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How to find glial cells in lymphatic tissue: The problems of desmin or GFAP expression in stromal cells and oligodendrocytes being out of their element

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Abstract

We confirmed the presence of wired antigen presenting cells (wAPC) in lymph nodes [1,2]. →We were able to identify Schwann cells in the hilum, medullary and capsule region of the lymph node (I). →We found oligodendrocyte-like cells in the lymph node regions mentioned above regions, myelinating more than one axon (II). →GFAP positive signals could be found densely distributed throughout the whole lymph node (III). →The used polyclonal GFAP antibody also marked desmin in stromal cells in cortical, paracortical and medullary regions (IV). →These questionable results (signal for desmin and GFAP) also appeared in many other non-lymphoid organs (V)-(table) →Taking the results together one question arises: Who takes over the glial function of accompanying axons in the lymph node B and T-cell regions? (VI)
Summary
We confirmed the presence of wired antigen presenting cells (wAPC) in lymph nodes [1,2].
We were able to identify Schwann cells in the hilum, medullary and capsule region of the lymph node (I).
We found oligodendrocyte-like cells in the lymph node regions mentioned above regions, myelinating more than one axon (II).
GFAP positive signals could be found densely distributed throughout the whole lymph node (III).
The used polyclonal GFAP antibody also marked desmin in stromal cells in cortical, paracortical and medullary regions (IV).
These questionable results (signal for desmin and GFAP) also appeared in many other non-lymphoid organs (V) (table)
Taking the results together one question arises: Who takes over the glial function of accompanying axons in the lymph node B and T-cell
regions? (VI)
How to find glial cells in lymphatic tissue: The problems of desmin or GFAP expression in
stromal cells and oligodendrocytes being out of their element
C. Wülfing1, H. Günther1, S. Henne1, J. Oehlmann1, J.Urban1, D. Pleizier1, N. Renevier1, C. Lohr2
1 Group for interdisciplinary neurobiology and immunology, Biocenter Grindel, University of Hamburg
2Division of Neurophysiology, University of Hamburg, Hamburg, Germany
Home page and contact info: www.ini-research.org
Results
Figure 1:Ultrastructure image by TEM of a nerve within the medulla of an inguinal lymph node.
Figure 2: Ultrastructure image by
TEM of a nerve within the medulla
of an inguinal lymph node. (A)In
addition to myelinated (white
arrowhead) and non-myelinating
Schwann cells (white arrow). (B)
Surrounded by the endoneurium
(black arrow), there are two nerve-
accompanying cells (a and b) that
obviously myelinate more than one
axon (red asterixes). This structure
is completely surrounded by a
basement membrane (black
arrowhead).
Figure 3: Left site (AD),
immunohistochemical staining
with polyclonal anti-GFAP
antibody (DAKO) in different
areas of the lymph node. Scale
bar for AD50 µm. HEV= High
endothelial venules (HEV). Right
site (E) silver staining of lymph
node cross section. C=
capsule,T= T-cell region, M=
medulla. GFAP signal crosses the
subcapsular sinus (arrowheads
below the capsule (arrows)
Figure 4: Mandibular lymph node co-
stained with polyclonal anti-GFAP
antibody (DAKO) in red and desmin in
orange. DAPI for nuclear staining in
blue. Desmin and GFAP signals are co-
located throughout the tissue. The
enlargements show the reticular
structures, typical for stromal cells,
distributing through the lymph node
parenchyma.
I Identification of Schwann cell like structures in murine lymph node
II Oligodendrocyte like cells myelinating several axons
III GFAP immunoreactivity is densely distributed throughout the lymph node
IV GFAP and Desmin costaining reveals stromal cells as target for GFAP antibody
V Binding of GFAP polyclonal antibody to desmin in other organs
Desmin GFAP poly GFAP mAB
WB IHC WB IHC WB IHC
Brain
- - x x x x
Heart
x x - x - -
Lymph
node x x x x - -
Liver
x x - x - x
Thymus
x x - x - -
Spleen
x x - x - -
Skin
x x - - - -
Lung
x x - - - -
WB= western blot; IHC= immunohistochemistry staining; GFAP=Glial fibrillary acidic protein; mAB= monoclonal
antibody
Conclusion
Further details see publication: Günther, H.S., Henne, S., Oehlmann, J. et al.GFAP and desmin expression in lymphatic
tissues leads to difficulties in distinguishing between glial and stromal cells. Sci Rep 11,13322 (2021).
Oligodendrocytes are so far exclusively described in the CNS and to detect them
in the periphery was a fascinating discovery which still has to be confirmed by
further experiments. One possibility is, that these oligodendrocytes are
responsible for taking over the axons from their accompanying Schwann cells in
the Hilum and medullary regions passing them over to the cells that have glial
function in the lymph node B- and T-cell area. As here Schwann cells are
obviously missing, the question remains: Who takes over the role of
accompanying axons in the lymph node Cortex and Paracortex ? The results of
a GFAP antibody marking seemingly desmin in stromal cells opens up another
question: Can stromal cells take over glial function and is there a new
intermediate filament type associated with that function ?
[1]: Wülfing, C., and Günther, H. Dendritic cells and macrophages neurally hard-wired in the lymph node. Sci Rep 5, 16866 (2015)
[2]: Wülfing, C., et al. (2018). Neural architecture in lymphoid organs: Hard-wired antigen presenting cells and neurite networks in antigen entrance areas. Immunity, inflammation and disease, 6(2), 354370.
VI Who is accompanying axons taking over the glial function in the lymph node?
Figure 5: Lymph node co-stained with GFAP in orange, Collagen type 3 (Coll 3) in red and neurofilament (NF) in green. A
GFAP and Coll 3 show the network of fibroblastic reticular cells and the conduit system in the T-cell region of a lymph
node. The orange signal for GFAP in the cytoplasm of the cell lines the margins of the tubular conduit filled with Coll 3
(red). B From the magnification (white framed) one can already see how a NF signal (green), tightly associated with a
wAPC (arrow), seems to break out of the conduit. C In the 3D view of a Z-stack, one can clearly see how this axonal
structure (green) moves out of the conduit and is connected to a cell that we would describe as wAPC (arrow) .
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