scholarly journals Differential Expression of Neurokinin-1 Receptor by Human Mucosal and Peripheral Lymphoid Cells

2000 ◽  
Vol 7 (3) ◽  
pp. 371-376 ◽  
Author(s):  
Triona Goode ◽  
Joe O'Connell ◽  
Wen-Zhe Ho ◽  
Gerald C. O'Sullivan ◽  
J. Kevin Collins ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Peripheral Blood Lymphocytes ◽  
Lymphoid Cells ◽  
Receptor Expression ◽  
Rt Pcr ◽  
Peripheral Blood Mononuclear ◽  
Neurokinin 1 Receptor ◽  
Neurokinin 1

ABSTRACT Substance P (SP) has been implicated in peripheral and mucosal neuroimmunoregulation. However, confusion remains regarding immunocyte expression of the receptor for SP, neurokinin-1 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-1R 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-1R 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.

scholarly journals Spurious detection of terminal deoxynucleotidyl transferase in phytohemagglutinin-stimulated lymphocytes

Blood ◽  
1986 ◽  
Vol 68 (1) ◽  
pp. 310-312
Author(s):  
JA Fletcher ◽  
R Bell ◽  
M Koekebakker ◽  
AA Dowers ◽  
RP McCaffrey ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Protein S ◽  
Peripheral Blood Lymphocytes ◽  
Lymphoid Cells ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Peripheral Blood Mononuclear ◽  
Biochemical Assays ◽  
Inducible Protein

Expression of terminal transferase (TdT) is believed to be restricted to primitive lymphoid cells; recently, however, indirect immunofluorescent (IF) assays have been used to demonstrate the apparent presence of TdT in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocytes and in various nonlymphoid malignancies. Using an IF assay, we found that a heteroantiserum to TdT reacted with cultured and PHA-stimulated human peripheral blood mononuclear cells, but we were unable to confirm the presence of TdT in these cells using immunoblotting and biochemical assays. We conclude that the IF results are spurious and most likely represent recognition by the heteroantiserum of inducible protein(s) other than TdT.

scholarly journals Retrovirus-mediated gene transduction into canine peripheral blood repopulating cells

Blood ◽  
1994 ◽  
Vol 83 (6) ◽  
pp. 1467-1473 ◽  
Author(s):  
HP Kiem ◽  
B Darovsky ◽  
C von Kalle ◽  
S Goehle ◽  
D Stewart ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Mhc Class Ii ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Peripheral Blood Lymphocytes ◽  
Lymphoid Cells ◽  
Peripheral Blood Mononuclear ◽  
Hematopoietic Reconstitution ◽  
Peripheral Blood Progenitor Cells

Abstract Genetically marked peripheral blood progenitor cells were used to investigate their contribution to long-term hematopoietic reconstitution after autologous marrow and peripheral blood cell transplantation. After autologous marrow harvest and cryopreservation, canine peripheral blood progenitor cells were mobilized in three dogs by treatment with recombinant canine stem cell factor for 8 days. Peripheral blood mononuclear cells were collected and enriched for major histocompatibility complex (MHC) class II antigen-positive cells by avidin-biotin immunoadsorption, thereby enriching for repopulating cells. Subsequently, the cells were cocultivated for 24 hours on irradiated vector-producing packaging cells (PA317/LN), followed by an 11-day incubation in a vector containing long-term marrow culture system. On the day of transplantation, the animals were irradiated with 9.2 Gy total body irradiation (TBI), and transduced peripheral blood cells and untransduced cryopreserved marrow cells were infused within 2 hours of TBI. All three dogs engrafted. Two dogs are long-term survivors showing intermittently G418-resistant marrow-derived colony- forming unit granulocyte-macrophage colonies at a median of 1% and 2%, respectively (range, 1% to 10%), for now up to 48 weeks after transplantation. Neo-specific sequences were detected by polymerase chain reaction in peripheral blood granulocytes for now up to 65 weeks and in peripheral blood lymphocytes for up to 75 weeks after transplantation. Peripheral blood samples of the dogs were free of helper virus and no side effects from the transduction were observed. One of the three dogs died from chronic canine distemper sclerosing encephalitis on day 84, whereas the other two dogs are alive at 15 and 17 months. Our data show successful retroviral transduction of canine peripheral blood repopulating cells. Long-term persistence of marked myeloid and lymphoid cells after transplantation suggests that peripheral blood contains repopulating cells that contribute to long- term hematopoietic reconstitution after otherwise lethal TBI.

scholarly journals Spurious detection of terminal deoxynucleotidyl transferase in phytohemagglutinin-stimulated lymphocytes

Blood ◽  
1986 ◽  
Vol 68 (1) ◽  
pp. 310-312 ◽  
Author(s):  
JA Fletcher ◽  
R Bell ◽  
M Koekebakker ◽  
AA Dowers ◽  
RP McCaffrey ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Protein S ◽  
Peripheral Blood Lymphocytes ◽  
Lymphoid Cells ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Peripheral Blood Mononuclear ◽  
Biochemical Assays ◽  
Inducible Protein

Abstract Expression of terminal transferase (TdT) is believed to be restricted to primitive lymphoid cells; recently, however, indirect immunofluorescent (IF) assays have been used to demonstrate the apparent presence of TdT in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocytes and in various nonlymphoid malignancies. Using an IF assay, we found that a heteroantiserum to TdT reacted with cultured and PHA-stimulated human peripheral blood mononuclear cells, but we were unable to confirm the presence of TdT in these cells using immunoblotting and biochemical assays. We conclude that the IF results are spurious and most likely represent recognition by the heteroantiserum of inducible protein(s) other than TdT.

scholarly journals Expression of Dopamine D1−4 and Serotonin 5-HT1A-3A Receptors in Blood Mononuclear Cells in Schizophrenia

2021 ◽  
Vol 12 ◽  
Author(s):  
Adam Wysokiński ◽  
Elżbieta Kozłowska ◽  
Ewa Szczepocka ◽  
Anna Łucka ◽  
Justyna Agier ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Rt Pcr ◽  
Peripheral Blood Mononuclear ◽  
Schizophrenia Group ◽  
Blood Mononuclear Cells ◽  
Mrna And Protein Expression

Introduction: The aim of this study was to determine the mRNA expression profile of dopamine D1, D2, D3, D4 and serotonin 5-HT1A, 5-HT2A, and 5-HT3A receptors in peripheral blood mononuclear cells (PBMCs) in schizophrenia and the in vitro effect of antipsychotics on the expression of these receptors in PBMCs of healthy subjects.Materials and Methods: Twenty-seven patients with schizophrenia and 29 healthy controls were recruited for the study. All study subjects underwent thorough clinical assessment, including anthropometric and body composition measurements. The expression of mRNA for dopamine D1−4 and serotonin 5-HT1A−3A receptors was measured using quantitative RT-PCR in peripheral blood mononuclear cells. In vitro mRNA and protein expression of these receptors was measured using quantitative RT-PCR and Western Blotting in PBMCs cultured with quetiapine, haloperidol, aripiprazole, risperidone, olanzapine or clozapine at IC50, half of IC50, and one-quarter of IC50 concentrations.Results: The key finding was that the schizophrenia group demonstrated significantly higher mRNA expression of D1, D2 and D4 receptors (p < 0.001), and significantly lower mRNA expression of 5-HT3A receptors (p < 0.01). After adjusting for smoking, the mRNA expression of D1 lost its significance, while that of D3, 5-HT1A, 5-HT2A became significant (all three were lower in the schizophrenia group). These receptors also demonstrated different ratios of mRNA expression in the schizophrenia group. The in vitro experiments showed that high concentrations of antipsychotics influenced the mRNA and protein expression of all studied receptors.Conclusion: Schizophrenia patients display a distinctive pattern of dopamine and serotonin receptor mRNA expression in blood mononuclear cells. This expression is little affected by antipsychotic treatment and it may therefore serve as a useful diagnostic biomarker for schizophrenia.

scholarly journals Retrovirus-mediated gene transduction into canine peripheral blood repopulating cells

Blood ◽  
1994 ◽  
Vol 83 (6) ◽  
pp. 1467-1473 ◽  
Author(s):  
HP Kiem ◽  
B Darovsky ◽  
C von Kalle ◽  
S Goehle ◽  
D Stewart ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Mhc Class Ii ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Peripheral Blood Lymphocytes ◽  
Lymphoid Cells ◽  
Peripheral Blood Mononuclear ◽  
Hematopoietic Reconstitution ◽  
Peripheral Blood Progenitor Cells

Genetically marked peripheral blood progenitor cells were used to investigate their contribution to long-term hematopoietic reconstitution after autologous marrow and peripheral blood cell transplantation. After autologous marrow harvest and cryopreservation, canine peripheral blood progenitor cells were mobilized in three dogs by treatment with recombinant canine stem cell factor for 8 days. Peripheral blood mononuclear cells were collected and enriched for major histocompatibility complex (MHC) class II antigen-positive cells by avidin-biotin immunoadsorption, thereby enriching for repopulating cells. Subsequently, the cells were cocultivated for 24 hours on irradiated vector-producing packaging cells (PA317/LN), followed by an 11-day incubation in a vector containing long-term marrow culture system. On the day of transplantation, the animals were irradiated with 9.2 Gy total body irradiation (TBI), and transduced peripheral blood cells and untransduced cryopreserved marrow cells were infused within 2 hours of TBI. All three dogs engrafted. Two dogs are long-term survivors showing intermittently G418-resistant marrow-derived colony- forming unit granulocyte-macrophage colonies at a median of 1% and 2%, respectively (range, 1% to 10%), for now up to 48 weeks after transplantation. Neo-specific sequences were detected by polymerase chain reaction in peripheral blood granulocytes for now up to 65 weeks and in peripheral blood lymphocytes for up to 75 weeks after transplantation. Peripheral blood samples of the dogs were free of helper virus and no side effects from the transduction were observed. One of the three dogs died from chronic canine distemper sclerosing encephalitis on day 84, whereas the other two dogs are alive at 15 and 17 months. Our data show successful retroviral transduction of canine peripheral blood repopulating cells. Long-term persistence of marked myeloid and lymphoid cells after transplantation suggests that peripheral blood contains repopulating cells that contribute to long- term hematopoietic reconstitution after otherwise lethal TBI.

Toll like receptor 9 (TLR 9) expression in peripheral blood mononuclear cell (PBMCs) from treated and untreated Grave’s disease patients

QJM ◽  
2021 ◽  
Vol 114 (Supplement_1) ◽  
Author(s):  
Ashraf M Okba ◽  
Rasha Y Shaheen ◽  
Gehan M. H Mostafa ◽  
Hanan M Ali ◽  
Sylvia W Abo El Fadle ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cell ◽  
Mononuclear Cell ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Peripheral Blood Lymphocytes ◽  
Graves Disease ◽  
Toll Like Receptor ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Abstract Background It is well known that Autoimmune thyroid disease is multifactorial with multiple genetic and environmental factors, immune malfunction also incriminated in the development of this disease, The exact pathogenesis of this disease remains unclear despite the fact that the production of autoantibodies destroys self-tolerance and agitate the adaptive immune system. Our study will answer the question is there a difference in Toll like receptor 9 (TLR 9) expression in peripheral blood mononuclear cell (PBMCs) from Grave’s disease patients. Objective to measure TLR9 percentage expression on peripheral blood mononuclear cells in patients with Graves’ disease. Methods 60 subjects were included in this study; 30 with Graves’ disease and 30 healthy individuals as control group. All the patients were subjected to the following: Full history, clinical examination, thyroid functions, Thyroid ultrasound, Radioisotope thyroid scan: to assess uptake of thyroid gland and Toll like receptor 9 (TLR 9) percentage expression on peripheral blood mononuclear cells will be analyzed using flow cytometry technique. Results The present study proved that patients with Graves’ disease had higher levels of percentage expression of TLR 9 on peripheral blood lymphocytes. Conclusion percentage expression of TLR9 on peripheral blood lymphocytes is higher in Graves’ patients.

Sign in / Sign up

Export Citation Format

Share Document