Autoantibodies and Autoimmunity
467
Cells are
permeabilized
and fixed
Cells are incubated
with an overlay of
diluted autoantibody
Cells are incubated
with an overlay of
fluorochrome-tagged
anti-antibody
Cells are viewed with a
fluorescence microscope
to visualize the antibody-
labeled autoantigen
Cells
Glass slide
Wash
Wash
Fig. 2
Diagrammatic representation of the steps involved in the indirect
immunofluorescence (IIF) test: see text for explanation.
have identifed unexpected protein dis-
tributions
such
as
the
localization
oF
the nucleolar protein fbrillarin to the
outer surFace oF the chromosomes dur-
ing cell division (±ig. 3e, arrowheads). The
localization oF some autoantigens during
the cell cycle has aided in their identifca-
tion. Detection oF proliFerating cell nuclear
an
t
igen(PCNA
)inS
-phasece
l
ls(±
ig
.3c
)
suggested its involvement in DNA synthe-
sis, while the distribution oF speckles along
the metaphase plate produced by other au-
toantibodies (±ig. 3F, arrowheads) was an
important clue in their identifcation as
autoantibodies to centromeric proteins A,
B, and C.
The II± test has also proved useFul
in the identifcation oF autoantibodies
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