Dendritic cell elimination as an assay of cytotoxic T lymphocyte activity in vivo

Ritchie, David; Hermans, Ian F.; Lumsden, Joanne M.; Scanga, Connie B.; Roberts, Joanna M.; Yang, Jianping; Kemp, Roslyn A.; Ronchese, Franca

doi:10.1016/s0022-1759(00)00300-8

Cited by 51 publications

(59 citation statements)

References 15 publications

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“…We have previously reported that antigen-loaded DC are eliminated by specific CD8 ϩ T cells in vivo and fail to accumulate in the draining lymph node (20). In vitro experiments (data not shown) also showed that CTL can kill DC in an antigen-specific fashion, providing further support for a direct killing mechanism in vivo.…”

Section: Resultsmentioning

confidence: 74%

“…DC were harvested from culture, washed, and labeled with carboxy-fluorescein diacetate succinimidyl ester (CFSE, Molecular Probes) or the orange f luorescent dye chloromethyl-benzoyl-aminotetramethyl-rhodamine (CMTMR, Molecular Probes) with the protocols described (20). The target DC populations used throughout this study were not treated with LPS, and treatment with 0.1-1 g͞ml LPS did not change their susceptibility to elimination in vivo (K. Andrew and F.R., unpublished work).…”

mentioning

confidence: 99%

“…In vivo elimination of antigen-loaded DC was assessed as described (20). Mice were injected s.c. in the forelimb or intradermally into the ear with 1-2 ϫ 10 6 cells containing equal numbers of CFSE-labeled DC loaded with 1 M peptide (DC ϩ ) and CMTMR-labeled control DC (DC Ϫ ).…”

mentioning

confidence: 99%

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Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8⁺T cellsin vivo

Yang

Huck²,

McHugh³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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170

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The lifespan and survival of dendritic cells (DC) in vivo are potentially critical to the expansion of T cell immune responses. We have previously reported that DC loaded with specific antigen are rapidly eliminated by cytotoxic T lymphocytes (CTL) in vivo, but the site, mechanism, and consequences of DC elimination were not defined. In this article we show that DC elimination in vivo occurs in a perforin-dependent manner and does not require IFN-␥ or the presence of CD4 ؉ CD25 ؉ regulatory T cells. Most importantly, failure to eliminate DC had profound consequences on the CTL immune response. Perforin-deficient mice showed a progressive increase in the numbers of antigen-specific CD8 ؉ T cells after repeated immunizations with DC. In contrast, in control mice the number of antigen-specific CD8 ؉ T cells did not notably increase with repeated immunizations. Lastly, we also show that CTLmediated elimination of DC occurs in peripheral tissues but not in the lymph node. Our data suggest that CTL act as ''gatekeepers'' that control access of antigen-loaded DC into the lymph node, thereby preventing continued expansion of antigen-specific T cells.cytotoxic T cells ͉ immunoregulation ͉ killing D endritic cells (DC) are powerful antigen-presenting cells that are critical for the initiation of CD4 ϩ and CD8 ϩ T cell responses. DC reside in peripheral tissues where they take up antigens from the external environment, then migrate to the lymph nodes where they interact with antigen-specific T cells and induce their activation to proliferation and effector function (1, 2). The lifespan of DC once in the lymph node is thought to be relatively brief, but experimental estimates have not yielded a consistent figure (3-5).The mechanism and regulation of DC survival and death (6, 7) are likely to be important in maintaining the homeostatic balance of the immune system. A few reports have linked extended survival of DC to enhanced or dysregulated T cell immune responses and lymphoproliferative disease (8-10). In contrast, reduced survival of DC has been associated with impaired immune responses (7). It is presently unclear whether T cells also influence the lifespan of DC in an antigen-specific fashion. In lymph nodes of mice adoptively transferred with CD4 ϩ T cell receptor (TCR) transgenic T cells, DC presenting specific antigen disappear more rapidly than DC not presenting antigen (11), suggesting that T cells may have a role in regulating DC numbers and survival. Experiments using infection with Listeria or malaria also suggest that the number of antigen-presenting cells becomes limiting during the early phases of CD8 ϩ immune responses and prevents the continued expansion of antigen-specific T cells (12, 13). Together, these experiments suggest the attractive hypothesis that T cells may be able to regulate their own responses in a feedback fashion, by affecting the survival of antigen-presenting DC.We have previously reported that DC loaded with antigen and injected s.c. into immune mice are rapidly eliminated by CD8 ϩ T cel...

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Section: Resultsmentioning

confidence: 74%

mentioning

confidence: 99%

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Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8⁺T cellsin vivo

Yang

Huck²,

McHugh³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

124

170

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“…In vivo cytotoxicity assay was performed as described by Ritchie et al, 16 with slight modifications. Briefly, a singlecell suspension of 10 7 naive Spc ml À1 was labeled with two different concentrations (0.5 or 5.0 mM) of the fluorescent dye CFSE (Molecular Probes, Leiden, The Netherlands).…”

Section: In Vivo Cytotoxicity Assaymentioning

confidence: 99%

DNA immunization using constant-current electroporation affords long-term protection from autochthonous mammary carcinomas in cancer-prone transgenic mice

Curcio

Khan²,

Amici

et al. 2007

Cancer Gene Ther

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A recently developed, adaptive constant-current electroporation technique was used to immunize mice with an intramuscular injection of plasmid coding for the extracellular and transmembrane domains of the product of the rat neu 664V-E oncogene protein.In wild-type BALB/c mice, plasmid electroporation at lower current settings elicits higher antibody titers, a strong cytotoxic response and completely protects all mice vaccinated with 10, 25 and 50 mg of plasmid against a lethal challenge of rat neu þ carcinoma cells. BALB/c mice transgenic for the transforming rat neu 664VÀE (ErbB-2, Her-2/neu) oncogene (BALB-neuT 664VÀE ) develop an invasive mammary gland carcinoma by 20 weeks of age. Remarkably, when transgenic BALB-neuT 664VÀE mice were vaccinated at a 10-week interval with 50 mg of plasmid with 0.2 A electroporation, mice remained tumor free for more than a year. A single administration of plasmid associated with electroporation was enough to markedly delay carcinogenesis progression in mice with multiple microscopic invasive carcinomas, and keep about 50% of mice tumor free at one year of age. Thus, vaccination using a clinically relevant dose of plasmid encoding the extracellular and transmembrane domains of the neu oncogene delivered by electroporation prevents long-term tumor formation. These improvements in the efficacy of this cancer vaccine regimen vastly increase its chances for clinical success.

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“…[17][18][19] Recently, it has been documented that DCs undergo rapid apoptosis during antigen-specific interaction with T cells both in vitro 20 and in vivo. [21][22][23][24] Several studies have shown that genetically modified DCs expressing TAA could effectively process and present antigenic peptides to T cells and trigger cytotoxic response against tumor cells expressing the target antigen. 12,25 It is important to consider that the TAA-modified DCs would also become a potential target and are at risk of being eliminated by the effector T cells they have activated.…”

Section: Introductionmentioning

confidence: 99%

Ex vivo generation of genetically modified dendritic cells for immunotherapy: implications of lymphocyte contamination

et al. 2005

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Genetically modified dendritic cell (DC) vaccines expressing tumor-associated antigens are currently used for cancer immunotherapy. Peripheral blood (PB) monocyte precursors are a relatively convenient source of DCs for use in clinical studies, but are often contaminated by lymphocytes. The current study was conducted to examine the impact of Tlymphocyte contamination on genetically modified DC product. PB monocyte-derived DCs were efficiently transduced (75-95%) with an HIV-1-based self-inactivating lentiviral vector encoding a model antigen, the enhanced green fluorescent protein (eGFP). The lymphocyte-free DC culture transduced with Lenti-eGFP showed stable expression of eGFP without measurable decline in viability. In contrast, the eGFP-positive DCs disappeared rapidly in transduced DC cultures containing lymphocyte contaminants, concurrent with detectable activation and expansion of T-lymphocytes. Upon antigen recall, these T cells elicited major histocompatability complex-restricted antigen-specific cytotoxicity against eGFP-positive autologous DCs and mitogen-stimulated T lymphoblasts, mainly through the perforin-mediated pathway. In summary, this study demonstrate that the relative purity of DC cultures could determine the persistence of gene-modified DC, which may affect the induction of effective immune responses by DC vaccination strategies. Gene Therapy (2005) 12, 259-271.

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Dendritic cell elimination as an assay of cytotoxic T lymphocyte activity in vivo

Cited by 51 publications

References 15 publications

Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8⁺T cellsin vivo

Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8⁺T cellsin vivo

DNA immunization using constant-current electroporation affords long-term protection from autochthonous mammary carcinomas in cancer-prone transgenic mice

Ex vivo generation of genetically modified dendritic cells for immunotherapy: implications of lymphocyte contamination

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Dendritic cell elimination as an assay of cytotoxic T lymphocyte activity in vivo

Cited by 51 publications

References 15 publications

Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8+T cellsin vivo

Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8+T cellsin vivo

DNA immunization using constant-current electroporation affords long-term protection from autochthonous mammary carcinomas in cancer-prone transgenic mice

Ex vivo generation of genetically modified dendritic cells for immunotherapy: implications of lymphocyte contamination

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Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8⁺T cellsin vivo

Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8⁺T cellsin vivo