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. 2009 Sep;127(5):251-8.
doi: 10.1590/s1516-31802009000500002.

Prevalence of chimerism after non-myeloablative hematopoietic stem cell transplantation

Azulamara da Silva Ruiz  1 , Maria de Lourdes Ferrari Chauffaille, Solivanda Trindade Alves, José Salvador Rodrigues de Oliveira
Affiliations

Prevalence of chimerism after non-myeloablative hematopoietic stem cell transplantation

Azulamara da Silva Ruiz et al. Sao Paulo Med J. 2009 Sep.

Abstract
in English, Portuguese

Context and objective: Non-myeloablative hematopoietic stem cell transplantation (NMA-HSCT) is performed in onco-hematological patients who cannot tolerate ablative conditioning because of older age or comorbidities. This approach does not completely eliminate host cells and initially results in mixed chimerism. Long-term persistence of mixed chimerism results in graft rejection and relapse. Involvement of graft-versus-host disease is concomitant with complete chimerism and graft-versus-tumor effect. The aim of this study was to evaluate the prevalence of chimerism in onco-hematological patients who underwent NMA-HSCT.

Design and setting: Observational clinical study on chimerism status after human leukocyte antigen-identical NMA-HSCT at the Discipline of Hematology and Hemotherapy of Universidade Federal de São Paulo.

Methods: We sequentially analyzed the amplification of APO-B, D1S80, DxS52, FVW, 33.6, YNZ-2 and H-ras primers using variable number of tandem repeats (VNTR) on 17 pairs and fluorescent in situ hybridization (FISH) with the XY probe and SRY primer on 13 sex-unmatched pairs.

Results: The informativeness of the primers using VNTR was 60% for APO-B, 75% D1S80, 36% DxS52, 14% FVW, 40% YNZ-22 and 16% H-ras. The SRY primer was informative in female receptors with male donors. The XY-FISH method was informative in 100% of the sex-unmatched pairs.

Conclusion: These methods were sensitive and informative. In VNTR, the association of APO-B with D1S80 showed 88% informativeness. The quantitative FISH method was more sensitive, but had the disadvantage of only being used for sex-unmatched pairs.

CONTEXTO E OBJETIVO:: O transplante de células hematopoiéticas não-mieloablativo é realizado em pacientes com doenças onco-hematológicas que não suportam condicionamentos ablativos devido à elevada idade ou ao acometimento por comorbidades. Esta abordagem não elimina completamente as células do hospedeiro, resultando, inicialmente, em quimerismo misto. A persistência do quimerismo misto na evolução de longo prazo resulta na rejeição ao enxerto e recaída. O acometimento pela doença do enxerto contra hospedeiro é concomitante ao quimerismo completo e ao efeito enxerto versus tumor. O objetivo deste estudo foi avaliar a prevalência do quimerismo em doenças onco-hematológicas tratadas com o transplante não-mieloablativo de células hematopoiéticas.

TIPO DE ESTUDO E LOCAL:: Estudo clínico observacional do estado de quimerismo após transplante antígenos leucocitários humanos-idêntico não-mieloabaltivo realizado na Disciplina de Hematologia e Hemoterapia da Universidade Federal de São Paulo.

MÉTODOS:: Analisamos sequencialmente a amplificação dos primers APO-B, D1S80, DxS52, FVW, 33,6, YNZ-22, H-ras pelo VNTR (variable number of tandem repeats) em 17 pares e FISH (fluorescent in situ hybridization) pela sonda XY e do primer SRY em 13 pares de não relacionados a sexo.

RESULTADO:: A informatividade dos primers pelo VNTR foi de 60% para APO-B; 75% D1S80; 36% DxS52; 14% FVW; 40% YNZ-22 e 16% H-ras. O primer SRY foi informativo em receptores femininos com doadores masculinos. O método XY-FISH foi informativo em 100% dos pares de não relacionado a sexo.

CONCLUSÃO:: Estes métodos foram sensíveis e informativos. No VNTR, a associação do APO-B com D1S80 mostrou 88% de informatividade. O FISH, método quantitativo, foi mais sensível, porém com desvantagem de ser usado somente nos pares não relacionados a sexo.

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Conflict of interest statement

Conflict of interest: Not declared

Figures

Figure 1
Figure 1. Evaluation using 2% agarose gel electrophoresis marker D1S80 following polymerase chain reaction (PCR) amplification (variable number of tandem repeats, VNTR technique) in case 7. (1) donor product; (2) patient product before transplantation; (3) and (4) donor and patient products after transplantation; (5) and (6) patient productsafter transplantation and after molecular relapse; (7) 100-base pair marker.
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