1996 Fiscal Year Final Research Report Summary
Gene therapy for adult T-cell leukemia using HIV vector
| Project/Area Number |
07671218
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Hematology
|
|---|
| Research Institution | Kumamoto University |
|---|
Principal Investigator |
MATSUSHITA Shuzo Kumamoto University Hospital Assistant Professor, 医学部・附属病院, 助手 (00199788)
|
|---|
| Project Period (FY) |
1995 – 1996
|
| Keywords | gene therapy / ATL / HIV vector / Thymidine kinase / ganciclovir |
|---|
| Research Abstract |
Adult T-cell leukemia/lymphoma (ATL) is derived from CD4-positive T-cells, and has a poor prognosis because of its resistance to chemotherapy. To evaluate the effectiveness of gene therapy for ATL,the effect of ganciclovir on ATL cell lines transfected with the thymidine kinase gene of herpes simplex type 1 (HSV-TK) was analyzed. To tranfer the HSV-TK gene to ATL cells, an HIV-vector that has specific infectivity to CD4-positive cells was used. The thymidine kinase gene of herpes simplex virus type 1 was inserted into the long terminal repeats of human immunodeficiency virus type-1 (HIV-1) and driven by the SL3 promoter (named HXBSL3TK). HXBSL3TK was cotransfected with HXBCAT as a reporter into MT2 or HUT102 cells by DEAE-dextran. The cells were incubated with ganciclovir, and chloramphenicol acetyltransferase (CAT) activity was analyzed. The CAT activity of the MT2 cells and HUT102 cells transfected with HXBSL3TK decreased dosedependently with ganciclovir. HXBSL3TK was also contransfe
… More
cted into Cos cells with an HIV-1 packaging vector that has gag, pol and env driven by a cytomegalovirus promoter. The supernatant was transferred to MT2 cells or Raji cells, and incubated with ganciclovir. Ninety percent of the MT2 cells transduced by HXBSL3TK and incubated with ganciclovir were killed, but Raji cells were not killed. In addition HXBTK that expresses the HSV-TK gene and Tat gene driven by the LTR of HIV-1 was constructed. HXBTK had a higher expression of the HSV-TK gene and higher sensitivity to ganciclovir than did HXBSL3TK.Furthermore, the packaging cell lines were established, effective transduction to the fresh ATL cells were demonstrated.
HIV vector has been shown to be one of the most promising new tools for human gene therapy strategy. In this study, the effect of ganciclovir on adult T-cell leukemia (ATL) cell lines transduced with HIV vector carrying the HSV-TK gene was evaluated. A high efficiency of killing of ATL cell lines and specificity for CD4-positive cells were demonstrated. These results suggest the effectiveness of HIV vector carrying the HSV-TK gene for gene therapy for ATL. Less
|
