Curing and preventing method

A technology of reagents and receptors, applied in the field of chronic immune-mediated inflammatory diseases, dynamics and/or screening of animal models of the reagents, can solve problems such as glomerulonephritis that cannot be prevented

Inactive Publication Date: 2005-11-02
WALTER & ELIZA HALL INST OF MEDICAL RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

G-CSF-deficient mice do not develop neutrophil-mediated glomerulonephritis, but do not prevent T cell/macrophag

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0112] Mouse

[0113] C57BL / 6 (B6; wild type, [WT]) mice were obtained from Walter and Eliza Hall Institute (WEHI) Animal Supplies (Victoria, Australia). G-CSF-defective (G-CSF - / -) Mice were obtained from Ludwig Institute for Cancer Research, Victoria, Australia, and were produced by directed destruction of the Cysf3 gene on 129 / OLA embryonic stem (ES) cells, and injected the gene into B6 blastocysts (Lieschke et al. , Blood 84: 1737-1746, 1994). The mice were backcrossed to the B6 background for more than 20 generations. All mice are: ≥8 weeks of age at the time of the experiment, fed standard rodent feed, and drank water ad libitum, and were housed (≤6 mice / cage) in a cage containing sawdust. All animal methods were approved by the Institutional Ethics Committee.

Embodiment 2

[0115] Induction of mBSA / IL-1-induced arthritis (acute arthritis)

[0116] This method is based on a previously described technique (Lawlor et al., Arthritis and Rheumatism 44:442-450, 2001). The mice were anesthetized, and 10 μl of 20 mg / ml mBSA (Sigma, St Louis, MO) was injected into the knee joint by the intra-articular route. The control joints received the same volume of vehicle (normal saline). Then 20μl of 12.5μg / ml recombinant human IL-1β (specific activity 5×10) in normal saline / 0.5% (v / v) normal mouse serum (vehicle) was injected subcutaneously (s.c.) 8 U / mg; Amgen, Thousand Oaks, CA) was injected into the hind paw pad of the mouse, and the injection was repeated on the second day.

[0117] On the 7th day (or at the indicated time point) the mice were slaughtered, and the knee joints removed, and fixed in 10% (v / v) neutral buffered formalin solution for at least 2 days, decalcified and performed Paraffin treatment. Frontal tissue sections (4 μm) were cut at four dep...

Embodiment 3

[0120] Induction of collagen-induced arthritis (CIA)

[0121] Chicken type II collagen (CII; Sigma) was dissolved in 10 mM acetic acid at a concentration of 2 mg / ml overnight at 4°C, and emulsified in an equal volume of Freund’s complete adjuvant (CFA). The agent was prepared to a concentration of 5 mg / ml by adding heat-inactivated Mycobacterium taberculosis (strain H37 Ra; Difco Laboratories, Detroit, MI, USA) to Freund's incomplete adjuvant (Difco). 100 μl of the emulsion was injected into several parts of the tail base of the mouse by intradermal (i.d.) injection, and the above injection was repeated 21 days later.

[0122] The animals were monitored for erythema and limb swelling, and each mouse was clinically scored 3 times a week until 40 days. The scoring system is as described above (Campbell et al., European Journal of Immunology 30:1568-1575), where 0=normal, 1=slight swelling, 2=severe swelling, and 3=joint deformity and / or stiffness, and each mouse The maximum ...

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Abstract

The present invention relates generally to a method for treating or preventing or otherwise ameliorating the effects of inflammatory conditions such as but not limited to chronic immune-mediated inflammatory diseases. The present invention further provides pharmaceutical compositions comprising agents which inhibit one or more inflammatory cytokines and/or which down-regulate expression of genes which encode inflammatory cytokines. Such compositions are useful in the treatment and prophylaxis of inflammatory conditions such as inflammatory arthritis amongst other chronic immune-mediated inflammatory diseases. The present invention further provides an animal model for studying the kinetics of and/or screening for agents useful in the treatment or prophylaxis of inflammatory conditions.

Description

Invention field [0001] The present invention generally relates to methods for treating or preventing or ameliorating the effects of inflammatory states, such as, but not limited to, chronic immune-mediated inflammatory diseases. The present invention also provides a pharmaceutical composition containing an agent capable of inhibiting one or more inflammatory cytokines and / or capable of down-regulating the expression of genes encoding inflammatory cytokines. The composition can be used to treat and prevent inflammatory conditions, such as inflammatory arthritis, and other chronic immune-mediated inflammatory diseases. The present invention also provides an animal model for studying the kinetics of an agent for treating or preventing inflammatory conditions and / or screening the agent. Background of the invention [0002] The bibliographic details of the references provided in this specification are listed at the end of this specification. [0003] In this specification, the referen...

Claims

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Application Information

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IPC IPC(8): A01K67/00
Inventor K·E·劳洛尔I·P·维克斯I·K·坎贝里A·W·罗伯特斯D·梅特卡夫
Owner WALTER & ELIZA HALL INST OF MEDICAL RES
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