Mouse CSF / Rat CSF serial sampling tables, Cytokines Classification (Interleukins / Chemokines / Interferons / TNF / Colony Stimulating / TGFβ), Interleukins (IL-1β / IL-6 / IL-9 NLRP3 narrative), Interferons (Type I / Type II / IFN-γ MHC class II), interferon-stimulated gene (ISG) MOA (JAK-STAT / IFNAR / ↑ ISG gene expression), Multiplex cytokine assay (Mouse / Rat / Human Luminex / Olink panels)

Serial sampling — Examples

MOUSE CSF

	Why	Olink can
ARNJ	Immunoassay needs minimum 25 uL, but mice usually can produce 10 uL, so the gene therapy project. AAV was injected into cisterna magna in mice now. Thereafter, the treated mice were alive before AHC study. By adapting this method, serial CSF can be obtained in mice. (Bergquist 1996, Nasal MS Pharmacol) 3. Also, as I mentioned, simulation around cisterna magna may allow us to do it. One of our collaborators in Osaka 1) possesses that technique. We have connection to Dr. Osaka Takeda from Osaka Univ who developed non-invasive overnight accumulative CSF collection from canulae from free moving mouse
Aging cuff	See SOP of CSC (When the animals are 2-3 weeks old at this time, and it is not feasible to collect CSF at that age) Mouse CSF is available and require 10-20 uL.
(Riddle, 2022 #2521)	Detailed protocol using glass capillary \| 8-15 uL...

Rat CSF


(Pacegi, 2009 #1817)	RAT (10 uL, 사용 가능) Anesthetized..., is yielded 100-120 uL of CSF per rat.

Cytokines

Classification

Cytokine family	Functional subgroups	Physiologic effect
Interleukins	IL-1, IL-7, FN-γ-related	stimulate hematopoiesis
	IL-2, IL-4	regulate cell-mediated immunity
	IL-3, IL-4, IL-5, IL-13, IL-15	regulate T and B cell cooperation
Chemokines	α CXC	- regulate migration of granulocytes and lymphocytes - promote angiogenesis and inflammation - regulate maturation of monocytes - enable medullar homing - regulate migration of lymphocytes
	β CC
	γ C-X-C
Interferons	Type I (IFN α, β, ω, etc)	antiviral immunity / anti-proliferative effect / antianttotic activity / response to intracellular pathogens
Interferons	Type II (IFN γ)
tumor necrosis factors	TNFα	proinflammatory propergy, accolyses non-specific immunity, adhesive molecule expression on endothelial cells, may cause apoptosis, overproduction causes SIRS
tumor necrosis factors	TNFβ	similar in effect, but produced by T- and B-lymphocytes
Colony Stimulating factors	GCSF, GMCSF, MCSF	stimulation of granulocytes and macrophages
	Erythropoietin	stimulation of erythrocyte production
	Thrombopoietin	stimulation of thrombocytes
TGFβ	- stimulates growth of fibroblasts and extracellular matrix production - lead to MMPs inhibition

Interleukins

Genus	upstream		Downstream
IL-1β	IL-1β precursor is cleaved by cytosolic caspase 1 (interleukin-1 β convertase) to form mature IL-1β		Various: CAPS, scleroso, cancers
(McGonagh, 2012 #2783) IL-6 is a known downstream target of IL-1β, consistently increased in serum from patients with NLRP3 inflammasome-mediated conditions. However, an IL-6 wall background failed to provide phenotypic rescue and did not significantly impact inflammatory cytokine levels. In a second model of IL-1β driven inflammation, NLRP3 activation by monosodium urate crystals similarly increased IL-6, Consistent with this evidence the with NLRP3 model, ongoing of IL-6 did not impact an acute neurophasic response in this in vivo evaluation of gouty arthritis. Taken together, our results indicate IL-6 is a reliable marker of inflammation, with no direct role in inflammasome-mediated disease.
(Renga, 2018 #2704) IL-9 has a pro-inflammatory activity at the onset of VVC by promoting NLRP3 inflammasome activity

Interferons

Definition: A group of signaling proteins

Classes of interferons

type	Produced by
IFN-α	Most cell types including lymphocytes (NK cells, B-cells and T-cells), macrophages, fibroblasts, endothelial cells, osteoblasts and others
IFN-β
IFN-κ, ε, ω, δ and ζ
IFN-Y
IFN-γ		some antiviral activity, but is more important in establishing cellular immunity through activating macrophages and promoting major histocompatibility complex (MHC) class II

interferon-stimulated gene (ISG)

definition
- a gene whose expression is stimulated by interferon
- cf) interferon (?) 다른 다른 interferon gene 과 혼동하지 말 것
Examples
- So many, 10% of whole genes
MOA

	trigger release of IFNs	Inside Outside cell ?	Interferon activates the JAK-STAT signaling pathway. Interferons bind to interferon receptors. All type I IFNs bind to a specific cell surface receptor complex known as the IFN-α receptor (IFNAR) that consists of IFNAR1 and IFNAR2 chains.	transcription of ISGs is induced expression of proteins that will prevent the virus from producing and replicating its RNA and DNA(?)	↑ ISG gene expression
...of molecules uniquely found in microbes (eg viral glycoproteins, viral RNA; bacterial proteins (LPS); bacterial flagella; CpG motifs); by item recognition receptors (eg membrane Toll-like receptors or the cytoplasmic receptors RIG-I or MDA5); can

Multiplex cytokine assay

	Mouse	Rat	Human
R&D system	https://www.rndsystems.com/target/multiplex-cytokine-assay/mouse-of-cytokine-luminex-performance-prarmised-kit_lcstm20 / 50 uL	https://www.rndsystems.com/target/multiplex-cytokine-assay-kits/species/Rat	https://www.rndsystems.com/target/multiplex-cytokine-assay-kits/species/Human
	Proteome Profiler Rat Cytokine Array	Olink inflammation panel: 96
Relative level, serum, 29 cytokines, 1 Kit / $605 Olink: ~1 uL Proteome Profiler Rat XL Cytokine Array: Olink cytokine panel: 48~40, Absolute, Contains 4 membranes each
20uL custom list, part vs full? Detectability: 18 mouse cytokines, only 1 uL?

Uncertain Spans

location	transcription	uncertainty
MOUSE CSF / ARNJ row	`ARNJ` row label	reads as written (likely an OCR rendering of a Korean / English mix); preserved verbatim.
ISG MOA cell	trailing tokens `...molecules uniquely found in microbes`	the row prose is partially clipped at the left edge and inferred from the visible suffix; preserved verbatim.
Multiplex tail	`20uL custom list, part vs full? Detectability: 18 mouse cytokines, only 1 uL?`	trailing OCR fragment with operator-side question; preserved verbatim.