Adenoviral-encoded antigens are presented efficiently by a subset of dendritic cells expressing high levels of alpha(v)beta3 integrins

J Leukoc Biol. 2006 Jun;79(6):1271-8. doi: 10.1189/jlb.1105694. Epub 2006 Mar 30.

Abstract

Dendritic cells (DC) play a central role in antigen presentation and are often targeted by adenoviral (Ad)-based gene therapy. However, DC lack the coxsackie-Ad receptor, and little is known about the process by which they acquire and present Ad-encoded antigens. We examined the expression of alpha(v)beta3 integrins (CD51/CD61) on mouse bone marrow-derived DC (BM-DC) and their susceptibility to transduction by Ad vectors. Less than 10% of BM-DC precursors expressed CD51, but expression increased over time in culture with granulocyte macrophage-colony stimulating factor (GM-CSF)/interleukin (IL)-4. After 7 days, 28 +/- 1.7% of CD11c+ DC expressed high levels of CD51 (CD51(hi)), and the remaining DC expressed low levels of CD51 (CD51(lo)). CD51(hi) CD express higher major histocompatibility complex type 1 (MHC I); however, both of the DC subsets expressed similar levels of MHC II and costimulatory molecules. When exposed to a first-generation Ad vector, transgene expression was restricted to the CD51(hi) DC subset and blocked by soluble peptides expressing an arginine, glycine, aspartic acid (RGD) sequence, confirming the role of integrins in viral entry. Consistent with this, a modified Ad expressing an RGD-binding sequence in its fiber knob (Ad-RGD) transduced the CD51(hi) DC subset with significantly higher efficiency. When BM-DC were transduced with an Ad-expressing ovalbumin (Ad-OVA), the CD51(hi) subset proved superior in activating OT-I (T cell receptor-OVA) T cells. Similar to in vitro effects, systemic administration of GM-CSF/IL-4 increased the expression of CD51 on splenic DC and rendered these cells susceptible to Ad transduction. These results suggest that a limited subset of DC expressing high levels of alpha(v)beta3 integrins is preferentially transduced by Ad vectors and activates CD8+ T cell responses against Ad-encoded antigens.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / immunology*
  • Animals
  • Antigen Presentation*
  • Antigens, Viral / immunology*
  • B7-1 Antigen / immunology
  • B7-2 Antigen / immunology
  • Bone Marrow Cells / chemistry
  • Bone Marrow Cells / classification
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / immunology
  • Dendritic Cells / chemistry
  • Dendritic Cells / classification
  • Dendritic Cells / drug effects
  • Dendritic Cells / immunology*
  • Genetic Vectors / immunology*
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • H-2 Antigens / immunology
  • Integrin alphaV / biosynthesis
  • Integrin alphaV / genetics
  • Integrin alphaVbeta3 / analysis*
  • Integrin beta3 / biosynthesis
  • Integrin beta3 / genetics
  • Interleukin-4 / pharmacology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Oligopeptides / pharmacology
  • Ovalbumin / immunology
  • Recombinant Proteins / pharmacology
  • Spleen / cytology
  • Transduction, Genetic
  • Transgenes

Substances

  • Antigens, Viral
  • B7-1 Antigen
  • B7-2 Antigen
  • Cd86 protein, mouse
  • H-2 Antigens
  • Integrin alphaV
  • Integrin alphaVbeta3
  • Integrin beta3
  • Oligopeptides
  • Recombinant Proteins
  • Interleukin-4
  • arginyl-glycyl-aspartic acid
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Ovalbumin