Ann Rheum Dis 2001;60:248-253 ( March )

Extended report

Lack of evidence for an increased microchimerism in the circulation of patients with Sjögren's syndrome I Toda^a, M Kuwana^a, K Tsubota^b, Y Kawakami^a

^a Institute for Advanced Medical Research, Keio University School of Medicine, Tokyo, Japan, ^b Department of Ophthalmology, Tokyo Dental College, Japan

Correspondence to: Dr M Kuwana, Institute for Advanced Medical Research, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan kuwanam{at}med.keio.ac.jp

Accepted for publication 15 May 2000

OBJECTIVETo examine the hypothesis that fetal microchimerism plays a part in the pathogenic process of Sjögren's syndrome (SS).
METHODSGenomic DNA samples were extracted from peripheral blood whole nucleated cells and the CD34+ cell enriched fraction of patients with SS and healthy women who had male offspring as well as nulliparous women. A Y chromosome-specific sequence was detected as a marker for fetal cells by a nested polymerase chain reaction (PCR) and by DNA hybridisation combined with PCR using specific primers and probes. All procedures were performed with great care to avoid the contamination of male DNA.
RESULTSA nested PCR and DNA hybridisation combined with PCR was established that can detect a single male cell out of 1.67×10⁵ female cells. It was not possible to increase the sensitivity further because the amount of template DNA held in the PCR was limited. When these methods were used, no fetal cells were detected in any samples from patients with SS, though they were detected in whole nucleated cells from two healthy women who had delivered sons previously.
CONCLUSIONSThe findings indicate that circulating fetal cells in patients with SS are uncommon (<1 in 1.67×10⁵), if they exist. With the conventional PCR based methods that were used, it is difficult to evaluate the quantitative difference in circulating fetal cells between patients with SS and healthy women.

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© 2001 by Annals of the Rheumatic Diseases
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