High-titer lentiviral vectors stimulate fetal calf serum-specific human CD4 T-cell responses

Implications in human gene therapy

L. Bao, H. Guo, X. Huang, S. Tammana, M. Wong, R S Mc Ivor, X. Zhou

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Human immunodeficiency virus-1-derived lentiviral vectors have been increasingly used for gene delivery in both pre-clinical and clinical models. Numerous studies have shown that dendritic cells (DC) transduced with concentrated lentiviral vectors can induce primary T-cell responses to viral and tumor antigens. In this study, we attempted to generate influenza hemagglutinin-specific CD4 T cells using lentiviral vectors containing the signal sequence and human lysosome-associated membrane protein to target hemagglutinin to the major histocompatibility complex class II processing pathway. Autologous dendritic cells were generated in serum-free medium and transduced with concentrated, high-titer lentiviruses to stimulate autologous T cells. Unexpectedly, we failed to generate influenza hemagglutinin-specific CD4 T cells rather than T cells specific for fetal calf serum (FCS). By limiting dilution, we established several FCS-specific CD4 T-cell clones restricted by human leukocyte antigen-DR1 and human leukocyte antigen-DR4. Lentiviruses produced in human serum-adapted 293 cells or in serum-free medium were unable to sensitize dendritic cells for recognition by FCS-specific CD4 T-cell clones. Our results indicate that residual FCS in concentrated lentiviral pellets is, in part, responsible for its immunogenicity. These FCS-specific CD4 T cells may be useful in testing clinical grade lentiviral vectors for the presence of contaminating FCS.

Original languageEnglish (US)
Pages (from-to)788-795
Number of pages8
JournalGene therapy
Volume16
Issue number6
DOIs
StatePublished - Apr 3 2009

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Genetic Therapy
T-Lymphocytes
Serum
Hemagglutinins
Dendritic Cells
Lentivirus
Serum-Free Culture Media
HLA Antigens
Human Influenza
Clone Cells
Lysosome-Associated Membrane Glycoproteins
Viral Tumor Antigens
Protein Sorting Signals
Major Histocompatibility Complex
HIV-1
Genes

Cite this

High-titer lentiviral vectors stimulate fetal calf serum-specific human CD4 T-cell responses : Implications in human gene therapy. / Bao, L.; Guo, H.; Huang, X.; Tammana, S.; Wong, M.; Mc Ivor, R S; Zhou, X.

In: Gene therapy, Vol. 16, No. 6, 03.04.2009, p. 788-795.

Research output: Contribution to journalArticle

Bao, L. ; Guo, H. ; Huang, X. ; Tammana, S. ; Wong, M. ; Mc Ivor, R S ; Zhou, X. / High-titer lentiviral vectors stimulate fetal calf serum-specific human CD4 T-cell responses : Implications in human gene therapy. In: Gene therapy. 2009 ; Vol. 16, No. 6. pp. 788-795.
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