Immuno - Lec 8
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Transcript of Immuno - Lec 8
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Antigen Processing and Presentation
Aya Al-Nobani
Ziad Al-Nasser
Tuesday, 12/7/2011
15
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Immunity Lecture #8
Tuesday 12/7/2011Done by: Aya al Nobani
Review of the last lecture
We have been talking about the antigen recognition molecules and the last
one is the major histocompatibility antigen (MHC) and by now you should be
able to know its structure, function, encoding genes which are present on theshort arm of chromosome #6 consisting of 3 loci; class 1 (B,A,C), class 2
(DP,DQ,DR), and class 3 which is not involved with antigen recognition (it
codes for complement, cytokines molecules and some enzymes needed for
steroid metabolism).
Class 1 MHC antigen: is present on all the nucleated cells in our body and
they are involved in antigen processing and presentation to T-cytotoxic cells
(the killing process requires class 1 MHC). Then we talked about the allelic
forms of the encoding genes and these alleles are inherited in a co-dominant
manner, so that in class 1 we have a maximum number of 6 different alleles
and a minimum number of 3 (depends on having heterozygous or
homozygous forms).
Class 2: is present on the surface of antigen presenting cells (APC are
nucleated >> they have both class 1 & 2). Their main function is to present
antigens to the T-helper cells, and regarding the alleles it has a maximum
number of 6 alleles and a minimum of 3.
And we talked about the binding between the non-self antigen and the self
MHC in order to bind with the TCR (restriction phenomena), and the
additional binding (matching) between class 1 MHC with CD8 and class 2 with
CD4 for the signalling process to take place.
Each one of us has a haplotype and this haplotype is like a finger print and
we use this haplotype in tissue transplantation (HLA antigens,
transplantation antigen) so we have to match the haplotype of the donor
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with the recipient before the transplantation in order to minimize the chance
of rejection, the higher the degree of matching the better the acceptance. In
auto-graft (same person)or isograft (identical twins) thematching is
100%, and in allo-graft, which is more common, it depends on the degree of
matching but still we have to supress the immune response of the recipient.
Also the xeno-graft (retro-graft) we take the tissue from a pig and then
transplant it into the patient, and here we must use immune-suppressants.
In the clinical serology lab (immunology lab), we can do many tests in order to
identify the tissue type (the tissue type means the haplotype) so we identify
the allelic form of the antigen for the donor and the recipient. Therere two
types of tests that can be used:
1. Micro-cytotoxicity: used to identify the allelic forms of class 1 (lociA,B,C). In order to do this test, you need to have antibodies against all
the allelic forms of the antigens that are known (the source of these
antibodies is a woman who has been pregnant a lot of times; so that
she would be exposed to her husbands antigen, or people that have
had blood transfusion more than once from different people; this person
develops antibodies against these antigens but this is not a problem;
only RBCs are of concern rather than WBCs) and we fix these antibodies
in plates (each in a separate plate) then we take part of the buffy coatfrom the donors blood (to take the lymphocytes) and put it in the
plates and we add a complement; now if the lymphocyte had the
antigen specified for that antibody it will react and the complement
will be fixed and the cell is going to be lysed otherwise it will not, and
we have a die (that only stains dead cells) that we add that will give a
blue colour if there was a reaction (reaction leads to cell lysis). Then you
see which plates reacted (had blue dots). To know the allelic forms of
the donor, you are supposed to have two of each kind (in case of
heterozygote) or one (in case of homozygote), and then we do the
same for the recipient.
* The degree of colour gives an idea about the reaction.
2. Mixed lymphocyte culture: the idea of this test that if you have twolymphocytes that are incompatible in antigens of class 2 MHC, they
react against each other and they multiply and increase in number. Sowe bring a lymphocyte with a known class 2 MHC and we fix that cell
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in a plate, then add a lymphocyte from the person that we want to
know his haplotype; if these cells are alike, then no reaction is going to
take place, and vice versa. And to know if a reaction took place, we
add triated thymidine (I think what he means is thymine = one of the
nitrogenous bases used in DNA duplication); so if the cells multiply,thymine is going to be used and you can measure the radioactivity, if
no radioactivity can be detected, that means there was noreaction.
*note: you can use the Micro-cytotoxicity test to detect class 2 but the
Mixed lymphocyte culture is the assigned one!!
Now lets start with chapter10
Antigen processing and presentation
Antigen processing and presentation is the sole function of the MHC, whether
the antigen is going to bind with class 1 or class 2 MHC depends on the way
the antigen is going to be trafficked inside the antigen presenting cell
(APC).
*APC for class 2 MHC: macrophages, dendritic cells, B cells (the doctor
mentioned another name but it was unclear and I couldnt find it) and itsused for the extracellular antigens so that the t-helper cells (CD4) will
recruit the B cells (by producing cytokines) to form the antibodies in order to
neutralize the antigens (e.g. toxins and bacteria) or that cytokines will induce
the phagocytic cells to kill the bacteria.
*APC for class 1 MHC: cells that are infected with a virus or a tumor cell (the
tumor antigen is a modified antigen intracellularly) and its used for the
intracellular antigens (cytosolic derived antigens) so that the cytotoxic cells
(CD 8) will be activated to kill the infected cell or the tumor cells.
The doctor said that the viral antigen is formed within the cell.
Remember the MHC restriction: it means that the TCR does notrecognise the antigen unless it was presented by MHC.
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*Endocytic(extracellular) pathway
Extracellular antigen will enter a vesicle by phagocytosis to be in the soluble
form, 95-99% of theses antigen will be killed by the phagocytosis and the rest
will require the formation of antibodies (adaptive immune response), these
antigen are the ones that go in the whole processing and presentation process
(to be presented with MHC class 2 to activate CD4).
The antigen (e.g. toxin or bacteria) will enter theAPC either by binding to the
surface or if antibodies will facilitate it by opsonisation to form the
phagosome (where the pH is low) then the lysosome will bind to form the
phagolysosome and then the toxins or the bacteria will be degraded into
simple polypeptides (by enzymeslike endosomalproteases) and here
comes the role of class 2 MHC which is formed in the endoplasmic reticulum
and processed through the Golgi apparatus then it will be covered with a
protective chain which is called the invariant chain, this chain will protect
the class 2 MHC when its not in use, but when needed a proteolytic enzyme
will lyse this chain so that the MHC will be free to bind with the antigen, MHC
will go into the vesicle and bind to the antigen so that when the vesicle comes
in contact with the cytoplasmic membrane they will be presented on the
surface. T-helper cells (CD4) will bind to the antigen MHC complex through
the TCR leading to the activation of t-helper cells which are going to produce
cytokines needed to continue with the immune response.
The nature of the antigen is important because its going to determine the
antibody which will neutralize the toxin or the cytokines that will activate the
macrophages which will kill the bacteria inside like mycobacterium
tuberculosis.
If there was a deficiency in one of the enzymes (proteases) needed to degrade
the antigen, processing will not take place and this will make the person
immune-compromised.
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*Cytosolic pathway
Suppose a person is infected with a virus, we know that viruses multiply inside
cells and utilize the protein synthetic apparatus and make new viral proteins
the cytosolic pathway will be activated.
**If the viruses were killed or inactivated so that they cant
multiply inside the cell they will trigger the extra-cellular pathway
When the viral proteins are formed within the cell, they are going to be
processed and degraded into small amino acids and then they enter the
endoplasmic reticulum through the transporter antigen presentation
where they will bind to MHC antigen class 1 and then they presented on the
surface of the cell so that the t- cytotoxic cells will recognize and bind to them
to be activated. This is one factor that is required to activate the t-cytotoxic
cells the other one is the cytokines from the t-helper cells.
Proteins (TAP 1 & 2) >> these proteins come from the MHC antigen class 2.
TAP1 & TAP2 are transporter proteins of viral antigens (proteins) into the
endoplasmic reticulum where the class 1 MHC is going to be formed. If TAP 1
&2 were missing (genetic mutation) , the antigen presentation of viruses willstop >> and the cell-mediated immunity is going to be defected (while the
humeral one wont be affected).
Certain viruses like Herpes virus are evasive; they can escape the cell-
mediated immunity by producing enzymes that will inactivate the TAP 1 &
TAP2 so that the viruses cannot be transported to the area where class 1 MHC
are made. The viruses then will survive because they cannot be detected by
the cytotoxic cells.
What really determines which pathway the antigen is going to
take (the cytosolic or the extracellular) is how the antigen is
going to be trafficked inside the cell not the antigen type.
Super-antigen/ T-cell activation independent of processing
We talked about it before , it doesnt need processing or presentation it binds
to the variable B-chain of the TCR and MHC class 2 (not in the groove) and it
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cause toxic shock syndrome because it activates massive number of
lymphocytes. This no specificity is an exception rather than the rule.
*B-cells have a dual function; it can act as an antigen presenting cell (they
have class 2 MHC) and it can develop into plasma cells to produce antibodies.
*T-helper cell activation leads to the production of cytokines and there
are many kinds of cytokines, one of them is the one that activates the
macrophages in order to enhance their ability of killing inside the
macrophage or help in the production of antibodies.
*t-cytotoxic cells: when activated, produce substances called perforin and
cytolysin (their function is to kill the cell by a biochemical pathway that will
convince the cell to commit suicide; apoptosis).
This table in the book lists the differences between the cytosolic, phagocytic,
and endocytic pathways: (the doctor read the whole table)
** Some bacteria multiply intracellualrly like chlamydia and rickettsia; these
bacteria require the activation of T-cytotoxic cells so it utilizes class1 MHC.
*Some of the bacteria can evade the immune response; these are very
successful pathogens. For example, mycobacterium tuberculosis that
causes TB disease. The cell wall of these organism is composed of a specialtype of lipid called mycolic aids and this kind of lipid is extremely resistant to
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phagocytosis, so they can escape the phagolysosome into the cytosol and stay
for a longer period of time, in fact the phagosome can take these bacteria to
other places in our body, so how can we get rid of them?
We require the macrophages to be activated which is done by the mechanismwe have been talking about (cytokines from the T-helper cells).
Another example is Adenoviruses (naked viruses) which interfere with the
transcription of class 1 MHC, so they can stay in our body for a longer period of
time.
In order for the body to kill those evasive bacteria or viruses, its going to
activate other mechanisms including the natural killer NK whereits way of
killing is completely different from that of the cytotoxic cells.
Vaccination
If I can get a virus and inactivate it (e.g. measles) and then inject it in our
body, since they are killed, they are going to be trafficked through the
extracellular pathway (phagocytic or endocytic pathways) and antibodies are
going to develop. But if I want to have memory cells against future infections
with the same virus, I require the activation of cytotoxic cells, so the antigen
must be trafficked through the cytosolic pathway. In order for this to happen,I can use live-attenuated vaccines (e.g. measles, mumps, polio and
rubella). These attenuated vaccines are very successful; the memory cells that
they make are from the T-cytotoxic cells. However, its not always possible to
attenuate viruses or to use it on human.
Another way to make memory cells is to use DNA vaccines ; if you can
get a DNA of a virus (if the virus has RNA instead of DNA, you can make a
complimentary copy) and then through a mechanism like retro-viruses youcan transfer this DNA into the cell then transcription and translation into viral
protein will occur. So the trafficking here simulates the cytosolic pathway. And
as we know this will activate the T-cytotoxic cells and memory cells develop.
The idea of these vaccines is to change the way the virus is trafficked in order
to form memory cells.
And the Doctor MUST give an introduction for the next lecture
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And in the next lecture we will talk more about the molecular changes that
happens in order for the lymphoid cells to be activated; this is important in
order for me to know what level I am going to affect in order to activate or
suppress the immune response.
In general, first you need a stimulus to activate the lymphocyte which is either
an infection or an extracellular antigen; when it enters the body, it has to bind
to receptors as we explained earlier, and then it sends signals through trans-
membrane cell surface receptors (many types) and these have to be occupied
or crossed over in order to send the signal inside after that transduction of
these signals takes place.
When we talked about the CD3 and we said they have protein tyrosine
kinases that get activated by the signals.After the signal passes through, ithas to amplified intracellularly through biochemical pathways (secondary
pathways) and hours after that youll have a response and the outcome of
this response is gene transcription and then translation into proteins > cytokine
production > cell division > clonal expansion > release of defensive molecules.
The most important cytokine is interleukin 2 which plays a major role in
the growth and activation of cells. Other interleukins play roles in isotype
switching (IgM > IgG, IgM > IgE ). Also, interleukins can recruit macrophagesor activate the bone marrow to produce more progenitor cells and call them
into the area and so on.
And we will talk about this in details in the next lecture
El 7amdella 3ala el salameh :D
hon nezlat ra7met rabna o 8arar el doctor ywa2ef :D
Thanx to all my friends who disturbed me while I was writing the
lecture :)
Thanx to you my friends for being such a wonderful people who
bring joy into my life
*love all , trust few , harm none *