TY  - JOUR
  - Goode, T.,O'Connell, J.,Ho, W. Z.,O'Sullivan, G. C.,Collins, J. K.,Douglas, S. D.,Shanahan, F.
  - 2000
  - May
  - Clinical And Diagnostic Laboratory Immunology
  - Differential expression of neurokinin-1 receptor by human mucosal and peripheral lymphoid cells
  - Validated
  - ()
  - 7
  - 3
  - 371
  - 376
  - Substance P (SP) has been implicated in peripheral and mucosal neuroimmunoregulation. However, confusion remains regarding immunocyte expression of the receptor for SP, neurokinin-l receptor (NK-1R), and whether there is differential NK-1R expression in the mucosal versus the peripheral immune system. In the same assay systems, we examined the expression of NK-1R in human lamina propria mononuclear cells (LPMC), peripheral blood mononuclear cells (PBMC), peripheral blood lymphocytes (PBL), monocytes, and monocyte-derived macrophages (MDM), Using standard reverse transcription (RT)-PCR, mRNA expression of both the long and the short isoforms of the NK-IR was evident in LPMC but not in PBMC, PBL, monocytes, or MDM, However, by using nested RT-PCR NK-1R mRNA expression was detected in PBMC, PBL, monocytes, and MDM, This level of expression was found to represent one NK-IR mRNA transcript in >1,000 cells. In contrast, by using competitive RT-PCR we demonstrate that LPMC express a more biologically significant level of eight NK-1R mRNA transcripts per cell. Flow cytometric detection of NK-1R expression at the protein level was evident in LPMC but not in PBMC, These findings illustrate the extreme sensitivity of nested RT-PCR and the advantages of competitive RT-PCR in comparative studies of receptor expression in different cell populations. This study suggests that, under normal conditions, readily detectable expression of NK-1R in human mononuclear cells occurs at the mucosal level rather than in the peripheral circulation.Substance P (SP) has been implicated in peripheral and mucosal neuroimmunoregulation. However, confusion remains regarding immunocyte expression of the receptor for SP, neurokinin-l receptor (NK-1R), and whether there is differential NK-1R expression in the mucosal versus the peripheral immune system. In the same assay systems, we examined the expression of NK-1R in human lamina propria mononuclear cells (LPMC), peripheral blood mononuclear cells (PBMC), peripheral blood lymphocytes (PBL), monocytes, and monocyte-derived macrophages (MDM), Using standard reverse transcription (RT)-PCR, mRNA expression of both the long and the short isoforms of the NK-IR was evident in LPMC but not in PBMC, PBL, monocytes, or MDM, However, by using nested RT-PCR NK-1R mRNA expression was detected in PBMC, PBL, monocytes, and MDM, This level of expression was found to represent one NK-IR mRNA transcript in >1,000 cells. In contrast, by using competitive RT-PCR we demonstrate that LPMC express a more biologically significant level of eight NK-1R mRNA transcripts per cell. Flow cytometric detection of NK-1R expression at the protein level was evident in LPMC but not in PBMC, These findings illustrate the extreme sensitivity of nested RT-PCR and the advantages of competitive RT-PCR in comparative studies of receptor expression in different cell populations. This study suggests that, under normal conditions, readily detectable expression of NK-1R in human mononuclear cells occurs at the mucosal level rather than in the peripheral circulation.
  - 1071-412X1071-412X
  - ://WOS:000086873100008://WOS:000086873100008
DA  - 2000/05
ER  - 

@article{V235379704,
   = {Goode,  T. and O'Connell,  J. and Ho,  W. Z. and O'Sullivan,  G. C. and Collins,  J. K. and Douglas,  S. D. and Shanahan,  F. },
   = {2000},
   = {May},
   = {Clinical And Diagnostic Laboratory Immunology},
   = {Differential expression of neurokinin-1 receptor by human mucosal and peripheral lymphoid cells},
   = {Validated},
   = {()},
   = {7},
   = {3},
  pages = {371--376},
   = {{Substance P (SP) has been implicated in peripheral and mucosal neuroimmunoregulation. However, confusion remains regarding immunocyte expression of the receptor for SP, neurokinin-l receptor (NK-1R), and whether there is differential NK-1R expression in the mucosal versus the peripheral immune system. In the same assay systems, we examined the expression of NK-1R in human lamina propria mononuclear cells (LPMC), peripheral blood mononuclear cells (PBMC), peripheral blood lymphocytes (PBL), monocytes, and monocyte-derived macrophages (MDM), Using standard reverse transcription (RT)-PCR, mRNA expression of both the long and the short isoforms of the NK-IR was evident in LPMC but not in PBMC, PBL, monocytes, or MDM, However, by using nested RT-PCR NK-1R mRNA expression was detected in PBMC, PBL, monocytes, and MDM, This level of expression was found to represent one NK-IR mRNA transcript in >1,000 cells. In contrast, by using competitive RT-PCR we demonstrate that LPMC express a more biologically significant level of eight NK-1R mRNA transcripts per cell. Flow cytometric detection of NK-1R expression at the protein level was evident in LPMC but not in PBMC, These findings illustrate the extreme sensitivity of nested RT-PCR and the advantages of competitive RT-PCR in comparative studies of receptor expression in different cell populations. This study suggests that, under normal conditions, readily detectable expression of NK-1R in human mononuclear cells occurs at the mucosal level rather than in the peripheral circulation.Substance P (SP) has been implicated in peripheral and mucosal neuroimmunoregulation. However, confusion remains regarding immunocyte expression of the receptor for SP, neurokinin-l receptor (NK-1R), and whether there is differential NK-1R expression in the mucosal versus the peripheral immune system. In the same assay systems, we examined the expression of NK-1R in human lamina propria mononuclear cells (LPMC), peripheral blood mononuclear cells (PBMC), peripheral blood lymphocytes (PBL), monocytes, and monocyte-derived macrophages (MDM), Using standard reverse transcription (RT)-PCR, mRNA expression of both the long and the short isoforms of the NK-IR was evident in LPMC but not in PBMC, PBL, monocytes, or MDM, However, by using nested RT-PCR NK-1R mRNA expression was detected in PBMC, PBL, monocytes, and MDM, This level of expression was found to represent one NK-IR mRNA transcript in >1,000 cells. In contrast, by using competitive RT-PCR we demonstrate that LPMC express a more biologically significant level of eight NK-1R mRNA transcripts per cell. Flow cytometric detection of NK-1R expression at the protein level was evident in LPMC but not in PBMC, These findings illustrate the extreme sensitivity of nested RT-PCR and the advantages of competitive RT-PCR in comparative studies of receptor expression in different cell populations. This study suggests that, under normal conditions, readily detectable expression of NK-1R in human mononuclear cells occurs at the mucosal level rather than in the peripheral circulation.}},
  issn = {1071-412X1071-412X},
   = {://WOS:000086873100008://WOS:000086873100008},
  source = {IRIS}
}