Lack of evidence for an increased microchimerism in the circulation of patients with Sjögren's syndrome

I. Toda, M. Kuwana, Kazuo Tsubota, Yutaka Kawakami

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Objective - To examine the hypothesis that fetal microchimerism plays a part in the pathogenic process of Sjögren's syndrome (SS). Methods - Genomic 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. Results - A 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. Conclusions - The 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.

Original languageEnglish
Pages (from-to)248-253
Number of pages6
JournalAnnals of the Rheumatic Diseases
Volume60
Issue number3
DOIs
Publication statusPublished - 2001

Fingerprint

Chimerism
Polymerase chain reaction
DNA
Polymerase Chain Reaction
DNA Contamination
Chromosomes
Contamination
Blood
Y Chromosome
Nuclear Family

ASJC Scopus subject areas

  • Rheumatology
  • Immunology

Cite this

Lack of evidence for an increased microchimerism in the circulation of patients with Sjögren's syndrome. / Toda, I.; Kuwana, M.; Tsubota, Kazuo; Kawakami, Yutaka.

In: Annals of the Rheumatic Diseases, Vol. 60, No. 3, 2001, p. 248-253.

Research output: Contribution to journalArticle

@article{526d42f921ba4e49951d96d3873007fa,
title = "Lack of evidence for an increased microchimerism in the circulation of patients with Sj{\"o}gren's syndrome",
abstract = "Objective - To examine the hypothesis that fetal microchimerism plays a part in the pathogenic process of Sj{\"o}gren's syndrome (SS). Methods - Genomic 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. Results - A 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. Conclusions - The 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.",
author = "I. Toda and M. Kuwana and Kazuo Tsubota and Yutaka Kawakami",
year = "2001",
doi = "10.1136/ard.60.3.248",
language = "English",
volume = "60",
pages = "248--253",
journal = "Annals of the Rheumatic Diseases",
issn = "0003-4967",
publisher = "BMJ Publishing Group",
number = "3",

}

TY - JOUR

T1 - Lack of evidence for an increased microchimerism in the circulation of patients with Sjögren's syndrome

AU - Toda, I.

AU - Kuwana, M.

AU - Tsubota, Kazuo

AU - Kawakami, Yutaka

PY - 2001

Y1 - 2001

N2 - Objective - To examine the hypothesis that fetal microchimerism plays a part in the pathogenic process of Sjögren's syndrome (SS). Methods - Genomic 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. Results - A 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. Conclusions - The 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.

AB - Objective - To examine the hypothesis that fetal microchimerism plays a part in the pathogenic process of Sjögren's syndrome (SS). Methods - Genomic 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. Results - A 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. Conclusions - The 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.

UR - http://www.scopus.com/inward/record.url?scp=0035089869&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035089869&partnerID=8YFLogxK

U2 - 10.1136/ard.60.3.248

DO - 10.1136/ard.60.3.248

M3 - Article

C2 - 11171687

AN - SCOPUS:0035089869

VL - 60

SP - 248

EP - 253

JO - Annals of the Rheumatic Diseases

JF - Annals of the Rheumatic Diseases

SN - 0003-4967

IS - 3

ER -