Peripheral blood mononuclear cell
A peripheral blood mononuclear cell (PBMC) is any peripheral blood cell having a round nucleus.[1] These cells consist of lymphocytes (T cells, B cells, NK cells) and monocytes, whereas erythrocytes and platelets have no nuclei, and granulocytes (neutrophils, basophils, and eosinophils) have multi-lobed nuclei. In humans, lymphocytes make up the majority of the PBMC population, followed by monocytes, and only a small percentage of dendritic cells.[2]
These cells can be extracted from whole blood or buffy coat samples[3] using a hydrophilic colloid and density gradient centrifugation,[4][5][6] which separates the blood into a top layer of plasma, followed by a layer of PBMCs and a bottom fraction of polymorphonuclear cells (such as neutrophils and eosinophils) and erythrocytes.[7] The polymorphonuclear cells can be further isolated by lysing the erythrocytes. Basophils are sometimes found in both the denser and the PBMC fractions.[5]
A high-quality PBMC isolate has a viability over 90% and a low concentration of erythrocytes and granulocytes when processed fresh.[8] Cell numbers and viability can be determined by manual or automated cell count or by using a flow cytometer.[9] The latter can also be used to determine the percentage of PBMCs compared with other cell types (which should be >85%).[9] If you do not have the capacity to isolate PBMCs, PBMC isolation can be outsourced to a central laboratory service provider.[10]
Cell Type | % | Number of cells isolated from 1mL of blood (x10^3) |
T Cells | 60 | 300-1200 |
T Helper Cells | 70 of T cells | 210-840 |
Cytotoxic T Cells | 30 of T cells | 90-360 |
Monocytes/Macrophages | 15 | 75-300 |
B Cells | 10 | 50-200 |
Natural Killer Cells | 15 | 75-300 |
Table 1: Cell types contained within the PBMC compartment and their typical distribution in healthy adults.[4]
Clinical significance
Infections
Recent studies indicate that PBMCs may be susceptible to pathogenic infections,[11] such as Ureaplasma parvum and urealiticum, Mycoplasma genitalium and hominis and Chlamydia trachomatis infections. PBMCs may be also susceptible to viral infections.[12][13] Indeed, footprints of JC polyomavirus and Merkel cell polyomavirus have been detected in PBMCs from pregnant women and women affected by spontaneous abortion.[12][13]
Research uses
Many scientists conducting research in the fields of immunology (including auto-immune disorders), infectious disease, hematological malignancies, vaccine development, transplant immunology, and high-throughput screening are frequent users of PBMCs. In many cases, PBMCs are derived from blood banks. PBMC fraction also contains progenitor populations, as demonstrated by methylcellulose based colony forming assays.
PBMCs have been thought to be an important route of vaccination. PBMCs from cancer patients can be extracted and cultured in vitro. Subsequently, PBMCs are challenged with tumor antigens such as tumor stem cell antigen. Inflammatory cytokines are usually added to aid in antigen uptake and recognition by PBMCs.
See also
References
- ^ Delves, Peter, et al. Roitt's Essential Immunology, 11th Ed. ISBN 978-1-4051-3603-7
- ^ The impact of food bioactives on gut health : in vitro and ex vivo models. Verhoeckx, Kitty, 1970-, Cotter, Paul (Paul D.),, European Cooperation in the Field of Scientific and Technical Research (Organization). Cham. 2015-05-19. ISBN 9783319157917. OCLC 908392100.
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: CS1 maint: location missing publisher (link) CS1 maint: others (link) - ^ Puckrin, Zara. "Protocol for PBMC isolation from buffy coat samples". www.reprocell.com. Retrieved 2022-08-09.
- ^ a b Bittersohl, Heike; Steimer, Werner (2016-01-01), Oellerich, Michael; Dasgupta, Amitava (eds.), "Chapter 9 - Intracellular concentrations of immunosuppressants", Personalized Immunosuppression in Transplantation, San Diego: Elsevier, pp. 199–226, ISBN 978-0-12-800885-0, retrieved 2022-08-09
- ^ a b Miyahira, Andrea (22 Nov 2012). "Types of immune cells present in human PBMC". sanguinebio.com. Archived from the original on 22 July 2016. Retrieved 23 Sep 2014.
- ^ Cottler-Fox, Michele; Montgomery, Matthew; Theus, John (2009-01-01), Treleaven, Jennifer; Barrett, A John (eds.), "CHAPTER 24 - Collection and processing of marrow and blood hematopoietic stem cells", Hematopoietic Stem Cell Transplantation in Clinical Practice, Edinburgh: Churchill Livingstone, pp. 249–256, ISBN 978-0-443-10147-2, retrieved 2022-08-12
- ^ "Isolating PBMCs from whole blood using density gradient centrifugation". www.reprocell.com. Retrieved 2022-10-24.
- ^ "Methods in Enzymology | Redox Cell Biology and Genetics Part B | ScienceDirect.com by Elsevier". www.sciencedirect.com. Retrieved 2022-08-12.
- ^ a b "Methods in Enzymology | Gene Transfer Vectors for Clinical Application | ScienceDirect.com by Elsevier". www.sciencedirect.com. Retrieved 2022-08-12.
- ^ Puckrin, Zara. "What is PBMC? The peripheral blood mononuclear cell compartment". www.reprocell.com. Retrieved 2022-08-12.
- ^ Contini C, Rotondo JC, Magagnoli F, Maritati M, Seraceni S, Graziano A (2018). "Investigation on silent bacterial infections in specimens from pregnant women affected by spontaneous miscarriage". J Cell Physiol. 34 (3): 433–440. doi:10.1002/jcp.26952. PMID 30078192.
- ^ a b Tagliapietra A, Rotondo JC, Bononi I, Mazzoni E, Magagnoli F, Maritati M (2019). "Footprints of BK and JC polyomaviruses in specimens from females affected by spontaneous abortion". Hum Reprod. 34 (3): 433–440. doi:10.1002/jcp.27490. PMID 30590693.
- ^ a b Tagliapietra A, Rotondo JC, Bononi I, Mazzoni E, Magagnoli F, Maritati M (2019). "Droplet-digital PCR assay to detect Merkel cell polyomavirus sequences in chorionic villi from spontaneous abortion affected females". J Cell Physiol. 235 (3): 1888–1894. doi:10.1002/jcp.29213. PMID 31549405.