Extraembryonic expression of the human MHC class I gene HLA-G in transgenic mice

Evidence for a positive regulatory region located 1 kilobase 5' to the start site of transcription

Cynthia M Schmidt, R. G. Ehlenfeldt, M. C. Athanasiou, L. A. Duvick, H. Heinrichs, C. S. David, H. T. Orr

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Trophoblast, the only fetal tissue in direct contact with maternal cells, fails to express the polymorphic HLA class I molecules HLA-A and -B, but does express the nonpolymorphic class I molecule HLA-G. It is thought that HLA-G may provide some of the functions of a class I molecule without stimulating maternal immune rejection of the fetal semiallograft. As a first step in identifying the cis-acting DNA regulatory elements involved in the control of class I expression by extraembryonic tissue, several types of transgenic mice were produced. Two HLA-G genomic fragments were used, 5.7 and 6.0 kb in length. These included the entire HLA-G coding region, 1 kb of 3' flanking sequence, and 1.2 or 1.4 kb of 5' flanking sequence, respectively. A hybrid transgene, HLA-A2/G, was produced by replacing the 5' flanking sequence, first exon, and early first intron of HLA-G with the corresponding elements of HLA-A. Comparison of transgene mRNA expression patterns seen in HLA-A2/G and HLA-G transgenic mice suggests that 5' flanking sequences are largely responsible for the differing patterns of expression typical of the classical class I and HLA-G genes. Studies comparing the extraembryonic HLA-G expression levels of founder embryos transgenic for either the 5.7- or 6.0- kb HLA-G transgene showed that the 6.0-kb transgene directed HLA-G expression far more efficiently than did the 5.7-kb HLA-G transgene, producing extraembryonic HLA-G mRNA levels similar to those seen in human extraembryonic tissues. The results of these studies suggest that the 250-bp fragment present at the extreme 5' end of the 6.0-kb HLA-G transgene and absent from the 5.7-kb HLA-G transgene contains an important positive regulatory element. This 250-bp fragment lies further upstream than any of the previously documented class I regulatory regions and may function as a locus control region.

Original languageEnglish (US)
Pages (from-to)2633-2645
Number of pages13
JournalJournal of Immunology
Volume151
Issue number5
StatePublished - Jan 1 1993

Fingerprint

HLA-G Antigens
MHC Class I Genes
Nucleic Acid Regulatory Sequences
Transcription Initiation Site
Transgenic Mice
Transgenes
5' Flanking Region
HLA-A2 Antigen
HLA-A Antigens
Locus Control Region
Mothers
3' Flanking Region
Messenger RNA
HLA-B Antigens

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Extraembryonic expression of the human MHC class I gene HLA-G in transgenic mice : Evidence for a positive regulatory region located 1 kilobase 5' to the start site of transcription. / Schmidt, Cynthia M; Ehlenfeldt, R. G.; Athanasiou, M. C.; Duvick, L. A.; Heinrichs, H.; David, C. S.; Orr, H. T.

In: Journal of Immunology, Vol. 151, No. 5, 01.01.1993, p. 2633-2645.

Research output: Contribution to journalArticle

Schmidt, Cynthia M ; Ehlenfeldt, R. G. ; Athanasiou, M. C. ; Duvick, L. A. ; Heinrichs, H. ; David, C. S. ; Orr, H. T. / Extraembryonic expression of the human MHC class I gene HLA-G in transgenic mice : Evidence for a positive regulatory region located 1 kilobase 5' to the start site of transcription. In: Journal of Immunology. 1993 ; Vol. 151, No. 5. pp. 2633-2645.
@article{840fa6e0f7f54013b0360bff6967ca03,
title = "Extraembryonic expression of the human MHC class I gene HLA-G in transgenic mice: Evidence for a positive regulatory region located 1 kilobase 5' to the start site of transcription",
abstract = "Trophoblast, the only fetal tissue in direct contact with maternal cells, fails to express the polymorphic HLA class I molecules HLA-A and -B, but does express the nonpolymorphic class I molecule HLA-G. It is thought that HLA-G may provide some of the functions of a class I molecule without stimulating maternal immune rejection of the fetal semiallograft. As a first step in identifying the cis-acting DNA regulatory elements involved in the control of class I expression by extraembryonic tissue, several types of transgenic mice were produced. Two HLA-G genomic fragments were used, 5.7 and 6.0 kb in length. These included the entire HLA-G coding region, 1 kb of 3' flanking sequence, and 1.2 or 1.4 kb of 5' flanking sequence, respectively. A hybrid transgene, HLA-A2/G, was produced by replacing the 5' flanking sequence, first exon, and early first intron of HLA-G with the corresponding elements of HLA-A. Comparison of transgene mRNA expression patterns seen in HLA-A2/G and HLA-G transgenic mice suggests that 5' flanking sequences are largely responsible for the differing patterns of expression typical of the classical class I and HLA-G genes. Studies comparing the extraembryonic HLA-G expression levels of founder embryos transgenic for either the 5.7- or 6.0- kb HLA-G transgene showed that the 6.0-kb transgene directed HLA-G expression far more efficiently than did the 5.7-kb HLA-G transgene, producing extraembryonic HLA-G mRNA levels similar to those seen in human extraembryonic tissues. The results of these studies suggest that the 250-bp fragment present at the extreme 5' end of the 6.0-kb HLA-G transgene and absent from the 5.7-kb HLA-G transgene contains an important positive regulatory element. This 250-bp fragment lies further upstream than any of the previously documented class I regulatory regions and may function as a locus control region.",
author = "Schmidt, {Cynthia M} and Ehlenfeldt, {R. G.} and Athanasiou, {M. C.} and Duvick, {L. A.} and H. Heinrichs and David, {C. S.} and Orr, {H. T.}",
year = "1993",
month = "1",
day = "1",
language = "English (US)",
volume = "151",
pages = "2633--2645",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "5",

}

TY - JOUR

T1 - Extraembryonic expression of the human MHC class I gene HLA-G in transgenic mice

T2 - Evidence for a positive regulatory region located 1 kilobase 5' to the start site of transcription

AU - Schmidt, Cynthia M

AU - Ehlenfeldt, R. G.

AU - Athanasiou, M. C.

AU - Duvick, L. A.

AU - Heinrichs, H.

AU - David, C. S.

AU - Orr, H. T.

PY - 1993/1/1

Y1 - 1993/1/1

N2 - Trophoblast, the only fetal tissue in direct contact with maternal cells, fails to express the polymorphic HLA class I molecules HLA-A and -B, but does express the nonpolymorphic class I molecule HLA-G. It is thought that HLA-G may provide some of the functions of a class I molecule without stimulating maternal immune rejection of the fetal semiallograft. As a first step in identifying the cis-acting DNA regulatory elements involved in the control of class I expression by extraembryonic tissue, several types of transgenic mice were produced. Two HLA-G genomic fragments were used, 5.7 and 6.0 kb in length. These included the entire HLA-G coding region, 1 kb of 3' flanking sequence, and 1.2 or 1.4 kb of 5' flanking sequence, respectively. A hybrid transgene, HLA-A2/G, was produced by replacing the 5' flanking sequence, first exon, and early first intron of HLA-G with the corresponding elements of HLA-A. Comparison of transgene mRNA expression patterns seen in HLA-A2/G and HLA-G transgenic mice suggests that 5' flanking sequences are largely responsible for the differing patterns of expression typical of the classical class I and HLA-G genes. Studies comparing the extraembryonic HLA-G expression levels of founder embryos transgenic for either the 5.7- or 6.0- kb HLA-G transgene showed that the 6.0-kb transgene directed HLA-G expression far more efficiently than did the 5.7-kb HLA-G transgene, producing extraembryonic HLA-G mRNA levels similar to those seen in human extraembryonic tissues. The results of these studies suggest that the 250-bp fragment present at the extreme 5' end of the 6.0-kb HLA-G transgene and absent from the 5.7-kb HLA-G transgene contains an important positive regulatory element. This 250-bp fragment lies further upstream than any of the previously documented class I regulatory regions and may function as a locus control region.

AB - Trophoblast, the only fetal tissue in direct contact with maternal cells, fails to express the polymorphic HLA class I molecules HLA-A and -B, but does express the nonpolymorphic class I molecule HLA-G. It is thought that HLA-G may provide some of the functions of a class I molecule without stimulating maternal immune rejection of the fetal semiallograft. As a first step in identifying the cis-acting DNA regulatory elements involved in the control of class I expression by extraembryonic tissue, several types of transgenic mice were produced. Two HLA-G genomic fragments were used, 5.7 and 6.0 kb in length. These included the entire HLA-G coding region, 1 kb of 3' flanking sequence, and 1.2 or 1.4 kb of 5' flanking sequence, respectively. A hybrid transgene, HLA-A2/G, was produced by replacing the 5' flanking sequence, first exon, and early first intron of HLA-G with the corresponding elements of HLA-A. Comparison of transgene mRNA expression patterns seen in HLA-A2/G and HLA-G transgenic mice suggests that 5' flanking sequences are largely responsible for the differing patterns of expression typical of the classical class I and HLA-G genes. Studies comparing the extraembryonic HLA-G expression levels of founder embryos transgenic for either the 5.7- or 6.0- kb HLA-G transgene showed that the 6.0-kb transgene directed HLA-G expression far more efficiently than did the 5.7-kb HLA-G transgene, producing extraembryonic HLA-G mRNA levels similar to those seen in human extraembryonic tissues. The results of these studies suggest that the 250-bp fragment present at the extreme 5' end of the 6.0-kb HLA-G transgene and absent from the 5.7-kb HLA-G transgene contains an important positive regulatory element. This 250-bp fragment lies further upstream than any of the previously documented class I regulatory regions and may function as a locus control region.

UR - http://www.scopus.com/inward/record.url?scp=0027247977&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027247977&partnerID=8YFLogxK

M3 - Article

VL - 151

SP - 2633

EP - 2645

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 5

ER -