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. 1999 Jun 1;59(11):2536-40.

Recombinant virus vaccination against "self" antigens using anchor-fixed immunogens

K R Irvine  1 , M R Parkhurst, E P Shulman, J P Tupesis, M Custer, C E Touloukian, P F Robbins, A G Yafal, P Greenhalgh, R P Sutmuller, R Offringa, S A Rosenberg, N P Restifo
Affiliations

Recombinant virus vaccination against "self" antigens using anchor-fixed immunogens

K R Irvine et al. Cancer Res. .

Abstract

To study the induction of anti-"self" CD8+ T-cell reactivity against the tumor antigen gp100, we used a mouse transgenic for a chimeric HLA-A*0201/H-2 Kb molecule (A2/Kb). We immunized the mice with a recombinant vaccinia virus encoding a form of gp100 that had been modified at position 210 (from a threonine to a methionine) to increase epitope binding to the restricting class I molecule. Immunogens containing the "anchor-fixed" modification elicited anti-self CD8+ T cells specific for the wild-type gp100(209-217) peptide pulsed onto target cells. More important, these cells specifically recognized the naturally presented epitope on the surface of an A2/Kb-expressing murine melanoma, B16. These data indicate that anchor-fixing epitopes could enhance the function of recombinant virus-based immunogens.

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Figures

Fig. 1
Fig. 1
Immunization with recombinant poxviruses expressing gp100 induced CD8+ T-cell lytic responses against gp100154–162 but not against gp100209–217. A2/Kb transgenic mice were immunized once i.v. with 107 pfu of rVV-FLgp100. Three to 6 weeks later, pooled splenocytes (two mice per group) were harvested and stimulated for 6 days with 2 μg of the indicated peptides per ml. Cultures were then assayed for specific lytic activity in a 51Cr-release assay against either T2 cells alone (□) or T2 cells pulsed with the relevant native peptides, gp100154–162 or gp100209–217 (■). A 100:1 E:T ratio is shown. This experiment was repeated two times with similar findings.
Fig. 2
Fig. 2
T-cell recognition of gp100-specific epitopes of rVV-infected HLA-A*0201+, gp100− breast cancer cell line. The MDA 231 HLA-A*0201+, gp100− breast cancer cell line was infected at an MOI of 10 for 1 h at 37°C with rVV-Tyrosinase (rVV-Tyr), rVV-FLgp100, rVV-FLgp100(210M), rVV-ESgp100209–217, or rVV-ESgp100209–217(210M). The infected MDA 231 cells were washed and plated at 105/well and coincubated with 2.5 × 104 human T-cell clones for 18 h. Clone 70 specifically recognizes gp100154–162, and clone DW2C8 recognizes either gp100209–217 or gp100209–217(210M). Supernatants were assayed for human IFN-γ release by ELISA.
Fig. 3
Fig. 3
Immune recognition of the gp100209–217 epitope naturally processed and presented by B16/A2/Kb. In A, A2/Kb transgenic mice were immunized once i.v. with 107 pfu of rVV-Tyrosinase (Tyr), rVV-ESgp100209–217 (ES209), or rVV-ESgp100209–217(210M) (ES210M). Three to 6 weeks later, pooled splenocytes (two mice per group) were harvested and stimulated in vitro for 7 days with 2 μg of gp100154–162 (154–162), gp100209–217 (209–217), or gp100210–217(210M) (210M) peptides per ml. Cultures were restimulated once with the same peptide. After 7 days in culture, splenocytes were assayed at a 25:1 E:T ratio for specific lytic activity in a 51Cr-release assay against T2 cells alone, T2 pulsed with the relevant gp100209–217, or a murine melanoma stably transfected with the chimeric A2/Kb molecule, B16/A2/Kb. In B and C, the rVV-ESgp100209–217(210M)-immune splenocyte culture stimulated in vitro with gp100209–217(210M) was further assayed for cytokine release in response to a panel of target cells. Specifically, 105 effector cells were admixed with 105 target cells in each well of 96-well plates. Eighteen to 24 h later, supernatants were harvested and assayed by ELISA for the presence of either murine IFN-γ (B) or murine GM-CSF (C).
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References

    1. Hellstrom I, Hellstrom KE. Tumor vaccines—a reality at last? J. Immunother. 1998;21:119–126. - PubMed
    1. Mitchell MS. Immunotherapy of melanoma. J. Investig. Dermatol. Symp. Proc. 1996;1:215–218. - PubMed
    1. Fenton RG, Longo DL. Danger versus tolerance: paradigms for future studies of tumor-specific cytotoxic T lymphocytes. J. Natl. Cancer Inst. 1997;89:272–275. - PubMed
    1. Rosenberg SA, Yang JC, Schwartzentruber DJ, Hwu P, Marincola FM, Topalian SL, Restifo NP, Dudley ME, Schwarz SL, Spiess PJ, Wunderlich JR, Parkhurst MR, Kawakami Y, Seipp CA, Einhorn JH, White DE. Immunologic and therapeutic evaluation of a synthetic peptide vaccine for the treatment of patients with metastatic melanoma. Nat. Med. 1998;4:321–327. - PMC - PubMed
    1. Rosenberg SA, Zhai Y, Yang JC, Schwartzentruber DJ, Hwu P, Marincola FM, Topalian SL, Restifo NP, Seipp CA, Einhorn JH, Roberts B, White DE. Immunizing patients with metastatic melanoma using recombinant adenoviruses encoding MART-1 or gp100 melanoma antigens. J. Natl. Cancer Inst. 1998;90:1894–1900. - PMC - PubMed

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