Neurofilament Light Chain ,Chitinase-3 Like-1 Proteins and Plasmacytoid Dendritic Cells in Multiple Sclerosis

Study Overview
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Brief Summary
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Multiple sclerosis (MS) is an autoimmune neurodegenerative disease characterized by demyelination and neurodegeneration of the central nervous system (CNS) . Current diagnostic criteria and management depend on MRI, clinical status, and oligoclonal immunoglobulin g bands . These markers often fail to predict relapse, progression and therapy response .There is an increased need to identify biomarkers for clinical endpoints . One of the hallmark features of MS is axonal damage which associated with brain and cervical atrophy.Nf levels indicate the extent of axonal damage. Neurofilaments are composed of four subunits: neurofilament light polypeptides (NfL) is the most abundant and soluble and it is highly sensitive to neurodegenerative processes . Chitinase 3-like 1 (CHI3L1) is expressed in astrocytes in the brain tissue of MS patients . CHI3L1 plays a role as prognostic biomarker in patients with MS. CHI3L1 cerebrospinal fluid levels were associated correlated with disease activity and neurological disability. Dendritic cells (DCs) are highly specialized antigen-presenting cells with a key role in activating and preventing CNS immune-mediated damage in MS . Dendritic cells express Human Leukocyte Antigen-antigen D Related (HLA-DR) . Plasmacytoid dendritic cells characterized by the expression of blood dendritic cells antigen-2 (BDCA-2) .Plasmacytoid dendritic cells are present in the cerebrospinal fluid (CSF), leptomeninges and demyelinating lesions of patients with MS . Plasmacytoid dendritic cells also exhibit up-regulation of chemokine (CCR7C) expression. It was demonstrated increased amounts of chemokine CCR7 in the CSF from MS patients during relapses .CCR7 controls migration and functional activity of regulatory T cells and plays an important role in the establishment of tolerance . Tolerogenic DCs (TolDCs) present an intermediate phenotype between immature dendritic cells (iDCs) and mature dendritic cells (mDCs) regarding costimulatory molecules, a pronounced shift toward anti-inflammatory . TolDCs exhibit tolerogenic molecules such as HLA-G and CD274 [programmed death-ligand 1 (PD-L1)] either in peripheral blood or in CSF. These characteristics lead to T cell clonal anergy and T cell unresponsiveness due to Ag presentation in the presence of low co-stimulation .We aim to investigate the role of NfL,(CHI3L1) and markers of plasmacytoid dendritic cells in MS.

Official Title
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Combined Neurofilament Light Chain ,Chitinase-3 Like-1 Protein Levels and Tolerogenic Plasmacytoid Dendritic Cells in Defining Disease Course in Multiple Sclerosis Patients

Conditions
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Multiple Sclerosis

Intervention / Treatment
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* Diagnostic Test: Neurofilament Light Chain ,Chitinase-3 Like-1 Protein Levels and Tolerogenic plasmacytoid Dendritic Cells


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: 1-MS patients diagnosis 2017 McDonald criteria. 2-Patients age 20-60 years 3-All males and female patients. 3-Willingness and ability to comply with the protocol. 4-Written informed consent given by the patient before the beginning of any study-related procedure, 5-Non-MS control matched age and sex persons Exclusion Criteria:1-patients with Intracranial space-occupying lesion. 2-Abnormal intracranial pressure due to increased CSF pressure or Arnold-Chiari malformation. 3-Anticoagulant medication, coagulopathies and uncorrected bleeding diathesis. 4-Congenital spine abnormalities. 5-Local skin infection at the puncture site. 6-Not fulfill Macdonald criteria.

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 60 Years

Sexes Eligible for Study
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All

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Multiple sclerosis patients<br>MS patients according to 2017 Macdonald criteria | Diagnostic Test: Neurofilament Light Chain ,Chitinase-3 Like-1 Protein Levels and Tolerogenic plasmacytoid Dendritic Cells<br>* Serum and CSF levels assay of NfL and CHI3L1 using the enzyme linked immunosorbent assay kits (ELISA). Peripheral blood mononuclear cells (PBMC) will be obtained by Ficoll-Hypaque gradient. plasmacytoid dendritic cells will be identified in (CSF) and (PBMN) based on their selective expression of surface antigen (cluster of differentiation 303) CD303, named blood dendritic cells antigen 2 (BDCA-2).Antihuman antibodies: BDCA-2, HLA-DR , CD274 , HLA-G and CCR7 for identification of tolerogenic plasmacytoid dendritic cells using flowcytometry.<br>|
| Non Multiple sclerosis persons<br>Control persons | Diagnostic Test: Neurofilament Light Chain ,Chitinase-3 Like-1 Protein Levels and Tolerogenic plasmacytoid Dendritic Cells<br>* Serum and CSF levels assay of NfL and CHI3L1 using the enzyme linked immunosorbent assay kits (ELISA). Peripheral blood mononuclear cells (PBMC) will be obtained by Ficoll-Hypaque gradient. plasmacytoid dendritic cells will be identified in (CSF) and (PBMN) based on their selective expression of surface antigen (cluster of differentiation 303) CD303, named blood dendritic cells antigen 2 (BDCA-2).Antihuman antibodies: BDCA-2, HLA-DR , CD274 , HLA-G and CCR7 for identification of tolerogenic plasmacytoid dendritic cells using flowcytometry.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Levels of different markers in MS diagnosis | The changes of levels of NfL and CHI3L1 using ELISA kits and its correlation with the expression of identification markers of plasmacytoid dendritic cells and its activity (HLA-DR, BDCA-2, ,CCR7C ,HLA-G and CD274) using flowcytometry in different clinical presentations of MS in consistent with MRI findings. | 2 years | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Different samples levels in MS diagnosis | Changes of levels of NfL and CHI3L1 using ELISA kits and markers of plasmacytoid dendritic using flowcytometry between blood and CSF samples of MS diagnosis. | 2 years |