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

Durán, Mari-Carmen; Willenbrock, Saskia; Carlson, Regina; Feige, Karsten; Nolte, Ingo; Escobar, Hugo Murua

doi:10.1111/j.2042-3306.2012.00616.x

Cited by 5 publications

(3 citation statements)

References 13 publications

(43 reference statements)

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“…The compositions of the three main PBMC subpopulations (mean Ϯ standard deviation [SD]) were determined by flow cytometry. The percentages were 88.82% Ϯ 0.84% CD3 ϩ T lymphocytes, 2.60% Ϯ 1.47% CD14 ϩ monocytes, and 8.57% Ϯ 1.6% IgM ϩ B lymphocytes, which are still within the ranges of previously obtained values (11,44). The rate of infection by parental EHV-1 and EHV-4 was determined for each PBMC subpopulation (mean Ϯ SD).…”

Section: Growth Analysis and Rates Of Infection By Ehv-1 And Ehv-4 Inmentioning

confidence: 69%

Role of gB and pUS3 in Equine Herpesvirus 1 Transfer between Peripheral Blood Mononuclear Cells and Endothelial Cells: a DynamicIn VitroModel

Spiesschaert

Goldenbogen

Taferner

et al. 2015

J Virol

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Infected peripheral blood mononuclear cells (PBMC) effectively transport equine herpesvirus type 1 (EHV-1), but not EHV-4, to endothelial cells (EC) lining the blood vessels of the pregnant uterus or central nervous system, a process that can result in abortion or myeloencephalopathy. We examined, using a dynamic in vitro model, the differences between EHV-1 and EHV-4 infection of PBMC and PBMC-EC interactions. In order to evaluate viral transfer between infected PBMC and EC, cocultivation assays were performed. Only EHV-1 was transferred from PBMC to EC, and viral glycoprotein B (gB) was shown to be mainly responsible for this form of cell-to-cell transfer. For addressing the more dynamic aspects of PBMC-EC interaction, infected PBMC were perfused through a flow channel containing EC in the presence of neutralizing antibodies. By simulating capillary blood flow and analyzing the behavior of infected PBMC through live fluorescence imaging and automated cell tracking, we observed that EHV-1 was able to maintain tethering and rolling of infected PBMC on EC more effectively than EHV-4. Deletion of US3 reduced the ability of infected PBMC to tether and roll compared to that of cells infected with parental virus, which resulted in a significant reduction in virus transfer from PBMC to EC. Taking the results together, we conclude that systemic spread and EC infection by EHV-1, but not EHV-4, is caused by its ability to infect and/or reprogram mononuclear cells with respect to their tethering and rolling behavior on EC and consequent virus transfer. IMPORTANCEEHV-1 is widespread throughout the world and causes substantial economic losses through outbreaks of respiratory disease, abortion, and myeloencephalopathy. Despite many years of research, no fully protective vaccines have been developed, and several aspects of viral pathogenesis still need to be uncovered. In the current study, we investigated the molecular mechanisms that facilitate the cell-associated viremia, which is arguably the most important aspect of EHV-1 pathogenesis. The newly discovered functions of gB and pUS3 add new facets to their previously reported roles. Due to the conserved nature of cell-associated viremia among numerous herpesviruses, these results are also very relevant for viruses such as varicella-zoster virus, pseudorabies virus, human cytomegalovirus, and others. In addition, the constructed mutant and recombinant viruses exhibit potent in vitro replication but have significant defects in certain stages of the disease course. These viruses therefore show much promise as candidates for future live vaccines. E quine herpesvirus type 1 (EHV-1) and EHV-4 are members of the Herpesviridae family and Alphaherpesvirinae subfamily (1, 2). After initial replication in the upper respiratory tract, EHV-1 infects immune cells and migrates past the epithelial basement membrane to the lymph nodes and bloodstream (1-4). As a result, EHV-1 is able to spread throughout the body, where it infects endothelial cells (EC), causing vascular lesions an...

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Section: Growth Analysis and Rates Of Infection By Ehv-1 And Ehv-4 Inmentioning

confidence: 69%

Role of gB and pUS3 in Equine Herpesvirus 1 Transfer between Peripheral Blood Mononuclear Cells and Endothelial Cells: a DynamicIn VitroModel

Spiesschaert

Goldenbogen

Taferner

et al. 2015

J Virol

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“…CD14 (cluster of differentiation 14) is a common monocyte marker used to identify these cells not only in humans, but also in many other species, e.g. dog, cattle or horse [7, 8, 9]. CD16 (cluster of differentiation 16; also Fcγ receptor III) is primarily known as a marker of natural killer cells.…”

Section: Introductionmentioning

confidence: 99%

CD4 and MHCII phenotypic variability of peripheral blood monocytes in dogs

et al. 2019

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In humans and mice, the detailed phenotypic and functional characterization of peripheral blood monocytes allows for identification of three monocyte subsets. There are also evidences of monocyte phenotypic heterogeneity in other species, including cattle, sheep, pig and horse. However, little is known about such variability in dogs. The aim of the study was to determine whether and how peripheral blood monocytes of healthy dogs differ in the presence of MHCII and CD4 and in the basal production of reactive oxygen species (ROS). Three distinct subsets of CD11b + CD14 + monocytes were found in peripheral blood samples of healthy dogs, based on the variations in the density of MHCII and CD4 surface molecules: MHCII + CD4 – (Mo1), MHCII + CD4 + (Mo2) and MHCII – CD4 + (Mo3). The Mo2 and Mo3 were significantly lower in percentage than Mo1 but their basal ROS production was higher. Within the Mo2 and Mo3 subsets, the percentage of cells producing ROS was significantly higher comparing to cells lacking this activity. Canine peripheral blood monocytes vary in the expression of MHCII and CD4 and in the activity suggesting that cells within the three identified subsets carry out different functions. The higher production of ROS in non-activated cells within small subsets of Mo2 and Mo3 monocytes might indicate their immunomodulatory potential.

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“…However, the isolation of these cell types is a time‐consuming process that restricts a rapid clinical application of biomaterial–cell constructs . In contrast, the isolation of monocytes could easily be realized within the time span of an operation from the peripheral blood of a patient . Monocytes have already been shown to support the healing of bone defects, but little is known about the underlying processes .…”

Section: Discussionmentioning

confidence: 87%

Monocyte preseeding leads to an increased implant bed vascularization of biphasic calcium phosphate bone substitutes via vessel maturation

Barbeck

Unger

Booms

et al. 2016

J. Biomed. Mater. Res.

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The present study analyzes the influence of the addition of monocytes to a biphasic bone substitute with two granule sizes (400-700 μm and 500-1000 μm). The majority of the added monocytes was detectable as mononuclear cells, while also low amounts of (chimeric) multinucleated giant cells (MNGCs) were found. No increase in the total number of MNGCs was established, but a significantly increased percent vascularization. Altogether, the results show that the added monocytes become involved in the tissue response to a biomaterial without marked changes in the overall reaction. Monocyte addition enables an increased implant bed vascularization especially via induction of vessel maturation and, thus intervenes positively in the healing reaction to a biomaterial. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2928-2935, 2016.

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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

Abstract: The number of DCs available is critical for further use of these cells and the herein described protocol will therefore help to improved DC generation for therapeutic approaches in horses.

Cited by 5 publications

References 13 publications

Role of gB and pUS3 in Equine Herpesvirus 1 Transfer between Peripheral Blood Mononuclear Cells and Endothelial Cells: a DynamicIn VitroModel

Role of gB and pUS3 in Equine Herpesvirus 1 Transfer between Peripheral Blood Mononuclear Cells and Endothelial Cells: a DynamicIn VitroModel

CD4 and MHCII phenotypic variability of peripheral blood monocytes in dogs

Monocyte preseeding leads to an increased implant bed vascularization of biphasic calcium phosphate bone substitutes via vessel maturation

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