Quantification of T-cell receptor excision circle DNA using fluorescence resonance energy transfer and the LightCycler system

Loeffler, Juergen; Bauer, R.; Hebart, Holger; Douek, Daniel C.; Rauser, Georg; Bader, Peter; Einsele, Hermann

doi:10.1016/s0022-1759(02)00337-x

Cited by 107 publications

(146 citation statements)

References 27 publications

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“…The Light Cycler PCR and detection system (Roche Diagnostic, Mannheim, Germany) was used for the amplification and quantification of signal joint TCR excision circles (sjTRECs) as previously described. 5 Primers for the amplification of the sjTRECs sequence were 5 0 -CAC-ATC-CCT-TTC-AAC-CAT-gCT-3 0 (forward) and 5 0 -CCC-Agg-ATg-gAA-AAC-ACA-gTg-3 0 (reverse). For the amplicon detection two different probes were used.…”

Section: Evaluation Of Signal Joint Tcr Excision Circlesmentioning

confidence: 99%

Immune reconstitution after cord blood transplants supported by coinfusion of mobilized hematopoietic stem cells from a third party donor

Martín-Donaire

Rico

Bautista

et al. 2009

Bone Marrow Transplant

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Low severity of GVHD, substantial graft vs tumor (GVT) and slow development of protective immunity are welldocumented features of cord blood transplants (CBT). We have evaluated the immune reconstitution of adult recipients of single-unit CBT supported by the coinfusion of third party donor (TPD) mobilized hematopoietic stem cells (MHSC), a procedure-'dual CB/TPD-MHSC transplant'-that results in early recovery of circulating granulocytes, high rates of CB engraftment and full chimerism. Cumulative recovery of natural killer and B cells at or above the median values of normal controls were 1.0 and 0.76 by the sixth and ninth months. Recovery of T cells was much slower, naive cells lagging behind those of memory and effector (committed) immunophenotypes. Serial analyses of signal joint TCR excision circles showed a general pattern of very low levels by the third month after CBT, followed by recovery to levels persistently similar or higher than those observed before transplantation and in normal controls. Our results are consistent with the clinical observations of substantial GVT effect together with low incidence of serious GVHD and slow development of protective immunity and suggest that thymic function contributes substantially to the recovery of T-cell populations in adults receiving dual CB/TPD-MHSC transplants.

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Section: Evaluation Of Signal Joint Tcr Excision Circlesmentioning

confidence: 99%

Immune reconstitution after cord blood transplants supported by coinfusion of mobilized hematopoietic stem cells from a third party donor

Martín-Donaire

Rico

Bautista

et al. 2009

Bone Marrow Transplant

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“…Briefly, two different oligonucleotides hybridize to an internal species-specific sequence of the 18S rRNA gene of C. albicans. One probe was labeled at the 5Ј end with the Light Cycler Red 640 fluorophore (5Ј-TGG CGA ACC AGG ACT TTT ACT TTG A) (Tibmolbiol), and the other was labeled at the 3Ј end with fluorescein (5Ј-AGC CTT TCC TTC TGG GTA GCC ATT; Tibmolbiol) (20). During fluorescence resonance energy transfer fluorescein is excited by the light source of the Light Cycler instrument.…”

Section: Quantification Of the Number Of Viable C Albicans In The Esmentioning

confidence: 99%

IL-13 Attenuates Gastrointestinal Candidiasis in Normal and Immunodeficient RAG-2−/− Mice via Peroxisome Proliferator-Activated Receptor-γ Activation

Coste

Lagane

Filipe

et al. 2008

The Journal of Immunology

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We recently demonstrated that in vitro peroxisome proliferator-activated receptor-γ (PPARγ) activation of mouse peritoneal macrophages by IL-13 or PPARγ ligands promotes uptake and killing of Candida albicans through mannose receptor overexpression. In this study, we demonstrate that i.p. treatment of immunocompetent and immunodeficient (RAG-2−/−) mice with natural and synthetic PPARγ-specific ligands or with IL-13 decreases C. albicans colonization of the gastrointestinal (GI) tract 8 days following oral infection with the yeast. We also showed that Candida GI infection triggers macrophage recruitment in cecum mucosa. These mucosal macrophages, as well as peritoneal macrophages, overexpress the mannose receptor after IL-13 and rosiglitazone treatments. The treatments promote macrophage activation against C. albicans as suggested by the increased ability of peritoneal macrophages to phagocyte C. albicans and to produce reactive oxygen intermediates after yeast challenge. These effects on C. albicans GI infection and on macrophage activation are suppressed by treatment of mice with GW9662, a selective PPARγ antagonist, and are reduced in PPARγ+/− mice. Overall, these data demonstrate that IL-13 or PPARγ ligands attenuate C. albicans infection of the GI tract through PPARγ activation and hence suggest that PPARγ ligands may be of therapeutic value in esophageal and GI candidiasis in immunocompromised patients.

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“…Primers described previously 32 were used to amplify a conserved region of the fungal 18 s rRNA gene. Panfungal oligonucleotide hybridisation probes (3FL-TTC AAC TAC gAg CTT TTT AAC Tg and LC Red640-AAC AAC TTT AAT ATA CgC TAT Tgg A (Tib MOLBIOL, Berlin, Germany)) (Loeffler, personal communication, 2000) were used to bind to a common sequence present in all fungal species.…”

Section: Pcr Assaymentioning

confidence: 99%

A prospective study of real-time panfungal PCR for the early diagnosis of invasive fungal infection in haemato-oncology patients

Jordanides

Allan

McLintock

et al. 2005

Bone Marrow Transplant

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Summary:A blinded prospective study was performed to determine whether screening of whole blood using a real-time, panfungal polymerase chain reaction (PCR) technique could predict the development of invasive fungal infection (IFI) in immunocompromised haemato-oncology patients. In all, 78 patients (125 treatment episodes) were screened twice weekly by real-time panfungal PCR using LightCyclert technology. IFI was documented in 19 treatment episodes (five proven, three probable and 11 possible), and in 12, PCR was sequentially positive. PCR positivity occurred in: 4/5 proven; 2/3 probable; 6/11 possible; and 29/106 with no IFI. In 8/12 with IFI and sequentially positive PCR results, PCR positivity occurred before (median 19.5 days) and in 4/12 (median 10.5 days) after the initiation of empirical antifungal therapy. Based on sequential positive results for proven/probable IFI sensitivity, specificity, positive predictive value and negative predictive value were 75, 70, 15 and 98%, respectively. Real-time panfungal PCR is a sensitive tool for the early diagnosis of IFI in immunocompromised haemato-oncology patients. It may be most useful as a screening method in high-risk patients, either to direct early pre-emptive antifungal therapy or to determine when empirical antifungal therapy can be withheld in patients with antibiotic-resistant neutropenic fever. However, these strategies require further assessment in comparative clinical trials.

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Quantification of T-cell receptor excision circle DNA using fluorescence resonance energy transfer and the LightCycler system

Cited by 107 publications

References 27 publications

Immune reconstitution after cord blood transplants supported by coinfusion of mobilized hematopoietic stem cells from a third party donor

Immune reconstitution after cord blood transplants supported by coinfusion of mobilized hematopoietic stem cells from a third party donor

IL-13 Attenuates Gastrointestinal Candidiasis in Normal and Immunodeficient RAG-2−/− Mice via Peroxisome Proliferator-Activated Receptor-γ Activation

A prospective study of real-time panfungal PCR for the early diagnosis of invasive fungal infection in haemato-oncology patients

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