CHAPTER 2: SELF-STUDY QUESTIONS

-What is the mucosal epithelium and which cells and barriers can you find there?

The mucosal epithelium is a one-cell layer thick barrier between the outside world and the host. For
example, the gut epithelial layer or the epithelium of the lung. The epithelial barrier is kept firmly
closed through so-called tight junction proteins. Different epithelial cell types protect the barrier
from breaching, such as: 1) Goblet cells; they make mucus and by peristalsis the bacterial trapped in
the mucus will be transported out of the gut, 2) Paneth cells (in mammals) in the bottom of the
crypts that make anti-microbial peptides, 3) absorptive enterocytes that also can secrete
antimicrobial peptides and are selectively taking up nutrients. 4) enteroendocrine cells that secrete
hormones important for satiety and bowel movements.

-How can the host distinguish between self and non-self?

Our immune system contains cells that are only specific for non-self. For example, innate cells such as
macrophages and neutrophils have receptors (TLRs etc.) that recognize foreign (bacterial) molecules
but not self-proteins. T and B cells have specific receptors that are only activated when they
recognize foreign peptides and proteins. T cells recognize peptides in the context of an MHC (Major
Histocompatibility Complex) molecule on the surface of host cells. We have two types of MHC: MHC
class I and MHC class II. MHC class I is present on the surface of all cells with a nucleus (so basically all
cells except red blood cells in mammals). The cell is a small factory and it has a lot of products. The
MHC class I molecule does nothing else than to show the products the cells is making on the outside
of the cell. It ‘presents’ it to the immune system. Now if the cell is healthy the peptides in the MHC
class I molecule on the cell are not ‘recognized’ by the surveying T cells. They see them, but do not
attach or respond with activation. If a cell is infected with an internal bacterium or a virus, also these
products are made inside the cell and they also end up in the MHC class I. These peptides the T cells
(CD8+ T cells or cytotoxic T cells) do recognize as ‘foreign’. They will bind and get activated and kill
the infected cell immediately, to prevent spread of the infection. The other MHC molecule is MHC
class II. This molecule is only present on professional antigen presenting cells such as macrophages, B
cells and dendritic cells. These cells can eat bacteria and other microbes/viruses when they infect the
host. The peptides of the broken down bacteria will end up in the MHC class II molecule of the
antigen presenting cell and this will be recognized by the T cells. These T cells (CD4+ cells) do not kill
the immune cells (that would be a waste!) but instead make cytokines to combat the infection and
provide help to B cells to make antibodies specific for the infectious agent.

-What DAMPs, PAMPs and MAMPs? Give examples

Conserved structural motifs only present in bacteria are pathogen-associated molecular patterns or
microbe-associated molecular pattern. We now tend to call them microbe-associated molecular
pattern since there are very little of those molecules that are only pathogen specific.

DAMPs are danger-associated molecular patterns that are associated with cell stress. These
molecules are normally not present outside of host cells and are therefore seen as ‘foreign’ if
detected.

Examples of MAMPs/PAMPs: LPS, peptidoglycan, flagellin

Example of DAMPs: Heat-shock proteins, S100 proteins, Uric Acid

-Describe molecules produced by the microbiota that can be recognized by the host as non-self

LPS, Lipotechoid acid, peptidoglycan.

-What are Pattern Recognition Receptors?

Receptors on host cells that can recognize PAMPs/MAMPs and DAMPs. When they bind their ligand a
signalling cascade starts that activates the cell and cytokines are induced to augment the immune
response.

-Which bacterial ligands are recognized by the most important Toll-like receptors (TLRs)?

Toll like receptor ligands recognized

TLR1/2 peptidoglycan and (triacyl) lipopeptides

TLR2 peptidoglycan and lipoteichoic acid

TLR2/6 peptidoglycan and lipoteichoic acid

TLR4 lipopolysaccharides (LPS)

TLR5 Flagellin

NOD1 D-glutamyl-meso-diaminopimelic acid (DAP); component of peptidoglycan

NOD2 muramyl dipeptide (MDP); component of peptidoglycan

-Why do some microbe associated molecular patterns (for instance LPS) trigger either a pro- or an
anti-inflammatory response?

The lipid A molecule of LPS, is perceived by the host Toll-like receptor (TLR4), a cell surface-located
pattern recognition receptor. TLR4 is the main animal sensor of microbial LPS. The lipids end in acyl
chains, and different numbers of phosphate groups or carbon atoms in the acyl chains makes the LPS
binding stronger or weaker to TLR4. Such differential binding to TLR4 leads to different chemical
signals being released in the immune cell, resulting in a different type of immune response. As an
example, although Bacteroides and Escherichia both belong to Gram-negative bacteria and produce
lipopolysaccharide (LPS), the chemical composition of the LPS of Bacteroides is different from E. coli
LPS. Moreover, there is a difference in how the human immune system responds to Bacteroides and
E. coli LPS. Whereas E. coli LPS is a potent innate immune activator, Bacteroides LPS is in fact
inhibiting innate immune signalling. Thus, chemical variation in common microbial structures (here,
the phosphate groups and/or acyl chains of LPS) makes it possible for immune cells to distinguish
between bacteria, and to adjust their immune response accordingly. Indeed, chemical variation is a
key feature of microbial structures and various small molecular decorations can be perceived and
used by a wide variety of animal immune receptors. Resulting immune responses can range from
excessively inflammatory to tolerant, and thereby will have substantial influence on host
homeostasis and health.