Difference between revisions of "2014 Group 1 Project"

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==Introduction==
 
==Introduction==
  
Phagocytosis is a specialised type of endocytosis where large (≥0.5 μm) solid particles are internalised through the receptor-mediated engulfment of membrane-derived vesicles called phagosomes. After the vesicles detach from the plasma membrane (scission), the phagosome matures by fusing with endosomes and lysosomes (which contain hydrolytic enzymes) to form a phagolysosome. The hydrolytic enzymes in the phagolysosome break down the internalised solid particles. The mechanism behind Phagocytosis is clathrin independent and usually requires actin polymerisation<ref><pubmed>14732160</pubmed></ref>. Phagocytosis is triggered by the interaction between ligands on the particle surface and receptors on the phagocytic cell. Particles may be directly recognised directly or may be tagged by opsonins before being recognised by specific receptors. Phagocytosis plays a central role in host defence against infective agents, and in tissue remodelling and maintenance, though some microorganisms use phagocytosis as a means to invade host cells and thus avoid direct destruction by serum antibodies or cytotoxic cells. Phagocytes can be divided into two groups; Professional Phagocytes (usually referring to Polymorphonucleocytes - PMNs), and Non-professional Phagocytes.
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Phagocytosis is a specialised type of endocytosis where large (≥0.5 μm) solid particles are internalised through the receptor-mediated engulfment of membrane-derived vesicles called phagosomes. After the vesicles detach from the plasma membrane (scission), the phagosome matures by fusing with endosomes and lysosomes (which contain hydrolytic enzymes) to form a phagolysosome. The hydrolytic enzymes in the phagolysosome break down the internalised solid particles. The mechanism behind Phagocytosis is clathrin independent and usually requires actin polymerisation<ref><pubmed>14732160</pubmed></ref>. Phagocytosis is triggered by the interaction between ligands on the particle surface and receptors on the phagocytic cell. Particles may be directly recognised or may be tagged by opsonins before being recognised by specific receptors. Phagocytosis plays a central role in host defence against infective agents, and in tissue remodelling and maintenance, though some microorganisms use phagocytosis as a means to invade host cells and thus avoid direct destruction by serum antibodies or cytotoxic cells. Phagocytes can be divided into two groups; Professional Phagocytes (usually referring to Polymorphonucleocytes - PMNs), and Non-professional Phagocytes.
  
 
http://www.sciencedirect.com/science/article/pii/S0960982211006117
 
http://www.sciencedirect.com/science/article/pii/S0960982211006117

Revision as of 14:41, 1 May 2014

UNSW ANAT3231 Course Coordinator Dr Mark Hill
2014 Projects: Group 1 | Group 2 | Group 3 | Group 4

Transport into the cell from the plasma membrane (Endocytosis)

Phagocytosis of yeast by a Leukocyte

Introduction

Phagocytosis is a specialised type of endocytosis where large (≥0.5 μm) solid particles are internalised through the receptor-mediated engulfment of membrane-derived vesicles called phagosomes. After the vesicles detach from the plasma membrane (scission), the phagosome matures by fusing with endosomes and lysosomes (which contain hydrolytic enzymes) to form a phagolysosome. The hydrolytic enzymes in the phagolysosome break down the internalised solid particles. The mechanism behind Phagocytosis is clathrin independent and usually requires actin polymerisation[1]. Phagocytosis is triggered by the interaction between ligands on the particle surface and receptors on the phagocytic cell. Particles may be directly recognised or may be tagged by opsonins before being recognised by specific receptors. Phagocytosis plays a central role in host defence against infective agents, and in tissue remodelling and maintenance, though some microorganisms use phagocytosis as a means to invade host cells and thus avoid direct destruction by serum antibodies or cytotoxic cells. Phagocytes can be divided into two groups; Professional Phagocytes (usually referring to Polymorphonucleocytes - PMNs), and Non-professional Phagocytes.

http://www.sciencedirect.com/science/article/pii/S0960982211006117

http://www.ncbi.nlm.nih.gov/pubmed/18085665

http://www.annualreviews.org.wwwproxy0.library.unsw.edu.au/doi/full/10.1146/annurev.immunol.17.1.593

http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0040190

http://www.nature.com/nri/journal/v8/n2/full/nri2240.html

History

Structure of Plasma Membrane

Plasma membranes surround the cell, that is composed of lipid bilayer. The lipid bilayer consists of double-layered amphipathic phospholipids that reflect each other; individually these phospholipids have a hydrophilic polar head and 2 uncharged, hydrocarbons, hydrophobic tails. The hydrophilic head faces outwards, i.e. articulates with the extracellular environment and internal cellular department. and the hydrophobic tails face towards each other, internally. This orientation is energetically favourable. This amphipothic property allows self-sealing, however this causes the plasma membrane to have no intrinsic strength.

Membrane proteins are specialized proteins that carryout specific functions for the membrane and are either embedded in the inner, outer or both phospholipid layers. Glycoproteins are proteins with polymers of oligosaccharides (sugar short chains) that coat the surface (non-cytosolic side) of all eukaryotic cells. This is called the carbohydrate layer of the cell, which functions to protect the cell surface from damage, provide lubrication for assisting with motility and cell-cell recognition and adhesion.

Cholesterol molecules are also abundant in the membrane and closely associate with the phospholipids and enhance permeability-barrier properties of the bilayer by decreasing the deformability regions of the phospholipids and prevent crystallization of hydrocarbon chains.

Alberts B, Johnson A, Lewis J, et al. Molecular Biology of the Cell. 4th edition. New York: Garland Science; 2002. The Lipid Bilayer. [1]

Receptors

In order for the innate immune system to function properly, there must be a mechanism where Phagocytes can differentiate host cells from foreign particles. Phagocytes can identify cells using "Pattern-recognition receptors" (PRRs) located on the plasma membrane, which interact with specific conserved motifs on pathogens called “Pathogen-associated molecular patterns” (PAMPs)[2]. Examples of Pathogen-associated motifs include mannans in the cell wall of yeast, formylated peptides in bacteria, and lipopolysaccharides and lipoteichoic acids on the surface of Gram negative and Gram positive bacteria.


http://www.annualreviews.org/doi/pdf/10.1146/annurev.immunol.17.1.593


Fc Receptor-Mediated Phagocytosis

Fc receptors (FcRs) are specialised receptors for different immunoglobulin isotypes (IgA, IgE, IgM, and IgG), and are involved in regulating and executing antibody‐mediated responses. FcRs fall into two general classes; those involved in effector functions, and those that transport immunoglobulins across epithelial barriers.

http://www.sciencedirect.com/science/article/pii/S0065277607960058

Complement Receptor-Mediated Phagocytosis

Complement proteins are a group of proteins present in serum that opsonise bacteria. The C3b or C3bi receptors (CRs) on macrophages can detect the marked bacteria and phagocytose them. CR1, CR3, and CR4 also participate in phagocytosis of complement-opsonized particles. CR1 is a single-chain transmembrane protein consisting of a large extracellular lectin-like complement-binding domain and a short 43 amino acid cytosolic domain.


http://www.jbc.org/content/267/4/2656.full.pdf

Mannose Receptor-Mediated Phagocytosis

Mechanism of Phagocytosis (how material is transported through the membrane and into the cell)

Phagocytosis

The mechanism by which relatively large particles (>~0.S µm) are taken into a cell is called Phagocytosis. Phagocytosis is a type of endocytosis that is clathrin independent and usually requires actin polymerisation.

Morphological Mechanisms

Firstly, the phagocytes attach to the particles that are IgG-opsinzed1,2.

Then the engulfment of particles by phagocytosis is demonstrated by the zipper mechanism, where plasma membrane receptors bind with ligands from particle surface, like a zipper, and then the pseudopods makes it way around the particle by attaching to each ligand (fig ) forming a phagocytic cup. The zipper model can be divided into active or passive. The passive zipper model is not supported by actin polymerization instead just by ligand-receptor bonds and is therefore slower and effective on small particles, but more variable cup shape. Active zipper model involves actin polymerization is faster then passive and can take large particles as well + irreversible.

The actin polymerization fills in gaps / stops pseudopods from forming back.

{The role of actin polymerization in phagocytosis is hence to stabilize ligand receptor bonds and to rectify membrane movements in a ratchet-like fashion, leading to unidirectional movement of the leading edge of the engulfing cell.}3

A 50% increase in tether force was observed in phagocytic cells, compared to the resting cells membrane tension, due to pseudopod extension.4 Additionally, during phagocytosis the cell membrane expands, to engulf particles, by cell spreading.

Once the phagocyte extends its pseudopodia around a particle, then a constriction occurs around at the pseudopod margin to close the phagosome. The pseudopod extension orientates around the phagosome/particle to envelope The pseudopod extension is caused by actin polymerization at the distal margin of the closing phagosome, beneath the plasma membrane. In fig 1, the fluorescent actin inside macrophages, actin concentrations were highest at the distal margins of closing phagosomes (at 1.5 minutes, 3.5 minutes and 4 minutes). Bar, 10 mm.

During contrition, Myosin is also detected during phagosome closure; with myosin II andIXb is detected in ruffles and early phagocytic cups, then myosin IV at the later stage of phagosome closure and myosin V when the phagosomes are fully internalized.

The phagsome is enclosed at the circular margin by constriction into a narrower apertature and then closes by membrane fusion forming an intracellular vacuole.5


Exocytosis activation completes the phagocytosis/ engulfment. Additionally, inward bead movement (engulfment) and ingestion is most probably due to contraction and exocytosis activation. This confirms that membrane tension is an exocytosis activator and that exocytosis is required for phagocytosis to complete.4


Engulfment can be affected by the particle shape and temperature.2,3

Dependence of cup shape on cell-membrane parameters, particle size and shape.

‘Shows that uniformly opsonized particles of various shapes are only ingested if the surface that contacts the macrophage membrane is less than a minimum tangent angle. This indicates a level of signal integration in forming phagocytic cups’

Fluorescent actin inside macrophages

The phagocyte extends its pseudopodia around a particle, then a constriction occurs around at the pseudopod margin to close the phagosome. The pseudopod extension orientates around the phagosome/particle to envelope. The pseudopod extension is caused by actin polymerization at the distal margin of the closing phagosome, beneath the plasma membrane. In fig 1, the fluorescent actin inside macrophages, actin concentrations were highest at the distal margins of closing phagosomes (at 1.5 minutes, 3.5 minutes and 4 minutes). Bar, 10 mm.

During contrition, Myosin is also detected during phagosome closure; with myosin II and IXb is detected in ruffles and early phagocytic cuos, then myosin IC at the later stage of phagosome closure and myosin V when the phagosomes are fully internalized.

The phagosome/particle is enclosed at the circular margin by constriction of the pseudopods, into a narrower aperture and then closes by membrane fusion forming an intracellular vacuole. 5


References.

1. F M. GRIFFIN, JR, J.A.GRIFFIN, J.E. LEIDER, AND S.C. SILVERSTEIN (1975) ‘STUDIES ON THE MECHANISM OF PHAGOCYTOSIS I . Requirements for Circumferential Attachment ofParticle-Bound Ligands to Specific Receptors on theMacrophage Plasma Membrane’

2. Champion, J. A. & Mitragotri, S. ‘Role of target geometry in phagocytosis.’ Proc. Natl Acad. Sci. USA .103, 4930–4934 (2006).

3. S. Tollis, A.E. Dart, G.Tzircotis & R.G. Endres, (2010) ‘The zipper mechanism in phagocytosis: energetic requirements and variability in phagocytic cup shape.’ BMC Systems Biology 4:149.

<pubmed>21059234</pubmed> | Pubmed

4. ‘Plasma membrane tension orchestrates membrane trafficking, cytoskeletal remodeling, and biochemical signaling during phagocytosis' Thomas A. Masters, Bruno Pontes, Virgile Viasnoffa, You Li, and Nils . Gauthier, (2013) Proc Natl Acad Sci U S A. 2013 Jul 16;110(29):11875-80. doi: 10.1073/pnas.1301766110. Epub 2013 Jul 2.

<pubmed>23821745</pubmed>| Pubmed

5. Joel A. Swanson, Melissa T. Johnson , Karen Beningo , Penny Post , Mark Mooseker and Nobukazu Araki (1999) ‘Contractile activity in macrophage phagosomes’ Journal of Cell Science 112, 307-316

6.‘Contemporaneous cell spreading and phagocytosis: Magneto-resistive real-time monitoring of membrane competing processes’ A. Shoshi, J.Schotter, P.Schroeder, M.Milnera, P.Ertl, R.Heer, G.Reiss, H.Brueckl (2012) Biosensors andBioelectronics40(2013)82–88. doi: 10.1016/j.bios.2012.06.028. Epub 2012 Jun 23.

<pubmed>22770907</pubmed>| Pubmed


Professional

Macrophages
Neutrophils

Non-Professional

<pubmed>10358769</pubmed>

<pubmed>14732172</pubmed>

<pubmed>14732160</pubmed>

<pubmed>16380325</pubmed>

<pubmed>14732161</pubmed>

<pubmed>11861619</pubmed>

Diseases related to endocytosis

Diseases linked to the mechanism of phagocytosis tend to lead to autoimmune disorders. In these diseases, there is a tendency of phagocytic activity again the immune system, hindering the basic action of elements such as neutrophils and macrophages. In certain circumstances, some of the problems associated with the phagocytic mechanism tend to be the hypoactive kind, wherein the different activities of phagocytes in the cells are impeded. Some of these examples are found below.

Chronic Granulomatous Disease

This is an autoimmune disease detected very early in life. Patients with such disease are found to take antibacterial and antimycotic drugs at a very early age. Severe infection in patients is the primary sign of the disease, although there are patients that can go years without showing any symptoms. In Chronic Granulomatous Disease, the activity of neutrophils are impaired and this there is an increase in infection as there is no acting mechanism acting against these foreign bacteria.

[3]

Chediak Higashi Syndrome

This is rather the opposite of CGD. In this disease, there is a hyperactivity in phagocytosis. It is caused by one autosomal recessive gene which can cause albinism, a decrease in resistance to infection and large granules of leukocytes. It can present as an accelerated lymphoma-like phase, with issues such as hepatosplenomegaly. The phagocytic cells in the body tend to become autologous for leukocytes, leaving to defense mechanism again foreign invaders. In an experiment conducted by Komiyama et al., there were instances of erythrocytes being engulfed by these phagocytes as well.

[4]

Current/future research

Infertility related to phagocytosis

A recent study has shown that infertile men have low testosterone, but have a heightened level of estrogen in the body. This heightened level of E2 has been seen to cause an increase in the macrophage activation factors in the blood, found to relate to the engulfment of Leydig cells. There is found to be an increase of phosphatidylserine, a substance which has a role in the signalling of macrophages to on go engulfment of another cell. Experiments showed that enhanced E2 levels tend to cause an increase in macrophage activating factors leading to Leydig Cell hyperplasia as well as an increased macrophage activity.

[5]

Complications

References

  1. <pubmed>14732160</pubmed>
  2. <pubmed>1739426</pubmed>
  3. <pubmed>18155283</pubmed>
  4. <pubmed>3764339</pubmed>
  5. <pubmed>24762434</pubmed>