Stromal Immunology
PHOL0008
Dr Sophie Acton
Summary
• Understand stromal architecture of lymphoid tissues
• Know which cell types contribute to lymphoid tissue structure and
what functions they perform
• Examples of the active involvement of stromal cells in immune
responses
Adaptive immunity
• Fast vs. slow
• General vs. specific
• One time vs. memory
• Requires antigen
• Requires B and T cells
• Long lasting protection
What is Stroma?
noun
ANATOMY•BIOLOGY
the supportive tissue of an epithelial organ, tumour, gonad, etc., consisting of connective tissues and blood
vessels.
The connective tissue forming the framework or matrix of an organ or part
The colorless framework of a red blood corpuscle or other cell
The Greek word "stroma" means "anything spread out for sitting or lying upon," essentially a mat.
The stroma in anatomy is thus the supporting tissue.
Stromal Cells
• In the context of Immunology, stromal cells = non-immune cells
• In lymphoid tissues, stromal supports immune cell function
• Thymus
• Bone marrow • Fibroblasts
• Lymph nodes • Lymphatic endothelial cells
• Spleen • Blood endothelial cells
• Payers patches • Pericytes
• Tertiary lymphoid tissue
Lymphoid Tissues
• Primary
bone marrow
thymus
• Secondary
lymph nodes
payers patches
spleen
tonsils
• Tertiary
pathology
Cells of the immune system
• Diversity of immune cell
types
• Requirements for survival
• Requirements of activation
• Requires specific niches in
tissues
• Immune responses require
specific interactions
between cell types
The need for a niche…
• What is a niche?
• How does stroma support a niche?
• How is immune function altered when stromal niches fail to function
properly?
Bone marrow
The bone marrow niche for haematopoietic stem cells
Sean J. Morrison & David T. Scadden
Nature volume 505, pages327–334(2014)
Thymus
DOI:10.1038/nri1883
The thymus and T-cell commitment: the right niche for Notch?
Eric J. Jenkinson, William E Jenkinson, +1 author Graham Anderson
Thymic selection
• Editing of naïve T cell
repertoire
• protection against
autoimmunity
• Killing of cells which
recognize self antigens
Spleen
• Similar organization to lymph
nodes
• Sites of clearance for ‘old’
immune cells
• Red pulp/white pulp
Peyers patches
• Formed along the intestines,
• Filters fluids from the gut
• Home to immune cells,
• First responders
Lymph node
• We will focus on lymph node as our model of secondary lymphoid
tissue.
• Best studied, but still much we don’t know
• Stromal immunology is an emerging field within immunology
Lymph node architecture
B cells
B cells
T cells
B cells
T cells
B cells
B cells
B cells
B cells
Lymph node architecture
Collagen
B cells
Lymph node architecture
• Organisation and Lymph flow
separation of cell types
Afferent lymphatics
Subcapsular sinus
• Cell trafficking
controlled by stromal High endothelial venules
cells
Fibroblastic stroma
• Fibroblasts
• Blood vessels Efferent lymphatics
• Lymph vessels
Lymph flow
Lymph node structure
PDPN/Collagen IV
Victor Martinez
Lymph node architecture
Subcapsular
sinus HEV’s
B-cell follicle T-cell zone
HEV’s
Tilescan 40x oil CD3 – T-cells PDPN
Lymph node structure
Lymph node development
Lymph node development
• Lymphatic endothelium are
required to initiate formation
• LTi – lymphoid tissue inducer cells
instruct the first stromal cells to
express FRC markers
• LTo – lymphoid tissue organizer
cells, start to express chemokines
and recruite immune cells
doi: 10.1242/dev.028456
Break
• What is stroma?
• Why is stroma important to immune function?
• What cell types act as stroma for immune function
See you in 10 minutes
Stromal cell types and their functions
• Endothelial cells Lymph flow
• Lymphatics Afferent lymphatics
• Blood
Subcapsular sinus
• Fibroblasts High endothelial venules
Fibroblastic stroma
• Pericytes Efferent lymphatics
Lymph flow
Blood vessels
• HEV – high endothelial venules
• Specialised for immune cell trafficking
• Control flow of lymphocytes in and out of lymphoid tissues, similarly
to blood vessels in our other tissues
• Express integrin ligands
• Express chemokines
• How to HEVs allow continuous trafficking of immune cells yet remain
an intact barrier to prevent bleeding in lymphoid tissue?
Regulation of HEV function
https://doi.org/10.1111/imr.12414
Lymph Node Architecture - FRCs
• Fibroblastic reticular cells
• Most abundant
• Span the full volume
• Provide structural support
• Highly contractile
FRC
FRC nuclei
FRC functions
• Routes of cell trafficking
• Source of growth factors
• Control conduit flow
• Control lymph node expansion
• Act as pericytes for HEVs
FRC subsets
• Phenotype and function
is adapted to specific
microenvironment
• Varying FRC functions
defines different niches
within lymphoid tissues
• Single cell RNAseq has
allowed us to define
different FRC subsets
https://doi.org/10.1016/j.immuni.2018.04.006
FRCs for dendritic cell trafficking
• Provide CCL19
• Provide podoplanin
• Provide physical route
https://doi.org/10.4049/jimmunol.1402520
Dendritic cells are the
immune sentinels of
our body
Chemokine signalling
• https://www.sciencedirect.com/topics/medicine
-and-dentistry/chemokine-receptor
Step 1 – Cell migration
Chemokines - a family of small cytokines, or signaling
proteins secreted by cells. induce directed chemotaxis in
nearby responsive cells; they are chemotactic cytokines.
CXCL13
CXCL13
CCL19
CXCL13
CCL21
CCL19
CCL21 CXCL13
CXCL13
CXCL13
CXCL13
CCR7 – for lymph node homing
• In CCR7 KO mice, organization is scrambled https://doi.org/10.1016/S0092-8674(00)80059-8
CLEC-2 / podoplanin interaction – DC migration
CLEC-2 / podoplanin interaction – LN expansion
CLEC-2 / podoplanin interaction – LN expansion
PDPN - Actomyosin contractility
CLEC-2 CLEC-2
CLEC-2
P P P P
D D D D
P P P P CD44 CD44
Cholesterol-rich
N N N N
domain
pS S
ezrin ezrin ezrin
actin
cytoskeleton z rin ez
rin
e
RhoA RhoA
Contraction Relaxation/stretching
ROCK
PDPN - Actomyosin contractility
** **
*** ***
FRCs + control DCs FRCs + CLEC-2 deleted
** DCs **
n.s. n.s. n.s. n.s.
14 80
12
** 60 **
LN cellularity (x10^6)
10
LN mass (mg)
8
n.s. n.s.
40
n.s. n.s.
6
4
20
0 0
_ _ _ _
CFA/OVA + + CFA/OVA + +
CLEC-2-Fc _ _ CLEC-2-Fc _ _
+ + + +
Lotte de Winde
Control Control
CD11c∆CLEC-2 CD11c∆CLEC-2
Acton et al. Nature 2014
FRCs associate with the conduit network
Ian White
Conduit flow in inflammation is fragmented
Dendritic cells inhibit matrix deposition by FRCs
ACUTE INFLAMMATION LN EXPANSION
CLEC-2+ DCS ECM
turnover
X Acute
Inflammation
Acute
Inflammation
ECM
turnover
Conduit
Flow?
Martinex et al. Cell reports 2019
FRC polarity
Microtubule
LL5b/a
Hotta A et al. J Cell Biol. 2010 Roozendaal R et al. Int Immunol. 2008
LL5beta associates with conduits in vivo
PDPN
Laminin
LL5b
Victor Martinez
Podoplanin
• Control DC trafficking
• Binds to CCL21
• Controls FRC contractility
• Upregulated in inflammation
Tertiary lymphoid tissue
• Develops during inflammation
• Cancer
• Autoimmune diseases
• Functions in similar way to secondary lymphoid tissue but structure is
less well defined
• Might get small areas or large areas developing, across multiple
regions of disease.
Ectopic lymphoid follicles
https://doi.org/10.1016/j.immuni.2011.10.015
Cancer associated fibroblasts
Zoom
PDPN MHCII PDPN WT PDPN KO.
Tumour microenvironment
Pancreatic cancer – highly stromal
What happens without functional stroma?
Viral infection
Eg. HIV
Summary
• Stromal immunology is a new and rapidly developing area if interest
for immunology research
• Its time to start understanding cell biology and immunology in the
context of tissues
• Cells don’t function in isolation, they adapt to their
microenvironment, and their microenvironment supports their
function.
Reading list
• Fibroblastic reticular cells in health and disease – Fletcher et al. 2015
• Fibroblastic Reticular Cells: Organization and Regulation of the T
Lymphocyte Life Cycle – Brown et al. 2015
• Single-Cell RNA Sequencing of Lymph Node Stromal Cells Reveals
Niche-Associated Heterogeneity. – Rodda et al. 2018
