The effect of monocytes in the peripheral blood CFU-C assay system

Abstract The effect of cellular interactions in the in vitro assay of myeloid progenitor cells in peripheral blood (PB CFU-C) was investigated. Ficoll-Paque-separated peripheral blood mononuclear cells (PB MNC) from 7 healthy subjects were cultured at cell concentrations from 10 to 0.625 X 10(5) MNC/plate in doubling dilutions. The number of colonies per 10(6) lymphocytes plated (corrected colony count, CC) was significantly higher when 2.5 X 10(5) or less PB MNC were cultured than when 5 or 10 X 10(5) cells were cultured. This decrease in CC when large numbers of cells were cultured was not present when the nonadherent cells only were cultured. The inhibition was reproduced when adherent cells were added back to the nonadherent cells. The inhibition appeared to be proportional to the number of monocytes present. A model depicting the role of monocytes in the PB CFU-C assay system is presented. The increased understanding of cellular interaction represents an important step towards the standardization of the PB CFU-C assay.

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The effect of monocytes in the peripheral blood CFU-C assay system

Blood ◽

10.1182/blood.v62.1.112.bloodjournal621112 ◽

1983 ◽

Vol 62 (1) ◽

pp. 112-117 ◽

Cited By ~ 1

Author(s):

LB To ◽

DN Haylock ◽

CA Juttner ◽

RJ Kimber

Keyword(s):

Peripheral Blood ◽

Mononuclear Cells ◽

Assay System ◽

Cellular Interaction ◽

Cellular Interactions ◽

Peripheral Blood Mononuclear ◽

Vitro Assay ◽

Myeloid Progenitor Cells ◽

Nonadherent Cells

The effect of cellular interactions in the in vitro assay of myeloid progenitor cells in peripheral blood (PB CFU-C) was investigated. Ficoll-Paque-separated peripheral blood mononuclear cells (PB MNC) from 7 healthy subjects were cultured at cell concentrations from 10 to 0.625 X 10(5) MNC/plate in doubling dilutions. The number of colonies per 10(6) lymphocytes plated (corrected colony count, CC) was significantly higher when 2.5 X 10(5) or less PB MNC were cultured than when 5 or 10 X 10(5) cells were cultured. This decrease in CC when large numbers of cells were cultured was not present when the nonadherent cells only were cultured. The inhibition was reproduced when adherent cells were added back to the nonadherent cells. The inhibition appeared to be proportional to the number of monocytes present. A model depicting the role of monocytes in the PB CFU-C assay system is presented. The increased understanding of cellular interaction represents an important step towards the standardization of the PB CFU-C assay.

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In vitro generation of antigen-specific hemolytic plaque-forming cells from human peripheral blood mononuclear cells.

Journal of Experimental Medicine ◽

10.1084/jem.154.4.1069 ◽

1981 ◽

Vol 154 (4) ◽

pp. 1069-1084 ◽

Cited By ~ 25

Author(s):

J Misiti ◽

T A Waldmann

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Specific Response ◽

Cellular Interactions ◽

Peripheral Blood Mononuclear ◽

Sheep Erythrocytes ◽

Blood Mononuclear Cells

We have described a culture and assay system for the sensitization of human peripheral blood mononuclear cells with a T cell-dependent antigen, sheep erythrocytes, in the absence of nonspecific stimulatory agents and with the subsequent generation of macroscopic hemolytic plaques. We have shown that the antibody produced by the plaque-forming cells generated in this culture system is specific for the sensitizing antigen, and that the plaques created are not false plaques because their formation is inhibited by cycloheximide. The success of this system can be attributed to several critical factors including large numbers of peripheral blood mononuclear cells (5 x 10(6) culture), a prolonged period of incubation (10-11 d), continuous rocking during the entire period of incubation, culturing in large (35-mm) flat-bottomed culture dishes in the presence of human plasma, and the appropriate antigen concentration (5 x 10(6) sheep erythrocytes/culture). Furthermore, the generation of macroscopic hemolytic plaques requires plaquing sensitized peripheral blood mononuclear cells in target cell monolayers fixed in an agarose matrix with an incubation period of 2-3 h. We have further shown that the antigen-specific response measured by this system is dependent on adherent cells and T lymphocytes. At least one population of the helper T cells is sensitive to 2,000 rad irradiation. This system is simple, sensitive, and should serve as an effective tool for the analysis of cellular interactions involved in the generation of human antigen-specific plaque-forming cells, the genetic control the human immune response, and the pathophysiology of altered immunoregulation in disease.

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The effects of Malassezia on pro-inflammatory cytokine production by human peripheral blood mononuclear cells in vitro

Medical Mycology ◽

10.1046/j.1365-280x.1998.00137.x ◽

1998 ◽

Vol 36 (2) ◽

pp. 97-106 ◽

Cited By ~ 6

Author(s):

S. KESAVAN ◽

C. E. WALTERS ◽

K. T. HOLLAND ◽

E. INGHAM

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Inflammatory Cytokine ◽

Mononuclear Cells ◽

Cytokine Production ◽

Human Peripheral Blood ◽

Peripheral Blood Mononuclear ◽

Inflammatory Cytokine Production ◽

Blood Mononuclear Cells

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CD8+ T cells from feline immunodeficiency virus (FIV) infected cats suppress exogenous FIV replication of their peripheral blood mononuclear cells in vitro

Archives of Virology ◽

10.1007/s00705-002-0827-1 ◽

2002 ◽

Vol 147 (8) ◽

pp. 1517-1529 ◽

Cited By ~ 14

Author(s):

T. Hohdatsu ◽

T. Sasagawa ◽

A. Yamazaki ◽

K. Motokawa ◽

H. Kusuhara ◽

...

Keyword(s):

T Cells ◽

Peripheral Blood Mononuclear Cells ◽

Cd8 T Cells ◽

Feline Immunodeficiency Virus ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Immunodeficiency Virus ◽

Blood Mononuclear Cells

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Potential Role and Impact of Peripheral Blood Mononuclear Cells in Radiographic Axial Spondyloarthritis-Associated Endothelial Dysfunction

Diagnostics ◽

10.3390/diagnostics11061037 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1037

Author(s):

Patricia Ruiz-Limon ◽

Maria L. Ladehesa-Pineda ◽

Clementina Lopez-Medina ◽

Chary Lopez-Pedrera ◽

Maria C. Abalos-Aguilera ◽

...

Keyword(s):

Endothelial Dysfunction ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Axial Spondyloarthritis ◽

Endothelial Injury ◽

In Vitro Studies ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

Endothelial dysfunction (ED) is well known as a process that can lead to atherosclerosis and is frequently presented in radiographic axial spondyloarthritis (r-axSpA) patients. Here, we investigated cellular and molecular mechanisms underlying r-axSpA-related ED, and analyzed the potential effect of peripheral blood mononuclear cells (PBMCs) in promoting endothelial injury in r-axSpA. A total of 30 r-axSpA patients and 32 healthy donors (HDs) were evaluated. The endothelial function, inflammatory and atherogenic profile, and oxidative stress were quantified. In vitro studies were designed to evaluate the effect of PBMCs from r-axSpA patients on aberrant endothelial activation. Compared to HDs, our study found that, associated with ED and the plasma proatherogenic profile present in r-axSpA, PBMCs from these patients displayed a pro-oxidative, proinflammatory, and proatherogenic phenotype, with most molecular changes noticed in lymphocytes. Correlation studies revealed the relationship between this phenotype and the microvascular function. Additional in vitro studies confirmed that PBMCs from r-axSpA patients promoted endothelial injury. Altogether, this study suggests the relevance of r-axSpA itself as a strong and independent cardiovascular risk factor, contributing to a dysfunctional endothelium and atherogenic status by aberrant activation of PBMCs. Lymphocytes could be the main contributors in the development of ED and subsequent atherosclerosis in this pathology.

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