Intravaginal immunization with HPV vectors induces tissue-resident CD8+ T cell responses
Nicolas Çuburu, … , Douglas R. Lowy, John T. Schiller
Nicolas Çuburu, … , Douglas R. Lowy, John T. Schiller
Published November 12, 2012
Citation Information: J Clin Invest. 2012;122(12):4606-4620. https://doi.org/10.1172/JCI63287.
View: Text | PDF
Research Article Immunology

Intravaginal immunization with HPV vectors induces tissue-resident CD8+ T cell responses

Abstract

The induction of persistent intraepithelial CD8+ T cell responses may be key to the development of vaccines against mucosally transmitted pathogens, particularly for sexually transmitted diseases. Here we investigated CD8+ T cell responses in the female mouse cervicovaginal mucosa after intravaginal immunization with human papillomavirus vectors (HPV pseudoviruses) that transiently expressed a model antigen, respiratory syncytial virus (RSV) M/M2, in cervicovaginal keratinocytes. An HPV intravaginal prime/boost with different HPV serotypes induced 10-fold more cervicovaginal antigen-specific CD8+ T cells than priming alone. Antigen-specific T cell numbers decreased only 2-fold after 6 months. Most genital antigen-specific CD8+ T cells were intra- or subepithelial, expressed αE-integrin CD103, produced IFN-γ and TNF-α, and displayed in vivo cytotoxicity. Using a sphingosine-1-phosphate analog (FTY720), we found that the primed CD8+ T cells proliferated in the cervicovaginal mucosa upon HPV intravaginal boost. Intravaginal HPV prime/boost reduced cervicovaginal viral titers 1,000-fold after intravaginal challenge with vaccinia virus expressing the CD8 epitope M2. In contrast, intramuscular prime/boost with an adenovirus type 5 vector induced a higher level of systemic CD8+ T cells but failed to induce intraepithelial CD103+CD8+ T cells or protect against recombinant vaccinia vaginal challenge. Thus, HPV vectors are attractive gene-delivery platforms for inducing durable intraepithelial cervicovaginal CD8+ T cell responses by promoting local proliferation and retention of primed antigen-specific CD8+ T cells.

Authors

Nicolas Çuburu, Barney S. Graham, Christopher B. Buck, Rhonda C. Kines, Yuk-Ying S. Pang, Patricia M. Day, Douglas R. Lowy, John T. Schiller

×

Figure 6

HPV Ivag prime/boost immunization induces polyfunctional CD8+ T cells with cytotoxic activity.

Options: View larger image (or click on image) Download as PowerPoint
HPV Ivag prime/boost immunization induces polyfunctional CD8+ T cells wi...
Depo-Provera–treated mice were immunized with 5 × 107 IU HPV16MM2 or HPV16 control Ivag, and 1 month later, mice were immunized with 5 × 107 IU HPV45MM2 or HPV45 control, respectively. (A) Two weeks after the last immunization, the production of IFN-γ, TNF-α, and IL-2 by CD8+ T cells was measured after in vitro stimulation with M282–90 peptide in cervicovaginal, spleen, and ILN cell suspensions. The percentage of CD8+ T cells in each quadrant is indicated in each plot. (B) Relative proportion of single-, double-, and triple-cytokine-producing CD8+ T cells restimulated with M282–90 peptide after subtraction of background cytokine production of unstimulated CD8+ T cells. (C) In vivo cytotoxic activity was assessed 2 weeks after the last immunization. Target cells consisting of an equal mixture of two splenocyte populations labeled with a high and a low concentration of CFSE were pulsed with M282–90 peptide or remained unpulsed, respectively, were injected Ivag and i.v. Twenty-four hours later, cervicovaginal, spleen, and ILN cell suspensions were analyzed by flow cytometry. Representative histograms gated on CFSE-positive cells. Mean percentage ± SD of M282–90 specific lysis is indicated.