Biomedical subjects

Cheong-Hee Chang

Publications and source records attributed to Cheong-Hee Chang.

22 records · Page 2Linked to original sources

PubMed2003

Gene transfection and expression in resting and activated murine CD4 T cell subsets.

It has been difficult to assess the role of specific genes in activation and differentiation of peripheral T cell subsets such as naive, effector and memory T cells due to the impairments in T cell development and immune pathologies often observed in genetically manipulated mouse models, and the lack of reliable methods for introducing genes into primary mouse T cells. In this study, we demonstrate transient transfection of genes into resting and activated mouse CD4 T cell subsets using "Nucleofection", a modified electroporation technique. Using this approach, cDNA encoding green fluorescent protein (GFP) is efficiently taken up and expressed by purified polyclonal and antigen-specific mouse naive, effector and memory CD4 T cells isolated from BALB/c or TCR-transgenic mice. The resultant transfected resting T cells are fully amenable to TCR-mediated activation. We also demonstrate that expression of endogenous gene can be turned on in resting T cells by transfection of a transcriptional transactivator. Our results demonstrate for the first time, the expression of exogenously transfected genes and the modulation of endogenous gene expression in primary mouse T cell subsets. This technology will enable a variety of mechanistic questions on T cell activation, function and signaling to be addressed in T cells that differ in activation history and functional capacities.

Animals↗

PubMed2003

Phosphorylation of class II transactivator regulates its interaction ability and transactivation function.

The MHC class II transactivator (CIITA) plays a central role in adaptive immune responses by controlling the expression of MHC class II genes. CIITA binds DNA-binding proteins and co-activator proteins to form an enhanceosome complex necessary for MHC class II gene expression. Here we demonstrate that CIITA interactions depend upon the phosphorylation status of CIITA. Hyper-phosphorylated CIITA interacts with co-activator p300, RFX5 and CIITA itself, which in turn results in induction of MHC class II promoter activity. Moreover, the C-terminal region of CIITA containing leucine-rich repeats (LRR) is a regulatory domain for CIITA self-association and LRR binding to CIITA is negatively regulated by phosphorylation. cAMP-dependent protein kinase (PKA) phosphorylates CIITA, and serine residues residing in a region between the proline/serine/threonine-rich domain and the GTP-binding domain are phosphorylated by PKA in vitro. The maximum transactivation potential of CIITA requires PKA phosphorylation as demonstrated by reduced transactivation activities of the mutant bearing substitutions of serine residues at the PKA site.

Amino Acid Sequence↗

PubMed2002

Aberrant expression of Fas ligand in mice deficient for the MHC class II transactivator.

The MHC class II transactivator (CIITA) is a critical regulator of MHC class II genes and other genes involved in the Ag presentation pathway. CIITA-deficient mice lack MHC class II expression on almost all APCs. In this study, we show that these mice also have aberrant Fas ligand expression on both CD4 T cells and B cells. We found that Fas ligand expression was greatly increased on CIITA-deficient CD4 T cells during the Th1 differentiation process. However, both CIITA-deficient and control Th1 effector cells up-regulated Fas ligand to similar levels if cells were reactivated. The introduction of CIITA into primary CD4 T cells via retroviral infection resulted in a reduction in the level of Fas ligand and delay in apoptosis after activation. Interestingly, activated B cells from the CIITA-deficient mice also showed increased levels of Fas ligand that could be to some degree inhibited by the introduction of IL-4.

Animals↗

PubMed2002

Function and regulation of class II transactivator in the immune system.

The class II transactivator (CIITA) is a potent and critical transcriptional regulator. It activates genes necessary for antigen presentation function. It also regulates cytokine gene expression in CD4 T cells. We recently found that CIITA prevents cell death by inhibiting Fas ligand (FasL) gene expression. Thus, CIITA regulates multiple immune responses. The activation and the repression function of CIITA are mediated by its interaction with other transcription factors. To activate the target gene, CIITA interacts with DNA binding proteins and recruits the coactivator CBP/p300 to the promoter forming an enhanceosome necessary for transcription. In addition, CIITA interacts with self. Inter- and intramolecular interactions of CIITA are essential for transactivation function. Each domain of CIITA has a distinct role and all domains are required for CIITA activity. However, the regulatory mechanisms of CIITA interaction with self and other proteins are poorly understood and remain to be investigated.

Animals↗