Quantitative assessment of hematopoietic chimerism after bone marrow transplantation by real-time quantitative polymerase chain reaction

Alizadeh, Mehdi; Bernard, Marc; Danic, Bruno; Dauriac, Charly; Birebent, Brigitte; Lapart, Christine; Lamy, Thierry; Prise, P. Y. Le; Beauplet, A.; Bories, Dominique; Sémana, G.; Quelvennec, Erwann

doi:10.1182/blood.v99.12.4618

Cited by 327 publications

(345 citation statements)

References 28 publications

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“…All SNPs used have been tested and validated at our laboratory. As the validation of the method [16] showed loss of precision at levels lower than 1/1,000, we have chosen to present values lower than this as <1/1,000. The sensitivity is equal to that of the nested PCR method.…”

Section: Methodsmentioning

confidence: 99%

“…The second method, which is quantitative, was based on single nucleotide polymorphisms (SNP) markers and the analysis involved real-time PCR using TaqMan technology (Applied Biosystems, Foster City, California, www.appliedbiosystems.com) as described by Alizadeh et al [16]. Patient, HSCT-, and MSC-donor DNA samples were screened with different SNPs.…”

Section: Methodsmentioning

confidence: 99%

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Analysis of Tissues Following Mesenchymal Stromal Cell Therapy in Humans Indicates Limited Long-Term Engraftment and No Ectopic Tissue Formation

et al. 2012

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Mesenchymal stromal cells (MSCs) are explored as a novel treatment for a variety of medical conditions. Their fate after infusion is unclear, and long-term safety regarding malignant transformation and ectopic tissue formation has not been addressed in patients. We examined autopsy material from 18 patients who had received human leukocyte antigen (HLA)-mismatched MSCs, and 108 tissue samples from 15 patients were examined by PCR. No signs of ectopic tissue formation or malignant tumors of MSC-donor origin were found on macroscopic or histological examination. MSC donor DNA was detected in one or several tissues including lungs, lymph nodes, and intestine in eight patients at levels from 1/100 to <1/1,000. Detection of MSC donor DNA was negatively correlated with time from infusion to sample collection, as DNA was detected from nine of 13 MSC infusions given within 50 days before sampling but from only two of eight infusions given earlier. There was no correlation between MSC engraftment and treatment response. We conclude that MSCs appear to mediate their function through a “hit and run” mechanism. The lack of sustained engraftment limits the long-term risks of MSC therapy.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Analysis of Tissues Following Mesenchymal Stromal Cell Therapy in Humans Indicates Limited Long-Term Engraftment and No Ectopic Tissue Formation

et al. 2012

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“…The level of 1% was chosen, as the microsatellites and variable numbers of tandem repeat methods have a sensitivity limit of 1-5%, whereas the real-time PCR method has a sensitivity of 0.01%. 15 MC was defined as having a recipient fraction of between 1 and 99%. Patients with MC were also divided into three subgroups: (1) those with high MC, defined as having 430% recipient DNA in two consecutive samples, (2) those with stable MC, with consistent recipient levels of o30% and (3) those with decreasing MC, when an initial stable MC turned into DC.…”

Section: Chimerism Definitionmentioning

confidence: 99%

Allogeneic hematopoietic SCT in patients with non-malignant diseases, and importance of chimerism

Svenberg

Mattsson

Ringdén

et al. 2009

Bone Marrow Transplant

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In this retrospective study, we evaluated the chimerism status and outcome in 58 patients (64 transplants) with non-malignant diseases. Reduced intensity conditioning (RIC) was given in half of the transplants. Mixed chimerism (MC) was defined as 41% recipient cells. Two consecutive samples showing 430% recipient cells were defined as high chimerism (high MC). Patients with high MC and the management of these patients were analyzed in greater detail. The overall survival rate was 87%. In total, 23 transplants were donor chimerism (DC) and 41 transplants showed some degree of MC. The incidence of MC was 78 and 50% after RIC and myeloablative conditioning, respectively (P ¼ 0.04). Acute GVHD of grades II-III was more common in patients with DC (39%) than in patients with MC (8%) (P ¼ 0.002). Owing to high MC, donor lymphocyte infusions were given in 17 cases. The level of MC was reduced in seven cases, unchanged in four cases, increased in one case and there was graft rejection in five cases. A second transplant was carried out in six cases with rejections, five are alive and in remission. We conclude that patients with non-malignant diseases, who develop MC after transplant have less acute GVHD. Despite the high incidence of MC, overall survival is promising.

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“…In addition, previous assessment of genomic copy number in cancer is performed either at a relatively low resolution by comparative genomic hybridisation (CGH) (5-10 Mb) 11,12 or at higher resolution by laborious individual locus specific techniques that can analyse only a few loci simultaneously. 13 Array-based CGH is a sensitive and a high-throughput technique but is technically challenging, requires expensive equipment and uses 1 sample at a time. 14,15 Multiplex Amplifiable Probe Hybridisation (MAPH) provides an alternative DNA-based approach for detecting and quantifying genomic copy number alterations in tumours.…”

mentioning

confidence: 99%

High‐resolution analysis of 16q22.1 in breast carcinoma using DNA amplifiable probes (multiplex amplifiable probe hybridization technique) and immunohistochemistry

Rakha

Armour

Pinder

et al. 2005

Intl Journal of Cancer

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Loss of the chromosomal material at 16q22.1 is one of the most frequent genetic aberrations found in both lobular and low-grade nonlobular invasive carcinoma of the breast, indicating the presence of a tumour suppressor gene (TSG) at this region in these tumours. However, the TSG (s) at the 16q22.1 in the more frequent nonlobular carcinomas is still unknown. Multiplex Amplifiable Probe Hybridisation (MAPH) is a simple, accurate and a high-resolution technique that provides an alternative approach to DNA copy-number measurement. The aim of our study was to examine the most likely candidate genes at 16q22.1 using MAPH assay combined with protein expression analysis by immunohistochemistry. We identified deletion at 16q22.1 that involves some or all of these genes. We also noticed that the smallest region of deletion at 16q22.1 could be delineated to a 3 Mb region centromeric to the P-cadherin gene. Apart from the correlation between E-cadherin protein expression and its gene copy number, no correlation was detected between the expression of E2F-4, CTCF, TRF2 or P-cadherin with their gene's copy number. In the malignant tissues, no significant loss or decrease of protein expression of any gene other than E-cadherin was seen in association with any specific tumour type. No expression of VE-cadherin or Ksp-cadherin was detected in the normal and/or malignant tissues of the breast in these cases. However, there was a correlation between increased nuclear expression of E2F-4 and tumours with higher histological grade (p ‫؍‬ 0.04) and positive lymph node disease (p ‫؍‬ 0.02), suggesting that it may have an oncogenic rather than a tumour suppressor role. The malignant breast tissues also showed abnormal cytoplasmic cellular localisation of CTCF, compared to its expression in the normal parenchymal cells. In conclusion, we have demonstrated that MAPH is a potential technique for assessment of genomic imbalances in malignant tissues. Although our results support E-cadherin as the TSG in invasive lobular carcinoma, they argue against the candidacy of E2F-4, CTCF, TRF2, P-cadherin, Ksp-cadherin and VE-cadherin as TSGs in breast cancer.

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Quantitative assessment of hematopoietic chimerism after bone marrow transplantation by real-time quantitative polymerase chain reaction

Cited by 327 publications

References 28 publications

Analysis of Tissues Following Mesenchymal Stromal Cell Therapy in Humans Indicates Limited Long-Term Engraftment and No Ectopic Tissue Formation

Analysis of Tissues Following Mesenchymal Stromal Cell Therapy in Humans Indicates Limited Long-Term Engraftment and No Ectopic Tissue Formation

Allogeneic hematopoietic SCT in patients with non-malignant diseases, and importance of chimerism

High‐resolution analysis of 16q22.1 in breast carcinoma using DNA amplifiable probes (multiplex amplifiable probe hybridization technique) and immunohistochemistry

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