T cells Flashcards

1
Q

• Describe the major steps of T cell development

A

BONE MARROW

  1. hematopoietic stem cells from Bone marrow go to the thymus

THYMUS

  1. they are now called thymocytes-
  2. in the thymus they are double negative cells - since they have CD4+ and CD8+ within the cytosol and NOT expressed on the cell membrane
  3. they begin to form preTBeta allele for the TCR ( within the double negative form)
    - during this process Beta Selection occurs - which is when RANDOM Gene rearrangements (VDJ recombination) ( DNA break and repair mechanisms occur) of the TCR B chain loci occur
    - this occurs by the expression of RAG1 and RAG2 genes - which expressed recombinase enzyme which induces the recombination of the Beta chain loci -

this process now creates the functional receptor complex called preTCR ( containing the TCR beta chain , preTalpha subunit and CD3 signalling subunits )

  1. this promoted cell survival and entry into the cell cycle
    this expression also downregulates the RAG gene expression to prevent a rearrangement of the 2nd TCR beta allele. ( this is why all T cells in the periphery express a TCR beta allele which has single specificity = allelic exclusion!)
    then the cells become Double positive - since the CD4+ and CD8+ have left the cytosol and now expressed on the cell membrane along with the preTCR complex
  • during this stage the signals which induce the preTalpha subunit to become rearranged are sent - due to the expression of the preTBeta subunit and the CD4+ and CD8+ on the membrane.
  • during this the RAG genes are re-expressed to induce the RANDOM rearrangement of the preTalpha subunit for the TCR.

T CELLS ARE THEN SELECTED-

  1. The next stage is the Positive selection stage - a thymic epithelial cell carrying either an MHC class 1 or Class 2 which has a ligand attached is exposed to the double positive preT cell -
    -it depends on which Co-receptor ( e.g. CD4+ or CD8+ )
    the T cell has which determines the binding in this case -
    - if the preTcell successfully reacts ( weak reactivity) then survival signals are sent to activate the T cell to survive by the CD3 complex ( this complex is always present - in ( when it is double negative) or on ( double positive or single positive) all T cells and is important for signal transduction.
    - if the preTcell does not show any reactivity towards the MHC/ligand complex- then no survival signals will be sent and so NO survival and the cell will die !!!
  2. the next stage is Negative selection stage - which is when a thymic epithelial cell carrying either one of the MHC classes which has a self peptide - e.g. insulin -( which was transcribed by the AIRE genes, ) binds too strongly to a TCR - in this case this would induce APOPTOSIS - since the preTcell has attacked a self peptide - which would lead to autoimmunity ! -
    so if the cell successfully is able to NOT create a strong reactive signal in this case it would survive - as it is important that it recognises the MHC complex - however even more important that it does NOT cause a reactive response to it - when it is a self peptide !
  3. The preTcell , before it leaves the thymus will now engage one coreceptor and lose the other ( either CD4+ or CD8+ depedning on the MHC class - MHCII binds with the coreceptor CD4+ and MHCI binds with the coreceptor CD8+

PERIPHERY

  1. now the T cell is ready to leave the Thymus to the periphery where it will go to the lymph nodes to encounter antigens and become activated etc.
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2
Q

Demonstrate a good understanding of the key extracellular signals that control T cell development

A
  • Cytokines …
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3
Q

What does the T cell do when it goes out into the periphery ?

A

it waits around until a TCR is engaged by a pathogen , the T cell will undergo clonal expansion and differentiation

this differentiation is shaped by Cytokines

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4
Q

What is a TCR?

A

TCR is a multisubunit complex - contains specific antigen binding subunits - alpha and Beta subunit , very variable produced as the products of gene rearrangements , that recognise peptide MHC complexes - these two chains are looped and connected to invarient signalling chains - CD3 complex , and these can activate many signalling pathways -e.g. cytosolic tyrosine pathways - through ITAM interaction ( with the phosphorylation of ITAM by zap70 , and this switches on a whole signalling cascade that can change the transcriptional program of the T cells ,
T cells will each have unique receptor - about 20,000 T cell receptors on T cells - all the same on an individual T cell , but different between different T cells

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5
Q

What family of cytokines is really important ?

A

Gamma chain Cytokines - e.g. IL2 and IL7

different cytokines are needed to differentiate the T cells

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6
Q

Place the events of antigen capture and presentation in an anatomical context

A
T cell activation often occurs
at anatomically distant sites
to an infection
Dendritic cells (DC) have 
remarkable capacity to capture 
antigen, process it, migrate to 
lymphoid organs and present it 
to T cells
On sensing pathogens, DC 
undergo a process of ‘maturation’
which transforms them from
antigen capturing cells to 
antigen presenting cells 

Summary of diagrams:

A pathogen adheres to EPITHELIUM , if a SKIN wound occurs then it allows the pathogens to penetrate the EPITHELIUM
local infection occurs which allows innate immunity to take place , where macrophages will attack the pathogen. In the case of macrophages they will use phagocytosis to further break down the antigen.

Dendritic cells contain PRR for example Toll Like receptors ( to recognise PAMPS) and so will recongnise the PAMP molecules.

Dendritic cells are a link between innate and adaptive immunity. ( since macrophages will NOT migrate !)

Dendritic cells take the infection to LYMPH NODES and stimulate the adaptive immunity. mature T cells are recruited to the LYMPH NODES and recognise the antigen and create an immune response at the SITE OF INFECTION.- this immune response would require the proliferation and differentiation of the T cells into cytotoxic T cells 9 killer cells) or T helper cells- (which are helpful to activate the cytotoxic T cells , the B cells to produce antibodies and for macrophages to break down microbes.)

Since Dendritic cells are APC ( antigen presenting cells) they are able to present the antigen to be attacked by T cells or Antibodies using the MHC molecules.

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7
Q

Explain why dendritic cells (DC) are critical for initiating T cell activation

A
  • Dendritic cells are known as antigen presenting cells - and are vital for initiating T cell activation because they contain MHC complexes which present the antigen to the TCR( T cell receptor ) , this is vital to activate naive T cells to proliferate and differentiate .
    they deliver 3 signals
  1. peptide/ MHC complexes
  2. costimulatory molecules
  3. cytokines that guide T cell differentiation

once these are activated by the DC , T cells can engage other cells e.g. B cells

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8
Q

Explain how pathogen-derived products signal to DC and how this links to adaptive immune responses?

A

pathogen sensing occurs through the reception of TLRs - which sense microbial products inside and outside the dendritic cell - this initiates several changes in the DC which would then result in the adaptive immune responses

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9
Q

Understand that DC provide additional signals besides MHC/peptide (e.g. costimulatory molecules and cytokines) that determine T cell responses?

A

Dendritic cells (DC) deliver at least 3 signals to T cells

  • peptide/MHC complexes
  • costimulatory molecules
  • cytokines that guide T cell differentiation

it is important that dendritic cells and T cells are glued together - before T cells can actually be effective ( takes days for T cell to make effector proteins in response to this…)

note: when dendritic cells are triggered to become matured when their PRR is encountered with the microbial product - they release NFKB , IRFs , MAP kinases - which are all important for promoting the response of maturation…

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10
Q

Outline how DC can be involved in graft or transplant rejection?

A

The donated organ contains dendritic cells - which if not fully histocompatible , will migrate to host lymphoid tissue and activate the host T cells.
- alloreactive- T cells have not been tolerised to these unexpected peptide /MHC pairs , these T cells once activated may go through blood to destroy the graft !

faster rejection - direct allorecognition pathway - allorecognition T cells will attack MHC varient ( allotypes) directly !

Chronic ( slower ) rejection is driven by an indirect pathway of recognition of MHC variants ( allotypes)-

  • the acute rejection wanes as donar DC die off but may first stimulate the indirect pathway of allorecognition - which would involve endocytosis by the dendritic cell of the recipient

Indirect allorecognition can lead to the induction of anti- HLA antibodies against the graft -

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11
Q

What is cross presentation?

A
Cross-presentation is the presentation of exogenous antigens on class I MHC, an 
important exception to the rule that only endogenous antigens appear on class I MHC
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12
Q

Dendritic cells take up and present antigens by various pathways - what are they ?

A

Virus’ are presented by MHC1 and the type of naive T cell activated is CD8 T cells. Virus’ can be taken up by :

  • viral infection
  • cross presentation after phagocytic or macropinocytic uptake
  • transfer from incoming dendritic cell to resident dendritic cell

extracellular Bacteria ( and soluble antigens and virus particles) are presented by MHC class 2 , and the naive T cell activated is CD4 T cells
these can be taken up by:
- receptor mediated endocytosis
- macropinocytosis

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13
Q

What drives dendritic cell maturation?

A
  • TLR signalling drives a progrmme of dendritic cell maturation

TLRs ( are types of Pattern recognition receptors )
TLR 4 –receptor for LPS !
TLR1 and TLR2 – senses other bacterial material
TLR3 – senses nucelic acid signatures – unique to pathogens !
TLR signalling from different types of TLR drives dendritic cell maturation

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14
Q

what changes are seen in DC following TLR signalling ?

A
  • increased antigen capture
  • increased biosynthesis of MHC ( and increased translocation of MHC to cell surface!)
  • increased expression of costimulatory molecules : CD80 and CD86 - ( these are very similar) and CD40 !
  • increaed expression of CCR7 ( a chemokine receptor that guides migration of DC towards CCL21 which is expressed in lymphoid tissues )
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15
Q

What do T cells do?

A

T cells respond to bacterial, antigens or viral antigens

and mount an immune response to clear the bacteria or virus from the body

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16
Q

Describe different T cell subpopulations in the peripheral immune system and understand T cell differentiations

A

TCR triggering induces clonal expansion,proliferation and differentiation

naive T cells differentiate into effector T cells and memory T cells.
effector T cells can be subdivided into Helper T cells and Cytotoxic T cells

17
Q

Describe the T cell response to antigen and cytokines

A

T cell responses to immune stimulation :
- a complex network of
transcriptional and post - transcriptional mechanisms-
TRANSCRIPTIONAL REPROGRAMMING!-

  1. Change migration pattern and move to site of infection
  2. Proliferate and clonally expand
  3. Induce expression of genes
    for cytokines,cytokine receptors etc
  4. Upregulate growth and protein synthesis to permit rapid
    cell division and cytokine production.
  5. Differentiate into killer cells or different types of helper T cells
    to make sure the immune response is appropriate
18
Q

What are the TCR regulated genes?

A
  1. Genes encoding cytokines eg Interferon gamma, IL-2, TNF alpha
  2. Genes encoding cytokine receptors.
  3. Genes encoding cell cycle regulators
  4. Genes encoding nutrient transporters eg amino acid, glucose and transferin transporters. This allows the T cell to match metabolism to metabolic demand.
  5. Genes encoding chemokine receptors and adhesion molecules – this allows the T cells to change the way they traffic around the body
19
Q

What are three responses of the T cell to immune activated situations?

A
  • metabolic - the naive T cell is non- proliferating and less energy dependent , using oxidative phosphorylation for ATP generation whereas immune activated cytotoxic T cells are high proliferating and are glycolytic !
  • motility - naive T cells are moving alot - recirulating in the blood - and so use cytokine receptor CD62L and CCR7- the adhesion molecule
    whereas these cytokine receptors - CD62L and CCR7 are downregulated in the immune activated cells
  • Cytokines - naive T cells express-IL 7 receptors to allow the T cells to survive in the lymphoid tissues - , whereas these are downregulated in the effector T cells - and instead there is an up regulation of inflammatory receptors e.g IL2 and IL12 receptors.
20
Q

What signalling pathway allows T cells to switch on Gene expression to make cytokines?

A

Immune responses are controlled by lymphocyte derived cytokines
e.g TNAa,IL2,IL3,IL4,IL-5.

The expression of genes encoding these cytokines is regulated by combinations of different transcription factors.

Key transcription factors for lymphocyte derived cytokines include members of the NFAT family. ( Nuclear Factor of Activated T cells)-

In quiescent T cells NFAT proteins are phosphorylated and cytosolic

T cell activation results in NFAT dephosphorylation and translocation to the nucleus

Nuclear NFAT forms a complex with AP-1 transcription factors to regulate cytokine gene expression

21
Q

How can antigen activated T cells differentiate to make different types of
effector cells?

A

The specification these different effector cells is controlled by signaling pathways induced by cytokines produced by activated T cells or cells of the innate immune system

The type of cytokine made by innate lymphoid cells depends on the pathogen

So T cell effector cell differentiation is initiated by the antigen receptor in a stop/go decision but the type of effector T cell you get is controlled by cytokine signaling.

22
Q

What are some Key cytokines for differentiation?

A
IL 2 - regulatory T cells
IL2 and IL12 - Th1
IL6, IL21, IL23 - TH17 
IL4 - Th2
IL7, IL15 - memory T cells
IL2, IL12 - cytotoxic effectors ( killers)
23
Q

What type of cytokines use the common gamma chain?

A

IL-2: ( made by T cells as well !) regulates T cell proliferation and differentiation of cytotoxic T cells and regulatory T cells

IL-4.IL21: crucial for Th2 cell differentiation and hence B cell function- important for T cells to help B cells

IL-7: crucial for survival and proliferation of T and B cell progenitors in the bone marrow and thymus ( when T cells first come into Thymus !)

IL-9:mast cell growth factor

IL-15: crucial for NK cell differentiation and T cell memory

Thymus development controlled by overlapping antigen receptor and cytokines
IL-7 one of the cytokines that uses the common gamma chain is essential for the survival of CD4/CD8 double negative cells in the thymus

all use common gamma chain and very critical !

this family are all controlled by TCR !

  • the receptor wont be expressed on Naive T cells , only on activated T cells !!! and all need this chain ! -
24
Q

What is an X - linked SCID? and why is the gamma chain so important for it ?

A

males: Mutation of the common gamma chain gene which is on the X chromosome

It binds and recruits a tyrosine kinase known as Jak3
( janus kinase 3) – important for signalling of all these cytokines !!!

  • Much more common since gamma chain on X chain – the mutant on gamma chain – on one x- if male gets it they will die without transplant

Non X-linked SCID: Females with mutations in Jak3