An improved method to generate equine dendritic cells from peripheral blood mononuclear cells: Divergent maturation programs by IL-4 and LPS

Dietze, Barbara; Cierpka, Eva; Schäfer, Marcel; Schill, Wolfgang; Lutz, Manfred B.

doi:10.1016/j.imbio.2008.07.024

Cited by 13 publications

(17 citation statements)

References 20 publications

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“…Considering that 82.6% of the CD14+ cells were expressing the surface marker eqMHCII 4 days after selection, using this percentage and the number of vital CD14+ cells, our DC outcome was 14.78 × 10 6 DCs from 70 ml of blood resulting in 21.1 × 10 6 DCs from 100 ml of blood (82.6% × [28.7 × 10 6 cells] × [89% vital cells]) = 14.78 × 10 6 DCs/70 ml blood) (Table 3). Results show that the herein DC outcome is 2‐fold higher than the previous outcome of 10 × 10 6 DCs/100 ml blood reported [10].…”

Section: Resultscontrasting

confidence: 65%

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Enhanced protocol for CD14+ cell enrichment from equine peripheral blood via anti‐human CD14 mAb and automated magnetic activated cell sorting

Durán¹,

Willenbrock

Carlson

et al. 2012

Equine Veterinary Journal

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Section: Resultscontrasting

confidence: 65%

“…Directly after PBMC isolation an additional cell selection step was introduced using a human CD14 mAb. The equine CD14 positive cells were selected and cultured applying the recommended cytokine concentrations for DC generation [10].…”

Section: Discussionmentioning

confidence: 99%

Enhanced protocol for CD14+ cell enrichment from equine peripheral blood via anti‐human CD14 mAb and automated magnetic activated cell sorting

Durán¹,

Willenbrock

Carlson

et al. 2012

Equine Veterinary Journal

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“…Peripheral blood mononuclear cells (PBMC) were isolated via 1077 Ficoll-Paque (Amersham Biosciences, Piscataway, NJ) density centrifugation at 700 ϫ g for 15 min. We modified previously described magnetic bead positive selection and plastic adherence protocols to perform monocyte isolation (12,46). Briefly, 10 8 PBMC were labeled with anti-equine CD14 monoclonal antibody (MAb; hybridoma clone 105, kindly provided by Bettina Wagner, Cornell University, Ithaca, NY) (26), washed with phosphate-buffered saline (PBS), incubated with anti-mouse IgG1 microbeads (Miltenyi Biotech, Bergisch Gladbach, Germany), and positively selected over an LS column (Miltenyi Biotech) according to the manufacturer's protocol.…”

Section: Methodsmentioning

confidence: 99%

“…Interestingly, the AMLR displays characteristics of a normal immune response (including specificity and memory) and is reduced in a variety of human disease states (23, 53). Furthermore, the self-reactive T cells in the AMLR demonstrate a capacity for immunosuppression and increased transcription of the Treg transcription factor FOXP3, suggesting that such DC-stimulated T cells are involved in immune regulation during the normal immune response in vivo (25,45,51).A protocol for generating monocyte-derived DCs has been described in the horse, and the phenotype of these cells has been characterized by several groups (9,12,19). However, relatively little is known about the ability of equine DCs to induce proliferation and differentiation of autologous T cells.…”

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confidence: 99%

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Activation-Induced FoxP3 Expression Regulates Cytokine Production in Conventional T Cells Stimulated with Autologous Dendritic Cells

Cavatorta

Erb

Felippe

2012

Clin. Vaccine Immunol.

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A defining feature of dendritic cells (DCs) is their ability to induce the proliferation of autologous T cells in the absence of foreign antigen-a process termed the "autologous mixed leukocyte reaction" (AMLR). We report that equine monocytederived DCs, but not macrophages, are potent inducers of the AMLR. The response is contact dependent and major histocompatibility complex class II dependent and primarily involves CD3؉ CD4 ؉ CD8 ؊ T cells. Upon stimulation with DCs or the mitogen concanavalin A, a subset of the proliferating T cells expresses the regulatory T-cell (Treg) transcription factor FoxP3. Although many of these FoxP3 ؉ T cells are capable of producing the effector cytokines interleukin-4 (IL-4) and gamma interferon (IFN-␥), they are more likely to produce IL-10 and less likely to produce IFN-␥ than equivalent FoxP3 DCs take up antigen from the peripheral tissues, process it, and present it to naive T cells in the draining lymph node. Depending on the local cytokine environment during DC-mediated T cell activation, the antigen-specific T cell can proliferate and differentiate into one of a variety of effector T cell phenotypes, including (i) gamma interferon (IFN-␥)-producing Th1 cells that are important for immunity to intracellular pathogens and tumors; (ii) interleukin-4 (IL-4)-, IL-5-, and IL-13-producing Th2 cells that promote antibody production, help prevent parasitic diseases, and are involved in allergic responses; and (iii) IL-17-producing Th17 cells that protect against extracellular infections (58). DCs also play a critical role in immune regulation because they are capable of both expanding thymus-derived FoxP3 ϩ (designated FOXP3 in humans) natural regulatory T cells (nTregs) and inducing naive T cells to develop into induced regulatory T cells (iTregs) in the periphery (55, 56). In turn, Tregs use a variety of mechanisms (such as the production of the anti-inflammatory cytokines IL-10 and transforming growth factor ␤ [TGF-␤]) to keep effector responses in check and to prevent immune-mediated disease (7).DCs also possess the unique ability to efficiently induce proliferation of autologous T cells in the absence of exogenous antigen. This process was first demonstrated with murine DCs several decades ago and was termed the "autologous mixed leukocyte reaction" (AMLR) (39). The AMLR has since been described with human cells and is thought to represent polyclonal activation of autoreactive T cells specific for self-antigens presented by DCs (2,3,10,38,44). Interestingly, the AMLR displays characteristics of a normal immune response (including specificity and memory) and is reduced in a variety of human disease states (23, 53). Furthermore, the self-reactive T cells in the AMLR demonstrate a capacity for immunosuppression and increased transcription of the Treg transcription factor FOXP3, suggesting that such DC-stimulated T cells are involved in immune regulation during the normal immune response in vivo (25,45,51).A protocol for generating monocyte-derived DCs has been described in the h...

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Impact of equine herpesvirus type 1 (EHV-1) infection on the migration of monocytic cells through equine nasal mucosa

Baghi

Nauwynck

2014

Comparative Immunology, Microbiology and Infectious Diseases

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An improved method to generate equine dendritic cells from peripheral blood mononuclear cells: Divergent maturation programs by IL-4 and LPS

Cited by 13 publications

References 20 publications

Enhanced protocol for CD14+ cell enrichment from equine peripheral blood via anti‐human CD14 mAb and automated magnetic activated cell sorting

Enhanced protocol for CD14+ cell enrichment from equine peripheral blood via anti‐human CD14 mAb and automated magnetic activated cell sorting

Activation-Induced FoxP3 Expression Regulates Cytokine Production in Conventional T Cells Stimulated with Autologous Dendritic Cells

Impact of equine herpesvirus type 1 (EHV-1) infection on the migration of monocytic cells through equine nasal mucosa

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