Cholera toxin induced novel genes in human lymphocytes and monocytes.
Cholera toxin (CT) is well known as an inducer of the accumulation of cellular cAMP through the ADP-ribosylation of the Gs protein by CT. CT is also one of the most powerful mucosal adjuvants. However, the molecular mechanisms of the CT adjuvanticity are not well understood. Here, the transcriptional responses of cultured human lymphocytes and monocytes in response to CT were analyzed using differential display-PCR. The full complement of cellular mRNA was examined by high resolution polyarylamide gel electrophoresis and sequence analyses of the PCR products of 240 primer sets. Over 100 genes with altered expression were initially identified. The expressions of 65 of these genes were further analyzed and confirmed using custom glass cDNA arrays, RT-PCR and real-time PCR. Immunomodulatory genes such as CD2, HIF1, CXCL2, L-plastin, LILR and IFI30 were affected by CT. In addition, 14 novel genes with previously unknown functions were found to be CT induced. These CT induced gene expression alterations provide more insight in the mechanisms of CT actions. The CT induced gene expressions alterations could contribute to the CT adjuvanticity.
Royaee
Atabak R
AR
Department of Chemistry, Georgetown University, 37th & 654 Reiss Science Bldg, Washington, DC 20057, USA.
Jong
Linda
L
Mendis
Chanaka
C
Das
Rina
R
Jett
Marti
M
Yang
David C H
DC
D018801:CD2 Antigens / Q000235:genetics; D002772:Cholera Toxin / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics; D008214:Lymphocytes / Q000187:drug effects* / Q000378:metabolism*; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018801:CD2 Antigens; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; D002772:Cholera Toxin