Comparação da utilização de Short Tandem Repeats e do gene da amelogenina na avaliação do quimerismo nos doentes pós transplante alogénico de progenitores hematopoiéticos
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2016-12-15
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A análise de quimerismo, nos doentes pós transplante alogénico de progenitores hematopoiéticos, tornou-se um procedimento de rotina para avaliar o sucesso do transplante e na deteção de falência do enxerto. Após alotransplante o doente apresenta quimerismo completo, quando apenas têm células do dador ou quimerismo misto, quando têm células próprias e do dador. Para a análise de quimerismo, as células do dador e do doente são distinguidas por marcadores genéticos designados como short tandem repeats informativos, que são utilizados para efetuar o seguimento dos doentes no período pós transplante.
Este trabalho teve como objetivos monitorizar o estado de quimerismo em doentes pós alotransplante, e comparar duas técnicas moleculares para avaliação do estado de quimerismo: amplificação de short tandem repeats, e amplificação do gene da amelogenina em pares dador/doente de sexos diferentes, pela técnica de reacção em cadeia da polimerase seguida de electroforese capilar. Os short tandem repeats utilizados foram: D11s, F131, THO, VWA, FES, FGA, HPRT. A análise foi realizada através de ADN extraído do sangue, nas subpopulações de leucócitos - células mononucleares e granulócitos.
No presente estudo, foi analisado o quimerismo de 24 doentes alotransplantados, o seguimento médio destes doentes foi de 6 meses. O quimerismo foi analisado pela amplificação de short tandem repeats (n=24) e do gene da amelogenina (n=14). Os resultados obtidos para a análise de quimerismo foram: 20 doentes apresentaram sempre quimerismo completo e 4 doentes quimerismo misto. Dos doentes com quimerismo misto, 2 evoluíram para quimerismo completo e 2 mantiveram o mesmo estado.
Foi possível concluir que, para a avaliação do quimerismo dos doentes, ambas as técnicas, short tandem repeats e gene da amelogenina, são informativas e podem ser realizadas separadamente ou de uma forma complementar.
Chimerism analysis after allogeneic hematopoietic stem cell transplantation has become a routine procedure, for monitoring the engraftment of donor cells, thus allowing early detection of graft failure. After allogeneic stem cell transplantation the patient has complete chimerism when only has donor cells, or mixed chimerism when he has his own and the donor’s cells. To the chimerism analysis, donor cells and patient’s cells are distinguished by genetic markers, named informative STR. They are used to perform the following of patients in the post-transplant period. The aim of this study were to monitor chimerism status in patients following allogeneic stem cell transplantation, and to compare two molecular techniques to assess the state of chimerism: amplification of short tandem repeats and amplification of amelogenin gene in pairs donor/patient of different gender by polymerase chain reaction technique. The analysis was performed using DNA extracted from blood leukocyte subpopulations - mononuclear cells and granulocytes. The short tandem repeats used were: D11s, F131, THO, VWA, FES, FGA, HPRT and the amelogenin gene, were amplified by the polymerase chain reaction, followed by capillary electrophoresis analysis. In this study, we performed follow up evaluation of 24 patients submitted to allogeneic hematopoietic stem cell transplantation. The median follow up of these patients was 6 months. The chimerism was analyzed by amplification of short tandem repeats (n=24) and amelogenin gene (n=14). Majority of patients (n=20) consistently showed complete chimerism and 4 patients showed mixed chimerism. From the patients with mixed chimerism, 2 progressed to complete chimerism and 2 remained in the same condition. In summary, both techniques, short tandem repeats and amelogenin gene, are informative and can be performed individually or in a complementary way to evaluate the chimerism status of allotransplanted patients.
Chimerism analysis after allogeneic hematopoietic stem cell transplantation has become a routine procedure, for monitoring the engraftment of donor cells, thus allowing early detection of graft failure. After allogeneic stem cell transplantation the patient has complete chimerism when only has donor cells, or mixed chimerism when he has his own and the donor’s cells. To the chimerism analysis, donor cells and patient’s cells are distinguished by genetic markers, named informative STR. They are used to perform the following of patients in the post-transplant period. The aim of this study were to monitor chimerism status in patients following allogeneic stem cell transplantation, and to compare two molecular techniques to assess the state of chimerism: amplification of short tandem repeats and amplification of amelogenin gene in pairs donor/patient of different gender by polymerase chain reaction technique. The analysis was performed using DNA extracted from blood leukocyte subpopulations - mononuclear cells and granulocytes. The short tandem repeats used were: D11s, F131, THO, VWA, FES, FGA, HPRT and the amelogenin gene, were amplified by the polymerase chain reaction, followed by capillary electrophoresis analysis. In this study, we performed follow up evaluation of 24 patients submitted to allogeneic hematopoietic stem cell transplantation. The median follow up of these patients was 6 months. The chimerism was analyzed by amplification of short tandem repeats (n=24) and amelogenin gene (n=14). Majority of patients (n=20) consistently showed complete chimerism and 4 patients showed mixed chimerism. From the patients with mixed chimerism, 2 progressed to complete chimerism and 2 remained in the same condition. In summary, both techniques, short tandem repeats and amelogenin gene, are informative and can be performed individually or in a complementary way to evaluate the chimerism status of allotransplanted patients.
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Dissertação de Mestrado em Bioquímica
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Bioquímica , Quimerismo , Amelogenina , Transplante alogénico , Progenitores hematopoiéticos