T-Cells – development and function

- T cell receptors are created by somatic recombination

o There’s somatic mutation at the CDR regions
o Like 2/3 are not functional
o WHAT HAPPENS TO THOSE?
o TCR have to recognize self MHC molecules
▪ Do not want to target self
o All this shit goes down in the thymus
- T cell precursors (CD34+, hematopoetic stem cells) in bone marrow, migrate to the thymus
o Thymuscyte (yah fuck spelling)
▪ Means this shit is in the thymus
o Thymus is highly structured
▪ Cortical part outside – afterwards will migrate into the medulla
• Cortex has developing T cells in various stages of development
• Thymic epithelial cells
• Some macrophages, fibroblasts (maintain structure), dendritic cells
▪ Medullary part inside
• Thymic epithelial cells in the medulla do other things
• Some macrophages and dendritic cells
o CD34+ will commit early to T cell
▪ CD2 is used as a marker for those
o Gene rearrangement begins
▪ Gamma delta first  gamma delta t cell
• 5% of T cells are gamma delta
• Cell will exit the thymus and enter the blood
• As CD4-/CD8- (double negative) T cell
• Rearrange receptor genes
o Delta subunit rearranges first = heavy chain
o Then the gamma chain and beta chain will rearrange
o Gamma delta receptor will complex with CD3
o This shit will signal
o Shit will be like ok cool this works guess im gonna be a delta gamma
o BOOM DONE WITH THAT SHIT
• That’s fine and shit except it takes helluh days so usually beta chain is faster
▪ Alpha beta first  alpha beta T cell
• Most of circulating
• 1/3 CD8 cytotoxic T cells
• 2/3 CD4 T helper
• These will express both receptors initially (double positive)
• Will undergo positive selection
o Check if the TCR can recognize MHC displaying self antigen
o Since it’s still in the thymus, just a bunch of our own shit is floating around in
there
o If this works, one of the two co-receptors will be co-selected
o The ones that bind self too strongly will actually FUCKIN DIE
• This is how this shit happens:
o Beta will rearrange
▪ Beta is the heavy chain (VDJC)
o The beta can signal with pTα – sufficient for “pausing” further gene
rearrangement **FIRST CHECKPOINT**
▪ The pTα is a surrogate α chain
 ▪ Those bitches dimerize
▪ Recruits CD3 complexes
▪ All that shit together can signal
▪ Antigen binding surfaces are touching each other LOL~~~~~
• Cell survival signal
▪ Still in the ENDOPLASMIC RETICULUM of thymus cortical cells
▪ Downregulates the RAG genes
• Prevent building of other beta chains
• Already have one that works, don’t need no more shit
• This is called ALLELIC EXCLUSION
• Need just one on the TCR
o Basically 4 attempts for beta subunit (chain) gene rearrangement
• 2 attempts per chromosome
• So fuckin up ain’t that bad
o Default signal is DEATH
▪ DEATH BY NEGLECT
o After it gets survival signal, cells will proliferate – EXPANSION to like 100 cells
with the same b cell subunit
▪ Can potentially become CD4 or CD8 (double positive cells)
▪ Anyway, then they’ll all try to rearrange their alpha chain
• So they’ll all have different alpha chains
▪ Alpha rearrangement is helluh simpler
• Just Variable and Joining and constant segments
▪ Both alleles are fair game for successful rearrangement
▪ If this bitch can’t make an α chain successfully IT FUCKIN DIES
o ALSO SIDE NOTE! The delta heavy chain stuff is in between the alpha shit
▪ So if the alpha happens then the delta shit won’t happen lol suckuh
o The alpha-beta will complex to CD3 AND THEN traffic to the surface
▪ Then it will go do positive selection
o POSITIVE SELECTION – A/B will be associated with CD3 and move to surface, as mentioned above
▪ Thymic epithelial cells strongly express MHC class I and II
• Typically MHC II only APC
• So the thymic epithelial cells are the only NON APC cells that display MHC II
• Testing the TCR to recognize MHC + self (since still in the thymus)
• 90% of the cells will not find the MHC molecule it can bind to with correct affinity
o Death by neglect
• 10% will bind correctly and receive survival signals
• This needs to happen rapidly or death by neglect
• Whatever MHC molecule is successful at the binding
o Will determine if CD4 or CD8
▪ Will upregulate whatever works
▪ Downregulate whatever doesn’t
▪ So if binds MHC II , will be CD4
▪ MCH I will be CD8
▪ RULE OF 8’S
o NEGATIVE SELECTION
▪ Get rid of anything that binds too tightly to self molecules
• Or to the MHC molecule
• Don’t want no autoimmune disease son
▪ Want cells that recognize and bind to MHC antigen
▪ TCR is not just touching peptide antigen but also the MHC molecule as well
▪ Moderate binding gets survival signal
 ▪ Too weak or too strong will not get any survival signal and just die
▪ THIS IS CALLED IMMUNOLOGICAL TOLERANCE
• BASICALLY this just means there’s no immune binding on self antigen
• Central tolerance
o Happening in a central lymphoid organ – the thymus
o This is key in not getting autoimmune disease
▪ This shit works because there is expression of AIRE
• Auto immune regulator
• Basically it’s shit that will let the thymic medullary cells express a bunch of different
kind of surface molecules “promiscuous gene expression”
o Transient expression during negative selection
• Tissue specific/ gender specific / disease specific expression of proteins that you
wouldn’t expect to find in ONE given cell
• This shit is done on purpose to show your immune system that might develop LATER
o Gender specific, tissue specific, disease state
• APS I
o Basically this is polyendocrinopathy
o Immune system is attacking a bunch of different endocrine/epithelial tissues
o Patients have T cells that have affinity for variety of self markers on multiple
tissues
o Can’t be compensated for by peripheral tolerance
▪ If cell has more affinity for self antigen than normal CD4 helper cell
• These are called T-REG CELLS
• Not enough to have been negatively selected and left to die
• Somewhat self reactive
• Has surface markers used to ID
o CD4+
o Upregulate subunits of IL-2 receptors
o May aid in autoimmune issues
o TXN repressor called FOXP3+
o Also has CTLA4+
o There are also peripheral tolerance shit
▪ To be discussed later
o Thymus declines with age
o Thymus is the only organ where you can generate new receptor specificities as discussed above
o After puberty your thymus goes downhill
▪ Ability to make new T cell receptor specificietes are like super reduced
• In elderly you do get some gene rearrangement because you still see TRECs
o The little excision circles after RAG gene rearrangement shit
o Cannot be measured in anyone who doesn’t have a thymus OR people who are
taking organ transplant rejection RX
o Only 2% of the cells that enter the thymus leave as functional naïve t cells
▪ The other 98% get eaten by macrophages
T CELL PRIMING
- Occurs in secondary lymphoid tissue
o Lymph nodes
o If it is able to find cognate antigen, it will clonally expand and differentiate into an effector cell
▪ Stay in lymph node
▪ Migrate to the site of infection
o This is basically the beginning of the immune response
- If antigen enters through the blood
o Going to be presented at the spleen
o Spleen only gets shit through the blood
T CELL TRAPPING
- Helluh small number of T cells will actually be able to recognize a given antigen during infection
o Enhance the chances of the T cells encountering antigen
o T cells will visit tiny little lymph nodes where a bunch of antigen is being presented
▪ Basically a thirsty AF college kid going to super small parties trying to git sum but from just one
person
- There are particular surface markers on surface of naïve t cells that allow them to enter the secondary lymphoid
tissue
▪ CCR7
▪ Selectins and addressins let the T cell into the endothelium
o Naïve t cell leaves the vasculature through the HEV (high endothelial venule) OR it’s come in through the
lymph
o Will aquire antigen through the blood or the lymph
o If the T cell finds cognate antigen, it will be activated and expand its population WHILE STILL STUCK TO
THE APC that successfully activated it
▪ All the daughter cells will also be activated
▪ Once that shit is done, the T cells are mature and can leave the lymph node and go out and be
effector T cells
- Note that the lymph node has specific areas for B and T cells, as well as macrophages and shit
o B cells in follicles
o T cells in paracortex
o Macrophages in medullary sinuses
- If the T cell doesn’t find shit it can bind it’ll just go back out and run around to try and find something it can bind
to
- Shit also needs costimulatory signals
o APC needs to have the costimulatory molecules
▪ They will express them only in the presence of antigen
▪ That way shit doesn’t just randomly get activated

- Once the T cell is activated, you want shit to stop

o Don’t want it to continually be costimulated
o That would be ridiculous
o Don’t just turn shit on gotta turn it off again
o Kinda like sex lolololol
o But not really
o The T cells have a ligand that can out compete the binding for costimulation
▪ CTLA4
• Higher binding affinity than the B7 costim.

Next lecture

- CD-25 is a part of IL-2 receptor

o IL-2 R needs to be upregulated
▪ At the point where we have made a pre-t cell receptor
• The successful B chain signaling with pTα surrogate chain
• **this is a proliferation signal – make more of this shit bro!
o IL-2 is t-cell growth factor
- Expression of RAG changes
o Increases to make the Beta
o Decreases because NO YOU DON’T NEED MORE BETAS
o Increases again to make the alpha
o Decreases because NO WE DON’T NEED MORE ALPHA!!
- Naïve T cell will encounter cognate antigen in the lymph node
- Small space improves chances it will come into contact with antigen it can bind
- Naïve t cells have surface receptors called CCR7
o Helps hone to secondary lymphoid tissue
o Comes out of vasculature like neutrophils do
- Once they are inside the lymph node, bind to dendritic cell carrying antigen it recognizes
o T cell trapping
o Multiplies
- Lymph node is densely crowded space
- If t cell does not encounter cognate antigen in one, it’ll go to another lymph node
- Needs 2 sources of signaling once encounters cognate antigen
o MHC and costimulatory signal
▪ Costim only from APC (dendritic cells, macrophage, B cells)
▪ Dendritic are the best at costim
o ONLY in presence of pathogen do APCs express costimulatory B7 molecules
- Once T cell is activated, want to turn off signal
o CTLA4 – cytotoxic t lymphocyte associated protein 4
o Higher binding affinity for costimulatory B7 than CD28 (CD28 on T cell)
o It will quench the costim process
- Lymph node has particular areas where particular cells hang out
o Dendritic cells are everywhere EXCEPT for the follicles
o Macrophages are basically everywhere also INCLUDING the follicles
o Dendritic cells present antigen in cortical area to T cells
o Macrophages are in the cortex and medulla of lymph node – some APC but mostly cleaning up dead cell
- Dendritic cells in periphery are immature and just phagocytose stuff
o If find antigen, upregulate MHC I and II AND B7
o Migrate to the lymph node, find a T cell, activate it
- Macrophages in periphery
o Innate immune response
o Killing shit
o Different population of macrophages LIVE in the lymph nodes
▪ Clean up messes
▪ Free antigen can come in with the lymph
▪ Upregulate MHC I and II and B7, activate T cells
o BUT most of the t cell activation is done by dendritic cells
- T cell activation requires like helluh signals
o HELLUH MCH and B7 activation/signaling, approx. 100 interactions needed to generate the str of signal
to activate it
o iTAM = immunoreceptor tyrosine base activation motif
o signals down to nucleus to active T cell
o NFKB and other txn factors will change gene expression pattern so that the T cell can proliferate and
differentiate
- Proliferation of T cell
o Depends on IL-2
▪ T cell growth factor
o Need costimulatory interaction in order to get to the point where the cell is making IL2
o Cell can stimulate itself with IL2
o Upregulates its IL2-R to add subunits to increase affinity at the IL-2 receptor
o Result is there’s helluh shit to deal with antigen/pathogen
- Costimulation only happens in the presence of pathogen
o APC only upregulate costim ligand when pathogen is present
- Antigen recognition and no costim
o ANERGY “t-cell anergy”
 ▪ State of non responsiveness
o This means it’s probs self or it’s not a dangerous situation
▪ So that’s why T cell doesn’t activate
▪ It actually becomes tolerant to this antigen
• Even if it meets the antigen WITH costim later, it won’t do shit since it’s already seen it
and goes like NAH
o So vaccines need both signals or shit will go downnnn son
- GUESS WHAT THERES MORE THAN 1 KIND OF T HELPER CELL
o Th1 + Th2
o As the t cell is maturing, cytokines around it while being activated drive it to be a TH1 or TH2 cell
▪ Most of the reactions to pathogens are MOSTLY one thing or another
▪ If there’s a lot of IL2 and gamma interferon
• Th1 response – more of a cellular response
• Cell to cell contact
▪ Lot of IL4 or IL5
• Th2 – more of a humoral response involving B cells also
• Presence of antibodies
▪ Shift majority of response to either Th1 or Th2
o CD8 t cells can be activated one of two ways
▪ Need enough signals to activate cytolytic T cells
• Viral antigen on MHC I molecule
▪ Can be activated if there’s enough interaction for the T cell to produce its own IL and autocrine
activation
▪ If CD8 t cell can’t activate itself, needs help from CD4 t cell
• CD4 will recognize same antigen that the CD8 cell recognizes
o Costimulatory interactions going on in both cases
o IL2 is produced by CD4 T cell
o The IL2 from CD4 activates the CD8
- Properties and functions of effector t cells
o Once a t cell has met cognate antigen, no longer naïve
▪ Primed
▪ Has rcvd stimulation
▪ Expanded population
o Primed T cells can attack any cell displaying the pathogen that primed it
▪ No longer needs costimulation to kill shit
▪ This shit is important
▪ These bitches are dangerous
▪ Don’t want them hanging out for too long
▪ Expresses/upregulates various surface adhesion molecules to go to where the pathogens are
o Express characteristic set of cytokines/cytotoxins
▪ Gamma interferon – activates macrophages
▪ Lymphotoxin
• TNF B
o Promotes development of lymphoid tissue
o If there’s excess it will transform tissues into lymphoid like tissue
▪ Most of the killing is done by degranulation
• Release of lytic molecules onto the target cells
▪ Th1 cells
• Gamma interferon – cell mediate response
▪ Th2
• Express IL4 and 5 – humoral mediated response
▪ Basically there is a characteristic set of molecules that each T h cell expresses
▪ T cell binding to macrophage
 • Macrophage has phagosomes with a bunch of crap in it
• Gonna activate it and be like BITCH DIGEST THAT SHIT
▪ B cell presents antigen to T cells
• Antigen from surface immunoglobulin
o Receptor mediated endocytosis
o Chewed up and presented to Th2 cell
▪ Th2 cell will activate the B cell
▪ B cell will proliferate and differentiate
• Produce antibodies
- Most of the time lytic granules is what CD8 uses to kill shit
o There’s also a fast mediated response to kill shit
o Targeted degranulation onto target cell
▪ T cell arranges its cytoskeleton to direct lytic particles onto the target cell
▪ Apototic death –NOT A NECROTIC DEATH (necrosis is messy af)
• Clean death
• Necrosis = messy = causes inflammation
o This is similar to what NK cells do in the innate immune response
- So basically all this shit will make
o Memory t cells
▪ Smaller pop
▪ Maintaining immune memory
• Independent of antigen presence
▪ Both CD4 and CD8 cells produce memory T cells
▪ This is because they tend to upregulate BCL2
• Promotes cell survival
• BCL 2 = antiapoptotic protein
o Helps memory t cells be fairly long lived
o Can recognize pathogens that body has previously seen
o Faster response than activating a naïve t cell all over again
o Effector t cells

B CELLS – mature ones live for like 40 days

- B cell development: basically 3 options
o Clonal deletion following unsuccessful receptor editing
o Successful receptor editing
o Anergy development and death
o Note that B cells need constant “reassurance” (survival signals) by going into the lymphoid tissues – if it
can’t get it, bitches die
- B cell development happens in 2 places
o Starts in bone marrow
o Later phases happen in the periphery
▪ In lymphoid tissue
o As with t cell development, helluh steps before b cell becomes activated effector cell
▪ Success rate is helluh low
• Have to build the receptor
• Check if it can bind shit
• If it can’t bind shit it fuckin dies
o Step 1 – rearrange genes in bone marrow
▪ Stromal cells stimulate B cell development
• B cells start as CD34 + cells
• Will commit to B cell lineage
 • If successfully passes the first checkpoint, stromal cells release IL-7
o Produce B cell that is able to leave the marrow
▪ Heavy chain rearranges first
• This is more complicated (VDJC)
• Diversity segment to Joining segment
• 1 locus on 2 chromosomes
• Happens on both chromosomes
• Variable segment will be joined to DJ on first chromosome
o If that shit doesn’t work, it will be done on the second chromosome
• So basically you get 2 chances to make that shit work
o 50% success rate
• Once the Heavy is made, can bind surrogate light chains  test the heavy chain
o FIRST CHECKPOINT @ ER – surrogate light chain
o Similar to T cell
o Join IG-Beta and IG-Alpha (costimulatory for the B cell R)
o Projecting into the ER – NOT ON THE SURFACE
o If it works, will create more of that shit
o Need 2 or more molecules to cross link at variable regions
o Need to signal through the iTAMs to indicate shit works and to proliferate
o Need a lot of them to signal at once to get the str of signal needed
o This signal will downregulate RAG
▪ Allows for ALLELIC EXCLUSION
▪ Therefore will only express one type of receptor in that cell so there’s
not a bunch of different molecular species of heavy chain receptors
o Need a receptor that will recognize homogenous antigen with high avidity
binding
▪ Avidity: binding the same, multiple/many antigens at receptor
▪ Affinity: binding at 1:1 (one molecule, one receptor)
▪ Example: IG-M (pentamer) – each binding site on pentamer has low
affinity, but has a high AVIDITY
• IGM has shitty affinity so it’s not really used that much except at
the start of the immune response
▪ Light chain afterwards (VJC)
• RAG genes turned back on
• Rearrange light chain
• Starts first with KAPPA gene segments
o If that shit works then IGM will become KAPPA LIGHT CHAIN
• If that shit doesn’t work, rearrange lambda light chain segments
o End up with IGM that has LAMBDA light chains
• More than 2/3 of IG are kappa
• Functionally shit doesn’t matter, bitches are the same
• RAG genes turn back off
o Step 2 – b cells undergo negative selection in bone marrow
▪ SECOND CHECKPOINT completed receptor signaling @ the surface with IgA and IgB
▪ Self reactive cells fuckin die – like 75%
▪ Bone marrow and shit offer self antigen for B cell testing
▪ Soluble antigen in the blood plasma in bone marrow also
▪ Successful B cell that doesn’t react with self, will upregulate IgD and leave the marrow
• We don’t know wtf IgD does BUT THAT SHIT IS THERE SO WHATEVER
• Marker for maturity of B cells
• ***THIS IS THE ONLY TIME THAT B CELLS HAVE 2 KINDS OF RECEPTORS***
▪ If the B cell binds to nonsoluble antigen (on something)
 • RECEPTOR EDITING
• If you have a B cell that just made, bind in multivalent manner to self antigen
• B cell receptors are cross linked b/c antigen is not soluble
o Lots of receptors binding to antigen
o Upregulate RAG
o Light chain gene segments will rearrange
o New light chain
o New receptor may/may not be self reactive
o If new one is self reactive, will die
▪ Clonal deletion of any cell that fails receptor editing
o Not all cells have the same amount of time to do receptor editing
▪ Depends how long it took to build the light chain in the first place
• B CELL ANERGY
o Binds soluble antigen in the blood plasma in bone marrow
▪ In contrast to binding to nonsoluble antigen
o B cell will be unresponsive
o Will leave the marrow and dies within 5 days
o Will not get survival signals – dies helluh faster than a healthy B cell
o Binds to self will also go out and die
o Step 3 – immature B cell leaves bone marrow and circulates blood/lymph
▪ POSITIVE SELECTION in the lymph nodes
▪ Enter lymphoid tissue to get survival signals
• This is the positive selection – if it can’t get into the lymph node, won’t get survival
signal
o Fuckin dies
• To enter secondary lymphoid tissue, need correct receptor on surface to follow the
cytokine gradient
• Terminology sucks balls
• B cell will enter the lymph node through the HEV (high endothelial venule)
o CCL21 attracts B cells to HEV
o CCL21 and CCL 19 attract B cell into the lymph node
• Will enter the 1’ lymphoid follicle
o Attracted by CXCL 13, released by dendritic cell in 1’ follicle
o Follicle secretes cytokine that “lures in” B cells
▪ COME HERE KIDS WE HAVE CANDY
o Follicle = collection of B cells
o There’s other shit in there too
o B cell is interacting with follicular dendritic cells  maturation of B cell
▪ Dendritic cell gives B cell survival signals
▪ 2 way street – follicular dendritic cell gets shit too
▪ B cell expresses more IgD than IgM
• Still naïve – has not met cognate antigen
• Needs to keep doing this shit or it will die
• At some point it will encounter cognate antigen
o B cell encounters cognate antigen – gets help from helper T cell
▪ Becomes activated, proliferates – does 2 different things:
▪ Goes into the medulla
▪ Some of the B cells will secrete IgM – PLASMA CELLS – takes care of shit
fast
• Recall the affinity of IgM is low but the avidity is high (binds
multiple shit, it’s a pentamer) – but it’s a problem because it
doesn’t bind with high affinity
o Temporary fix only
 • Membrane bound IgM vs the secreted IgM
o Alternative processing of the primary transcript
o For secreted it’s clipped at a different part, lose bits of
gene product that would place the IgM into the
membrane
▪ Some will move back out to the secondary follicles and do a bunch of
shit
• 2’ follicle where the germinal center reactions occur
o B cells somatic hypermutation to increase hyperaffinity for antigen at hand
o Class switching
▪ IgM  IgG / IgA etc
o These cells will leave the germinal center and become plasma cells (live in bone
marrow, secrete antigens)
o OR can become isotyped switched memory cells in circulation that have less of a
need for survival stimulation from follicular cells
▪ Long lived
▪ Rapid secondary response if pathogen is encountered again
o With t cells, “survival check points” is the opposite
▪ First thing it does is to see if it can recognize the self presenting MHC molecules
▪ B cells don’t need this shit because they recognize intact antigen
• Just wanna get rid of the bitches that recognize self antigen
o B cell development has helluh different names for each stage
▪ Shit doesn’t matter
- NOTE THAT if we’re moving genes and shit around with RAG genes
o DNA has to be sterically accessible
o Various TXN factors will hold the DNA open so that RAG and shit can get in and put shit together
o TXN factors that open the DNA for T cell are different from TXN factors that open DNA for B cells
- B cell types
o B2 cells are the conventional B cells
o B1 arise early in fetal liver
▪ Primitive – not a lot of receptor diversity
• Limited gene rearrangement patterns
▪ CD5 surface molecules
▪ Don’t really know wtf they do in humans – mice CD5 do something else entirely
▪ Only 5 % of circulating B cells
- Can activate b cell in several different ways depending on what the antigen is
o B cell binds to bacterial cell many times
▪ Multi-valent
▪ Aka cross linking – that is how you can cross the threshold
▪ Generates enough signal through iTAMs to activate B cells
• Strength of signal matters
o Can also use co-stimulation for B cells
▪ Requires the residual/fragment of complement system
▪ “C3” opsonized
▪ CD19 provides additional signal transduction
- Most antigen = protein
o If the antigen is a protein, need T-cell help
▪ Recognition of antigen
▪ Co stimulation from T cell
▪ B cell will differentiate, proliferate, and produce antibodies
- Non-protein antigen – TI Antigen
o Shit like LPS, lipoteichoic acid
o Thymus independent antigen
 ▪ Not T cell independent
o There are several ways to do this
o TI2 is simpler
▪ Cross linking of the same antigen, large extent= sufficient to activate the B cell
o Antigen recognition and costimulatory and TLR  activate B cell
o Can activate the B cell without T cell help
▪ As long as the stimulation is enough
- WHAT HAPPENS IF THERE IS A PROTEIN ANTIGEN
o Naïve T cell will enter the lymph node and encounter a dendritic cell with antigen
▪ Recognizes antigen  proliferate
o Dendritic cell picked up pathogen from periphery
o Naïve B cell enters the primary follicle and interacts with dendritic cell
▪ Picks up antigen from follicular dendritic cell AND the survival signals
• B cells can also get antigen from macrophages in there but mostly it’s dendritic cells
• Dendritic cells also release cytokines to recruit B cells – that’s how they get there in the
first place
▪ May react to the same antigen as the expanding population the T cell above does
▪ Then the T/B cells form a cognate area
• “TFH” cells = Th2 cells
• End up with antibody production
• Basically the T cell will give help to the B cell, B cell will produce antibodies
- Cognate pairs will move to the medulla
o Population of B cells will expand
▪ Some of them differentiate and produce IgM (antigen specific)
• secrete IgM  circulation
• Short term solution to infection/pathogen
▪ Some will go back to the cortex and join the secondary follicle
- Secondary follicle = secondary focus of B-cell expansion
o Lymph nodes swell up because B cells are multiplying
o B cells are in various stages of doing shit
o Macrophages in there also
o T cells stay on the outside
o GERMINAL CENTER REACTIONS (affinity and class switching)
▪ Affinity maturation of B cells
• Somatic hypermutation – randomly introducing mutations in the B cell receptor
• Some of the mutated receptors will bind the antigen more tightly
o Increase binding affinity by 10x-100x
• Since this is random, most of the time it’s not successful
o You’ll get a receptor with shitty binding
o Bitches will die and get eaten by macrophages
• There’s a lot of room for fuckups which is why you can get b-cell cancer helluh easy
• In the hypervariable region (CDR loops) – VDJ
• Receptors bind more strongly
• Lots of cell division
o Recall the DNA must be open since protein and shit is being made
▪ Class switching
• IgM  IgA/IgG etc
o IgG (gamma) / IgE (epsilon) / IgA (alpha)
o AFTER GERMINAL CENTER REACTIONS
▪ You will end up with 10% memory cells – no longer IgM; class switched
▪ 90% of them will be plasma cells and secrete antibodies
• Plasma cells are TERMINALLY DIFFERENTIATED
 o Plasma cell 4 lyfe, wut wuttttt~~
o Antibodies can neutralize shit
▪ Can opsonize shit
• Enhance phagocytosis of whatever the Ig is stuck on
▪ With or without participation of complement
▪ Antibodies neutralize bacterial/viral pathogens
• Bind to the toxin / outside of virus
• Such that virus can’t enter cell because there’s a bunch of shit on it and it can’t bind
receptors
▪ Different classes of antibodies work with complement with differing efficacy
▪ IgM / IgG3 are helluh good at complement fixation
o Classical complement
▪ CRP or IgM
▪ Binding configuration is pretty similar
▪ C1 can bind to opsonized IgG on surface of pathogen
▪ Binding of Ig to antigen = immune complex
• This can also start the classical complement pathway
▪ Basically just need the spacial config to activate C1
o Immune complexes are bad
▪ U don’t want that shit floating around
▪ It gets taken to the liver to get taken care of
▪ If stays in system will clog up small blood vessels and fuck up your kidneys
- Antibody Dependent Cell mediated Cytotoxicity (ADCC)
o This is the property of NK cells
o Involves recognition by binding (by NK cells) to Fc portion of antibodies
o Enough cross linking will cause the NK cell to degranulate onto the target cell and fuck its shit up
o This mechanism is helluh important for various therapeutic Rx – use monoclonal antibodies
▪ Rituximab
▪ Anti CD20 monoclonal antibody
▪ CD20 is a B cell specific surface marker
▪ So Rituximab will go and bind to ALL THE B CELLS, which will then get destroyed by NK cells
▪ But then the people will be super immune compromised
- VACCINATION
o All this shit
o Primary immune response – start off with IgM response
o Some B cells will move to the secondary follicle
o Undergo germinal reactions
o Class switch and affinity maturation
o Can undergo several rounds of affinity maturation
o May occur within the same or subsequent infections
o With the second encounter of the same pathogen, HELLUH SHIT WILL GO DOWN
▪ Much faster
▪ IgM is inhibited
▪ Class switched antibody
▪ Affinity for antigen will be HELLUH HIGHER (or just moderately higher)
o So when we do vaccinations, we do the secondary response
▪ Swift and powerful response