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

Study Overview
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Brief Summary
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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
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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.

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

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

Intervention / Treatment
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* Genetic: polymerase chain reaction
* Other: flow cytometry
* Other: immunologic technique
* Procedure: biopsy


Participation Criteria
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Eligibility Criteria
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DISEASE CHARACTERISTICS: Diagnosis of refractory cytopenia (RC) including any of the following: Severe aplastic anemia (SAA) Fanconi's anemia Shwachman Diamond syndrome Dyskeratosis congenita Pearson syndrome All RC patients included in the EWOG MDS 2006 protocol irrespective of therapy Patients who have undergone hematopoietic stem cell transplantation (HSCT) may be enrolled on EWOG-MDS SCT RC RIC 06 or EWOG-MDS SCT MDS 06 protocol PATIENT CHARACTERISTICS: Not specified PRIOR CONCURRENT THERAPY: No prior immunosuppressive therapy for refractory cytopenia

Ages Eligible for Study
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Maximum Age: 17 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Arms and Interventions

| Intervention/Treatment |
| --- |
|Genetic: polymerase chain reaction|nan|
|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|nan|
|Procedure: biopsy|nan|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Number of patients with TCR V beta oligoclonality at diagnosis |  | 96 months | 
| Immunophenotype of patients with oligoclonal T-cell expansion |  | 96 months | 
| Number of patients with glycophosphatidylinositol (GPI) deficient clones |  | 96 months | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Number of patients with molecular response as compared to hematological response after IST |  | 96 months | 
| Number of patients with HLA-DR15 antigen expression and molecular response as compared to number of patients with other HLA-DR antigens and molecular response |  | 96 months | 
| Overall survival |  | 96 months | 
| Failure-free survival |  | 96 months | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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refractory cytopenia with multilineage dysplasia, aplastic anemia, Fanconi anemia, dyskeratosis congenita, Shwachman-Diamond syndrome, Pearson marrow-pancreas syndrome