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MIMM387 Lecture 5 Exercise 20: Macrophage Assays

- This document discusses macrophage assays and properties of macrophages and other immune cells. It covers topics like macrophage morphology, functions like phagocytosis, and experimental techniques for isolating and studying macrophages.

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Jared Berger
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49 views14 pages

MIMM387 Lecture 5 Exercise 20: Macrophage Assays

- This document discusses macrophage assays and properties of macrophages and other immune cells. It covers topics like macrophage morphology, functions like phagocytosis, and experimental techniques for isolating and studying macrophages.

Uploaded by

Jared Berger
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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MIMM387 Lecture 5

Exercise 20: macrophage assays

- Dendrites stretch out between the lymphocytes


- In the macrophage, you can see a kidney shaped or indent nucleus (Do not look like that in vivo,
only when fixed)
- Blood smear
- You can see the different morphologies of cells

- All lymphocytes and monocytes are called mononuclear cells because they have a single round
nucleus

- Polymorphonuclear leukocyte – lumpy segmented nucleus

o The granules do not stain blue or red --- Neutrophil = phagocytic


- Monocyte
o Kidney-shaped nucleus
o Double the size of RBC
o They have a reasonable amount of cytoplasm
o Phagocytic
- If we take a blood smear from a normal person, what do we find out?
o If you have normal levels of RBCs, WBC, etc
- Granulocytes:
o Neutrophils have a releatively short half-life
o Basophils are a transport form of Mast Cells
o The granules are stain with a blue dye /basic
- Agranulocytes
 Mononuclear cells
 Th, Tc, B cells
 NK cells
 Monocytes
- Non-nucleated blood cells
 RBCs and platelets
- Latex beads that are coated with C3b
- The anaphylaxin toxin creates a gradient that allows the APcs to sense that the bacteria is there
- The macrophage/dendritic cell contains CR1 receptor for opsonisation

- Swallow and dispose senescent cells


- Tissue reorganization: The tissue being broken down/etc during development
- Indices inflammation – release cytokines (IL1)
- Anti-bacterial compounds
- Tissue repair and wound healing
- Lymphocyte activation and immunity
- Tumorcidal responses

- M-CSF = colony stimulatory factor to get the bone marrow to make more macrophage
- G-CSF = colony stimulator factor to get the bone marrow to make more macrophage
- Nitric oxide and peroxinitride (NO, ONOO) can travel through cell membranes and directly
damage the DNA of cells
- We inject a mild irritant into the peritoneal cavity of the mouse in order to increase cell yield
o Can use TsA
- Every time you have an irritation, there is some activation of IFNa
o Can slightly activate the cells
- We wait four days by the time we do the experiment so the mononuclear cells will be mostly
present in the peritoneal cavity
- You pop the needle through the two membranes
o There is the skin and the abdominal wall
- The fluid is now inside the abdominal cavity
- You injected the medium in red (ph = 7), and it came out yellow why?
o The mouse is not breathing, thus glucose will be converted into lactic acid

- SL = Small Lymphocytes
- LL= Large lymphocytes
- Monocyte: Indent in the nucleus
- You do the injections directly below the spleen
- Punch cells in the agarose
- Place macrophages in the hole
- Put a control on the laeft and an attractant on the right
- We can now do 96 migrations at a time

- The cells will migrate into the membrane


- Measure how many um go into the membrane after their fixation and staining

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