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Hemoflagellate kinetoplasts and the diagnosis of SLE.

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REFERENCES
1. Pearson CM: Relapsing polychondritis, Arthritis and Al-
lied Conditions. Eighth edition. Edited by JL Hollander
and DJ McCarty, Jr. Philadelphia, Lea & Febiger, 1971, pp
866-873
2. Mannik M, Gilliland BC: Miscellaneous arthritides and ex-
traarticular rheumatism, Harrison’s Principles of Internal
Medicine. Eighth edition. Edited by GW Thorn, et al. New
York, McGraw-Hill, 1977, pp 2077-2080
3. Christian CL: Diseases with which arthritis is frequently
associated, Textbook of Medicine. Fourteenth edition. Edited by PB Beeson and W McDermott. Philadelphia, Saunders, 1975, pp 154-155
4. Martin J, Roenigk HH, Lynch W, Tingwald FR: Relapsing
polychondritis treated with dapsone. Arch Dermatol
112:1272-1274, 1976
5 . Barranco VP: Inhibition of lysosomal enzymes by dapsone.
Arch Dermatol 110563-566, 1974
Hemoflagellate kinetoplasts and the diagnosis
of SLE
To the Editor:
Recently, Aarden et a1 described an immunofluorescent method capable of detecting antibodies to
dsDNA by using the kinetoplast of the flagellate, Cri-
thidia luciliae as a substrate (1). The kinetoplast of these
organisms is a modified mitochondrion containing a localized concentration of dsDNA, which has been shown
by others to be exclusively double stranded DNA (2,3).
By use of well characterized sera specific for dsDNA,
the observations of Aarden et a1 were further confirmed
by several investigators ( 4 4 .
Since in several developing countries Trypanosoma organisms are used as substrate for the diagnosis
of Chagas’ disease, we have compared the Crithidia
method with two other hemoflagellates, namely Trypanosoma and Leishmania. The organisms were maintained in Roithman’s nondefined liquid media at room
temperature. To perform the test the organisms were air
dried on glass slides and fixed in 95% ethanol to serve as
a substrate. Lupus sera were serially diluted until the
highest reactive titer.
In Figures 1 to 3 we present the indirect immunofluorescence preparation of SLE patients with high
levels of antibodies to DNA using as organisms Crithidia, Trypanosoma, and Leishmania, respectively.
Strong staining of the kinetoplasts is evident with Crithidia whereas with the two other preparations the staining is less evident since the kinetoplast lies close to the
nuclei and both appear equally fluorescent, which is not
the case with Crithidia luciliae.
Figure 1. Crithidia organisms incubated with lupus sera and stained with fluorescein labeled antihuman IgG. Note the
strong staining of the kinetoplasts when compared to the nuclei (X IOOO).
206
Figure 2. Trypanosoma organisms incubated with lupus sera and stained with fluorescein labeled antihuman IgG. The kinetoplast and the nuclei are equally fluorescent and close one to each other (X 1OOO).
Figure 3. Leishmania organisms incubated with lupus sera and stained with fluorescein labeled antihuman IgG. The kinetoplast and the nuclei are strongly fluorescent and in close relation to each other (X IOOO).
207
It is our impression that of the three hemoflagellates tested, only the Crithidia organisms (as was
previously suggested by Aarden) are a substrate that
with indirect immunofluorescence will provide a
method readily reproducible in most clinical laboratories.
MORTONA. SCHEINBERG,
MD, PHD
ERNEYPLESSMANN DE CAMARGO, MD
NELSONF. MENDES,MD
Divisions of Immunology
and Parasitology
Escola Paulista de Medicina
Siio Paulo, Brazil
Supported by grants from the Wellcome Trust, Heisen
Fellowshipfor Leprosy Research, and CNPq.
REFERENCES
1. Aarden LA, de Groot ER, Feltkamp RE: Immunology of
DNA. 111. Crithidia luciliae, a simple substrate for the detection of anti dsDNA with the immunofluorescent technique. Ann NY Acad Sci 254:505-515, 1975
2. Fouts DL, Manning JE, Wolstenholme DR: Physicochemical properties of kinetoplast DNA crithidia acantocephalic. Crithidia luciliae and Trypanosoma lewisi. J Cell
Biol 67:378-399, 1975
3. Rion C, Delain E: Electron microscopy of the circular kinetoplast DNA from Trypanosoma cruzi: occurrence of attenuated forms. Proc Nat Acad Sci USA 62:2 10-2 17, 1969
4. Davis P, Christian B, Russell AS: Immunofluorescent technique for the detection of antibodies to n-DNA. Comparison with radioimmunoassay. J Rheumatol 4 12-20,
1977
5. Crowe W, Kushner I: An immunofluorescent method using
Crithidia luciliae to detect antibodies to double-stranded
DNA. Arthritis Rheum 20231 1-814, 1977
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