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, Robert G. Ehlenfeldt, Maria C. Athanasiou, Lisa A. Duvick, Hubert Heinrichs, Chella S. David, Harry T. Orr

Research output: Contribution to journalArticle

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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 - 1993

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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

Cite this

Schmidt, C. M., Ehlenfeldt, R. G., Athanasiou, M. C., Duvick, L. A., Heinrichs, H., David, C. S., & Orr, H. T. (1993). 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. Journal of Immunology, 151(5), 2633-2645.

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, Robert G.; Athanasiou, Maria C.; Duvick, Lisa A.; Heinrichs, Hubert; David, Chella S.; Orr, Harry T.

In: Journal of Immunology, Vol. 151, No. 5, 1993, p. 2633-2645.

Research output: Contribution to journalArticle

Schmidt, Cynthia M. ; Ehlenfeldt, Robert G. ; Athanasiou, Maria C. ; Duvick, Lisa A. ; Heinrichs, Hubert ; David, Chella S. ; Orr, Harry 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.
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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.",
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AU - Ehlenfeldt, Robert G.

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AU - Orr, Harry T.

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