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MITOTIC CELL (Metaphase)
Peripheral vs. nuclear membrane antibody: Antibody
that shows a peripheral pattern is generally associated
with DNA/DNP nuclear antigens. High titers of these
antibodies are suggestive of SLE. In substrates that do
not include mitotic cells, the peripheral pattern can be
difficult to distinguish from nuclear membrane antibody.
By using Immuno Concepts' mitotic cells, these patterns
can be differentiated, because the chromosome
region of the mitotic cells will be stained intensely in a
peripheral pattern, but will not be stained by nuclear
membrane antibody. This distinction is clinically impor-
tant because nuclear membrane antibody does not
have DNA/DNP specificity and is not associated with
CELLULE MITOTIQUE
SLE (35).
(Métaphase)
anti-centromere antibody (aca) vs. atypical speckled
antibody resembling centromere: In order to verify
anti-centromere antibody, the chromosome region
of the mitotic cells should stain brightly with discrete
speckles. If the chromosome region does not stain,
the antibody has neither kinetochore specificity nor as-
sociation with the CREST variant of scleroderma (36). If
the chromosome region does not stain, the antibody is
not anti-centromere, and should be reported as "atypi-
cal speckled."
cYtoPlaSmIc FluorEScEncE
Although autoantibodies to cytoplasmic antigens
CELLULA MITOTICA
are not commonly associated with connective tissue
disease, these antibodies may be detected using
(Metafase)
epithelial cell culture substrates (37). Mitochondrial and
smooth muscle antibodies are the two most commonly
detected antibodies and are generally associated
with mononucleosis, chronic active hepatitis, and liver
disease (38- 39). Using the HEp-2 cell substrate, smooth
muscle antibody has also been demonstrat-ed in
patients with warts (40).
anti-mitochondrial antibody (ama): Discrete speckles
concentrated in the perinuclear region of the cell and
extended in lower density to the outer regions of the
cytoplasm. This should be distinguished from anti-Golgi
antibody, which generally stains only one side of the
CÉLULA MITÓTICA
perinuclear region, and from anti-ribosomal antibody,
which demonstrates finer speckles with a strandlike
(Metafase)
appearance consistent with the location of the endo-
plasmic reticulum within the cell.
NOTE: Perinuclear speckles can most easily be dis-
tinguished from peripheral nuclear staining by noting
that the mitochondrial speckles form an interrupted
speckled staining around the outside of the nuclear
membrane, while peripheral sera form a solid smooth
staining inside the nuclear membrane.
REPORT SERA AS NEGATIVE FOR ANTINUCLEAR ANTI-
BODIES AND VERIFY POSITIVE FOR ANTIMITOCHONDRIAL
ANTIBODY ON AMA SPECIFIC SUBSTRATE.
anti-Smooth muscle antibody (aSma): Very fine
fibrous staining over the entire cytoplasm of cells with
MITOTISCHE ZELLE
a "spiderweb" appearance. Unlike mitochondrial
antibody, smooth muscle antibody staining is uniform
(Metaphase)
over the entire cytoplasm and may also extend over
the nucleus. Mitotic cells generally show large discrete
speckles outside the chromosome region. Smooth
muscle antibody has been shown to have a high speci-
ficity to actin (41-42).
REPORT SERA AS NEGATIVE FOR ANTINUCLEAR ANTIBODY
AND VERIFY POSITIVE FOR ANTI-SMOOTH MUSCLE ANTI-
BODY ON ASMA SPECIFIC SUBSTRATE.
lImItatIonS oF tHE tESt
1. Diagnosis cannot be made on the basis of anti-
nuclear antibody detection alone. The physician
must interpret these results in conjunction with the
MITOTISK CELL
patient's history and symptoms, the physical find-
(Metafas)
ings, and other diagnostic procedures.
2. Treatment should not be initiated on the sole basis
of a positive test for antinuclear antibodies. Clini-
Chromosome Region
Cytoplasm/
Nucleoplasm
Région chromosomique
Cytoplasme /
Nucléoplasme
La Zona Cromosomica
Citoplasma /
Nucleoplasma
Región Cromosómica
Citoplasma /
Nucleoplasma
Chromosomenbereich
Zytoplasma /
Nukleoplasma
ZELLE AUSSERHALB DER MITOSEPHASE
Kromosomområde
Cytoplasma /
Nukleoplasma
NON-MITOTIC CELL (Interphase)
cal indications, other laboratory findings, and the
physician's clinical impression must be considered
before any treatment is initiated.
3. Certain drugs, including procainamide and
hydralazine, may induce a lupus erythemato-
sus-like disease (43). Patients with drug-induced
LE may demonstrate positive homogeneous
or homogeneous/peripheral ANAs commonly
directed against nuclear histones (44).
4. A small percentage of patients with SLE may not
demonstrate ANAs by indirect immunofluorescence
but may have ANAs by other techniques (45).
5. Although a high-titered ANA may be highly sugges-
CELLULE NON MITOTIQUE
tive of connective tissue disease, it should not be
(Interphase)
considered diagnostic but rather viewed as a part
of the overall clinical history of a patient.
6. Staining patterns often change with progressive
titration of sera. This phenomenon is generally
due to the presence of more than one nuclear
antibody.
7. Because of the many options available in fluores-
cent microscopes, it is recommended that light
sources, filters, and optics be standardized when
comparing patient titers between laboratories.
8. Positive ANAs are also seen in a small percent-
age of patients with infectious and/or neoplastic
diseases (9).
EXPEctED ValuES
CELLULA NON IN MITOSI
In a large university medical center, using HEp-2 cell
(Interfase)
ANA substrate, the following data were generated over
a two-year period (46). Table 1.
PErFormancE cHaractErIStIcS
The Immuno Concepts ANA test system was evaluated
in comparison with two other fluorescent antibody tests
in commercial distribution (46). The study employed 97
serum samples from normal individuals as well as from
diagnoses including systemic lupus erythematosus (SLE),
mixed connective tissue disease (MCTD), Raynaud's-
progressive systemic sclerosis-CREST variant (PSS-CREST),
rheumatoid arthritis (RA), juvenile rheumatoid arthritis
CÉLULA NO MITÓTICA
(JRA), as well as other connective tissue disease. Sera
were tested at the recommended screening dilutions
(Interfase)
for each manufacturer. Study results are summarized
in table 2.
The patients positive in the "other connective tissue
disease" categories on IC's substrate had temporal
arthritis (1), undifferentiated connective tissue disease
(1), polymyositis (2), monoarthritis (2), polyarthritis (3),
and not further classifiable.
The hospitalized controls with positive ANAs on IC's
substrate had diabetes (2), unclassifiable arthritis (3),
hypothyroidism (1), and immune complex renal disease
(1) which did not meet criteria for diagnosis of SLE.
(Interphase)
ICKE-MITOTISK CELL
(Interfas)
6
Cytoplasm
Nuclear Membrane
Nucleoplasm
Nucleoli
Cytoplasme
Membrane Nucléaire
Nucléoplasme
Nucléoles
Citoplasma
Membrana Nucleare
Nucleoplasma
Nucleoli
Citoplasma
Membrana Nuclear
Núcleo
Nucleólos
Zytoplasma
Zellkermembran
Nukleoplasma
Nucleoi
Cytoplasma
Nukleärt Membran
Nukleoplasma
Nukleol
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