSGL-1 expressed on skin-homing T cells. Nature. 1997; 389:978–81.

Fig. 1.

The cumulative incidence of non-relapse mortality (A) and occurrence of gut-involved chronic GVHD (B,C) according to the D1S80 disparity.

Table 1.

Characteristics of the human long tandem repeat markers

Loci	Allele size (bp)	Allele number	PCR primer sequence
D1S80	224～6400	＞29	5'-GTC TTG TTG GAG ATG CAC GTG CCC CTT GC-3'
			5'-GAA ACT GGC CTC CAA ACA CTG CCC GCC G-3'
D1S111	500～1000	＞13	5'-TGT GAG TAG AGG AGA CCT CAC-3'
			5'-AAA GAC CAC AGA GTG AGG AGC-3'
D17S5	200～2000	＞11	5'-GGT CGA AGA GTG AAG TGC ACA G-3'

5'-CAC AGT CTT TAT TCT TCA GCG-3

Table 2.

The transplant outcomes in terms of overall survival, non-relapse mortality or relapse according to the VNTR disparities

Loci	2Y-OS (%)	2Y-NRM (%)	2Y-Relapse (%)
D1S80	0.0179	0.0305	0.2317
Fully matched (n=24, 29%)	72±10	17±9	25±10
Partially matched (n=27, 32%)	37±11	52±12	41±11
Mismatched (n=33, 39%)	38±9	50±10	43±11
D1S111	0.6327	0.6334	0.8000
Fully matched (n=24, 29%)	50±11	35±11	36±11
Partially matched (n=38, 46%)	50±8	40±9	31±8
Mismatched (n=21, 25%)	45±12	46±13	57±16
D17S5	0.8470	0.9814	0.9953
Fully matched (n=37, 47%)	48±9	43±9	37±9
Partially matched (n=17, 22%)	52±12	33±12	40±14
Mismatched (n=24, 31%)	36±12	50±14	39±13

Abbreviations: OS, overall survival; NRM, non-relapse mortality.

Table 3.

Transplantation outcomes according to VNTR disparity for D1S80 loci

Disparity of D1S80	Fully matched pair (n=24, 29%)	Partially matched pair (n=27, 32%)	Mismatched pair (n=33, 39%)	P-value
Follow-up duration (days)	347 (15～2181)	232 (17～1290)	479 (15～2181)
Engraftment
Myeloid	13.0 (10～24)	13.5 (10～25)	16 (10～30)	0.01
Platelet	14 (9～161)	14 (10～56)	17 (0～42)	0.26
Acute GVHD (n=81)	(24)	(26)	(31)
Overall	21 (88)	23 (89)	22 (71)	0.19
≥grade 2	17 (71)	20 (77)	20 (65)	0.59
≥grade 3	4 (17)	7 (27)	8 (26)	0.64
Skin,≥stage 2	12 (50)	14 (54)	15 (48)	0.92
Liver,≥stage 1	4 (17)	9 (31)	13 (42)	0.14
Gut,≥stage 1	14 (58)	16 (62)	17 (55)	0.88
Chronic GVHD (n=71)	(23)	(23)	(25)
Limited+extensive	16 (70)	19 (83)	20 (80)	0.60
Extensive	9 (39)	11 (48)	11 (44)	0.87
Skin involvement	11 (48)	13 (57)	11 (44)	0.68
Hepatic involvement	11 (48)	12 (52)	14 (56)	0.85
Gut involvement	0 (0)	5 (22)	4 (16)	0.05
Infectious events
CMV reactivation	15 (63)	15 (56)	17 (52)	0.71
Infectious events	11 (46)	10 (37)	18 (55)	0.40
Bacterial infections	6 (25)	7 (26)	10 (30)	0.91
Viral infections	6 (25)	3 (11)	10 (30)	0.21
Fungal infections	3 (13)	3 (11)	4 (12)	1.00
Survival
Relapse	8 (33)	10 (26)	6 (29)	0.84
Deaths	7 (29)	15 (56)	21 (64)	0.03
Causes of death
Non-relapse mortalities	4 (17)	11 (41)	15 (45)	0.06^‡
GVHD+/-infection	4 (17)	10 (37)	13 (39)	0.15^§
Others	0 (0)	1 (4)^∗	2 (6)^†	0.78
Progression	3 (12)	4 (15)	6 (18)	0.93

^∗ Others denote veno-occlussive disease (n=1)

^† Others denote veno-occlussive disease (n=1) and hemorrhagic uremic syndrome/thrombotic thrombocytopenic purpura (n=1), respectively.

^‡ P=0.02 when analyzed between D1S80 fully matched pairs versus others including partially matched and mismatched pairs by chi-square test,

^§ P=0.05 when analyzed between D1S80 fully matched pairs versus others including partially matched and mismatched pairs by chi-square test. Abbreviation: GVHD, graft-versus-host disease.

Table 4.

Multivariate survival analysis of the prognostic factor for overall survival and the cumulative incidence of non-relapse mortality or relapse in overall patients

	Risk factor	2-yr rate (%)	HR [95% CI]	P-value
Overall Survival
D1S80 disparity	Fully matched	72±10	1.0	0.03
	Others	38±7	2.217 [1.128～4.826]
Non-Relapse Mortality
D1S80 disparity	Fully matched	17±9	1.0	0.05
	Others	50±8	2.551 [1.000～7.021]
Acute GVHD, ≥gr 3	Grade 0～2	40±7	1.0	0.004
	Grade 3, 4	61±13	3.802 [1.538～9.346]
Relapse
Disease risk	Standard risk	28±8	1.0	0.01
	Advanced risk	57±10	3.155 [1.319～7.576]

For the analysis, the D1S80 disparity (fully matched pair vs partially matched or mismatched pairs), the disease status (standard vs advanced risk), transplanted dose of CD34+ cells (6×10⁶/kg), the donor type (sibling vs unrelated donors) and GVHD (acute GVHD grade 0～2 vs 3,4, and the development of chronic GVHD) were included.

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