Lab 2 Serum Proteins

Question 1

What 2 major groups are serum proteins divided into?

Answer 1

Albumin and globulins

Question 2

Which serum protein is synthesized by the liver? This serum protein is the _________ and easily dissolves in ____________ Albumin has a vast capacity of non selective binding of many ____________ Albumin is primary serum plasma responsible for the transport of _________________, ______________, ___________ and ______________ Some albumin accounts for _____% of total plasma protein and it provides ____% of colloidal osmotic pressure What does colloid osmotic pressure prevent? Values for albumin in serum are……

Answer 2

Albumin Smallest serum protein and easily dissolves in water Non selective binding of many ligands Transport of hydrophobic free fatty acids, unconquered bilirubin, some hormones, trace metal drugs 60% of total plasma protein and 80% of colloid osmotic pressure. Prevent edema Albumin in serum is 3.2-5.6 g/dl

Question 3

What are globulins composed of?

Answer 3

alfa-1 globulins , alfa 2 globulins, beta-globulins and gamma globulins

Question 4

What can be evaluated and expressed as serum total protein concentration and with a reference value of?

Because______ is quantitatively the most important serum protein its concentration will influence what?

Serum protein concentration reflects _______, _______ and _______.

Answer 4

The concentration of serum proteins. The reference value is 6-8g/dL(60-80g/L)

Albumin, concentration will influence the serum total protein concentration.

The serum protein concentration reflects the rate of synthesis, the degradation and volume of distribution.

Question 5

Define Hypoproteinemia

Answer 5

terms the serum protein concentration lower han 6g/dl(60g/L). It may reflect chronic liver diseases(eg. liver cirrhosis), nephrotic syndrome, protein-losing enteropathy, extensive burns and malnutrition.

Question 6

Hyperproteinemia

Answer 6

terms the serum protein concentration greater than 8g/dL(80 g/L). It usally reflect diverse malignant pathologies such as myeloma and Waldenstrom’s macroglobulinemia. A transient hyperproteinemia may be caused by dehydration

Question 7

What is the most important method for sepeation of plasma proteins in clinical laboratory?

This method is usually perfomed at alkaline pH on…

This method seperates the proteins by what ratio? How many fractions can be identified by staining and densitometric scanning?

Answer 7

Electrophoresis

at alkaline pH on cellulose acetate foil or on agarose gel

Their charge/mass ratio. 5 fractions

Question 8

Answer 8

Image 1: Gel with serum protein electrophoresis

Image 2: Densitometric scanning of electrophoresis gel

(albumin, a1 globulin, a2 globulin, B globulin, y globulin)

Question 9

Chart with proteins, concentration and % content

Answer 9

Highest to lowest concentration:

Albumin, y(gamma globulin), B globulin, a2 globulin, a1 globulin

Normal serum prtoein fractions from fastest to slowest electrophoretic mobility:

Albumins, a1 globulins, a2 globulins, B globulin and Y globulin

Question 10

What do Alpha-1 globulins contain?

What do Alpha 2 globulins contain?

What do Beta-globulins contain?

What are gamma globulins made of?

Answer 10

Alpha-1 globulins contain: retiniol-binding protein, a1-antiprotease(termed also a1-antiyrpsin), thyroxine-binding globulin, transcortin and a-fetoprotein

Alpha-2 globulins contain ceruloplasmin, a2-macroglobulin and haptoglobin

Beta-globulins contain transferin, hemopexin and fibrinogen. FIbrinogen is present at plasma but not at serum

Gamma globulins are composed mainly of immunoglobulins and C-reactive protein

Question 11

Immunglobulins aka________ are synthesized by_________, a component of the immune system

Immunglobulins have a defined specificity for a foreign particles like what that stimulate their synthesis?

Describe immunogen

Wha are immunoglobins?

Describe Antigens

Answer 11

Immunglobulins aka antibodies are synthesized by B lymphocytes.

Foreign particles like immunogen, antigen

Immunogen is a foreign particle entering the body which can elicit the immunologic response.

Immunoglobulins are a uniquely diverse group of molecules recognizing and reacting with a wide range of specific antigens and giving rise to a series of effects that result in the eventual elimination of the presenting antigen.

Antigen describes any agent that can be bound by the antibody

Question 12

The immunoglobulin is a ____ shaped molecule containing 2 identical units called _______ and 2 identical smaller units called_____.

What are the 5 heavy chains?

The molecules are characterized by 2 functional areas?

Answer 12

Y shaped molecule, 2 identical heavy chains(H) and 2 identical smalled light chains(L).

The 5 heavy chains are γ, μ, α, δ, and ε and the nature of heavy chain determines the class of immunoglobulin IgG, IgM, IgA, IgD and IgE

The 2 functional areas include:

• The Fab, or variable end is the area that recognizes and binds to the antigen
• The Fc end is responsible for the interaction with other componenets of the immunne system eg, complement ad T-helper cells

Question 13

What is the most common immunoglobun that protects tissue spaces and can freely cross the placenta?

It circulates in plasma in ___ concentration, accounting for ___% of immunlobulin present in ______.

IgG is the main _____ in second (delayed) response to an ______.

Where is IgG also present in? What does it appear to eliminate? How? By what system?

Answer 13

IgG.

high concentration, 75%, in adults

main antibody in second (delayed) resonse to an antigen

IgG is also present in all extracellular fluids and appears to eliminate small soluble antigenic proteins through aggregation and enhanced phagocytosis by reticuloendothelial system

Question 14

IgA has a ______ form and is widely found in _________ and presents an __________ barrier which protects_________surfaces

IgA represents ____% of plasma immunoglobulins

Answer 14

Dimerised form, widely found in secretions and presents an antiseptic barrier which protects mucosal surfaces

7-15%

Question 15

Structure of immunoglobulin G image

Answer 15

VL and VH are antigen binding site

Question 16

The pentameric structure of IgM image

Answer 16

Question 17

What is IgM confined to? What does IgM help with?

What % of serum immunoglobulins does IgM account for?

Which immunoglobulin is the first immunoglobulin to be synthesized after an antigenic challenge?

What structure does IgM have and because of this structure it does not cross….

Answer 17

Confined to the intravascular space and helps eliminate circulating antigens and microorganisms.

5-10% of serum immunoglobulins

IgM

Pentameric, IgM does not cross the placenta

Question 18

What are the minor immunoglobulins?

How much % does IgD account for and therefore it has a role that remains elusive. IgD is thought to be a surface receptor for antigen in ___ lymphocytes.

IgE is present in ____ only in trace amounts and acts to bind ______ and promote a release of _______ amines from _____cells.

What does IgE mediate immunity to and by activating what?

IgE has the lowest concentration in______

Answer 18

IgD and IgE

less than 0.5%. B lymphocytes

serum, bind antigen, promote a release of vasoactive amines from mast cells

It mediates immunity to worms(parasites) and by activating eosinophils

IgE has lowest concentration in serum

Question 19

What are the product of a single B cell and arise from benign or malignant transformation of B cells?

The proliferation of a single B cell clone results in production of __________

Usually monoclonal antibodies are structurally ______molecules but sometimes they may be in some way _______

The absolute physical identity of the monoclonal immunoglobulins leads to a single, dense band in gamma region called_________

Answer 19

Monoclonal antibodies

Production of identical antibodies

Structurally normal molecues, but sometimes they may be in some way fragmented or truncated.

Single dense band in gamma region called paraprotein band

Question 20

What 2 diverse malignant pathologies are monoclonal antibodies associated with? What benign transformations are monoclonal antibodes associated with?

Answer 20

Diverse malignant pathologies are such myeloma and Waldenstrom’s macroglobulinemia. Benign transformations that are monoclonal gammopathies of uncertain significance(MGUS)

Question 21

What are monoclonal gammopathies(paraproteinemias or dystoproteinemias)?

Serum protein electrophoresis in the monoclonal gammopathy image

Answer 21

Group of disorders characterized by the proliferation of a single clone of plasma cells, which produces an immunologically homogenous protein commonly referred to a as a paraprotein or monoclonal protein(M-protein)

Question 22

What is polyclonal gammopathy?

Why is there usually attributable to persistent, high level exposure to antigens?

Answer 22

Polyclonal gammopathy is a hypergammaglobulinemia resulting from an increased production of several different immunoglobulins and usually attributable to persistent, high level exposure to antigens

This is because the immunoglobulins produced by individual cell lines are slightly different from each other in terms of size and charge and so do not migrate to the same place on electrophoresis. This situation ocurs in a variety of infectious, inflammatory and immune-mediated diseases. In electrophoresis one can see a wide band composed of few smaller peaks

Question 23

What can occur as a result of infection, genetic abnoralities or effects of therapy?

Where the situation is irreversible what therapy has been used? By the addition of either _______ or ________.

Answer 23

Deficiencies or absence of immunoglobulins

Replacement therapy

immunoglobulin rich plasma or by the transplantation of bone marrow containing competent plasma cells

Question 24

What is acute phase response?

During acute phase response there is characterised marked increase in the _________ and where? There is also decrease in _______

There is an incease in the synthesis of proteins such as __________(eg. a1-antiprotease or a1-antitrypsin), ___________(fibrinogen, prothrombin) ,___________, __________(CRP) and ceruloplasmin which is of obvious clinical benefit

Answer 24

It is a nonspecific response to tissue injuy or infection and it affect several organs and tissues.

An increase in the synthesis of some proteins predominantly in the liver along with a decrease in the plasma concentration of some others.

Proteins such as proteinase inhibitors, coagulation proteins, complement proteins, C-reactive protein and ceruloplasmin

Question 25

The synthesis of albumin, prealbumin and transferrin decreases during the acute phase response and is therefore called________.

Answer 25

Negative acute phase reactants

Question 26

What is C-reactive protein a major component of and a marker of? Where is it synthesized?

In healthy individuals the serum concentration of CRP does not exceed_____.

What is an essential test in diagnosis and monitoring of infection and sepsis?

CRP rises within ___ hours of the start of inflammation, allowing the inflmmation to be confirmed much sooner than through the use of _________(ESR), which usually_______about a week after inflammation begins

Answer 26

Major component of acute phase response and marker of bacterial infection. Synthesized in liver.

Does not exceed 1 mg/L

Measurement of CRP concentration in plasma is an essential test

Within 6 hours, use of erythrocyte sedimentation rate, usually increases

Question 27

What may one detect using an assay that is approximately 100 times more sensitive than the conventional CRP measurement method? What is this measurement called?

Very small increases in CRP concentration, which can be detected only by hs-CRP measurement seem to reflect a state of what? This is associated with increased risk for what?

Answer 27

One may detect minimal fluctuations in the concentration of this protein. This measurement is called high senstivity CRP (hs-CRP)

Reflect a state of chronic low grade inflammation, which is associated with a increased risk of coronary heart disease(CHD)

Question 28

Define Ceruloplasmin

_____ carries the 10% of the plasma copper but binds the metal less tighty than ceruloplasmin does.

Ceruloplasmin exhibits ________ activity. The amount of ________in plasma is _______ in liver disease.

Answer 28

An a2-globulin and an acute-phase protein of hepatic origin that has a blue color because of its high copper content and carries 90% of copper present in plasma. Each ceruloplasmin molecule binds copper very tightly so that the copper is not readily exchangeable.

Albumin carries 10% of plasma copper and it donates its copper to tissues more readily than ceruloplasmin.

Exhibits a copper-dependent oxidase activity. The amount of ceruloplasmin in plasma is decreased in liver disease.

Question 29

Low levels of ceruloplasmin is found in what disease?

Answer 29

Wilson’s disease(hepatolenticular degeneration), a disease due to abnormal metabolism of copper.

Question 30

What is Wilson disease?

What leads to low levels of ceroplasmin levels in plasma?

As amount of copper accumulates patients may develop what? Where does the copper accumulate?

Answer 30

A genetic disease in which copper fails to be excreted in bile and accumulates in liver, brain, kidney and RBCs. It can be regarded as an inability to maintain a near-zero copper balance, resulting in copper toxicosis.

The increase of copper in liver cells appears to inhibit the coupling of copper to apoceruloplasmin nd leads to low levels of ceruloplasmin in plasma.

As amount of copper accumulates, patients may develop a hemolytic anemia, chronic liver disease(cirrhosis, hepatitis) and a neurologic syndrome owing to accumulation of copper in the basal ganglia and other centers

Question 31

What is a frequent clinical finding of Wilson disease? Describe it.

Treatment for Wilson disease

Answer 31

Kayser-Fleischer ring is a frequent clinical finding of Wilson disease, This is a green or golden pigment ring around the cornea due to deposition of copper in Descemet’s membrane

Treatment includes diet low in copper along with lifelong administration of penicillamine which chelates copper. Copper is excreted in the urine and thus depletes the body of excess of this mineral

Question 32

What is Menkes disease?

What was reported as basis of Menkes disease?

Answer 32

A disorder of copper metabolism which is X linkes(only male infants) and involes nervous system, connective tissue and vasculature and is fatal in infancy.

Mutations in the gene for a copper-binding P-type ATPase was reported as basis of Menkes disease. This ATPase is thought to be responsible for directing the efflux of copper from cells. When altered by mutation, copper is not mobilized normally from the intestine in which it accumulates, as it does in a variety of cells and tissues, from which it cannot exit.

Question 33

Menkes disease

Despite the accumulation of copper, the activites of what are decreased and what might be a reason for this?

Normal liver expresses very little ______ which explains…..

Answer 33

The activites of many copper-dependent enzymes are decreased, perhaps because of a defect of its incorporation into the apoenzymes.

Normal liver expresses very little ATPase which explains the absence of hepatic involvement in Menkes disease. This work led to the suggestion that licver might contain a different copper binding ATPase which could be involved in causing Wilson’s disease

Question 34

_____(ESR) is the only non specific index of the presence of disease, comparable to ____CRP and what does this reflect?

The rate of change of ESR is much less than that of CRP and it reflects the clinical status of the patient ______

Also affected by number and morphology of ______

ESR as a non-specific indicator of disease may be helpful in for ex the investigation of ___________ which do not provoke an acute phase reponse

Answer 34

Erythrocyte sedimentation rate , C-reactive protein. CPR is preferred over ESR in most pathological conditions

Clinical status of the patient less well

Number and morphology of RBCs

Paraproteinaemias, which do not provoke an acute phase reponse

Question 35

Define Cytokines

What do cytokines act as? What are they involved in?

Answer 35

A group of non antibody proteinaceous molecules produced by chiefly T lymphocytes on contact with antigen, so they are produced by another cell. They have small molecular weight.

Cytokines act as intercellular mediators of immune resonse. They are also involved in inflammation, tissue repair, hemopoiesis, cancer

Question 36

How many interleukins are identified? List them. Does each one has unique origin and function?

What are interleukins produced by?

What is their function?

IL-4 is a regulator of ________ and IL-16 acts as a __________

Answer 36

20 interleukins are identified. IL-1, IL-2, IL3, IL-4, IL5 to IL-20. Yes.

Interleukins produced by T-lymphocytes, antigen presenting cells(APCs), macrophages, natural killer (NK) cells, monocytes, mast cells, basophils, eosinophils, epithelial cells and other types of cells.

Function: Interleukins support growth and proliferation of T-cells, B-cells, they are involved in release of hormones and also act and promote development of cells in hemopoietic system

IL-4 is a regulator of allergic reactions specifically intermediate hyper sensitivity and IL-16 acts as a chemoattractant

Question 37

Define Interferons

What are the 3 classes?

What are they produced by?

What triggers their production?

Answer 37

Refers to group of glycoprotein molecules that interfere with viral infections of cells

IFN-α, IFN-β and IFN-γ classes. There are also several subclasses.

Produced by lymphocytes, macrophages, monocytes, fibroblasts and epithelial cells.

Viral infections trigger their production

Question 38

Anti-viral action is the most important property of interferons.

Other 4 important functions of interferons are:

Answer 38

Inhibition of T cells, B cells proliferation

Inhibition of hemopoiesis

Anti-tumor agents

Increased expression of MHC class proteins

Anti-viral action is the most important property of interferons.

Question 39

What are the 2 Tumor necrosis factors (TNFs)?

They are produced by? They are released in response to?

What is the most important property of TNFs?

Functions of TNFs

Answer 39

TNF-α, TNF-β

Produced by activated macrophages, monoctes, antigen presenting cells, T cells, B cells, natural killer cells and endothelial cells. They are released in response to pathogens

Tumor necrosis is the most important property of TNFs

Function: They are involved in immune response. They act as modulators of immune response. They induce apoptosis in certain cells. They are associated with septic shock and inflammation