Summary Class notes - Klinische I&I

Course
- Klinische I&I
- Knol, Denzer
- 2018 - 2019
- Universiteit Utrecht
- Biomedische Wetenschappen
329 Flashcards & Notes
1 Students
  • This summary

  • +380.000 other summaries

  • A unique study tool

  • A rehearsal system for this summary

  • Studycoaching with videos

Remember faster, study better. Scientifically proven.

PREMIUM summaries are quality controlled, selected summaries prepared for you to help you achieve your study goals faster!

Summary - Class notes - Klinische I&I

  • 1517785200 Refresh your Immunological Memory

  • What are the characteristics of the immune system?
    Adaptive, memory, specific (self-/non-self recognition), diversity (antigen recognition), acquired (education of lymf), systemic.
  • The innate immune systeme reacts first - What is the role of the phagocytic barrier? Which cells form this barrier?
    Phagocytes break down pathogens (removing and destroying) and present the peptides on their own MHC molecules (getting activated and signalling). 

    Phagocytic cells are: macrophages, monocytes, neutrophiles (granulocytes), and dendritic cells (in tissue).
  • How can neutrophiles manage phagosomal killing? More options
    Lysosomal proteases: killing and digestion
    Lactoferrine: Fe downgrade; Fe is essential for bacteria.
    Reactive oxygen
    Lysozyme: breakdown of cell walls.
    Defensine: Pore formation  

    Nets; Neutrophil extracellular traps; release of DNA and capture of the microorganism.
  • How do phagocytes recognize pathogens?
    Due to PAMP-PRR interaction. Different PRRs (often Toll like receptor) can recognize specific groups of pathogens.
  • What are the most important functions of the inflammatory barrier?
    Attracting immune cells and the complement system (local) and production of cytokines (systemic).
  • How do neutrofiles migrate trough the layer of endothelial cells and what is their function?
    The blood flow changes and endothelial cells are activated after injury. This causes influx of neutrofiles.; fagocytosis and formation of "pus".
  • Why is clotting important in the inflammatory respons?
    For producing networks to "capture" the pathogen; isolating injury site.
  • Why is the complement system important in the inflammatory respons?
    The complement system is able to:

    • Activate/attract phagocytes (killing bacteria)
    • Lysis target cell
    • Opsonization (marking bacteria)
    • Clearance immune complex (co-working with antibodies)
  • What is the strenght of the complement system?
    Immediate reaction, high quantity, tightly regulated, killing and "call for help" by chemokines (double action), working together with adaptive system.
  • Explain how the innate and adaptive immune systeme interact (direct and indirect)
    • Direct: via Fc-receptor (antigen bound antibody binds macrophage).
    • Indirect: via complement receptor (antigen bound antibody binds complement).
  • Explain the 3 possible ways for opsonization (=marking the pathogen for phagocytosis)
    Pic.
  • Which cells form a bridge between the innate and the adaptive system? How?
    Pic.
  • What is the difference between an antibody and a TCR (binding a pathogen)?
    • TCR: epitope, linear ("produced and presented")
    • Antibody: epitope, 3D, on pathogen
  • What types of antigen presentation do you know?
    • MHCII: presenting exogen antigens by proff APCs.
    • MHCI: presenting endogen antigens by all cells.
  • What are the three ways of antigen uptake?
    Pic.
  • What is essentiel for antigen processing after uptake by an APC?
    Acid millieu
  • What are the three important signals that a dendritic cell recieves?
    • What? Pattern recognition
    • Where? First contact, tissue factor
    • How much? Too little; ignore. Too much; down regulation.
  • Summary of the humoral and cell mediated response
    Pic.
  • How does a cytotoxic T-cell contribute to the immune respons?
    • Attacks cells infected by a virus and tumor cells by inducing lysis.
    • They release granule content into self-made pores.
  • Explain the structure of an antibody (short)
    Pic.
  • How is diversity of antibodies achieved?
    Hypervariable parts are formed by recombination of VDJ gensegments during development of de Bcel.
  • How do bcells develop in the bonemarrow and in the lymphoid tissue?
    Bonemarrow:
    • Gene recombination
    • Different types of mature Bcells
    Lymphoid tissue:
    • Antigen recognition
    • Proliferation and differentiation into plasma and memmory cells.
  • What are the four signals in APC CD4+T cell interaction?
    • Signal 1: TCR and MHC+peptide ánd CD4 costimulation is signal 1 (TCR MHC interaction is too weak).
    • Signal 2: Costimulation by CD28 on the Tcell and CD80/B7 on the APC.
    • Signal 3: Polarizing factors from the APC.
    • Signal 4: Adressing
  • What is the main difference between CD4 and CD8 costimulation?
    CD8 T cells need IL2 (licence to kill) for activation.
  • What is the site of T cell development?
    Production of thymocytes in bone marrow, development in thymus, activation/stimulation in the lymph nodes.
  • What is the site of B cell development?
    Production and development in the bonemarrow
  • What determines the isotype of the BCR and why is the isotype important?
    The heavy chain determines tyhe isotype of an Ig molecule. The isotype determines the effector function (neutralisation, opsonisation, complement activation).
  • What is a characteristic of immunoglobuline structures?
    A variable and a constant domain.
  • What is the difference between MHCI and MHCII?
    • MHCI:
Read the full summary
This summary. +380.000 other summaries. A unique study tool. A rehearsal system for this summary. Studycoaching with videos.

Latest added flashcards

Explain how valve based microfluid controls articficial micro-environments.
Due to valves, cytokines, hormones and other growth factors can be secreted in bursts. This highly influences the output of single cells.
Explain what a beadline capture antibodies is and how this contributes to multiplexed and longitudinal cytokine detection.
In a droplet, a beadline of capture antibodies in the middle. Produced cytokines are fluorescent.
Positive for cyt1: beadline shown. etc.
How can the best pDCs for pDC vaccination be selected? Mention microfluid droplets, microscopy and flow cytometry.
The interaction of the pDC and CTL needs to be investigated.
Input of CpG --> output of pDC (on CTL) --> output of CTL (tumor). 

De cellular interactions between pairs of single cells can be also measured in droplets (tip loading; controlled pairing of CTL + pDC). 

Microscopy for longitudinal effect; killing and cytokine secretion.
Flow cytometry for measuring the end-point; high throughput and multicoloar analysis.
Why is pDC heterogenticity important in pDC vaccination
The quality of the CTLs dictated by the pDC is very important. The best pDCs should be selected.
What can be concluded about (the heterogeneity in) the pDC derived type I IFN response?
Very heterogeneous; the (amount of) cytokine production differs, also when pDCs are stimulated the same.

The higher the amount of CpG, the more cytokine production. But not in all cells!
Primed pDCs (with CpG) show a increase in IFNalfa secretion.
How can be investigated how the type I IFN response is regulated on the single cell event?
With a droplet based microfluidic and with flow cytometry platform the amount of IFN I can be determined.

Droplet formation with IFN I capture beads.
Incubation (?) and cooling afterwards to stabilize droplets.
Secreted cytokines bind capture beads confined within the same droplet. 
Labeling with antibodies against IL-2, TNF-α, and IFN-gamma
Staining of the beads was measured by flow-cytometry.
pDC IFN I production can be measured with droplet based microfluidics. What do you already know about pDCs?
Plasmacytoid DC's are antigen presenting cells (less tha 0,1% of all leukocytes), major IFN I producers and critical for anti viral immunity.
They link innate and adaptive immunity. 
pDCs are a heterogeneous family (some have innate functions and secrete IFN I, some adaptive immune functions etc). 

When disregulated; hyperactivated; auto immune disease.
Which aspects of immunology can be measured with droplet based microfluidics?
Signalling:to detect secretion of cytokiness; droplets contain capture antibodies against the cytokine of interest and fluorescent detection antibodies. 

TCR/BCR: co-encapsulate single cells and poly(dT) magnetic beads together with lysis buffer. In individual droplets, cells are lyzed and mRNAs are annealed to poly(dT) beads which are subsequently recovered and emulsified for cDNA synthesis. PCR, sequencing. 

Antibody screening: cells are co-encapsulated together with fluorescent ab capture. 

(Same as nanowells)
How does droplet-based microfluidic technology works?
  • Cell isolation and modification: Cells of interest are isolated and form droplets together with cytokine capture beats/cytokine catch reagens. Stimulated with CpG (DNA) to stimulate cytokine production.
  • Encapsulation of single cells in droplets.
  • Incubation in droplets


Cytokines are captured inside the droplets with cells and can after this be measured with fluerescent antibodies....? 
What are the con's of microfluid based nanowell technology?
Still limited by the size of the plate
Pipetteren is lastig
Bij was en incubatie stappen moet statistisch worden berekend hoeveel wellen er zijn gevuld enz.