OMIP‐047: High‐Dimensional phenotypic characterization of B cells

Liechti, Thomas; Günthard, Huldrych F.; Trkola, Alexandra

doi:10.1002/cyto.a.23488

Cited by 18 publications

(19 citation statements)

References 30 publications

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“…We used 16-color flow cytometry to define main B cell subsets (transitional, naive, marginal zone [MZ], memory B cells, and plasmablasts) and memory B cell subsets in blood (Fig. S1, A and B; Liechti et al, 2018a ). By monitoring changes in B cell subsets during HIV-1 infection, we found increased frequencies of transitional B cells and plasmablasts in HIV-1–infected individuals ( n = 19) compared with healthy donors ( n = 29; Fig.…”

Section: Resultsmentioning

confidence: 99%

“…To monitor B cell populations by flow cytometry, PBMCs were stained using an 18-parameter panel as described in detail in Liechti et al (2018a) . Briefly, cells were thawed and washed with FACS buffer containing PBS (Thermo Fisher Scientific), 2% heat-inactivated FCS (Thermo Fisher Scientific), 2 mM EDTA (Sigma-Aldrich), and 20 µg/ml DNase (Sigma-Aldrich) and stained with live/dead fixable near-infrared dye (Life Technologies) for 30 min, enabling dead cell exclusion.…”

Section: Methodsmentioning

confidence: 99%

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Widespread B cell perturbations in HIV-1 infection afflict naive and marginal zone B cells

Liechti

Kadelka

Braun

et al. 2019

Journal of Experimental Medicine

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Perturbations in B cells are a hallmark of HIV-1 infection. This is signified by increased numbers of exhausted CD21neg memory B cells, driven by continuous antigen-specific and bystander activation. Using high-dimensional flow cytometry, we demonstrate that this exhausted phenotype is also prevalent among peripheral antigen-inexperienced naive and marginal zone (MZ) B cells in acute and chronic HIV-1 infection. A substantial fraction of naive and MZ B cells exhibit down-regulated CD21 levels and diminished response to B cell receptor (BCR)–dependent stimulation. Compared with CD21pos subsets, the CD21neg naive and MZ B cells differ in the expression of chemokine receptors and activation markers. Effective antiretroviral treatment normalizes peripheral naive and MZ B cell populations. Our results emphasize a more widely spread impairment of B cells in HIV-1 infection than previously appreciated, including antigen-inexperienced cells. This highlights the importance of monitoring functional capacities of naive B cells in HIV-1 infection, as exhausted CD21neg naive B cells may severely impair induction of novel B cell responses.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Widespread B cell perturbations in HIV-1 infection afflict naive and marginal zone B cells

Liechti

Kadelka

Braun

et al. 2019

Journal of Experimental Medicine

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“…Three additional dot plots were added to recognize the following B cell subsets: CD24 hi CD38 hi Breg, CD24 hi CD27 hi Breg and CD5 + CD1d + Breg. Gate strategy was reported in g.3a (26).…”

Section: Flow Cytrometric Analysismentioning

confidence: 99%

Follicular T helper and Breg cell balance in severe allergic asthma before and after omalizumab therapy

Bergantini

d’Alessandro

Cameli

et al. 2021

Preprint

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Background Severe allergic asthma (SAA) is based on type 2 (T2-high) immune responses to allergens promoting type 2 T helper (Th2) cell cytokine responses and production of IgE antibodies. Omalizumab was the first biological drug licensed for clinical use in the management of IgE-mediated SAA. Despite, emerging evidence supporting the prominent role of follicular T cells (Tfh), Breg and Treg subsets, in the development and progression of SAA, no data is available on the impact omalizumab therapy. Methods Ten SAA patients monitored at the Respiratory Diseases Unit of Siena University Hospital and 10 healthy sex- and age-matched controls were enrolled in the study. Clinical and functional parameters were collected at baseline (T0) and after 6 months of therapy (T6). Cellular population analysis were determined through multi-color flow cytometry. Results SAA patients showed higher percentages of Th17.1, Tfh and Tfh2 while CD24 hi CD27 hi Breg cell, Treg and Tfr percentages were significantly lower than controls. Higher percentages of Tfh2 in patients with nasal polyps than in those without and in controls were observed. At T6, significant decreases of Tfh and Tfh2 than T0 were observed. A slightly significant increase in Teff was reported at T6 with respect to T0. ΔIgE levels in serum were correlated with ΔCD19 + CD24 + CD27 + Breg cell percentages (r=-0.86, p=0.0022). Conclusions Our data explored the changes of Tfh cells, Tregs and Bregs in severe asthma. The restoration of immunological imbalance in SAA patients after omalizumab is surely intriguing and represents a glimpse of light in the comprehension of immunological effects of treatment.

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“…Considering this and the above-mentioned drawbacks, the notorious use of a dump gate seems to base more on historical grounds rather than actual need or experimental logic and should be scrutinized by the individual researcher on a case-by-case basis. In other words, knowledge about the cells of interest under physiological and pathological conditions and the potential pitfalls provided, dump gating can still be productive, for instance, to better demarcate other markers (17), to avoid loss of resolution due to excessive residual spillover from multiple channels following compensation (18), or to reduce background in specific applications such as multimer staining protocols (19). Despite the highest number of available channels, these examples should also be considered in the case of mass cytometry, which may face particular challenges with differential isotope transmission (20) and channel cross-talk (21) as well.…”

Section: Flow Cytometry: To Dump or Not To Dumpmentioning

confidence: 99%