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Specific Neuropilins Expression in Alveolar Macrophages among Tissue-Specific Macrophages.

Aung NY, Ohe R, Meng H, Kabasawa T, Yang S, Kato T, Yamakawa M - PLoS ONE (2016)

Bottom Line: The expression of both NRPs was detected in AMs, BMs and IVMs by IHC.The frequency of NRPs+ AMs in lung tissue adjacent to the cancer margin was significantly higher than the frequencies in inflamed and normal lung tissue.Furthermore, the possible origin of AMs from blood monocytes could be suggested from a co-expression of NRPs and DC-SIGN.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathological Diagnostics, Yamagata University Faculty of Medicine, Yamagata, Japan.

ABSTRACT
In the immune system, neuropilins (NRPs), including NRP-1 and NRP-2, are expressed in thymocytes, dendritic cells, regulatory T cells and macrophages. Their functions on immune cells around the neoplastic cells vary into pro-angiogenesis, tumor progression and anti-angiogenesis according to their ligands. Even though NRPs expression on malignant tumors and immune system has studied, a PubMed-based literature query did not yield any articles describing NRPs expression on tissue-specific macrophages. The aims of this study were (i) to detect NRPs expression on tissue-specific macrophages in the brain, liver, spleen, lymph node and lung; (ii) to observe NRPs expression in classes of macrophages, including alveolar macrophages (AMs), bronchial macrophages (BMs), interstitial macrophages (IMs), intravascular macrophages (IVMs) and macrophage subsets (M1, M2 and Mox) in lung; and (iii) to detect the co-expression of NRPs and dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN) in AMs. Both NRPs were specifically detected in AMs among tissue-specific macrophages by immunohistochemistry (IHC). NRPs mRNA expression levels were characterized in normal lung by reverse transcriptase polymerase chain reaction (RT-PCR) and in situ-polymerase chain reaction (in situ-PCR). The expression of both NRPs was detected in AMs, BMs and IVMs by IHC. The frequency of NRPs+ AMs in lung tissue adjacent to the cancer margin was significantly higher than the frequencies in inflamed and normal lung tissue. Double and triple IHC demonstrated that NRPs are expressed on all macrophage subsets in lung. Double IHC showed co-expression of DC-SIGN and NRPs in AMs. This study demonstrated for the first time the specific expression of both NRPs in AMs among tissue-specific macrophages and their expression on M1, M2 and Mox macrophages. Furthermore, the possible origin of AMs from blood monocytes could be suggested from a co-expression of NRPs and DC-SIGN.

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NRPs expression in tissue-specific macrophages compared to immunostaining with a cocktail of anti-CD68 and anti-CD163 antibodies.Tissue-specific macrophages were recognized by immunostaining with a cocktail of anti-CD68 and CD163 antibodies in serial sections. NRP-1 and NRP-2 expression was detected in alveolar macrophages in lung, but not in lymph node (sinus macrophages). And NRP-1 and NRP-2 also expressed on bronchial macrophages. Green arrow indicates NRP-2 expression on lymphatic vascular endothelium, used as positive control. Serial sections were counterstained with hematoxylin. NRP-1, neuropilin 1; NRP-2, neuropilin 2.
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pone.0147358.g002: NRPs expression in tissue-specific macrophages compared to immunostaining with a cocktail of anti-CD68 and anti-CD163 antibodies.Tissue-specific macrophages were recognized by immunostaining with a cocktail of anti-CD68 and CD163 antibodies in serial sections. NRP-1 and NRP-2 expression was detected in alveolar macrophages in lung, but not in lymph node (sinus macrophages). And NRP-1 and NRP-2 also expressed on bronchial macrophages. Green arrow indicates NRP-2 expression on lymphatic vascular endothelium, used as positive control. Serial sections were counterstained with hematoxylin. NRP-1, neuropilin 1; NRP-2, neuropilin 2.

Mentions: To investigate NRP-1 and NRP-2 expression in tissue-specific macrophages, single IHC for NRP-1 and NRP-2 and immunostaining with cocktail antibodies for CD68 and CD163 in serial tissue sections were performed. NRP-1 and NRP-2 were specifically expressed in AMs among tissue-specific macrophages (microglia in the brain, Kupffer cells in the liver, red pulp macrophages in the spleen and sinus histiocytes in the lymph node) (Figs 1 and 2). In addition, NRP-1 and NRP-2 expression was detected on neurons in brain tissue (Fig 1). In lung tissue, NRP-1 expression was observed in the nuclei and cytoplasm of neutrophils, some reactive lymphocytes, plasma cell-like lymphocytes and respiratory epithelial and glandular epithelial cells. NRP-2 expression in lung tissue was observed in respiratory epithelial and glandular epithelial cells. Weak NRP-1 expression was observed in the cytoplasm of hepatocytes in liver tissue. In spleen and lymph node, NRP-1 expression was observed in some lymphocytes in the marginal and mantle zones of lymphoid follicles, plasma cell-like lymphocytes and some dendritic cell-like cells. NRP-1 and NRP-2 expression was observed on lymphatic and vascular endothelial cells in brain, lung, liver, spleen and lymph nodes.


Specific Neuropilins Expression in Alveolar Macrophages among Tissue-Specific Macrophages.

Aung NY, Ohe R, Meng H, Kabasawa T, Yang S, Kato T, Yamakawa M - PLoS ONE (2016)

NRPs expression in tissue-specific macrophages compared to immunostaining with a cocktail of anti-CD68 and anti-CD163 antibodies.Tissue-specific macrophages were recognized by immunostaining with a cocktail of anti-CD68 and CD163 antibodies in serial sections. NRP-1 and NRP-2 expression was detected in alveolar macrophages in lung, but not in lymph node (sinus macrophages). And NRP-1 and NRP-2 also expressed on bronchial macrophages. Green arrow indicates NRP-2 expression on lymphatic vascular endothelium, used as positive control. Serial sections were counterstained with hematoxylin. NRP-1, neuropilin 1; NRP-2, neuropilin 2.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4764655&req=5

pone.0147358.g002: NRPs expression in tissue-specific macrophages compared to immunostaining with a cocktail of anti-CD68 and anti-CD163 antibodies.Tissue-specific macrophages were recognized by immunostaining with a cocktail of anti-CD68 and CD163 antibodies in serial sections. NRP-1 and NRP-2 expression was detected in alveolar macrophages in lung, but not in lymph node (sinus macrophages). And NRP-1 and NRP-2 also expressed on bronchial macrophages. Green arrow indicates NRP-2 expression on lymphatic vascular endothelium, used as positive control. Serial sections were counterstained with hematoxylin. NRP-1, neuropilin 1; NRP-2, neuropilin 2.
Mentions: To investigate NRP-1 and NRP-2 expression in tissue-specific macrophages, single IHC for NRP-1 and NRP-2 and immunostaining with cocktail antibodies for CD68 and CD163 in serial tissue sections were performed. NRP-1 and NRP-2 were specifically expressed in AMs among tissue-specific macrophages (microglia in the brain, Kupffer cells in the liver, red pulp macrophages in the spleen and sinus histiocytes in the lymph node) (Figs 1 and 2). In addition, NRP-1 and NRP-2 expression was detected on neurons in brain tissue (Fig 1). In lung tissue, NRP-1 expression was observed in the nuclei and cytoplasm of neutrophils, some reactive lymphocytes, plasma cell-like lymphocytes and respiratory epithelial and glandular epithelial cells. NRP-2 expression in lung tissue was observed in respiratory epithelial and glandular epithelial cells. Weak NRP-1 expression was observed in the cytoplasm of hepatocytes in liver tissue. In spleen and lymph node, NRP-1 expression was observed in some lymphocytes in the marginal and mantle zones of lymphoid follicles, plasma cell-like lymphocytes and some dendritic cell-like cells. NRP-1 and NRP-2 expression was observed on lymphatic and vascular endothelial cells in brain, lung, liver, spleen and lymph nodes.

Bottom Line: The expression of both NRPs was detected in AMs, BMs and IVMs by IHC.The frequency of NRPs+ AMs in lung tissue adjacent to the cancer margin was significantly higher than the frequencies in inflamed and normal lung tissue.Furthermore, the possible origin of AMs from blood monocytes could be suggested from a co-expression of NRPs and DC-SIGN.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathological Diagnostics, Yamagata University Faculty of Medicine, Yamagata, Japan.

ABSTRACT
In the immune system, neuropilins (NRPs), including NRP-1 and NRP-2, are expressed in thymocytes, dendritic cells, regulatory T cells and macrophages. Their functions on immune cells around the neoplastic cells vary into pro-angiogenesis, tumor progression and anti-angiogenesis according to their ligands. Even though NRPs expression on malignant tumors and immune system has studied, a PubMed-based literature query did not yield any articles describing NRPs expression on tissue-specific macrophages. The aims of this study were (i) to detect NRPs expression on tissue-specific macrophages in the brain, liver, spleen, lymph node and lung; (ii) to observe NRPs expression in classes of macrophages, including alveolar macrophages (AMs), bronchial macrophages (BMs), interstitial macrophages (IMs), intravascular macrophages (IVMs) and macrophage subsets (M1, M2 and Mox) in lung; and (iii) to detect the co-expression of NRPs and dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN) in AMs. Both NRPs were specifically detected in AMs among tissue-specific macrophages by immunohistochemistry (IHC). NRPs mRNA expression levels were characterized in normal lung by reverse transcriptase polymerase chain reaction (RT-PCR) and in situ-polymerase chain reaction (in situ-PCR). The expression of both NRPs was detected in AMs, BMs and IVMs by IHC. The frequency of NRPs+ AMs in lung tissue adjacent to the cancer margin was significantly higher than the frequencies in inflamed and normal lung tissue. Double and triple IHC demonstrated that NRPs are expressed on all macrophage subsets in lung. Double IHC showed co-expression of DC-SIGN and NRPs in AMs. This study demonstrated for the first time the specific expression of both NRPs in AMs among tissue-specific macrophages and their expression on M1, M2 and Mox macrophages. Furthermore, the possible origin of AMs from blood monocytes could be suggested from a co-expression of NRPs and DC-SIGN.

Show MeSH
Related in: MedlinePlus