In a broad sense, immunology is the study of autoimmune (AID), inflammatory and allergic diseases. The Immunology department at biolab performs highly automated, high-volume assays with rapid throughput to help in the diagnosis and follow-up of numerous pathologies such as connective tissue diseases, vasculitis, celiac disease, inflammatory bowel diseases (IBD), as well as food or respiratory allergies.

Furthermore multiple pathologies are diagnosed and monitored using protein studies: monoclonal gammopathies, inflammatory syndromes, immune deficiencies, kidney diseases, etc, the screening, identification, and follow-up are performed by capillary electrophoresis and immunofixation of serum and urinary proteins.

The prevalence of allergic symptoms has shown a steady increase in the past few years; the department’s has developed customized panels (e.g. Jordan Food and Respiratory Panel) to detect specific serum IgE antibodies to the common allergenic products.

Also the department has developed customized panels for human antibodies of the IgG and IgM classes against to five different EBV antigens: VCA gp125, VCA p19, EBNA-1, p22 and EA-D for diagnosis of infections with Epstein-Barr virus ( infectious mononucleosis, Burkitt’s lymphoma, nasopharyngeal carcinoma)and enable differentiation between acute and past EBV infections in one reaction, and a panel customized for human antibodies of the IgG and IgM classes against Borrelia antigens for the diagnosis of Lyme borreliosis and associated diseases: Erythema chronicum migrans, lymphadenosis cutis benigna, acrodermatitis chronica atrophicans, arthritis, carditis, lymphocytic meningoradiculoneuritis and neuroborreliosis.

Autoimmune diseases (AID) occur when an autoimmune reaction develops against many autoantigens present in our body. The Immunology department employees several techniques to detect over 100 autoantibodies to assist in the biological diagnosis of AID such as indirect immunofluorescence, the Luminex technology, immunoassays, immunoblots, immunochemistry and enzyme-linked immunosorbent assay (ELISA).

Autoimmunity:

Celiac Disease: is a long-term autoimmune disorder that primarily affects the small intestine and induced by a food antigen known as gliadin. Classic symptoms include gastrointestinal problems such as chronic diarrhoea, abdominal distention, malabsorption, loss of appetite and among children failure to grow normally.

First-line serology testing can be requested for reliable diagnosis by doing specific tests such as:

• Anti-endomysial antibodies IgG and IgA: These antibodies can be seen using sections of monkey esophagus by indirect immunofluorescence (IF).
• Anti-tissue transglutaminase antibodies and anti-gliadin antibodies IgG/IgA:

Screening by immunoblotting method using BlueDiver Dot instrument which is customized for the detection in human sera of IgG and IgA autoantibodies against Deamidated Gliadin and t-Transglutaminase antigens.

Anti- dsDNA antibodies:

The anti-double stranded DNA test is a blood test used to help diagnose lupus (systemic lupus erythematosus, SLE). This test is very specific for this disease because these antibodies aren't found in any other autoimmune disorders.

They are screened by using indirect immunofuorescence (IF) on Crithidia luciliae.

Anti-nuclear antibodies (ANA):

Antinuclear antibodies are autoantibodies that bind to contents of the cell nucleus. In normal individuals, the immune system produces antibodies to foreign proteins but not to human proteins. In some individuals, antibodies to human antigens are produced.

These antibodies are screened by using indirect immunofluorescence (IIF) on HEp-2 cells.

There are many fluorescence aspects for this test and each pattern can lead us to the diagnosis, for example: you can see:
1- The homogeneous and the coarse speckled fluorescence with SLE, systemic scleroderma and chronic juvenile arthritis patients.
2-  The fine speckled fluorescence with Sjogren's syndrome patients.
3- The centromere fluorescence with patients who have CREST syndrome (calcinosis, Raynaud disease, esophageal dysmotility, sclerodactyly and telangiectasia)
4- Cytoplasmic fluorescence which is indication for autoimmune hepatitis or primary biliary cirrhosis.

Extractable Nuclear Antigens (ENAs):

An extractable nuclear antigen panel, or an ENA Panel, tests for presence of autoantibodies in the blood that react with proteins in the cell nucleus. Usually done as a follow up to a positive antinuclear antibody (ANA) test and one is showing symptoms of an autoimmune disorder.

They are over 100 different soluble cytoplasmic and nuclear antigens. They are known as “extractable” because they can be removed from cell nuclei using saline.

Anti-ENA is a grouping of antibodies often used to screen for mixed connective tissue disease (MCTD), Sjögren's syndrome and systemic lupus erythematosus and commonly is composed of several tests:

• anti-Nucleosme, anti-DNA, anti-histones , and anti-Sm (for SLE)
• anti-RNP (for MCTD)
• anti-La/anti-SS-B (for Sjögren's)
• anti-Ro/anti-SS-A(for Sjögren's)
• anti-Scl70 (for Scleroderma)
• anti-Jo (for Dermatomyositis)
• anti-M2 native/recombinant and F-actin (for primary biliary cirrhosis)

These antigens can be screened using Western blotting technique on the EUROBlotMaster and BlueDiver Dot instruments. 

ANCA (anti-neutrophil cytoplasmic antibodies):

Are autoantibodies directed against antigens found in the cytoplasmic granules of neutrophils and monocytes. ANCA testing is usually performed to help diagnose or exclude: Vasculitis, Wegener's granulomatosis, microscopic polyangiitis, and chronic inflammatory bowel diseases.

There are two types of the ANCA test called the p-ANCA and the c-ANCA.

p-ANCA, or MPO-ANCA, or perinuclear anti-neutrophil cytoplasmic antibodies, are antibodies that stain the material around the nucleus of a neutrophil. They are a special class of anti-neutrophil cytoplasmic antibodies.

c-ANCAs, or PR3-ANCA, or Cytoplasmic antineutrophil cytoplasmic antibodies, are a type of autoantibody, an antibody produced by the body that acts against one of its own proteins. These antibodies show a diffusely granular, cytoplasmic staining pattern under microscopy.

These antibodies can be screened by indirect immunofluorescence (IIF) and the immunoblotting method using the BlueDiver Dot instrument.

For further information, please contact:

• Dr. Issa Abu-Dayyeh, Lab Director-Immunologist, i.abudayyeh@biolab.jo
• Jumana Abu Kwaik Specialist technologist,  j.kwaik@biolab.jo
• Ihab Khoury, Specialist technologist, i.khoury@biolab.jo