Phenotypic and functional characterization of porcine granulocyte developmental stages using two new markers

Pérez, Carlos; Revilla, Concepción; Álvarez, Belén; Chamorro, Sonia; Correa, Carlos; Doménech, Nieves; Alonso, Fernando; Ezquerra, Ángel; Domı́nguez, Javier

doi:10.1016/j.dci.2006.06.002

Cited by 15 publications

(15 citation statements)

References 22 publications

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“…Staining profiles were analyzed by FlowJo (Tree Star Inc., Oregon, USA). Gating strategy was performed as previously described4748 and as demonstrated in Fig. 1.…”

Section: Methodsmentioning

confidence: 99%

“…In brief, gates for major subsets were set based on forward scatter (FSC), side scatter (SSC) and CD172a47: granulocytes with FSC high SSC high CD172a med , monocytes with FSC high SSC med CD172a high , and lymphocytes with FSC med SSC low CD172a − . Further, immature neutrophils (6D10 + 2B2 − neutrophils) and mature neutrophils (6D10 + 2B2 + neutrophils) were gated48. NK cells were identified as CD16 + lymphocytes.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Delayed development of systemic immunity in preterm pigs as a model for preterm infants

Nguyen

Jiang

Frøkiær

et al. 2016

Sci Rep

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Preterm neonates are highly sensitive to systemic infections in early life but little is known about systemic immune development following preterm birth. We hypothesized that preterm neonates have immature systemic immunity with distinct developmental trajectory for the first several weeks of life, relative to those born at near-term or term. Using pigs as a model, we characterized blood leukocyte subsets, antimicrobial activities and TLR-mediated cytokine production during the first weeks after preterm birth. Relative to near-term and term pigs, newborn preterm pigs had low blood leukocyte counts, poor neutrophil phagocytic rate, and limited cytokine responses to TLR1/2/5/7/9 and NOD1/2 agonists. The preterm systemic responses remained immature during the first postnatal week, but thereafter showed increased blood leukocyte numbers, NK cell proportion, neutrophil phagocytic rate and TLR2-mediated IL-6 and TNF-α production. These immune parameters remained different between preterm and near-term pigs at 2–3 weeks, even when adjusted for post-conceptional age. Our data suggest that systemic immunity follows a distinct developmental trajectory following preterm birth that may be influenced by postnatal age, complications of prematurity and environmental factors. Consequently, the immediate postnatal period may represent a window of opportunity to improve innate immunity in preterm neonates by medical, antimicrobial or dietary interventions.

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“…Staining profiles were analyzed by FlowJo (Tree Star Inc., Oregon, USA). Gating strategy was performed as previously described4748 and as demonstrated in Fig. 1.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Delayed development of systemic immunity in preterm pigs as a model for preterm infants

Nguyen

Jiang

Frøkiær

et al. 2016

Sci Rep

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“…Purified mouse anti-CD4+ antibodies (Biosciences, Inc.) [24], which label porcine cells of monocyte/macrophage linage [25] were used to identify macrophages. Anti-pig 6D10 unconjugated antibodies (Abcam, Inc.) [26] were used to label granulocytes [27]. The effectiveness of these markers and their usefulness to identify specific porcine cells was confirmed by a study described in the Appendix.…”

Section: Methodsmentioning

confidence: 99%

Permeability of subcutaneous tissues surrounding long-term implants to oxygen

et al. 2014

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Certain types of implanted medical devices depend on oxygen supplied from surrounding tissues for their function. However, there is a concern that the tissue associated with the foreign body response to implants may become impermeable to oxygen over the long term and render the implant nonfunctional. We report oxygen flux recordings from electrochemical oxygen sensor devices with wireless telemetry implanted in subcutaneous porcine tissues. The devices remained implanted for up to 13 weeks and were removed with adjacent tissues at specified times for histologic examination. There are four main observations: (1) In the first few weeks after implantation, the oxygen flux to the sensors, or current density, declined to a sustained mean value, having unsynchronized cyclic variations around the mean; (2) The oxygen mass transfer resistance of the sensor membrane was negligible compared to that of the tissue, allowing for a sensitive estimate of the tissue permeability; (3) The effective diffusion coefficient of oxygen in tissues was found to be approximately one order of magnitude lower than in water; and (4) Quantitative histologic analysis of the tissues showed a mild foreign body response to the PDMS sensor membrane material, with capillaries positioned close to the implant surface. Continuous recordings of oxygen flux indicate that the tissue permeability changes predictably with time, and suggest that oxygen delivery can be sustained over the long term.

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“…The mDCs were differentiated following a standard protocol, which has been well determined for their morphology and surface phenotype of major histocompatibility complex class I-positive (MHC-I ϩ ), MHC-II ϩ , and CD11c ϩ cells as well as low expression of CD16, CD14, and CD80/86 (36). Porcine blood monocytes were recently grouped based on the surface expression of CD14, CD163, and swine leukocyte antigen DR (SLA-DR) (38); clearly, monocytes used in this study contain predominant CD14 ϩ CD163 low cells (Ͼ90%) and few CD14 ϩ CD163 high (6%) cells (see Fig. S2 in the supplemental material).…”

Section: Discussionmentioning

confidence: 99%

“…PBMCs were isolated using a 60% Ficoll-Paque Plus gradient (GE Healthcare, Piscataway, NJ). Monocytes were isolated from PBMCs with an anti-CD14 antibody (Ab), using magnetic beads conjugated with the corresponding secondary Ab (Miltenyi Biotec, Auburn, CA), and further phenotyped by a two-color immunofluorescence facilitated flow cytometric analysis (38). The monocyte-derived DCs (mDCs) were generated by culturing monocytes in the presence of IL-4 and granulocyte-macrophage colony-stimulating factor (GM-CSF) for 7 days as previously described (36,37).…”

Section: Animals and Isolation Of Primary Cellsmentioning

confidence: 99%

Antiviral Regulation in Porcine Monocytic Cells at Different Activation States

Sang

Rowland

Blecha

2014

J Virol

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IMPORTANCE Activation statuses of monocytic cells, including monocytes, macrophages (Ms), and dendritic cells (DCs), are critically important for antiviral immunity. Unfortunately, the activation status of porcine monocytic cells or how cell activation status functionally interacts with antiviral immunity remains largely unknown. This is a significant omission because many economically important porcine viruses are monocytotropic, including our focus, PRRSV, which alone causes nearly $800 million economic loss annually in the U.S. swine industries. PRRSV is ideal for deciphering how monocytic cell activation statuses interact with antiviral immunity, because it directly infects subsets of monocytic cells and subverts overall immune responses. In this study, we systematically investigate the activation status of porcine monocytic cells to determine the intricate interaction of viral infection with activation statuses and functionally regulate antiviral immunity within the framework of the activation paradigm. Our findings may provide a means of potentiating antiviral immunity and leading to novel vaccines for PRRS prevention.

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Phenotypic and functional characterization of porcine granulocyte developmental stages using two new markers

Cited by 15 publications

References 22 publications

Delayed development of systemic immunity in preterm pigs as a model for preterm infants

Delayed development of systemic immunity in preterm pigs as a model for preterm infants

Permeability of subcutaneous tissues surrounding long-term implants to oxygen

Antiviral Regulation in Porcine Monocytic Cells at Different Activation States

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