Restricted accessResearch articleFirst published online 1999-9
Adenovirus and Retrovirus Mediated Interferon Alpha Gene Transfer into CD34+ Cells Maintains Regeneration Capacity and Enhances Adhesion Molecules in K562 Cells
Systemic administration of interferon-α (IFN-α) results in cytogenetic remissions and enhanced survival in a significant percentage of patients with chronic mylogenous leukemia (CML) and lymphoma. However, this treatment is associated with deleterious toxic effects. Gene transfer of the IFN-α gene into hematopoietic progenitors represents a novel strategy to deliver high concentrations of IFN-α to a local area.
Methods
We compared the effect of the transfer of the IFN-α gene on the cell growth and differentiation of several CD34+ cells in culture and in a NOD/SCID animal model, using adenovirus and retrovirus constructs.
Results
Transient local expression of the IFN-α gene using an adenovirus vector was associated with normal proliferation of CD34+ progenitors as measured by a colony forming unit of granulocyte-macrophage (CFU-GM) growth. Flow cytometric determination revealed that there was no significant difference in viability of these cells for 24-hour transduction periods. Reverse transcriptase/polymerase chain reaction (RT-PCR) analysis of RNA from CD34+ harvested CFU-GM progenitors demonstrated expression of IFN-α mRNA; radioimmunoassay (RIA) revealed that transduced cells secreted substantial levels of IFN-α protein. Furthermore, we constructed a retroviral vector in which IFN-α cDNA was driven by a viral LTR promoter to evaluate the effect of permanent IFN-α gene expression on cell growth. Retroviral packaging cells PA317 with high titers of retrovirus were produced and used to infect CD34+ and K562 cells. RIA showed that IFN-α-transduced CD34+ cells (with the aid of fibronectin fragment CH-296) produced approximately 400 units of IFN-α protein compared to CD34+ cells, or cells transduced with empty vector. IFN-α transduced CD34+ generated similar numbers of CFU-GM colonies as compared to control CD34+ cells. Engraftment of CD34+ cells transduced with IFN-α gene in NOD/SCID mice was successful for the first 30 days. Additionally, we studied the effect of local IFN-α expression on the cellular adhesion molecules, VLA-4, Mac-1, ICAM-1, and L-selectin in K562 cells, and human umbilical endothelial vein cells. K562 cells transduced with the IFN-α gene expressed a significantly elevated level of VLA-4, Mac-1, and ICAM-1.
Conclusion
We conclude that expression of the IFN-α gene using retrovirus vectors results in an adequate localized expression of IFN-α mRNA and protein.
FialkowPJ, JacobsonRJ, PapayannopoulouT.Chronic myelocytic leukemia: clonal origin in a stem cell common to the granulocyte, erythrocyte, platelet and monocyte/macrophage. Am J Med1977;63:125–30.
2.
MartinPJ, NajfeldV, HansenJA, PenfoldJK, JacobsonRJ, FialkowPJ. Involvement of the B-lymphoid system in chronic myelogenous leukemia. Nature1980;287:49–50.
3.
NittaM, KatoY, StrifeA, WachterM, FiredJ, PerezA, JhanwarS, Duigou-OstenfdorfR, ChangantiRSK, ClarksonB.Incidence of involvement of the B and T lymphocyte lineages in chronic myelogenous leukemia. Blood1985;66:1053–61.
4.
McWhirterJR, WangJYI. Activation of tyrosine kinase and microfilament-binding functions by c-able by ber sequences in bcr/abl fusion protein. Mol Cell Biol1991;11:1553–65.
5.
RowleyJD. The Philadelphia chromosome translocation. A paradigm for understanding leukemia. Cancer1990;65:2178–84.
6.
GordonMY, DowdingCR, RileyGP, GoldmanJM. Altered adhesive interactions with marrow stroma of haematopoietic cells in chronic myeloid leukemia. Nature1987;328:342–4.
7.
VerfaillieCM, McCarthyJB, McGlavePB. Mechanisms underlying abnormal trafficking of malignant progenitors in chronic myelogenous leukemia. Decreased adhesion to stroma and fibronectin but increased adhesion to the basement membrane components laminin and collagen type IV. J Clin Invest1992;90:1232–41.
8.
WetzlerM, KurzrockR, LoweDG, KantarjianH, GuttermanJU, TalpazM.Alteration in bone marrow adherent layer growth factor expression: a novel mechanism of chronic myelogenous leukemia progression. Blood1991;78:2400–6.
9.
BhatiaR, McCarthyJB, VarfaillieCM. Interferon-a restores normal bl integrin-mediated inhibition of hematopoietic progenitor proliferation by the marrow microenvironment in chronic myelogenous leukemia. Blood1996;87:3883–91.
10.
BhatiaR, WaynerEA, McGlavePB, VerfaillieCM. Interferon-a restores normal adhesion of chronic myelogenous leukemia hematopoietic progenitors to bone marrow stroma by correcting impaired ß1 integrin receptor function. J Clin Invest1994;94:384–91.
11.
Italian Cooperative Study Group on Chronic Myeloid Leukemia. Interferon alfa-2a as compared with conventional chemotherapy for the treatment of chronic myeloid leukemia. N Engl J Med1994;330:820–5.
12.
BroxmeyerHE, LuL, PlatzerE, FeitC, JulianoL, RubinBY. Comparative analyses of the influences of human gamma, alpha and beta inteferons on human multipotential (CFU-GEMM), erythroid (BFU-E) and granulocyte macrophage (CFU-GM) progenitor cells. J Immunol1983;131:1300–5.
13.
GeisslerD, GastlG, AulitzkyW, TilgH, GagglS, KonwalinkaG, HuberC.Recombinant interferon-alpha-2C in chronic myelogenous leukemia: relationship of sensitivity of committed haematopoietic precursor cells in vitro (BFU-E, CFU-GM, CFU-Meg) and clinical response. Leuk Res1990;14:629–36.
14.
UpadhyayaG, GubaSC, SihSA, FeinbergAP, TalpazM, KantarjianHM, DeisserothAB, EmersonSG. Interferon-alpha restores the deficient expression of the cytoadhesion molecule lymphocyte function antigen-3 by chronic myelogenous leukemia progenitor cells. J Clin Invest1991;88:2131–6.
15.
DowdingC, GuoAP, OsterholzJ, SickowskiM, GoldmanJ, GordonM.Interferon-alpha overrides the deficient adhesion of chronic myelogenous leukemia primitive progenitor cells to bone marrow stromal cells. Blood1991;78:499–505.
16.
BhatiaR, McGlavePB, VerfaillieCM. Treatment of marrow stroma with interferon-a restores normal bl integrin-dependent adhesion of chronic myelogenous leukemia hematopoietic progenitors. Role of MIP-1 alpha. J Clin Invest1995;96:931–9.
17.
AmanMJ, KellerU, DerigsG, MohamadzadehM, HuberC, PeschelC.Regulation of cytokine expression by interferon alpha in human bone marrow stroma cells: inhibition of hematopoietic growth factors and induction of IL-1 receptor antagonist. Blood1994;84:4142–50.
18.
BhatiaR, McGlavePB, VerfaillieCM. Interferon-a treatment of marrow stroma results in enhanced adhesion of chronic myelogenous leukemia progenitors via mechanism involving MIP-la and TGF-b [abstract]. Exp Hematol1994;22:797a.
19.
ZoumbosN, GasconP, YoungNS. The function of lymphocytes in normal and suppressed hematopoiesis. Blut1984;48:1–9.
20.
EmersonSG, AntinJH. Bone marrow progenitor cells induce a regulatory autologous proliferative T lymphocyte response. J Immunol1989;142:766–72.
21.
EavesCJ, CashmanJD, KayRJ, DoughertyGJ, OtsukaT, GabouryLA, HoggeDE, LansdorpPM, EavesAC, HumphriesRK. Mechanisms that regulate the cell cycle status of very primitive hematopoietic cells in long-term human marrow cultures. II. Analysis of positive and negative regulators produced by stromal cells within the adherent layer. Blood1991;78:110–7.
22.
DunbarCE, FoxM, O'ShaughnesseyJ, DorenD, EmmonsRVB, SomaT, YuJM, CarterC, SellerS, HinesK, . Gene transfer to hematopoietic progenitor and stem cells: progress and problems. Hematology Branch, National Heart, Lung and Blood Institute, National Institute, National Institutes of Health, Bethesda, MD USA.
23.
MulliganRC. The basic science of gene therapy. Science1993;260:926–932.
24.
HananiaEG, KavanaghJ, HortobagyiG, GilesRE, ChamplinR, DeisserothAB. Recent advances in the application of gene therapy to human disease. Am J Med1995;99:537–552.
25.
CrystalRG, McElvaneyNG, RosenfeldMA, ChuC, MastrangeliA, HayJG, BrodySL, JaffeH, EissaNT, DanelC.Administration of an adenovirus containing the human CFTR cDNA to the respiratory tract of individuals with cystic fibrosis. Nat Genet1994;8:42–51.
26.
AhmedT, LuttonJD, FeldmanE, TaniK, AsanoS, AbrahamNG. Gene transfer of alpha interferon into hematopoietic stem cells. Leuk Res1998;22:119–24.
27.
FeldmanE, AhmedT, LuttonJD, FarleyT, TaniK, FreundM, AsanoS, AbrahamNG. Adenovirus mediated alpha interferon (IFN-α) gene transfer into CD34- cells and CML mononuclear cells. Stem Cells1997;15:386–95.
28.
WagenerF, FeldmanE, de WitteT, AbrahamNG. Heme induces the expression of adhesion molecules ICAM-1, VCAM-1, and E selectin in vascular endotheial cells. Proc Soc Exp Biol Med1997;216:456–463.
29.
AusubelFM, BrentR, KingstonRE, MooreDD, SeidmanJG, SmithJA, StruhlK.Current protocols in molecular biology. New York, NY: John Wiley … Sons, 1987.
30.
OguraH, TaniK, OzawaK, NagataS, AsanoS, TakakuF.Implantation of genetically manipulated fibroblasts into mice as antitumor a-interferon therapy. Cancer Res1990;50:5102–6.
31.
MarkowitzD, GoffS, BankA.Construction and use of a safe and efficient amphotropic packaging cell line. Virology1988;167:400–6.
32.
GengY, YuD, BlattLM, TaylorMW. Tumor suppressor activity of the human consensus type I interferon gene. Cytokines Mol Ther1995;1:289–300.
33.
TalpazM, KantarjianH, KurzrockR, TrujilloJMGuttermanJU. Interferon-alpha produces sustained cytogenetic responses in chronic myelogenous leukemia. Philadelphia chromosome-positive patients. Ann Intern Med1991;114:532–8.
34.
AustruyE, BagnisC, DubreuilP, MannoniP, ChabannonC.Retroviral gene transfer of the human interferon alpha-2 gene in human hematopoietic cell lines. Blood1996;88:434a.
35.
AustruyE, BagnisC, CarbucciaN, MarocC, BirgF, DubreuilP, MannoniP, ChabannonC.A defective retroviral vector encoding human interferon-a can transduce human leukemic cell lines. Cancer Gene Ther1998;5:247–56.
36.
BhatiaR, VerfaillieCM. Inhibition of BCR-ABL expression with antisense oligodeoxynucleotides restores ßl integrin-mediated adhesion and proliferation inhibition in chronic myelogenous leukemia hematopoietic progenitors. Blood1998;91:3414–22.
37.
ChertkovJL, JiangS, LuttonJD, HarrisonJ, LevereRD, TiefenthalerM, AbrahamNG. The hematopoietic stromal microenvironment promotes retrovirus-mediated gene transfer into hematopoietic stem cells. Stem Cells1993;11:218–27.
38.
CookPC, JiangS, ChertkovJL, FanY, LevereRD, AbrahamNG. Transfer of human adenine deaminase gene into murine hematopoietic stem cells: sequential study of spleen colon-forming units from bone marrow of living mice and the requirement of the microenvironment. Acta Haematol1996;96:57–63.
39.
NoltaJA, SmogorzewskaEM, KohnDB. Analysis of opitmal conditions for retroviral-mediated transduction of primitive human hematopoietic cells. Blood1995;86:101–10.
40.
MooreKA, DeisserothAB, ReadingCL, WilliamsDE, BelmontJW. Stromal support enhances cell-free retroviral vector transduction of human bone marrow long-term culture-initiating cells. Blood1992;79:1393–9.
41.
CasselA, Cottler-FoxM, DorenS, DunbarCE. Retroviral-mediated gene transfer into CD34-enriched human peripheral blood stem cells. Exp Hematol1993;21:585–91.
42.
HanenbergH, XiaoXL, DillooD, HashinoK, KatoI, WilliamsDA. Colocalization of retrovirus and target cells on specific fibronectin fragments increases genetic transduction of mammalian cells. Nature Med1996;2:876–82.
43.
MoritzT, DuttP, LiX, CarstanjecD, VikT, HanenbergH, WilliamsDA. Fibronectin improves transduction of reconstituting hematopoietic stem cells by retroviral vectors: evidence of direct viral binding to chymotryptic carboxy-terminal fragments. Blood1996;88:855–62.
44.
CashmanJD, LapidotT, WangJCY, DoedensM, ShultzLD, LansdorpP, DickJE, EavesCJ. Kinetic evidence of the regeneration of multilineage hematopoiesis from primitive cells in normal human bone marrow transplanted into immunodeficient mice. Blood1997;89:4307–16.
