Stable transfection of a human lymphoma line by sub-genomic fragments of Epstein-Barr virus DNA to measure humoral and cellular immunity to the corresponding proteins
✍ Scribed by Christina Welinder; Nils-Göran Larsson; Robert Szigeti; Barbro Ehlin-Henriksson; Gertrude Henle; Werner Henle; George Klein; Anne Ricksten; Lars Rymo; Dov Sulitzeanu
- Publisher
- John Wiley and Sons
- Year
- 1987
- Tongue
- French
- Weight
- 957 KB
- Volume
- 40
- Category
- Article
- ISSN
- 0020-7136
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✦ Synopsis
An Epstein-Barr virus (EBV)-negative human lymphoid Bcell line, DG75, was stably transfected with recombinant selection vectors that carry a subfragment of the BamHl WYH region (nucleotides 44664 to 50628). the BamHl K fragment, or a subfragment of the EcoRl D region (nucleotides 166614 to 170149) of 895-8 EBV DNA. These fragments contain the coding exons for the EBV-determined nuclear antigens EBNA2 and EBNA I, and the membrane antigen LMP, respectively. Antigen expression of the cells was detected by immunofluorescence. EBNAZ was expressed in 80-100% of the transfected cells, in contrast to EBNAI which was expressed in only 251, and LMP in only about 5% of the cells. Humoral antibody responses were measured by immunofluorescence and compared to cellular immunity as determined by the leukocyte migration inhibition (LMI) technique. Extracts from transfected cell lines expressing EBNAI, EBNA2 or LMP elicited an LMI response with cells from healthy EBV-seropositive individuals whereas the extract from the parental DC75 cell line did not. The results demonstrate the value of stably transfected cell lines expressing a defined EBV antigen for the monospecific analysis of host responses to the EBV-encoded antigen complex in growth-transformed cells.