Regulatory T cells (Tregs) are a type of T cell that suppresses immune responses by secreting negative regulatory (anti-inflammatory) cytokines. Let’s find out about the Regulatory T-cells separation technologies.
T cells that regulate or suppress other immune system cells are known as regulatory T cells (also known as Tregs). Tregs help to prevent autoimmune disease by regulating the immune response to self and foreign particles (antigens). Tregs are produced naturally by a healthy thymus. ‘Adaptive’ Treg are created by differentiating naive T cells outside of the thymus, i.e. the periphery, or in cell culture.
CD4+ T cells are commonly classified as regulatory T (Treg) cells or T helper (Th) cells. The cells regulate adaptive immunity against pathogens and cancer by activating other effector immune cells. Treg cells are CD4+ T cells that are in charge of suppressing potentially harmful Th cell activities.
We’ll look at how Pluribead cascade straining aids in Regulatory T Cells isolation.
Identification of Treg cells remains difficult because evidence suggests that all of the currently used Treg markers (CD25, CTLA-4, GITR, LAG-3, CD127, and Foxp3) are general T-cell activation markers rather than Treg-specific.
Treg-cell activation is antigen-specific, implying that Treg cells’ suppressive activities are antigen-dependent. Self-reactivity of Treg cells has been proposed, but extensive TCR repertoire analysis suggests that self-reactivity may be the exception rather than the rule. Because the ability to suppress is not unique to Treg cells, their classification as a distinct lineage remains debatable. Suppressive activities attributed to Treg cells may be exerted by conventional Th cell subsets such as Th1, Th2, Th17, and T follicular cells, at least in some experimental settings.
Natural Tregs are distinguished by the expression of both the CD4 T cell co-receptor and CD25, a component of the IL-2 receptor. Tregs are therefore CD4+CD25+. The expression of the nuclear transcription factor Forkhead box P3 (FoxP3) determines natural Treg development and function.
FoxP3 is required to keep the immune system suppressed. Self-reactive lymphocytes caused by naturally occurring mutations in the FOXP3 gene cause the rare but severe disease IPEX (Immune Dysregulation, Polyendocrinopathy, Enteropathy, X-Linked) in humans and scurfy in mice.
Tregs are thought to suppress CD4+ and CD8+ T cell activation, proliferation, and cytokine production, as well as B and dendritic cells. Tregs can produce soluble messengers with suppressive properties, such as TGF-beta, IL-10, and adenosine. CD152 (CTLA-4) and GITR (glucocorticoid-induced TNF receptor) are additional markers of natural Tregs, but it should be noted that these are also expressed by other T-cell types on a regular basis (e.g. activated T cells), so they are not conclusively diagnostic. However, the function of these markers on other T cells is unclear. T cells lacking specialized regulatory capacity may compete for resources such as growth factors and MHC class II stimulation, and thus play a regulatory role through this general mechanism.
What Are The Functions Of Treg Cells?
Treg cells’ primary function was originally defined as the prevention of autoimmune diseases through self-tolerance. Several additional functions have been proposed over the years, and it will be critical to determine what Treg cells actually do in the immune system.
- Prevention of autoimmune diseases through the development and maintenance of immunologic self-tolerance.
- Allergic and asthmatic symptoms are reduced.
- Oral tolerance is the induction of tolerance against dietary antigens.
- Induction of maternal-fetal tolerance.
- Pathogen-induced immunopathology is suppressed.
- Regulation of the immune response’s effector class.
- T-cell activation is suppressed in response to weak stimuli.
- Effector T cells regulate the magnitude of the immune response.
- Protection of commensal bacteria from immune system elimination.
- T cells that have been activated by a true high-affinity agonist ligand are prevented from killing cells that only express low-affinity T-cell receptor (TCR) ligands, Like the self peptide-major histocompatibility complex (MHC) molecule, which positively selected the T cell.
How To Identify Treg Cells?
Molecular markers are critical tools for defining and analyzing immune cell subpopulations. The failure to identify specific markers for suppressor T cells contributed significantly to the decline of suppressor T cells at the end of the 1980s.
The most commonly used Treg cell markers are:
- CD25.
- cytotoxic T lymphocyte-associated antigen 4 (CTLA-4).
- TNF receptor family-related gene induced by glucocorticoids (GITR).
- lymphocyte activation gene-3 (LAG-3).
- P3 forkhead/winged-helix transcription factor box CD127 (Foxp3).
Upon activation, all T cells express CD25, the -chain of the interleukin-2 (IL-2) receptor, IL-2 is a T-cell growth factor important for T-cell clonal expansion. CTLA-4 is a negative regulator of T-cell activation that is upregulated on all CD4+ and CD8+ T cells 2-3 days after activation. Similarly, T-cell activation induces the expression of GITR and LAG-3.
It has been proposed that CD127, the IL-7 receptor chain, could be used to distinguish CD127low Treg cells from CD127high conventional Th cells in humans. However, it has recently been discovered that most CD4+ T cells downregulate CD127 when activated. Furthermore, loss of CD127 is a characteristic feature of T follicular helper (Tfh) cells in human tonsils, which help B cells.
It has been reported that when activated, naive, CD25-negative mouse CD4+ T cells do not upregulate Foxp3. However, it is now well established that most human CD4+ and CD8+ T cells express Foxp3 transiently upon activation.
Finally, all of the Treg markers currently in use (CD25, CTLA-4, GITR, CD127, LAG-3, and Foxp3) appear to be general T-cell activation markers. This finding strongly implies that T-cell activation is required for T-cell suppression. However, it also implies that current Treg markers are not truly Treg-specific and, as a result, are ineffective at distinguishing Treg cells from activated conventional Th cells.
Pluribeads For Regulatory T Cells Separation
Pluribead Antibody Cell Separation helps in the gentle and safe isolation of Dendritic Cells and operates without the use of any magnetic components.
The method is simple: your pluriBeads (which contain bound target cells) are sieved through a strainer, with your target cells remaining on top and unwanted cells passing through. You are now ready to proceed with your target cells after detaching.
Key features of Pluribead
- Using a sample volume ranging from 200 l to 45 ml, no erythrolysis, gradient centrifugation, or other techniques should be used.
- Any type of sample material, such as PBMC, secretion/excretion material, liver, spleen, buffy coat, brain homogenate, whole blood, and so on, can be used.
- Isolate from a Variety of Species: Isolate from sheep, mice, rats, cows, dogs, and other animals.
- PluriBead Cascade Straining for Simultaneous Cell Isolation: Separate two different cell types from the same sample material at the same time.
- You can isolate up to six different targets from a single sample using sequential cell isolation.
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Reference:
Pubmed
Nature