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Immunologic thrombocytopenic purpura (ITP) in children is a pre-lupus condition if associated with the presence of anti-nuclear antibodies (ANA), providing a unique model for understanding the natural history of autoimmunity, particularly that of systemic lupus erythematosus (SLE). The investigators will describe the shared and/or unique immunological pathways involved at diagnosis in 70 children with ITP and in 20 children with SLE, and compare them between ITP-ANA- (more often transient), ITP-ANA+ (pre-lupus condition, more often persistent) and SLE
The development of an autoimmune disease (AID) requires several years. In systemic lupus erythematosus (SLE), anti-nuclear antibodies (ANA) appear several years before the onset of the disease. However, it is difficult to identify the early mechanisms of autoimmunity in humans as most patients' samples are obtained at the time of diagnosis.
The investigators hypothesize that immune thrombocytopenic purpura (ITP) in children provides a unique model for understanding the initial mechanisms leading to autoimmunity. Children with ITP have common immune system dysfunctions leading to the production of anti-platelet antibodies. However, despite this supposedly common dysfunction, the course and degree of loss of tolerance of the immune system are very heterogeneous: the disease is transient in 75 to 80% of cases, and 15 to 20% of children with ITP have ANA and their course is usually persistent or chronic. This subgroup of children with hematological AID, ITP ANA+ meets the criteria for a broader form of AID: pre-lupus. The investigators have recently shown that 16% of these children with pre-lupus / ITP-ANA+ progressed to complete SLE within a median of 3.8 years. Why do some children have transient or persistent ITP? limited or systemic ITP? The investigators aim to describe the shared and/or unique immunological pathways involved at diagnosis in children with ITP ANA- (transient or persistent), with ITP ANA+ (pre-lupus condition, transient or persistent), and in patients with SLE (persistent by definition) In this exploratory, prospective, bi-centric cohort study, the investigators will include 70 children with ITP and 20 children with SLE newly diagnosed, over 36 months with a 3-month follow-up for children with ITP-ANA- and SLE and a 48-month follow-up for children with ITP-ANA+. ANA positivity will be defined by a titer ≥ 1/160. Blood samples will be collected at inclusion and at 3 months (bio-collection), and further each 6 months for children with ITP-ANA+. ITP status will be defined at 3 months: transient or persistent. The investigators will analyze the signaling pathways (B and T lymphocytes, cytokines, disruption of tolerance and inactivation of X) involved in these different pathophysiological distinct subgroups
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| ITP-ANA- | Other |
| |
| ITP-ANA+ | Other |
| |
| SLE | Other |
| |
| Controls | Other |
|
| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Blood sampling baseline and M3 | Procedure | Blood sampling is a routine biological procedure performed under the same conditions as during a follow-up consultation. |
|
| Measure | Description | Time Frame |
|---|---|---|
| Characterisation of B and T lymphocyte populations | Characterisation of B and T lymphocyte populations through the study of surface markers and intracellular factors. This panel will, in particular, enable the identification of effector B cells such as plasma blasts, subsets of auto-reactive 'double-negative' (DN) B cells, regulatory B cells, and follicular T helper (Tfh) and peripheral extra-follicular T helper (Tph) cells. Each population will be compared between the patients and the control group. | Baseline, Month 3 visit. |
| Plasma markers | Soluble cytokines and factors regulating B-cell survival (BAFF, TACI, IL-6, IL-12p70, IFN-γ, IFN-β, TGF-β, and the IFN-regulated chemokine CXCL10) will be quantified in plasma using a customised multiplex Luminex assay. Interferon alpha levels will be specifically measured using ultra-sensitive SIMOA (Single Molecule Array) technology. | Baseline, Month 3 visit. |
| Plasma markers | Circulating plasma autoantigens - soluble P-selectin and CD40L derived from platelet activation - will be measured by ELISA. The levels of circulating and mitochondrial DNA will be measured by quantitative RT-PCR in collaboration with V. Sisirak (DR CNRS, Immunoconcept). The overall activity of plasma DNases will be determined by an in vitro DNA degradation assay. | Baseline, Month 3 visit. |
| Measure | Description | Time Frame |
|---|---|---|
| Single-cell transcriptomic analysis | Using single-cell RNA sequencing, a single-cell transcriptomic analysis, we will be able to quantify and analyse the transcriptomes of lymphocytes B This will enable us to characterise heterogeneous cell populations, as well as to reconstruct cell development pathways and model transcriptomic dynamics. This technique will enable the modelling of signalling pathways involving B lymphocytes in the subgroup of ITP patients with positive AAN titres, and the development of an algorithm and a mathematical model of the progression of these cellular and molecular markers |
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Inclusion criteria:
For patients :
For controls :
Exclusion criteria:
For patients :
For controls :
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| Name | Role | Phone | Extension | |
|---|---|---|---|---|
| Jérôme GRANEL, MD | Contact | +33 5 57 82 02 55 | jerome.granel@chu-bordeaux.fr | |
| Nathalie ALADJIDI, MD | Contact | nathalie.aladjidi@chu-bordeaux.fr |
| Name | Affiliation | Role |
|---|---|---|
| Jérôme GRANEL, MD | University Hospital, Bordeaux | Principal Investigator |
| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| Chu de Bordeaux- Groupe Hospitalier Pellegrin - Hôpital des Enfants | Bordeaux | 33076 | France |
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| Monitoring for SLE | Other | Monitoring for Systemic Lupus Erythematosus according to SLICC 2012 classification criteria |
|
| Procedure/Surgery: Blood sampling baseline and M3 then each 6 months until 48 months | Procedure | Blood sampling is a routine biological procedure performed under the same conditions as during a follow-up consultation. |
|
| moniktoring for SLE : baseline and M3 then each 6 months until 48 months | Other | Monitoring for Systemic Lupus Erythematosus according to SLICC 2012 classification criteria |
|
| Baseline, Month 3 visit , Month 6 visit , Month 12 visit, Month 18 visit, Month 24 visit , Month 30 visit, Month 36 visit, Month 42 visit and Month 48 visit. |
| Single cell epigenetic analysis | Using single-cell ATAC sequencing, we will be able to analyse the regions of DNA accessible to the transcription machinery. This will enable us to identify regulatory programmes such as signalling pathways. The analysis will also reveal heterogeneity amongst B-cell subpopulations, which may respond differently to signals. This will provide insight into the epigenetic status of genes, or at least into which factors regulate this signalling pathway and how the cell adjusts its response depending on its situation or stage of maturity, or on the signal itself. | Baseline, Month 3 visit , Month 6 visit , Month 12 visit, Month 18 visit, Month 24 visit , Month 30 visit, Month 36 visit, Month 42 visit and Month 48 visit. |
| CHU Toulouse - Hôpital des Enfants | Toulouse | 31026 | France |
|
| ID | Term |
|---|---|
| D016553 | Purpura, Thrombocytopenic, Idiopathic |
| D008180 | Lupus Erythematosus, Systemic |
| ID | Term |
|---|---|
| D011696 | Purpura, Thrombocytopenic |
| D011693 | Purpura |
| D001778 | Blood Coagulation Disorders |
| D006402 | Hematologic Diseases |
| D006425 | Hemic and Lymphatic Diseases |
| D057049 | Thrombotic Microangiopathies |
| D013921 | Thrombocytopenia |
| D001791 | Blood Platelet Disorders |
| D000095542 | Cytopenia |
| D006474 | Hemorrhagic Disorders |
| D001327 | Autoimmune Diseases |
| D007154 | Immune System Diseases |
| D006470 | Hemorrhage |
| D010335 | Pathologic Processes |
| D013568 | Pathological Conditions, Signs and Symptoms |
| D012877 | Skin Manifestations |
| D012816 | Signs and Symptoms |
| D003240 | Connective Tissue Diseases |
| D017437 | Skin and Connective Tissue Diseases |
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