충북대학교 종양연구소에서 다음과 같이 세미나를 개최하오니 참여하여 주시기 바랍니다.
- 다 음 -
1. 연 자 : 권 희 충 박사
(원자력의학원 방사건 의학연구센터 분자종양학실)
2. 일 시 : 2004년 6월 30일 오후 4시
3. 장 소 : 종양연구소 106호(세미나실)
4. 주 제 : Targeting HSV infection to gD-Receptor-Negative Cells Using a Soluble Receptor
5. 발표내용 :
HSV-1 initially attaches to cells through the binding of glycoproteins C and B (gC and gB) to glycosaminoglycans (GAGs) such as heparan sulphate (HS) moieties. Subsequent entry requires interaction between gD and one of the cell surface receptors, HveA or HveC. Virus-cell fusion requires the presence of gD, gB, gH/gL in the virion, but details of the process are not fully understood. A more detailed understanding of the process involved in virus-cell fusion may assist in the generation of targeted gene delivery vectors for experimental and therapeutic applications.
The overall goal of our studies is to develop methods for target cell-specific HSV infection by directing the virus to novel receptors. One lead we have followed is the use of soluble receptor molecules. We previously reported that a soluble V-domain fragment (Nec1123) of the natural HSV gD receptor nectin1 enabled HSV-1 entry into CHO-K1 cells lacking gD receptors and that this phenomenon was dependent on virus binding to cellular glycosaminoglycans (GAGs). We have performed optimization studies and now routinely obtain 75% CHO cell transduction using a replication-defective virus at an MOI of 3, comparable to the entry efficiency of this virus on CHO cells bearing authentic nectin1. We have extended these studies to test additional soluble receptor constructs. We find that a gD binding-deficient mutant version of Nec1123 fails to mediate virus entry while a nectin2 (HveB) counterpart of Nec1123 stimulates CHO cell infection by HSV-1 Rid1 but not wild-type virus. Curiously, soluble HveA as well as soluble nectin1 fragments consisting of the V domain along with one or both C2-like domains have thus far failed to mediate high levels of CHO cell transduction. Despite these limitations, our observations may be used to direct gD-receptor-independent infection of viruses attachable to cell-specific receptors via novel ligands replacing the GAG-binding regions of gC and gB. We have also shown that Nec1123 inhibits virus entry via cell-surface nectin1, suggesting that this approach may provide considerable cell-type specificity.