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Failure to detect functional neutrophil B helper cells in the human spleen.

Nagelkerke SQ, aan de Kerk DJ, Jansen MH, van den Berg TK, Kuijpers TW - PLoS ONE (2014)

Bottom Line: A novel role for human neutrophilic granulocytes was recently described, showing that these cells, upon entering the spleen, can be reprogrammed into a distinct B cell-helper neutrophil phenotype that is capable of eliciting B cell responses such as immunoglobulin secretion, class switch recombination and somatic hypermutation.Using similar protocols, we detected a homogeneous population of CD15(high)CD16(high) neutrophils in fresh human spleen samples, which did not differ in phenotype and function from blood neutrophils.Independent confirmation of a role for NBH cells is required.

View Article: PubMed Central - PubMed

Affiliation: Department of Blood Cell Research, Sanquin Research and Landsteiner Laboratory, Amsterdam, The Netherlands.

ABSTRACT
A novel role for human neutrophilic granulocytes was recently described, showing that these cells, upon entering the spleen, can be reprogrammed into a distinct B cell-helper neutrophil phenotype that is capable of eliciting B cell responses such as immunoglobulin secretion, class switch recombination and somatic hypermutation. Using similar protocols, we detected a homogeneous population of CD15(high)CD16(high) neutrophils in fresh human spleen samples, which did not differ in phenotype and function from blood neutrophils. No phenotypic characteristics of costimulatory nature were detected on splenic or circulating neutrophils, nor could we reproduce the immunoglobulin production of splenic B cells in the presence of splenic neutrophils, although B cell function and neutrophil activity were normal. Independent confirmation of a role for NBH cells is required.

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Sorting strategy for sorting Neutrophils and Naïve and Marginal Zone B Cells from splenocytes.Cells were sorted directly from splenocytes stained for CD19, IgD and CD27. Percentages indicate percentage of that population compared to the parent population. FN B Cells: follicular naïve B cells, MZ B cells: marginal zone B cells.
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pone-0088377-g001: Sorting strategy for sorting Neutrophils and Naïve and Marginal Zone B Cells from splenocytes.Cells were sorted directly from splenocytes stained for CD19, IgD and CD27. Percentages indicate percentage of that population compared to the parent population. FN B Cells: follicular naïve B cells, MZ B cells: marginal zone B cells.

Mentions: Using fresh spleen sample from healthy organ donors, we isolated MZ B cells (CD19+IgD+CD27+) and Follicular Naïve (FN) B cells (CD19+IgD+CD27−) by FACS-sorting (Figure 1). We isolated splenic neutrophils in two ways: by the EasySep-Neutrophil-Enrichment-Kit as reported by Puga et al.[10], or by FACS-sorting from splenocytes as based on their FSC/SSC profile (Figure 1). When co-culturing these B cells with neutrophils, neither of the two splenic neutrophil isolates induced any IgM, IgG or IgA production after 7 days (Figure 2), nor did these splenic neutrophils induce differentiation of B cells to plasmablasts (Figure S1 in File S1) [12]. The MZ B cells were viable and able to produce significant amounts of IgM, IgG and IgA upon stimulation with CpG (Figure 2), concomitant with plasmablast formation (Figure S1 in File S1). Similarly, viability and function of neutrophils was normal, as indicated by a normal production of reactive oxygen species in response to various stimuli (Figure S2 in File S1).


Failure to detect functional neutrophil B helper cells in the human spleen.

Nagelkerke SQ, aan de Kerk DJ, Jansen MH, van den Berg TK, Kuijpers TW - PLoS ONE (2014)

Sorting strategy for sorting Neutrophils and Naïve and Marginal Zone B Cells from splenocytes.Cells were sorted directly from splenocytes stained for CD19, IgD and CD27. Percentages indicate percentage of that population compared to the parent population. FN B Cells: follicular naïve B cells, MZ B cells: marginal zone B cells.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3921168&req=5

pone-0088377-g001: Sorting strategy for sorting Neutrophils and Naïve and Marginal Zone B Cells from splenocytes.Cells were sorted directly from splenocytes stained for CD19, IgD and CD27. Percentages indicate percentage of that population compared to the parent population. FN B Cells: follicular naïve B cells, MZ B cells: marginal zone B cells.
Mentions: Using fresh spleen sample from healthy organ donors, we isolated MZ B cells (CD19+IgD+CD27+) and Follicular Naïve (FN) B cells (CD19+IgD+CD27−) by FACS-sorting (Figure 1). We isolated splenic neutrophils in two ways: by the EasySep-Neutrophil-Enrichment-Kit as reported by Puga et al.[10], or by FACS-sorting from splenocytes as based on their FSC/SSC profile (Figure 1). When co-culturing these B cells with neutrophils, neither of the two splenic neutrophil isolates induced any IgM, IgG or IgA production after 7 days (Figure 2), nor did these splenic neutrophils induce differentiation of B cells to plasmablasts (Figure S1 in File S1) [12]. The MZ B cells were viable and able to produce significant amounts of IgM, IgG and IgA upon stimulation with CpG (Figure 2), concomitant with plasmablast formation (Figure S1 in File S1). Similarly, viability and function of neutrophils was normal, as indicated by a normal production of reactive oxygen species in response to various stimuli (Figure S2 in File S1).

Bottom Line: A novel role for human neutrophilic granulocytes was recently described, showing that these cells, upon entering the spleen, can be reprogrammed into a distinct B cell-helper neutrophil phenotype that is capable of eliciting B cell responses such as immunoglobulin secretion, class switch recombination and somatic hypermutation.Using similar protocols, we detected a homogeneous population of CD15(high)CD16(high) neutrophils in fresh human spleen samples, which did not differ in phenotype and function from blood neutrophils.Independent confirmation of a role for NBH cells is required.

View Article: PubMed Central - PubMed

Affiliation: Department of Blood Cell Research, Sanquin Research and Landsteiner Laboratory, Amsterdam, The Netherlands.

ABSTRACT
A novel role for human neutrophilic granulocytes was recently described, showing that these cells, upon entering the spleen, can be reprogrammed into a distinct B cell-helper neutrophil phenotype that is capable of eliciting B cell responses such as immunoglobulin secretion, class switch recombination and somatic hypermutation. Using similar protocols, we detected a homogeneous population of CD15(high)CD16(high) neutrophils in fresh human spleen samples, which did not differ in phenotype and function from blood neutrophils. No phenotypic characteristics of costimulatory nature were detected on splenic or circulating neutrophils, nor could we reproduce the immunoglobulin production of splenic B cells in the presence of splenic neutrophils, although B cell function and neutrophil activity were normal. Independent confirmation of a role for NBH cells is required.

Show MeSH