Allergy Asthma Immunol Res.  2020 Mar;12(2):274-291. 10.4168/aair.2020.12.2.274.

Evidence for the Presence of Long-Lived Plasma Cells in Nasal Polyps

• Zhang YN
• Song J
• Zhai GT
• Wang H
• Luo RZ
• Li JX
• Liao B
• Ma J
• Wang H
• Lu X
• Liu DB
• Liu Z

• KMID: 2468462
• DOI: http://doi.org/10.4168/aair.2020.12.2.274

Abstract

PURPOSE
Plasma cells and immunoglobulins (Igs) play a pivotal role in the induction and maintenance of chronic inflammation in nasal polyps. During secondary immune responses, plasma cell survival and Ig production are regulated by the local environment. The purpose of the present study was to investigate the presence of long-lived plasma cells (LLPCs) and specific survival niches for LLPCs in human nasal polyps.
METHODS
Nasal mucosal samples were cultured with an air-liquid interface system and the Ig levels in culture supernatants were analyzed by enzyme-linked immunosorbent assay. The characteristics of LLPCs in nasal polyps were determined by immunohistochemistry and immunofluorescence. The expression of neurotrophins as well as their receptors was detected by quantitative real-time polymerase chain reaction, immunohistochemistry, immunofluorescence, and Western blotting.
RESULTS
The numbers of CD138⁺ total plasma cells and BCL2⁺ plasma cells were increased in both eosinophilic and non-eosinophilic nasal polyps compared with those in normal tissues. The production of IgG, IgA, and IgE was detected in culture supernatants even after a 32-day culture of nasal polyps. Although the total numbers of plasma cells were decreased in nasal polyps after culture, the numbers of BCL2⁺ plasma cells remained stable. The expression of nerve growth factor (NGF) as well as tropomyosin receptor kinase (Trk) A, a high-affinity receptor for NGF, was upregulated in both eosinophilic and non-eosinophilic nasal polyps. In addition, BCL2⁺ plasma cell numbers were positively correlated with NGF and TrkA mRNA expression in nasal mucosal tissues. Polyp plasma cells had the expression of TrkA.
CONCLUSIONS
Human nasal polyps harbor a population of LLPCs and NGF may be involved in their prolonged survival. LLPCs may be a novel therapeutic target for suppressing the local Ig production in nasal polyps.

Keyword

Immunoglobulins; plasma cells; nasal polyps; survival; nerve growth factor

MeSH Terms

Blotting, Western
Enzyme-Linked Immunosorbent Assay
Eosinophils
Fluorescent Antibody Technique
Humans
Immunoglobulin A
Immunoglobulin E
Immunoglobulin G
Immunoglobulins
Immunohistochemistry
Inflammation
Mucous Membrane
Nasal Polyps*
Nerve Growth Factor
Nerve Growth Factors
Phosphotransferases
Plasma Cells*
Plasma*
Polyps
Real-Time Polymerase Chain Reaction
RNA, Messenger
Tropomyosin
Immunoglobulin A
Immunoglobulin E
Immunoglobulin G
Immunoglobulins
Nerve Growth Factor
Nerve Growth Factors
Phosphotransferases
RNA, Messenger
Tropomyosin

Figure

• 

  Fig. 1 Increased accumulation of BCL2+CD138+ plasma cells in nasal polyps. (A) Representative photomicrographs showing BCL2+CD138+ plasma cells in control tissues, and in eosinophilic and non-eosinophilic nasal polyp tissues. Red arrows indicate representative CD138+ plasma cells; green arrows indicate representative BCL2+ cells; yellow arrows indicate representative BCL2+CD138+ plasma cells (original magnification ×400). (B) Quantification of CD138+ total plasma cells and BCL2+CD138+ plasma cells in lamina propria. BCL2, B cell lymphoma 2; Eos CRSwNP, eosinophilic chronic rhinosinusitis with nasal polyps; Non-Eos CRSwNP, non-eosinophilic chronic rhinosinusitis with nasal polyps.

• 

  Fig. 2 Ig production from ex vivo cultured sinonasal mucosal samples. Eosinophilic and non-eosinophilic nasal polyps and inferior turbinate mucosal samples were cultured ex vivo for 32 days. Culture supernatants were collected at days 2, 4, 8, 16, and 32, and Ig levels in the culture supernatants were measured by enzyme-linked immunosorbent assay. Ig, immunoglobulin; Eos CRSwNP, eosinophilic chronic rhinosinusitis with nasal polyps; Non-Eos CRSwNP, non-eosinophilic chronic rhinosinusitis with nasal polyps.

• 

  Fig. 3 BCL2+CD138+ plasma cells are retained in nasal polyps after a 32-day ex vivo culture. (A) Representative photomicrographs showing CD138+ plasma cells in nasal polyp tissues before and after culture and the quantification of CD138+ plasma cells. Arrows indicate representative CD138+ plasma cells. (B) Representative photomicrographs and quantitative analysis of BCL2+CD138+ plasma cells in nasal polyp tissues before and after culture. Red arrows indicate representative CD138+ plasma cells; green arrows indicate representative BCL2+ cells; yellow arrows indicate representative BCL2+CD138+ plasma cells. (C) Representative photomicrographs show that CD138+ plasma cells in nasal polyp tissue do not have Ki-67 expression before and after culture. Red arrows indicate representative CD138+ plasma cells; green arrows indicate representative Ki67+ cells; white arrows indicate representative Ki67−CD138+ plasma cells (original magnification ×400). BCL2, B cell lymphoma 2.

• 

  Fig. 4 The mRNA expression of NTs (A) as well as their receptors (B), and correlations between BCL2+CD138+ plasma cell count and the expression level of NTs and their receptors (C) in sinonasal mucosa samples as detected by quantitative real-time polymerase chain reaction. NT, neurotrophin; Eos CRSwNP, eosinophilic chronic rhinosinusitis with nasal polyps; Non-Eos CRSwNP, non-eosinophilic chronic rhinosinusitis with nasal polyps; NGF, nerve growth factor; BDNF, brain derived neurotrophic factor; Trk, tropomyosin receptor kinase; BCL2, B cell lymphoma 2; P75, p75 neurotrophin receptor.

• 

  Fig. 5 Increased protein expression of NGF in epithelial cells in eosinophilic and non-eosinophilic nasal polyps. Representative photomicrographs showing NGF expression in nasal epithelial cells and infiltrating cells in control tissues and eosinophilic and non-eosinophilic nasal polyp tissues as detected by immunohistochemistry. Isotype control staining is also shown. Arrows indicate representative NGF+ cells. The expression intensity of NGF in epithelial cells was quantified (original magnification ×400). NGF, nerve growth factor; Eos CRSwNP, eosinophilic chronic rhinosinusitis with nasal polyps; Non-Eos CRSwNP, non-eosinophilic chronic rhinosinusitis with nasal polyps; IOD, integrated optical density.

• 

  Fig. 6 Increased protein expression of NGF in lamina propria in eosinophilic and non-eosinophilic nasal polyps. (A) Representative photomicrographs showing NGF+ cells in the lamina propria of control tissues, and eosinophilic and non-eosinophilic nasal polyp tissues as detected by immunohistochemistry. Arrows indicate representative NGF+ cells. The numbers of NGF+ cells in the lamina propria were quantified. (B) Representative immunostaining of consecutive tissue sections from a patient with eosinophilic chronic rhinosinusitis with nasal polyps (Eos CRSwNP) showing the expression of NGF by ECP+ eosinophils. Arrows with the same direction indicate the same cells in consecutive serial sections. (C) Representative double immunofluorescence staining of a tissue section from a patient with Eos CRSwNP showing the expression of NGF by tryptase-positive mast cells. Red arrows indicate representative NGF+ cells; green arrows indicate representative tryptase+ cells; yellow arrows indicate representative NGF+tryptase+ cells (original magnification ×400). NGF, nerve growth factor; Eos CRSwNP, eosinophilic chronic rhinosinusitis with nasal polyps; ECP, eosinophilic cationic protein; Non-Eos CRSwNP, non-eosinophilic chronic rhinosinusitis with nasal polyps. HPF, high power field.

• 

  Fig. 7 Increased protein expression of TrkA in eosinophilic and non-eosinophilic nasal polyps. (A) Representative photomicrographs showing TrkA expression in infiltrating cells in the lamina propria of control tissues, and eosinophilic and non-eosinophilic nasal polyp tissues as detected by immunohistochemistry. Arrows indicate representative TrkA+ cells. The numbers of TrkA+ cells in the lamina propria were quantified. (B) Representative double immunofluorescence staining of a tissue section from a patient with eosinophilic chronic rhinosinusitis with nasal polyps (Eos CRSwNP) showing the expression of TrkA by CD138+ plasma cells. Green arrows indicate representative TrkA+ cells; red arrows indicate representative CD138+ cells; yellow arrows indicate representative TrkA+CD138+ plasma cells (original magnification ×400). Trk, tropomyosin receptor kinase; Non-Eos CRSwNP, non-eosinophilic chronic rhinosinusitis with nasal polyps. HPF, high power field.

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  Fig. 8 Increased protein expression of NGF and TrkA in eosinophilic and non-eosinophilic nasal polyps detected by western blot analysis. (A) Representative blots of NGF and TrkA in whole—cell lysates of nasal tissues are shown. (B) NGF relative protein expression was quantified and normalized to β-actin expression. (C) TrkA relative protein expression was quantified and normalized to β-actin. NGF, nerve growth factor; Trk, tropomyosin receptor kinase; Eos CRSwNP, eosinophilic chronic rhinosinusitis with nasal polyps; Non-Eos CRSwNP, non-eosinophilic chronic rhinosinusitis with nasal polyps.

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