NCT00499070

Assessing Immune Function in Young Patients With Cytopenia That Did Not Respond to Treatment

TCR Vbeta Repertoire and PNH Clones in Children With Refractory Cytopenia (RC). An Open Nonrandomised Multi-Center Prospective Study

• Status: Completed
• Phase: —
• Study type: Observational
• Enrollment: 119 (actual)
• Sponsor: University Hospital Freiburg · Academic / Other
• Sex: All
• Age: 17 Years
• Healthy volunteers: Not accepted

Summary

RATIONALE: Studying biopsy, bone marrow, and blood samples from patients with cytopenia that did not respond to treatment may help doctors learn more about the disease and plan the best treatment. PURPOSE: This laboratory study is assessing immune function in young patients with cytopenia that did not respond to treatment.

Detailed description

OBJECTIVES: Primary * To evaluate the value of TCR V beta repertoire analysis for the determination of autoimmunity in refractory cytopenia (RC). * To evaluate which immunophenotypic hematopoietic subclones are associated with oligoclonal T-cell expansion in RC. * To evaluate the presence of paroxysmal nocturnal hemoglobinuria (PNH) clones in RC. Secondary * To compare the molecular response with the hematologic response in patients with RC after treatment with immunosuppressive therapy (IST). * To compare the molecular response with human leukocyte histocompatability antigen (HLA) expression in patients with RC after treatment with IST. OUTLINE: This is an open-label, multicenter, nonrandomized, prospective study. Patients undergo biopsy, bone marrow, and blood sample collection periodically for immunological studies. Samples are analyzed for TCR V beta repertoire and paroxysmal nocturnal hemoglobinuria (PNH) clone analysis via PCR heteroduplex analysis and immunophenotyping of CD14, CD16 , CD55, CD59, and CD24 expression via flow cytometry.

Conditions

• Dyskeratosis Congenita
• Fanconi Anemia
• Myelodysplastic Syndromes
• Pearson Marrow-pancreas Syndrome
• Shwachman-diamond Syndrome

Interventions

Type	Name	Description
GENETIC	polymerase chain reaction
OTHER	flow cytometry	For analyzing GPI deficient clones full blood will be analyzed by phenotyping using flowcytometry. For that purpose CD14, CD16 and CD24 expression will be evaluated in CD45 positive cells. Erythroid cells will be evaluated for CD55 and CD59 expression searching for clear populations with a lack of GPI-linked molecules. In addition, immunophenotyping using flowcytometry will be performed to evaluate which differentiation stages of the major hematopoietic lineages in BM and PB are associated with TCRVβ repertoire skewing. Comparison between BM and PB will identify which is the optimal compartment to analyze the responsible hematopoietic clones.
OTHER	immunologic technique
PROCEDURE	biopsy

Timeline

• Start date: 2007-01-01
• Primary completion: 2012-08-01
• Completion: 2012-08-01
• First posted: 2007-07-11
• Last updated: 2015-01-16

Locations

10 sites across 10 countries: Austria, Belgium, Czechia, Denmark, Germany, Ireland, Italy, Netherlands, Spain, Switzerland