AEGiS-13IAC: In vivo survival of Ul/anti-HIV-1 antisense RNA-expressing CD4+ cells from transduced CD34+ PBSC in HIV-1 positive individuals.

13th International AIDS Conference

Durban, South Africa - July 9-July 14, 2000

In vivo survival of Ul/anti-HIV-1 antisense RNA-expressing CD4+ cells from transduced CD34+ PBSC in HIV-1 positive individuals.

Int Conf AIDS 2000 Jul 9-14; 13:40 (abstract no.. LbPeA7033)

Engelhardt DL, Liu D, Wit Q, Ho J, Dunn E, Ng J, Cowan M, Conant M, Eden C, Laurence J, Thalenftld B
Enzo Biochem, New York, NY 10022. Fax: +1 212-583-0150, E-mail: Info@corp.enzobio.com.

OBJECTIVE: To restore immunocompetency in HIV-1-infected individuals by establishing a sustained population of peripheral blood mononuclear cells (PBMC expressing anti-HIV-1 antisense RNA and resistant to HIV-1 infection.

METHODS: A routine retrovirus- based transducing vector, HGTV-43, was developed to deliver three U1/anti-HIV-1 antisense RNA genes into CD34+ cells. When U937 cells expressing these antisense genes were challenged with HIV-1 strains, virus production was blocked and no integration of the HIV-1 viral DNA was detected. HGTV-43 was pseudotyped with human T cell leukemia virus II envelope protein. Titers of greater than 107/ml and transduction rates of 25-55% of CD34+ cells from colonies grown in agar were observed. Of particular note, the transduction was completed in 18-hours, in the absence of both stromal cell "feeder" layers and added cytokines.

RESULTS: Approximately 108 PBSC (from mobilized HIV-1 infected patients after leukopheresis and enrichment of the CD34+ cell population) were transduced with HGTV-43 followed by autologous infusion without ablation. A progressive increase over a 6-month period of autologous, transduced CD34+ PBSC into PBMC was observed by measuring the antisense RNA using real-time RT-PCR of PBMC RNA. (The amplicon produced in this assay was confirmed by restriction enzyme digestion.) Antisense RNA was observed in CD4+ cells at 6 months.

CONCLUSION: This observation is an unusual and perhaps unique example of the survival of transduced hematopoietic cells expressing continued transgene activity within nonablated adult human subjects. The results from bone marrow taps will also be presented.

Keywords: AEGIS, HIV-1, CD4-Positive T-Lymphocytes, Antigens, CD34, HIV Infections, Retroviridae, RNA, Antisense, Leukapheresis, Adult, Human, immunology, AIDS
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