Improved recovery of functionally active eosinophils and neutrophils using novel immunomagnetic technology

Son, Kiho; Mukherjee, Manali; McIntyre, Brendan A.S.; Eguez, Jose Cano; Radford, Katherine; LaVigne, Nicola; Ethier, Caroline; Davoine, Francis; Janssen, Luke J.; Lacy, Paige; Nair, Parameswaran

doi:10.1016/j.jim.2017.06.005

Cited by 31 publications

(27 citation statements)

References 29 publications

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“…Studies of eosinophils in and from mouse models of Th2‐mediated allergy have ultimately led to our current understanding of phenotype‐specific asthma and provide targets for novel therapies . However, while human eosinophils can be easily isolated by negative‐selection technologies, there are no readily‐available tools that facilitate isolation of live, untouched eosinophils from wild‐type (WT) mice. To date, the major source of mouse eosinophils has been peripheral blood and tissues from IL‐5 transgenic ( IL5 tg) strains in which profound eosinophilia results from cytokine overproduction in vivo.…”

Section: Introductionmentioning

confidence: 99%

Frontline Science: Cytokine-mediated developmental phenotype of mouse eosinophils: IL-5-associated expression of the Ly6G/Gr1 surface Ag

Limkar

Mai

Sek

et al. 2019

Journal of Leukocyte Biology

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Eosinophils have broad and extensive immunomodulatory capacity; recent studies have focused on the roles of distinct eosinophil subsets in specific tissue microenvironments. Ly6G is a GPI‐linked leukocyte surface Ag understood primarily as a marker of mouse neutrophils, although its full function is not known. Here, we show that Ly6G/Gr1, detected by mAbs 1A8 (anti‐Ly6G) and RB6‐8C5 (anti‐Gr1), is detected prominently on a significant fraction of eosinophils from mouse bone marrow and bone marrow‐derived culture, with fractions expressing this Ag increasing in IL‐5‐enriched microenvironments. Among our findings, we identified SiglecF+Gr1+ eosinophils in bone marrow from naïve, allergen‐challenged and IL‐5 transgenic mice; SiglecF+Gr1+ eosinophils were also prominent ex vivo in bone marrow‐derived eosinophils (bmEos) in IL‐5‐enriched culture. Reducing the IL‐5 concentration 20‐fold had no impact on the rate of generation of SiglecF+ bmEos but did result in a marked increase in the Gr1− fraction (from 17.4 ± 2% to 30 ± 2.3%, ***P < 0.005). Reducing the IL‐5 concentration also enhanced chemotaxis; SiglecF+Gr1− bmEos were considerably more responsive to eotaxin‐1 than were their SiglecF+Gr1+ counterparts. These results suggest that (i) IL‐5 regulates the expression of Ly6G/Gr1, either directly or indirectly, in cells of the eosinophil lineage, (ii) eosinophils generated in response to high concentrations of IL‐5 can be distinguished from those generated under homeostatic conditions by expression of the Ly6G/Gr1 cell surface Ag, and (iii) expression of Ly6G/Gr1 may have an impact on function, directly or indirectly, including the potential to undergo chemotaxis in response to eotaxin‐1.

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Section: Introductionmentioning

confidence: 99%

Frontline Science: Cytokine-mediated developmental phenotype of mouse eosinophils: IL-5-associated expression of the Ly6G/Gr1 surface Ag

Limkar

Mai

Sek

et al. 2019

Journal of Leukocyte Biology

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“…To date, isolation of viable human eosinophils from blood can be completed by density gradient or by antibody mediated depletion (e.g., magnetic depletion) of other leukocytes to avoid excessive manipulation of eosinophils 12 – 14 . These methods, while favorable for blood-derived eosinophil isolation, remain problematic for tissue-derived eosinophils, which require enzymatic degradation of tissue and inclusion of more diverse cell compositions within the digested sample.…”

Section: Introductionmentioning

confidence: 99%

A FACS-based approach to obtain viable eosinophils from human adipose tissue

Hernandez

Tew

et al. 2020

Sci Rep

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Eosinophils have been widely investigated in asthma and allergic diseases. More recently, new insights into the biology of these cells has illustrated eosinophils contribute to homeostatic functions in health such as regulation of adipose tissue glucose metabolism. Human translational studies are limited by the difficulty of obtaining cells taken directly from their tissue environment, relying instead on eosinophils isolated from peripheral blood. Isolation techniques for tissue-derived eosinophils can result in unwanted cell or ribonuclease activation, leading to poor cell viability or RNA quality, which may impair analysis of effector activities of these cells. Here we demonstrate a technique to obtain eosinophils from human adipose tissue samples for the purpose of downstream molecular analysis. From as little as 2 g of intact human adipose tissue, greater than 10 4 eosinophils were purified by fluorescence-activated cell sorting (FACS) protocol resulting in ≥ 99% purity and ≥ 95% viable eosinophils. We demonstrated that the isolated eosinophils could undergo epigenetic analysis to determine differences in DNA methylation in various settings. Here we focused on comparing eosinophils isolated from human peripheral blood vs human adipose tissue. Our results open the door to future mechanistic investigations to better understand the role of tissue resident eosinophils in different context.

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“…Son et al . 24 reported that the MACSxpress method consistently allowed high-purity isolation of eosinophils (95.0 ± 1.7%) and neutrophils (94.2 ± 10.1%). Then the cells were deposited on slides by cytocentrifugation (150× g , 4 min) and fixed in paraformaldehyde (4%).…”

Section: Methodsmentioning

confidence: 99%

“…The neutrophils and eosinophilic polynuclear cells were purified with the MACSxpress Whole Blood Neutrophil Isolation Kit and the MAC-Sxpress Whole Blood Eosinophil Isolation Kit (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the supplier's recommendations, respectively. Son et al 24 reported that the MACSxpress method consistently allowed high-purity isolation of eosinophils (95.0 ± 1.7%) and neutrophils (94.2 ± 10.1%). Then the cells were deposited on slides by cytocentrifugation (150×g, 4 min) and fixed in paraformaldehyde (4%).…”

Section: Purification Of Human Neutrophils and Eosinophilsmentioning

confidence: 99%

APRIL-producing eosinophils are involved in gastric MALT lymphomagenesis induced by Helicobacter sp infection

Blosse

Peru

Lévy

et al. 2020

Sci Rep

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The roles of the inflammatory response and production of a proliferation-inducing ligand (APRIL) cytokine in gastric mucosa-associated lymphoid tissue (MALT) lymphomagenesis induced by Helicobacter species infection are not clearly understood. We characterized the gastric mucosal inflammatory response associated with gastric MALT lymphoma (GML) and identified APRIL-producing cells in two model systems: an APRIL transgenic mouse model of GML induced by Helicobacter infection (Tg-hAPRIL) and human gastric biopsy samples from Helicobacter pylori-infected GML patients. In the mouse model, polarization of T helper 1 (tbet), T helper 2 (gata3), and regulatory T cell (foxp3) responses was evaluated by quantitative PCR. In humans, a significant increase in april gene expression was observed in GML compared to gastritis. APRIL-producing cells were eosinophilic polynuclear cells located within lymphoid infiltrates, and tumoral B lymphocytes were targeted by APRIL. Together, the results of this study demonstrate that the Treg-balanced inflammatory environment is important for gastric lymphomagenesis induced by Helicobacter species, and suggest the pro-tumorigenic potential of APRIL-producing eosinophils.

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Improved recovery of functionally active eosinophils and neutrophils using novel immunomagnetic technology

Cited by 31 publications

References 29 publications

Frontline Science: Cytokine-mediated developmental phenotype of mouse eosinophils: IL-5-associated expression of the Ly6G/Gr1 surface Ag

Frontline Science: Cytokine-mediated developmental phenotype of mouse eosinophils: IL-5-associated expression of the Ly6G/Gr1 surface Ag

A FACS-based approach to obtain viable eosinophils from human adipose tissue

APRIL-producing eosinophils are involved in gastric MALT lymphomagenesis induced by Helicobacter sp infection

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