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Donor Chimerism of B Cells and Nature Killer Cells Provides Useful Information to Predict Hematologic Relapse following Allogeneic Hematopoietic Stem Cell Transplantation.

Jiang Y, Wan L, Qin Y, Wang X, Yan S, Xie K, Wang C - PLoS ONE (2015)

Bottom Line: A total of 33 patients (33/153, 21.6%) had recurrent disease.Only the donor chimerism in BMB significantly decreased in the group with hematologic relapse as compared with the group without hematologic relapse (P=0.00, Independent-samples T test) in patients with B-ALL.Therefore, our results reveal a more effective path to individually predict for hematologic relapse by dynamic monitoring different cell lineages in different disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Shanghai First People's Hospital, Medical College, Shanghai Jiaotong University, Shanghai, China.

ABSTRACT
In this study we investigated the correlation between donor chimerism status and disease relapse following allogeneic hematopoietic stem cell transplantation (allo-HSCT). The chimerism of Fluorescence-activated cell sorter (FACS) sorted CD3+T lymphocytes of 153 cases, CD56+CD16+NK lymphocytes of 153 cases and CD19+B lymphocytes of 31 cases with acute B lymphoblastic leukemia (B-ALL) was analyzed post-transplant utilizing polymerase chain reaction amplification of short tandem repeats (PCR-STR). A total of 33 patients (33/153, 21.6%) had recurrent disease. The positive predictive values of declining donor chimerism for hematologic and isolated extramedullary relapse were 58.8% and 10% (P=0.018, Chi-Square). The positive predictive values of declining donor chimerism in BMB, BMT, BMNK and PBB for hematologic relapse were 11.6%, 0%, 0% and 0% under close monitoring in patients with B-ALL. Only the donor chimerism in BMB significantly decreased in the group with hematologic relapse as compared with the group without hematologic relapse (P=0.00, Independent-samples T test) in patients with B-ALL. The median drop of donor chimerism in PBT, BMT, PBNK and BMNK were 0%, 0%, 5.9% and 2.8% one or two weeks prior to hematologic relapse in patients with non-B-ALL. The donor chimerism in PBNK significantly decreased prior to hematologic relapse in the group with hematologic relapse as compared with the group without hematologic relapse (P=0.022, Independent-samples T test).These data suggest donor chimerism of BMB can be used to predict the occurrence of hematologic relapse in patients with B-ALL. Donor chimerism decrease in PBNK was associated with a somewhat increased risk of hematologic relapse in patients with non-B-ALL. Therefore, our results reveal a more effective path to individually predict for hematologic relapse by dynamic monitoring different cell lineages in different disease.

No MeSH data available.


Related in: MedlinePlus

Calculation of peak area in chimerism.The first line is the samples of the donor. The second line is the samples of the recipient prior to transplantation. The third line is the samples examined following transplantation. The fourth line is the calculation formula of peak area in chimerism.
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pone.0133671.g001: Calculation of peak area in chimerism.The first line is the samples of the donor. The second line is the samples of the recipient prior to transplantation. The third line is the samples examined following transplantation. The fourth line is the calculation formula of peak area in chimerism.

Mentions: We did operate according to the protocols of commercial kits. Genomic DNA which isolated from all cell subsets sorted by FACS was extracted by ReadyAmp Genomic DNA Purification System Kit (Promega). The final concentration of DNA was 0.5ng/ul. One day was the maximum time between FACS sorting and DNA extraction. Amplifications were performed with the AmpFlSTR Profiler Plus Kit (Applied Biosystems, USA) on an ABI 7300 Fast Real-Time PCR System. All Multiplex PCR reactions were completed in 20μl volume. The volume/quantity of DNA was 5ul/2.5ng. Cycle conditions were as follows: 95°C for 11 minutes followed by 28 cycles of 94°C for 1 minute,58°C for 1 minute,72°C for 1 minute, and a final step consisting of 45 minutes at 60°C. The PCR products were analyzed using capillary electrophoresis on ABI PRISM 3130 Genetic Analyzer. The GeneMapper v4.0 provided a complete STR genotyping. Serial samples were analyzed for degrees of donor-recipient chimerism using PCR of informative minisatellite regions [13,14]. PCR products were detected on ten alleles: D3S1358、vWA、FGA、Amelogenin、D8S1179、D21S11、D18S51、D5S818、D13S317 and D7S820. The loci on ten alleles of every patient were detected in donor and recipient samples before transplantation. The informative loci were selected from which had the difference between donor and recipient. Calculation of peak area in chimerism is as follows (Fig 1).


Donor Chimerism of B Cells and Nature Killer Cells Provides Useful Information to Predict Hematologic Relapse following Allogeneic Hematopoietic Stem Cell Transplantation.

Jiang Y, Wan L, Qin Y, Wang X, Yan S, Xie K, Wang C - PLoS ONE (2015)

Calculation of peak area in chimerism.The first line is the samples of the donor. The second line is the samples of the recipient prior to transplantation. The third line is the samples examined following transplantation. The fourth line is the calculation formula of peak area in chimerism.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4520605&req=5

pone.0133671.g001: Calculation of peak area in chimerism.The first line is the samples of the donor. The second line is the samples of the recipient prior to transplantation. The third line is the samples examined following transplantation. The fourth line is the calculation formula of peak area in chimerism.
Mentions: We did operate according to the protocols of commercial kits. Genomic DNA which isolated from all cell subsets sorted by FACS was extracted by ReadyAmp Genomic DNA Purification System Kit (Promega). The final concentration of DNA was 0.5ng/ul. One day was the maximum time between FACS sorting and DNA extraction. Amplifications were performed with the AmpFlSTR Profiler Plus Kit (Applied Biosystems, USA) on an ABI 7300 Fast Real-Time PCR System. All Multiplex PCR reactions were completed in 20μl volume. The volume/quantity of DNA was 5ul/2.5ng. Cycle conditions were as follows: 95°C for 11 minutes followed by 28 cycles of 94°C for 1 minute,58°C for 1 minute,72°C for 1 minute, and a final step consisting of 45 minutes at 60°C. The PCR products were analyzed using capillary electrophoresis on ABI PRISM 3130 Genetic Analyzer. The GeneMapper v4.0 provided a complete STR genotyping. Serial samples were analyzed for degrees of donor-recipient chimerism using PCR of informative minisatellite regions [13,14]. PCR products were detected on ten alleles: D3S1358、vWA、FGA、Amelogenin、D8S1179、D21S11、D18S51、D5S818、D13S317 and D7S820. The loci on ten alleles of every patient were detected in donor and recipient samples before transplantation. The informative loci were selected from which had the difference between donor and recipient. Calculation of peak area in chimerism is as follows (Fig 1).

Bottom Line: A total of 33 patients (33/153, 21.6%) had recurrent disease.Only the donor chimerism in BMB significantly decreased in the group with hematologic relapse as compared with the group without hematologic relapse (P=0.00, Independent-samples T test) in patients with B-ALL.Therefore, our results reveal a more effective path to individually predict for hematologic relapse by dynamic monitoring different cell lineages in different disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Shanghai First People's Hospital, Medical College, Shanghai Jiaotong University, Shanghai, China.

ABSTRACT
In this study we investigated the correlation between donor chimerism status and disease relapse following allogeneic hematopoietic stem cell transplantation (allo-HSCT). The chimerism of Fluorescence-activated cell sorter (FACS) sorted CD3+T lymphocytes of 153 cases, CD56+CD16+NK lymphocytes of 153 cases and CD19+B lymphocytes of 31 cases with acute B lymphoblastic leukemia (B-ALL) was analyzed post-transplant utilizing polymerase chain reaction amplification of short tandem repeats (PCR-STR). A total of 33 patients (33/153, 21.6%) had recurrent disease. The positive predictive values of declining donor chimerism for hematologic and isolated extramedullary relapse were 58.8% and 10% (P=0.018, Chi-Square). The positive predictive values of declining donor chimerism in BMB, BMT, BMNK and PBB for hematologic relapse were 11.6%, 0%, 0% and 0% under close monitoring in patients with B-ALL. Only the donor chimerism in BMB significantly decreased in the group with hematologic relapse as compared with the group without hematologic relapse (P=0.00, Independent-samples T test) in patients with B-ALL. The median drop of donor chimerism in PBT, BMT, PBNK and BMNK were 0%, 0%, 5.9% and 2.8% one or two weeks prior to hematologic relapse in patients with non-B-ALL. The donor chimerism in PBNK significantly decreased prior to hematologic relapse in the group with hematologic relapse as compared with the group without hematologic relapse (P=0.022, Independent-samples T test).These data suggest donor chimerism of BMB can be used to predict the occurrence of hematologic relapse in patients with B-ALL. Donor chimerism decrease in PBNK was associated with a somewhat increased risk of hematologic relapse in patients with non-B-ALL. Therefore, our results reveal a more effective path to individually predict for hematologic relapse by dynamic monitoring different cell lineages in different disease.

No MeSH data available.


Related in: MedlinePlus