By

S.DHARANIYA,
3rd year Biotechnology.
Autoimmune diseases can generally be classified into two groups: those that are
systemic in nature (i.e. systemic lupus erythematosus, Sjogren's syndrome,
scleroderma, rheumatoid arthritis, autoimmune vasculitis, mixed connective tissue
disease, and various other overlap syndromes); and those that are more organ or
tissue directed (autoimmune thyroid disease, myasthenia gravis, and certain skin
diseases such as bullous pemphigoid). symptoms range in severity . The
diagnosis of an autoimmune disease is based on an individual's symptoms,
findings from a physical examination, and results from laboratory tests.

Fever
Cough
CNS signs
Haemoptysis
Cough
Wheezing
Rash
Diarrhoea
Joint pain
Skin disorders
BASIC APPROACH TOWARDS THE
DIAGNOSTIC TESTS FOR
AUTOIMMUNE DISEASES

A number of different tests can help diagnose

autoimmune diseases. A common feature of the
tests is the detection of antibodies that react with
host antigens. Such tests involve measuring the
level of antibodies found in the blood. An elevated
amount of antibodies indicates that a humoral
immune reaction is occurring.A useful approach is
to determine the class of antibody that is present.
The most useful antibody tests involve introducing
the patient's antibodies to samples of his or her
own tissue. If the antibodies bind to the host tissue
it is diagnostic for an autoimmune disorder.
Antibodies from a person without an autoimmune
disorder would not react to "self" tissue. Tissues
from the thyroid, stomach, liver, and kidney are
used most frequently in this type of testing.
 ANA (Antinuclear Antibody) Test
To perform the ANA (antinuclear antibody) test, sometimes called FANA
(fluorescent antinuclear antibody test), a blood sample is drawn from the patient
and sent to the lab for testing. Serum from the patient's blood specimen is added to
microscope slides which have commerically prepared cells on the slide surface. If
the patient's serum contains antinuclear antibodies (ANA), they bind to the cells
(specifically the nuclei of the cells) on the slide. A second antibody, commercially
tagged with a fluorescent dye, is added to the mix of patient's serum and
commercially prepared cells on the slide. The second (fluorescent) antibody
attaches to the serum antibodies and cells which have bound together. When
viewed under an ultraviolet microscope, antinuclear antibodies appear as
fluorescent cells.
If fluorescent cells are observed, the ANA (antinuclear antibody) test is positive.
If fluorescent cells are not observed, the ANA (antinuclear antibody) test is
negative.
A titer is determined by repeating the positive test with serial dilutions until the test
yields a negative result. The last dilution which yields a positive result
(flourescence) is the titer which gets reported. For example, if a titer performed for
a positive ANA test is:
1:10 positive,1:40 positive,1:160 positive,1:320 negative
What is the significance of the ANA pattern?

Homogeneous - total nuclear fluorescence due to

antibody directed against nucleoprotein. Common in
SLE (lupus).
Peripheral - fluorescence occurs at edges of
nucleus in a shaggy appearance. Anti-DNA
antibodies cause this pattern. Also common in SLE
(lupus).
Speckled - results from antibody directed against
different nuclear antigens.
Nucleolar - results from antibody directed against a
specific RNA configuration of the nucleolus or
antibody specific for proteins necessary for
maturation of nucleolar RNA. Seen in patients with
systemic sclerosis.
Subsets of the ANA (antinuclear antibody) test are
sometimes used to determine the specific autoimmune
disease. For this purpose, a doctor may order anti-dsDNA,
anti-Sm, Sjogren's sydrome antigens(SSA, SSB), Scl-70
antibodies, anti-centromere, anti-histone, and anti-RN.
 What does a positive ANA result mean?
ANAs are found in patients who have various autoimmune diseases, but not
only autoimmune diseases. ANAs can be found also in patients with infections,
cancer, lung diseases, gastrointestinal diseases, hormonal diseases, blood
diseases, skin diseases, and in elderly people or people with a family history of
rheumatic disease. ANAs are actually found in about 5% of the normal
population.
The ANA results are just one factor in diagnosing, and must be considered
together with the patient's clinical symptoms and other diagnostic tests.
Medical history also plays a role because some prescription drugs can cause
"drug-induced ANAs".

What is the incidence of ANA in various diseases or conditions?

Statistically speaking the incidence of positive ANA (in percent) per conditon is:
Systemic lupus erythematosus (lupus or SLE) - over 95%
Progressive systemic sclerosis (scleroderma) - 60-90%
Rheumatoid Arthritis - 25-30%
Sjogren's syndrome - 40-70%
Felty's syndrome - 100%
Juvenile arthritis - 15-30%
Microplate ELISA: ANA Screen, Anti-ENA PoolPlus
                                                            
Incubate ELISA ANA Screen (antigen mixture of dsDNA, histones, nRNP/Sm,
Sm, SS-A, SS-B, Scl-70, Jo-1, ribosomal P-proteins, centromere).
•Screening test for predifferentiation of antibodies against cell nuclei (ANA) and
cytoplasm components.
•Indications: Sharp syndrome (MCTD), systemic lupus erythematosus, Sjögren's
syndrome, progressive systemic sclerosis, polymyositis/dermatomyositis.
•Serum dilution 1 : 200, conjugate class anti-human IgG, POD-labelled.
•One microplate well incubated per patient.
•1-point calibration, semiquantitative.
•Native antigens (exception: centromere, recombinant).
•The ANA Screen ELISA supplements the gold standard immunofluorescence. It is
based on a mixture of 10 highly purified antigens, which provide higher sensitivity and
specificity than the undefined cell extracts used by other manufacturers.
•Two ELISAs with different antigen combinations, adapted to particular indications or
for follow-up of immunofluorescence patterns, are available.
Erythrocyte Sedimentation Rate (ESR)
The erythrocyte sedimentation rate is a test which involves placing a blood sample in a
tube and determining how fast the red blood cells settle to the bottom in one hour. When
inflammation occurs the body produces proteins in the blood which make the red cells
clump together. Heavier cell aggregates fall faster than normal red cells. For healthy
individuals, the normal rate is up to 20 millimeters in one hour. Inflammation increases the
rate significantly.
Rheumatoid Factor (RF)
Rheumatoid factor is an antibody found in unusually large amounts of patients with
rheumatoid arthritis.
HLA Typing
White blood cells are typed for the presence of HLA-B27
C-Reactive Protein (CRP)
C-Reactive Protein measures the concentration in blood serum of a special type of protein
produced in the liver that is present during episodes of acute inflammation or infection.
Lupus Erythematosus (LE)
Lupus erythematosus (LE) cell test measures the presence of a special cell found mostly
in patients with systemic lupus erythematosus.
Anti-DNA and Anti-Sm
Lupus patients have antibodies to the heredity material DNA (deoxyribonucleic acid) and
sm,another substance in the cell’s nucleus.
Acute disseminated encephalomyelitis
Residual motor deficits are estimated to remain in about 8 to 30% of cases, the range in
severity from mild clumsiness to ataxia and hemiparesis. Patients with demyelinating
illnesses, such as MS, have shown cognitive deficits even when there is minimal
physical disability .
Addison's disease-chronic endocrine disorder
Hyponatraemia (low blood sodium levels), due both to a deficiency in Aldosterone (a
mineralocorticoid) dependant Sodium retention and also the effect of Corticotropin
-releasing hormone to stimulate secretion of ADH
Hypogammaglobulinemia
Reduction in gamma globulins,
Atopic dermatitis
Epidermal Barrier Dysfunction, Histamine intolerance .
IgA nephropathy(deposition of the IgA antibody in the glomerulus)

For an adult patient with isolated hematuria, tests such as ultrasound of the kidney
and cystoscopy are usually done first to pinpoint the source of the bleeding. These
tests would rule out kidney stones and bladder cancer, two other common urological
causes of hematuria. In children and younger adults, the history and association
with respiratory infection can raise the suspicion of IgA nephropathy. A kidney
biopsy is necessary to confirm the diagnosis. The biopsy specimen shows
proliferation of the mesangium, with IgA deposits on immunofluorescence and
electron microscopy. However, all patients with isolated microscopic hematuria (i.e.
without associated proteinuria and with normal kidney function) are not usually
biopsied since this is associated with an excellent prognosis. A urinalysis will show
red blood cells, usually as red cell urinary casts. Proteinuria, usually less than
2 grams per day, also may be present. Other renal causes of isolated hematuria
include thin basement membrane disease and Alport syndrome, the latter being a
hereditary disease associated with hearing impairment and eye problems.
Other blood tests done to aid in the diagnosis include CRP or ESR, complement
levels, ANA, and LDH. Protein electrophoresis and immunoglobulin levels can show
increased IgA in 50% of all patients.
Cancer
Most cancers are initially recognized either because signs or symptoms
appear or through screening. Neither of these lead to a definitive
diagnosis, which usually requires the opinion of a pathologist, a type of
physician (medical doctor) who specializes in the diagnosis of cancer and
other diseases. People with suspected cancer are investigated with
medical tests. These commonly include blood tests, X-rays, CT scans and
endoscopy.
POEMS syndrome
It is defined as the combination of a plasma-cell proliferative disorder (typically
myeloma), polyneuropathy, and effects on many other organ systems.
In addition to tests (such as EMG for neuropathy or CT scan and
bone marrow biopsy for myeloma), two other tests give abnormal results. These
are: a raised level of VEGF in blood; and a raised CSF protein level, as in some
other neuropathies. Even when myeloma is present, anaemia and
thrombocytopaenia are rare; on the contrary, polycythaemia and thrombocytosis
are often found.
Vitiligo
Tinea versicolor (also known as "Dermatomycosis furfuracea," "Pityriasis
versicolor," and "Tinea flava") is a condition characterized by a rash on the trunk
and proximal extremities.
 Treatment List for Autoimmunediseases
Hormone replacement therapy

Enzyme replacement therapy

Targeting Disease Mechanisms

Some drugs may also be used to target how the disease works. In other words,
they can suppress the immune system. Autoimmune disorder medications include
cyclophosphamide (Cytoxan®) and cyclosporine (Neoral® and Sandimmune®).
The same immune-suppressing drug may be used for many diseases.

New Treatment for Celiac Disease?

Research Shows Blocking a Protein May Reverse Celiac Disease Symptoms
Blocking an inflammatory protein called interleukin-15 (IL-15) may help
treat the symptoms of celiac disease and prevent the development of celiac
disease in certain at-risk peopleCeliac disease is an autoimmune and
inflammatory condition that is triggered by gluten, the protein found in
wheat, barley, and rye. When people with celiac disease eat gluten, it
triggers an inflammatory response that damages the lining of the small
intestine.