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PMID: 3279151 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Rescue of Daudi cell HLA expression by transfection of the mouse beta 2-microglobulin gene.

The Journal of experimental medicine ·Vol. 167 ·No. 2 ·1988-02-01 ·Pages 288-99

Seong RH, Clayberger CA, Krensky AM, Parnes JR

Abstract

The Daudi cell line is a B-lymphoblastoid line derived from a Burkitt lymphoma. Daudi cells lack cell surface expression of class I HLA molecules despite the presence of intracellular class I heavy chains. They have a defect in the gene encoding beta 2-microglobulin (beta 2m), resulting in lack of translatable mRNA for this protein. It has been thought that this deficiency is responsible for the lack of cell surface class I expression. However, data have recently been presented demonstrating that at least one mouse class I heavy chain can be expressed on the cell surface in the absence of beta 2m. These results raised the questions of whether the lack of beta 2m is the only defect in Daudi and whether transfer of this single gene could restore surface class I expression. We found that transfection of the mouse beta 2m gene into Daudi indeed rescued cell surface expression of class I HLA molecules, and that these molecules could be recognized both by monomorphic and allospecific mAbs. CTL clones specific for HLA-B17 or a determinant shared by HLA-B17 and HLA-A2 killed the Daudi cells transfected with the beta 2m gene, but not untransfected Daudi or Daudi transfected with vector alone. Mouse beta 2m on the transfected Daudi cells could exchange with human beta 2m when the cells were incubated in human serum. This exchange did not alter the ability of the cells to be killed by the specific CTLs. These results demonstrate that the lack of beta 2m is the sole reason for lack of surface class I molecules in Daudi cells, and that beta 2m is required for cell surface expression of the specific class I heavy chains of Daudi.

MeSH Terms
Animals Antigens, Neoplasm/analysis,isolation & purification Antigens, Surface/analysis Burkitt Lymphoma/analysis,genetics,immunology Cell Line Cytotoxicity, Immunologic Extracellular Space/immunology Genes, MHC Class I HLA Antigens/analysis,isolation & purification Humans Mice Mice, Inbred C57BL RNA, Messenger/analysis Transfection beta 2-Microglobulin/genetics,isolation & purification
Chemicals
Antigens, Neoplasm Antigens, Surface HLA Antigens RNA, Messenger beta 2-Microglobulin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Seong R H
Department of Medicine, Stanford University Medical Center, California 94305.
Clayberger C A
Krensky A M
Parnes J R
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Article Info
Journal
The Journal of experimental medicine
Abbr.
J Exp Med
ISSN
0022-1007
Published
1988-02-01
Pages
288-99
Language
English
Region
United States
NLM ID
2985109R
PMCID
PMC2188848
Subset
IM
Grants
NIGMS NIH HHS · GM-34991 · United States
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