Differential Effects of Viral Vectors on Migratory Afferent Lymph Dendritic Cells<i>In Vitro</i>Predict Enhanced Immunogenicity<i>In Vivo</i>

Cubillos‐Zapata, Carolina; Guzman, Efrain; Turner, Alison; Gilbert, Sarah C.; Prentice, Helen; Hope, Jayne; Charleston, Bryan

doi:10.1128/jvi.05127-11

Cited by 31 publications

(50 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We have recently described the differential effects of viral vectors on bovine dendritic cells draining from the skin (17). We reported that, similar to murine BMDC and human moDC, MVA induces apoptosis in DC draining from the skin.…”

Section: Tuberculosis Is a Highly Infectious Disease Caused In Humans Bymentioning

confidence: 99%

“…Recombinant antigen 85A (Ag85A) was obtained from Lionex GmbH (Germany) and used at 2 g/ml in all assays. , with the recombinant viruses using optimal multiplicities of infection (MOI) described previously (17).…”

Section: Methodsmentioning

confidence: 99%

“…Afferent lymphatic DC (ALDC) represent a major population of migrating DC (14,16,28,81) with functional and phenotypic heterogeneity. We and others have described ALDC as being high forward scatter (FSC high ) DEC-205 (17,28,34); within this population, subpopulations expressing various levels of SIRP␣ (CD172a), CD11a, CD26, and CD13 have also been described (5,28,29,40). These populations have been shown to interact with and stimulate T cells differently and have differential cytokine secretion profiles (35,40,52,72).…”

Section: Tuberculosis Is a Highly Infectious Disease Caused In Humans Bymentioning

confidence: 99%

“…To test the hypothesis that antigens expressed by recombinant vectors are efficiently presented by afferent lymph dendritic cells (ALDC), we infected ALDC in vitro with recombinant MVA (rMVA) or recombinant adenovirus 5 (rAd5) expressing mycobacterial antigen 85A (Ag85A) or loaded cells with recombinant Ag85A or PPD-B to stimulate T cells from BCG-vaccinated animals. We have previously shown that by infecting ALDC with MVA-GFP at an MOI of 1 to 5 PFU/cell, the number of GFP ϩ cells is similar to that of Ad5-GFP infected ALDC at an MOI of 100 IU/cell (17). We therefore used rMVA-Ag85A at an MOI of 3 PFU/cell and rAd5-Ag85A at 100 IU/cell.…”

Section: Mva-expressed Ag85a Is Not Efficiently Presented By Aldcmentioning

confidence: 99%

“…The cells were incubated for 24 h at 37°C, and the assay continued as previously described (36). In some assays, proliferation was assessed by labeling cells with carboxyfluorescein diacetate succinimydyl ester (CFDA-SE) as previously described (17). The results are shown as percent proliferation, determined as the percentage of CFSE LOW cells.…”

Section: Infectionmentioning

confidence: 99%

See 4 more Smart Citations

Modified Vaccinia Virus Ankara-Based Vaccine Vectors Induce Apoptosis in Dendritic Cells Draining from the Skin via both the Extrinsic and Intrinsic Caspase Pathways, Preventing Efficient Antigen Presentation

Guzman¹,

Cubillos‐Zapata²,

Cottingham

et al. 2012

J Virol

Self Cite

View full text Add to dashboard Cite

Dendritic cells (DC) are potent antigen-presenting cells and central to the induction of immune responses following infection or vaccination. The collection of DC migrating from peripheral tissues by cannulation of the afferent lymphatic vessels provides DC which can be used directly ex vivo without extensive in vitro manipulations. We have previously used bovine migrating DC to show that recombinant human adenovirus 5 vectors efficiently transduce afferent lymph migrating DEC-205 + CD11c + CD8 − DC (ALDC). We have also shown that recombinant modified vaccinia virus Ankara (MVA) infects ALDC in vitro , causing downregulation of costimulatory molecules, apoptosis, and cell death. We now show that in the bovine system, modified vaccinia virus Ankara-induced apoptosis in DC draining from the skin occurs soon after virus binding via the caspase 8 pathway and is not associated with viral gene expression. We also show that after virus entry, the caspase 9 pathway cascade is initiated. The magnitude of T cell responses to mycobacterial antigen 85A (Ag85A) expressed by recombinant MVA-infected ALDC is increased by blocking caspase-induced apoptosis. Apoptotic bodies generated by recombinant MVA (rMVA)-Ag85A-infected ALDC and containing Ag85A were phagocytosed by noninfected migrating ALDC expressing SIRPα via actin-dependent phagocytosis, and these ALDC in turn presented antigen. However, the addition of fresh ALDC to MVA-infected cultures did not improve on the magnitude of the T cell responses; in contrast, these noninfected DC showed downregulation of major histocompatibility complex class II (MHC-II), CD40, CD80, and CD86. We also observed that MVA-infected ALDC promoted migration of DEC-205 + SIRPα + CD21 + DC as well as CD4 + and CD8 + T cells independently of caspase activation. These in vitro studies show that induction of apoptosis in DC by MVA vectors is detrimental to the subsequent induction of T cell responses.

show abstract

Section: Tuberculosis Is a Highly Infectious Disease Caused In Humans Bymentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%