Neuroimmunology

Methodological accuracy and the reproducibility of laboratory data are fundamental, in the fields of both an efficient laboratory diagnostics, and high-profile research, especially if carried out within national and international networks. Pursuing such diagnostic and research activities is among the pre-eminent objectives of medical scientific institutes, such as the Scientific Institutes for Research, Hospitalization and Health Care (IRCCS) in Italy, specifically those mainly devoted towards translational research.

The determination of serum and cerebrospinal fluid biomarkers that can be used for different clinical purposes, including diagnosis, prognosis, patient stratification, personalization and monitoring of therapies, and rehabilitative pathways, represents a fundamental aim in the management of neuroinflammatory and neurodegenerative diseases.

In recent years, the neuroimmunology diagnostics sector has been revitalized by the discovery of conformational antibodies, which, to be measured, require innovative methods (cell-based assays) and specific laboratory expertise. The availability of commercial kits has extended the diagnostics to generalist laboratories, which have been absorbing the production of neuroimmunological tests for years. Insufficient expertise in non-specialized laboratories and suboptimal performance of commercial kits risk factors for false negative or false positive results, with risks of misdiagnoses and wrong therapies. Specialized laboratories can also use additional techniques, such as immunohistochemistry and tests on live rat neurons, which strengthen analytical specificity and support the search for new autoantibodies.

In addition to the new area of conformational antibodies, there is an urgent need to standardize more traditional methods, such as isoelectric focusing for the detection of oligoclonal bands, immunohistochemistry for the detection of onconeural antibodies, and the mainly commercial ELISAs, blots, and RIAs used for the diagnosis of other autoimmune neuroimmunological diseases, and of dementias.

The project has two main objectives:

• To optimize laboratory neuroimmunological diagnostic procedures and protocols, and to rationalize their requests in accordance with clinical guidelines (medical objective)

• To optimize and implement new laboratory tests for the determination of biomarkers in the field of neuroimmunological diagnostics (scientific objective)

• To share and implement laboratory procedures and protocols that guarantee both current neuroimmunological diagnostics of excellence, and the devising of new tests in the field of neuroimmunological and neurodegenerative disorders

• To establish common bases for collaborative research activities that increase the scientific competitiveness of the RIN at an international level

The laboratory tests included in the project are:

• Cerebrospinal fluid examination

  • • biochemical determinations of albumin and IgG in serum and cerebrospinal fluid
    • isoelectric focusing determination of IgG oligoclonal bands in serum and cerebrospinal fluid

• Onconeural antibodies

• Antibodies to surface / synaptic neuronal antigens

• Ganglioside antibodies

• MAG (myelin associated glycoprotein) antibodies

• AChR (acetylcholine receptor) antibodies

• MuSK (muscle-specific kinase) antibodies

The diagnostic part of the project is divided into the following steps:

• Identification of the methods used for the neuroimmunological diagnostics in the RIN centers

• Identification of clinical activities (number of patients attending each center) correlated to the main neurological pathologies that benefit from the neuroimmunological diagnostics

• Sending biological samples to test the analytical performances of the various laboratories involved (external quality assessment scheme, EQAS)

• Critical sharing of the EQAS results and start of the standardization process

• Verification of the standardization process through a new EQAS

• Production of documents relating to the various standardized tests in the form of consensus reports

The part of the project concerning the validation aspects of new markers (initially, neurofilaments and IgM oligoclonal bands) is divided into the following phases:

• Identification of the participating centers

• Presentation of the project to the Ethics Committee of the proposing center and subsequent submission to the Ethics Committees of the participating centers

• Collection of biological samples and clinical data by filling a specific database

• Carrying out determinations and evaluating results

• Drafting of documents in the form of consensus reports and data publication