Abstract: in the Saudi population and to document

Abstract:

Detection of Antinuclear
antibody (ANA) is first step in the diagnosis of autoimmune connective tissue
disorder (CTD). Gold standard Laboratory assay for the detection of antinuclear
antibodies (ANA) is indirect immunofluorescence (IIF) performed on HEp-2 cells
(cultured human epithelial cell. Presence of antinuclear antibodies (ANA) is
reflected by positive immunofluorescence staining, but since antinuclear
antibodies are further divided into subtypes, precise identification of these
autoantibody subtypes and their specific target antigens is not possible with indirect
immunofluorescence. Identification of these specific antibodies provides
valuable information in the diagnostic evaluation, prognosis and monitoring of
patients with autoimmune connective tissue disorder (CTD). Many known association
of these specific autoantibodies with the indirect immunofluorescence (IIF) staining
pattern of ANA in CTD can be found in western literature and is considered as
reference guide over the world.

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Since Immune
response to disease, antibody profile and individual immune status differs from
person to person and also from population to population, therefore, present study has been designed to
evaluate the definite
association between ANA patterns and specific antibodies in the serum in the Saudi
population and to document differences / similarities with other populations. To
the best of my knowledge, no such research work or data correlating the
autoantibodies and their ANA patterns is found in Saudia
Arabia.

In this study, we will analyze serum samples
from the eastern province of Saudia, referred to immunology laboratory,
providing services to a tertiary health center and teaching hospital (KFUH) for
ANA testing by Indirect Immunofluorescence method and samples further processed
for identification of the specific antibodies by line immunoassay in that
population. Later the two will be correlated with one another to establish any
definite link between the two.

 

Literature
Review:

 

A systemic autoimmune response is
hallmark of the autoimmune connective tissue disease (CTD) and is characterized
by the presence of antinuclear antibodies (ANA). Indirect immunofluorescence
(IIF) on human epithelial cell tumor line (Hep-2 cells) is the reference
technique used for the detection of antinuclear antibodies (ANA), and is also
used as a screening test. Positive immunofluorescence staining indicates the
presence of ANA and resulting
staining pattern depends on the location of the target antigen. However, it does not, allows precise
identification of these specific antibodies against nuclear antigens.

Identification of specific antibodies
is performed by specialized techniques such as enzyme linked immunosorbant
assay (ELISA), Western blotting or line immunoassay 1-3.  Literature review reveals some known
associations between a broad spectrum of specific antibodies and each specific autoimmune
rheumatic disease entity.  Most of these associations have been identified
with data obtained from various studies on Western population. It is need of
time to recognize that individual response to disease, immunity status and type
of antibodies, all, depend on genetic makeup and therefore varies from person
to person, population to population and place to place. Hence associations
between ANA pattern and specific antibodies known to us from research on
samples of western population cannot be applied to patients of CTD from some
other population.

There is neither any data nor any
research work correlating antinuclear antibody (ANA) immunofluorescence patterns
with the specific antibody immunoprofile in the Saudi population to date. 

The study that most closely resembles ours, reported in 2010, by
Sebastian et al 4.  This study reports
the results of sera tested for ANA using HEp-2010/ liver biochip and a
screening dilution of 1:100. In another study by Slater and Shmerling, ANA was
performed on HEp-2 cells at a titre of 1: 40 5. In Albania, Sulcebe and
Morcka also reported a similar study in 1992 6. They observed the results of
sera tested for ANA using rat liver substrate and a screening dilution of
1:100.

Indirect Immunofluorescence on HEp-2 cell is the standard approach
for detecting ANAs, and the staining patterns reveals location of the target
antigen. These patterns correspond to the presence of autoantibodies against
different nuclear antigens 7, 8. 
Although some staining patterns strongly suggest distinct antibody specificities,
additional tests are required to demonstrate antibody reactivities against
specific nuclear and cytoplasmic antigens. Identification
of the fine specificity may provide valuable assistance in diagnosis, prognosis
and monitoring of patients suffering from rheumatic connective tissue diseases.

 

MATERIALS AND METHODS;

In this study, we will analyze serum
samples from the eastern province of Saudia, referred to Immunology laboratory,
providing services to a tertiary health center and teaching hospital (KFUH) for
ANA testing by Indirect Immunofluorescence method and samples further processed
for identification of the specific antibodies by line immunoassay in that
population and the two will be correlated with one another to establish any
definite link between the two.

Data will be analyzed anonymously. The
review of medical records will be performed retrospectively on serological
tests (IIF and Line immunoassay) and patient data that are performed as part of
routine laboratory work.

 

Aims: To understand a definite
association between ANA Immunofluorescence staining patterns and specific autoantibodies
in the serum in the Saudi patients suffering from CTD and to document
differences and similarities with other populations.

 

Expected benefits:

 

Absence of data correlating the auto
antibodies and their antinuclear antibody (ANA) patterns with the immunoprofile
in the Saudi population represents a serious gap in our knowledge.

Data obtained from this study would,
therefore, provide a reference database for the Saudi population. In case a
definite correlation is found between the ANA staining patterns and the specific
antibodies identified by line immunoassay, one could restrict performing line
immunoassays which are expensive and use ANA-IIF fluorescent patterns to
predict presence of auto antibodies to precisely diagnose a CTD. This would reduce
the cost of laboratory investigations for the patient and would save the
resources of Laboratory and hospital used in diagnosis of Patients suffering
from Rheumatic Connective tissue diseases.