COMPLEMENT SYSTEM

Waqar Sarwar and Amna Shabbir ( College of Pharmacy, Govt .College University, Faisalabad) ( College of Pharmacy, Govt .College University, Faisalabad)

Abstract: Its a system that helps antibodies and phagocytic cells to clear pathogens. It consists of over 25 proteins
heat labial proteins, glycoprotein and proteins fragments. It is considered as a bridge between innate immunity and acquired immunity. The proteins and glycoprotein that compose the complement system are synthesized mainly by liver hepatocytes, although significant amounts are also produced by blood monocytes, tissue macrophages, and epithelial cells of the gastrointestinal tract. These components constitute 5% (by weight) of the serum globulin fraction. Most circulate in the serum in functionally inactive forms as proenzymes, or zymogens, which are inactive until proteolytic cleavage, which removes an inhibitory fragment and exposes the active site.

KeywordsComplement system, Serum proteins, Pro-proteins, Immune complex, Opsonization.

1. Introduction Its a system that helps antibodies and phagocytic cells to clear pathogens. It consists of over 25 proteins heat labial proteins, glycoprotein and proteins fragments. It is considered as a bridge between innate immunity and acquired immunity.The complement proteins circulate in blood as inactive precursor and are known are pro-proteins or proenzymes. Like serum proteins, serosal proteins and cell membrane receptors. The complement proteins are synthesized by liver hepatocytes, by macrophages, blood monocytes and epithelial cells of the genitourinal tract and gastrointestinal tract. The action of complement is nonspecific. Complement proteins[1] cannot bind to free antibodies or antigens but only to the antibody which has combined with its antigen. C binding site is on F c portion of IgM & IgD molecule only. These site are exposed only when abs are combined with antigens. Complement system acts as a cascade. Many of the components are enzymes (proteins) become activated when cleaved into two peptides with the action of proteases enzyme[2]. One peptide binds to the immune complex becomes a functional part of it while the other peptide diffuses away and can become an inflammatory mediator.

2. FUNCTIONS
Following are four main functions of complement function: 2.1 Inflammation: The 2nd peptide portion after cleavage binds to vascular endothelial cells and lymphocytes. These cells then produce cytokines which stimulate inflammation and initiate response to antigen. 2.2 Lysis: Lysis of bacteria and viruses is carried out by formation of polymers on the bacterial cell membrane and envelop of viruses. This leads to the formation of pores thus lipid bilayer of the cell or virus is disrupted. 2.3 Opsonization: The antigens presented by complement proteins bind to the receptors on phagocytic cells thus promote phagocytosis. This is called opsonization[3]. 2.4 Immune clearance:

Sometimes pathogens when enter in body they dont cause disease, antibodies surround these antigens and form antigen-antibody complex. This antigen-antibody complex sometimes cause problem in body like if it accumulates in kidney then glomerulonephritis. The complement proteins help removing immune complexes from the circulation and deposit them in the spleen and liver.

3. The Complement Components

The proteins and glycoprotein that compose the complement system are synthesized mainly by liver hepatocytes, although significant amounts are also produced by blood monocytes, tissue macrophages, and epithelial cells of the gastrointestinal tract. These components constitute 5% (by weight) of the serum globulin fraction. Most circulate in the serum in functionally inactive forms as proenzymes[4], or zymogens, which are inactive until proteolytic cleavage, which removes an inhibitory fragment and exposes the active site. Complement components are designated by: i. Numerals (C1C9) ii. By letter symbols (e.g., factor D) iii. Or by trivial names (e.g., homologous restriction factor). iv. Peptide fragments formed by activation of a component are denoted by small letters. In most cases, the smaller fragment resulting from cleavage of a component is designated a and the larger fragment designated b (e.g., C3a, C3b), The larger fragments bind to the target near the site of activation, and the smaller fragments diffuse from the site and can initiate localized inflammatory responses by binding to specific receptors.The complement fragments interact with one another to form functional complexes. Those complexes that have enzymatic activity are designated by a bar over the number or symbol e.g., C4b2a, C3bBb

4. Classical Pathway
i. ii. iii. iv. Activated by antigen antibody complex. Opsonization Cell lysis through MAC[5]. Cause inflammation by activating cells.

5. Alternative Pathway
Alternative pathway is faster than classical pathway because it does not require antibody for its initiation rather certain pathogens have ability to directly activate this pathway such as cell wall constituents of gram +ve and gram ve bacteria, yeast cell wall, viral envelops etc. This pathway is started by the hydrolysis of C3 serum protein. Its hydrolysis occurs because it contains the weak thioester bond. As a result of hydrolysis C3 is converted to C3b.this C3b has ability to get attached to microbial membranes. So it attaches to the pathogenic membrane. It also has ability to get attach with factor B. As a result of this attachment a site is exposed on factorB which acts as substrate for factorD. So factor D gets attach to it through this site and does cleavage of factor B into Ba and Bb. Bb gets attach to C3 and Ba is removed. So a complex is formed called C3bBb.this complex has C3 convertase activity which has half life of only 5 minutes unless properdin gets attach to it and stabilizes it and increases its half life from 5 minutes to 30 minutes. So this complex activates more unhydrolysed C3 molecules and as a result the initial steps are repeated and amplified so much so that 2106 molecules get deposited on the membrane of pathogen in less than 5 minutes, and as a result C3bBb3b complex[6] is formed. This complex has C5 convertase activity. Its enzymatic portion binds the C5 serum protein and non-enzymatic portion does cleavage of C5 into C5a and C5b.this C5b then again gets attach to the microbial surface and does lysis of pathogenic cell through membrane attack complex.

6. Mannan- binding Lectin Pathway

In recent years a new pathway has been discovered other than classical and alternative pathway. 6.1 Mode of action: When macrophages ingest bacteria, viruses and other foreign matter by phagocytosis, they release chemicals that stimulate the liver to produce lectins, proteins that bind to carbohydrates. i. One such lectin, mannose binding lectin (MBL) [7] binds to carbohydrate mannose.MBL binds to many pathogens because the MBL molecule recognize a distinctive pattern of carbohydrate that include mannose, which is found in bacterial cell wall and on some viruses. As a result of binding, MBL function as an apsonin to enhance phagocytosis. It activates C2 and C4. C2a and C4b activate C3.

ii. iii.

7. Regulation of Complement System by Regulatory Proteins

Following proteins help in regulation of complement system: 7.1 C1inb: This protein cause breakdown of C1q from C1r2s2. C1inb is basically a serine protease inhibitor[8]. These are those enzymes which breakdown the peptide bonds in protein and seine act as nucleophilic amino acid at the enzyme active site. This also prevents the further activation of C2a and C4b. 7.2 C4bBP: When C2a and C4b combines together in classical pathway they are converted to C3 convertase enzyme but when they combines with C4bBP[9] it alters this pathway and did not change into C3 convertase. 7.3 CR1: This protein also alters the above pathway and did not change into C3 convertase. 7.4 MCP: This protein also alters the above pathway and did not change into C3 convertase. 7.5 Factor 1: This protein converts the C4b into C4c and C4d. 7.6 Factor H: When Factor B and C3b combine together it forms C3 convertase. But when Factor H combines with it alters this pathway and did not change into C3 convertase. 7.7 CR1: This protein also alters the above pathway and did not change into C3 convertase. 7.8 MCP: This protein also alters the above pathway and did not change into C3 convertase. 7.9 Factor 1: This protein changes C3b into C3f and iC3b. Then, this factorB also cleaves iC3b into C3c and C3dg. 7.10 Decay accelerating Factor(DAF): It is basically a glycoprotein which is covalently bounded with glycophospholipids; it is also called as CD55. It combines with C4b2a and breakdown it into C4b and C2a. Then Factor 1 cleaves C4b into C4c and C4d.

7.11 S-protein: The C5b67 has ability to combines with membrane bounded cells. If they combine with nearby cells they produce an innocent bystander lysis of healthy cells. But when S-protein combines with it produces hydrophilic transition and does not combine with nearby cells. 7.12 HRF or MIRL: When it combines with C5b67 then it prevents the polymerization of C9. When it combines with C8 when it prevents the cell mediated lysis. MIRL also called as CD59. 7.13 RCA: C4bBP, Factor H, CR1, MCP, DAF and CD59[10] are collectively called as RCA. RCA is made up of short 60 amino acids which are called as short consensus repeats and in human encoded on chromosome number.

8. Role in Diseases
If complement system is not working properly it causes following some diseases: 8.11 Lupus Erythematous: When S-protein combines with nearby cells it produces innocent bystander lysis of cells and becomes the immune system hyperactive and lessens the ability of immune system to fight against pathogens resulting inflammation, damage to joints, skin, kidneys, blood, heart and lungs. 8.12 Hereditary Angioedema: In this disease C1inb deficiency increased and C1 complexes does not work properly and breakdown of C2 and C4 increased which leads to accumulation of edema fluid in skin, mucosa, abdominal cramp, vomiting, diahorrea and airway obstruction. 8.13 Barrquer Simon Syndrome: In this disease C3 level decreased and C1 and C4 levels also decreased and level of autoantibody C3NeF is increased which leads to renal diseases. 8.14 Paroxysmal Nocturnal Hemoglobinuria: DAF and HRF are glycophosphatidylinositol linked membrane proteins which are present on endothelial cells and erythrocytes. Deficiency of these proteins causes this disease resulting recurrent bouts of intravascular hemolysis, chronic hemolytic anemia, venous thrombosis and oliguria. Reference Mayilyan KR, Complement genetics, deficiencies, and disease associations, Institute of Molecular Biology, Armenian National Academy Sciences; 2012 Jul;3(7):487-96. Cavalli L, M Division of Mineral and Bone Metabolism Diseases, Department of Internal Medicine, University of Florence azzotta C, Brandi ML, Phosphatonins: physiological role and pathological changes, 2012 Jan;9(1):9-12 den Dunnen J, Vogelpoel LT, Wypych T, IgG opsonization of bacteria promotes Th17 responses via synergy between TLRs and FcRIIa in human dendritic cells, Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Jul 5;120(1):112-21.

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