A comparison of rainbow trout cell lines for their expression of the major histocompatibility complex genes and the induction of beta-2-microglobulin by dsRNA.
Western blotting with polyclonal antisera to polypeptides of the rainbow trout major histocompatibility (MH) genes and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to compare expression of MH genes in rainbow trout cell lines. One line was the spleen monocyte/macrophage-like RTS11, which grew loosely on plastic surfaces. Adherent cell lines were fibroblast-like RTG-2 from gonads and four epithelial-like cell lines from gill, intestine, liver and hepatoma: RTgill-W1, RTgutGC, RTL-W1, and RTH-149 respectively. All cell lines expressed a 45 kDa MHC class I alpha chain. All cell lines expressed beta-2-microglobulin (beta2m), which was at 11 kDa, but detection was abrogated following trypsinization prior to cell collection. All cell lines expressed transcripts for MH class II alpha and MH class II beta genes; however, MH class II polypeptides were expressed only in RTS11, the only cell line from a lineage of antigen-presenting cells. We report here that double stranded RNA up regulates beta2m and that these cell lines and antisera can be employed for studying MH regulation.
Kawano
Atsushi
A
Department of Biology, University of Waterloo, 200 University Ave., Waterloo, ON, Canada N2L 3G1.
Kales
Stephen C
SC
Fujiki
Kazuhiro
K
DeWitte-Orr
Stephanie J
SJ
Dixon
Brian
B
Lee
Lucy E J
LE
Bols
Niels C
NC
D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D008285:Major Histocompatibility Complex / Q000276:immunology*; D017686:Oncorhynchus mykiss / Q000276:immunology*; D011070:Poly I-C / Q000494:pharmacology; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D001613:beta 2-Microglobulin / Q000276:immunology*
D000276:Adjuvants, Immunologic; D012330:RNA, Double-Stranded; D001613:beta 2-Microglobulin; D011070:Poly I-C