Antinuclear province of Saudia, referred to immunology

Antinuclear
antibody (ANA) detection is first step in the diagnosis of connective tissue
disorder (CTD). The indirect immunofluorescence (IIF) performed on HEp-2 cells
(cultured human epithelial cell) is considered as a gold standard Laboratory
assay for the detection of antinuclear antibodies (ANA). Positive immunofluorescence
staining indicates presence of antinuclear antibodies (ANA) but does not reveal
precise identification of these autoantibodies and their target antigens. Identification
of these specific antibodies may provide valuable information in the diagnostic
evaluation, prognostic assessment and monitoring of patients with autoimmune
connective tissue disorder (CTD). Many known association of these specific
autoantibodies with the immunofluorescence staining pattern of ANA in CTD can
be found in western literature and has been considered as reference guide over
the world.

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 similarities / differences 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.

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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

 

Literature
Review:

 

A systemic autoimmune response is
hallmark of the 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 refernce technique used
for the detection of antinuclear antibodies (ANA), and is also used as
screening test. Though positive fluorescence staining indicates the presence of
ANA and staining patterns depends on location of the target antigen, it does not, however, allow precise
identification of these 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
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 on 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 cell substrate 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 on ANA using rat liver substrate and a screening dilution of 1:100 on
rheumatic as well as non-rheumatic diseases.

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 IIF patterns strongly suggest
distinct 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. 

 

 

References:

1.     
Von
Mühlen CA, Tan EM. Autoantibodies in the diagnosis of systemic rheumatic
diseases. Semin Arthritis Rheum 1995; 24:323–58.

2.     
 Meheus L, Van Venrooij WJ, Wiik A, Charles PJ,
Tzioufas AG, Meyer O, et al. Determination of the fine specificity of
antinuclear antibodies in connective tissue diseases using a multiparameter
line immunoassay (LIA) based on recombinant proteins. Clin Exp Rheumatol 1999;
17:205–14.

3.     
 Tan EM, Smolen JS, Mc Dougal JS, Butcher BT,
Conn D, Dawkins R, et al. A critical evaluation of enzyme immunoassays for
detection of antinuclear antibodies of defined specificities. Arthritis Rheum
1999; 42: 455–64.

4.     
Sebastian
W, Roy A, Kini U, Mullick S. Correlation of antinuclear antibody
immunofluorescence patterns with immune profile using line immunoassay in the
Indian scenario. Indian J Pathol Microbiol 2010; 53:427-32.

5.     
 Slater CA, Davis RB, Shmerling RH. Antinuclear
antibody testing: A study of clinical utility. Arch Intern Med 1996;
156:1421-25.

6.     
Sulcebe
G, Morcka K. Diagnostic and prognostic significance of different antinuclear
antibodies in more than 1000 consecutive Albanian patients with rheumatic
disease. Clin Exp Rheumatol 1992; 10: 255-61.

7.     
Arbuckle
MR, McClain MT, Rubertone M, et al. Development of autoantibodies before the
clinical onset of systemic lupus erythematosus. N Engl J Med 2003; 349:1526-33.

8.     
 Emlen W, O’Neill L. Clinical significance of
antinuclear antibodies: Comparison of detection with immunofluorescence and
enzyme-linked immunosorbent assays. Arthritis Rheum 1997; 40: 1612-18