OMIP‐023: 10‐Color, 13 antibody panel for in‐depth phenotyping of human peripheral blood leukocytes

Bocsi, József; Melzer, Susanne; Dähnert, Ingo; Tárnok, Attila

doi:10.1002/cyto.a.22505

Cited by 32 publications

(63 citation statements)

References 12 publications

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“…For a precise measurement of monocyte subpopulations, a combination of several markers might be necessary, as shown elsewhere. 34 Nevertheless, for the current approach, the CD11b staining showed to be sufficient, as also showed by others. 35 Once the presence of MΦ was proven, the cells were incubated with GNP for 24 hours and the GNP uptake was detected using FCM scatter analyses ( Figure 2).…”

supporting

confidence: 66%

Detection of gold nanorods uptake by macrophages using scattering analyses combined with diffusion reflection measurements as a potential tool for in vivo atherosclerosis tracking

Ankri¹,

Melzer²,

Tárnok³

et al. 2015

IJN

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In this study, we report a potential noninvasive technique for the detection of vulnerable plaques using scatter analyses with flow cytometry (FCM) method combined with the diffusion reflection (DR) method. The atherosclerotic plaques are commonly divided into two major categories: stable and vulnerable. The vulnerable plaques are rich with inflammatory cells, mostly macrophages (MΦ), which release enzymes that break down collagen in the cap. The detection method is based on uptake of gold nanorods (GNR) by MΦ. The GNR have unique optical properties that enable their detection using the FCM method, based on their scattering properties, and using the DR method, based on their unique absorption properties. This work demonstrates that after GNR labeling of MΦ, 1) the FCM scatter values increased up to 3.7-fold with arbitrary intensity values increasing from 1,110 to 4,100 and 2) the DR slope changed from an average slope of 0.196 (MΦ only) to an average slope of 0.827 (MΦ labeled with GNR) ( P <0.001 for both cases). The combination of FCM and DR measurements provides a potential novel, highly sensitive, and noninvasive method for the identification of atherosclerotic vulnerable plaques, aimed to develop a potential tool for in vivo tracking.

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confidence: 66%

Detection of gold nanorods uptake by macrophages using scattering analyses combined with diffusion reflection measurements as a potential tool for in vivo atherosclerosis tracking

Ankri¹,

Melzer²,

Tárnok³

et al. 2015

IJN

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“…After thawing, we incubated (37°C, 5 h, 5% CO 2 ) the cells (0.5 × 10 6 ) in culture medium containing RPMI 1640 with 10% FCS, 1× nonessential amino acids, 10 mM HEPES, 2 mM L-glutamine (all from Invitrogen) and 1% penicillin/streptomycin with 1:1,000 Golgi Stop (554724, BD Biosciences), 10 ng/ml phorbol 12-myristate 13-acteate (PMA) and 500 ng/ml calcium ionophore A23187. We immunophenotyped the cells in BAL fluid as described before (91). We stained cells with anti-human CD3 antibody (SP34-2), anti-human CD8 antibody (B9.11), anti-human CD14 antibody (RMO52), anti-human CD19 antibody (J3-119), anti-human CD45 antibody (J.33), anti-human CD56 antibody (NK901), anti-human CD69 antibody (TP1.55.3) and anti-human NKp46 antibody (9E2) (all diluted 1:1000, Biolegend).…”

Section: Methodsmentioning

confidence: 99%

Intestinal commensal bacteria mediate lung mucosal immunity and promote resistance of newborn mice to infection

et al. 2017

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Immature mucosal defenses contribute to increased susceptibility of newborn infants to pathogens. Sparse knowledge of age-dependent changes in mucosal immunity has hampered improvements in neonatal morbidity due to infections. Here, we report that exposure of neonatal mice to commensal bacteria immediately after birth is required for a robust host defense against bacterial pneumonia, the leading cause of death in newborn infants. This crucial window was characterized by an abrupt influx of interleukin (IL)-22 producing group 3 innate lymphoid cells (IL22+ILC3) into the lungs of newborn mice. This influx was dependent on sensing of commensal bacteria by intestinal mucosal dendritic cells. Disruption of postnatal commensal colonization or selective depletion of dendritic cells interrupted the migratory program of lung IL-22+ILC3 and made the newborn mice more susceptible to pneumonia, which was reversed by transfer of commensal bacteria after birth. Thus, the resistance of newborn mice to pneumonia relied on commensal bacteria-directed ILC3-influx into the lungs, which mediated IL-22-dependent host resistance to pneumonia during this developmental window. These data establish that postnatal colonization by intestinal commensal bacteria is pivotal in the development of lung defenses in mice.

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“…Samples were collected in the early morning (6:00 to 8:59: 62%) or in the late morning (9:00 to 12:00: 38%) and were processed within 2 h after drawing. Sample preparation was performed as described previously . In short, erythrocytes of were lysed by adding to 1 mL PB 25 mL lysis buffer (8.3 g L −1 NH 4 Cl, 1 g L −1 KHCO 3 and 0.04 g L −1 EDTA) for 10 min.…”

Section: Methodsmentioning

confidence: 99%

“…Cells were then labeled with an Optimized Multicolor Immunofluorescence Panel (OMIP) specifically developed for the LIFE study. The cocktail consisted of 13 antibodies labeled to 10 different fluorochromes . Optimal antibody concen trations were determined by titration.…”

Section: Methodsmentioning

confidence: 99%

Reference intervals for leukocyte subsets in adults: Results from a population‐based study using 10‐color flow cytometry

Melzer

Zachariae

Bocsi

et al. 2015

Cytometry Part B Clinical

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Background: Reference intervals for leukocyte subsets from peripheral blood are helpful for the understanding of disease states and therapy effects.Methods: We performed in-depth immunophenotyping for 608 healthy German adults from the Leipzig region from 40 to 79 years by 10-color flow cytometry (FCM) to gain reference information for various leukocyte subsets including subsets of granulocytes, monocytes and lymphocytes.Results: First, we derived gender-and age-specific reference intervals for males and females from 40 to 59 and from 60 to 79 years, respectively. Second, we further investigated the influence of gender and age on leukocyte counts. We found significantly higher cell counts for monocytes (P < 0.001) and NK cells (P < 0.001) in men, whereas women had higher counts for B cells (P < 0.001), Th cells (P < 0.001) and regulatory T cells (P 5 0.008). Furthermore, with increasing age, a decrease in Tc cells (about 8% within 5 years) and an increase in NK cells (<4% within 5 years) were observed.Conclusion: In future research, it should be investigated whether these are real ageing effects that can be confirmed in longitudinal studies. Furthermore, it is important to understand if the Tc cell count drop is functionally compensated by the increase of NK cells. V C 2015 International Clinical Cytometry Society

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OMIP‐023: 10‐Color, 13 antibody panel for in‐depth phenotyping of human peripheral blood leukocytes

Cited by 32 publications

References 12 publications

Detection of gold nanorods uptake by macrophages using scattering analyses combined with diffusion reflection measurements as a potential tool for in vivo atherosclerosis tracking

Detection of gold nanorods uptake by macrophages using scattering analyses combined with diffusion reflection measurements as a potential tool for in vivo atherosclerosis tracking

Intestinal commensal bacteria mediate lung mucosal immunity and promote resistance of newborn mice to infection

Reference intervals for leukocyte subsets in adults: Results from a population‐based study using 10‐color flow cytometry

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