Background Information

The causal link between Group A streptococcal infection and rheumatic fever is firmly established, but the underlying mechanisms of the disease remain a subject of debate. Moreover, the range of pathologic lesions induced by streptococci remains inadequately defined. One approach to the study of these issues is to inject Group A streptococcal components into experimental animals and observe their effects. The effects of injecting SCW fragments have been studied in several species. Injection of...

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Divide rats into groups as required for experiment and label appropriately (e.g., using cage cards, ear tags, or ear notches see unit 1.5). Label one group as control rats (to be injected with saline) and another group as experimental (to be injected with 10S PG-PS). 2. Weigh rats and determine average body weight of each group. 3. If arthritis severity is to be monitored by measuring ankle diameter (also see step 8), individually measure maximal lateral ankle diameter of each rat with a...

Induction And Clinical Evaluation Of Arthritis

In this protocol, arthritis is induced in Lewis rats by an intraperitoneal injection of an aqueous suspension of Group A PG-PS (SCW), prepared as described in Support Protocol 1. Disease development is monitored by visual inspection or measurement of ankle diameters for periods of 6 to 8 weeks. Incidence of disease should approach 100 . Lewis rats, female, 6- to 12-weeks-old or 100 to 200 g, specific-pathogen-free (Harlan Bioproducts) 3 to 6 mg rhamnose ml 10S PG-PS suspension in sterile 0.85...

Induction Of Collageninduced Arthritis In The Mouse

The mouse model of collagen-induced arthritis (CIA) is particularly advantageous for study, given the vast number of inbred strains available and their corresponding immunologic reagents. Disease induction is more complex than that in other autoimmune disease models e.g., experimental autoimmune encephalomyelitis (EAE unit 15.1) in that CIA requires both the activation of T cells and the production of antibodies specific for both the immunogen and the autoantigen, murine CII. This protocol is...

Collagen Induced Arthritis

Collagen-induced arthritis (CIA) is an experimental autoimmune disease that can be elicited in susceptible strains of rodents (rat and mouse) and nonhuman primates by immunization with type II collagen (CII), the major constituent protein of articular cartilage. Following immunization, these animals develop an autoimmune-mediated polyarthritis that shares several clinical, histological, and immunological features with the human autoimmune disease rheumatoid arthritis. As with rheumatoid...

Induction Of Adjuvant Arthritis

In this protocol, adjuvant arthritis (AA) is induced in Lewis rats in the classical manner using ground, heat-killed Mycobacterium tuberculosis H37Ra suspended in incomplete Freund's adjuvant (IFA see Support Protocol). Disease development is monitored daily during the critical period (days 10 to 35 post immunization) using one or more possible techniques. The success rate of the procedure should approach 100 . 10 mg ml heat-killed Mycobacterium tuberculosis (strain H37Ra) suspended in...

Adjuvant Arthritis in the

Adjuvant arthritis (AA) is an induced form of (sub)chronic arthritis. Strains of rats have a varying genetic susceptibility to AA, whereas mice generally are not susceptible. AA is most easily induced with mycobacteria suspended in oil, although in some strains of rats it can be induced with oily adjuvants in the absence of mycobacteria. The disease is a T cell-mediated autoimmune arthritis that is frequently used to study immunological aspects of rheumatoid arthritis (RA) and other arthritic...

Preparation Of Group A Scw 10s Pgps

This protocol describes the preparation of Group A streptococci and the isolation and fragmentation of 10S streptococcal cell wall peptidoglycan-polysaccharide polymers (SCW PG-PS) to be used for inducing SCW arthritis. Streptococcus pyogenes, Group A, D58 strain, (ATCC 43202) Todd Hewett broth (Difco), pH 6.8 to 7.0, supplemented with 3 g dextrose liter, autoclaved 0.006 M PBS, pH 7.0 dilute PBS, pH 7.0 (appendix2) with H2O (final NaCl concentration 0.006 M) Dry ice ethanol bath 2 (w v) SDS in...

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APG-2, Hspl 10, Hsp70RY, Hspa4, Hspl05a, and Hspl05 3 Protein folding, clathrin 11 (q24.1) 9 Involved in antigen 5 (q31.1) 18 presentation and radioresistance. Also oncogenic (overexpression of Mot-2 leads to p53 inactivation and cell transformation). Binds to Hsc70 to form high-molecular-weight complex involved in protein folding, thermotolerance, and embryogenesis Constitutively expressed also induced by metabolic stress such as glucose starvation Leukemia-derived Hsc70 is now being tested as...

Support Purification Of Cyanogen Bromide Fragments From Protocol 3 Collagen a1Ii Chains

Because the a1(II) chains contain a relatively small number of methionyl residues (e.g., 10 residues per a chain in bovine CII), cyanogen bromide (CNBr) cleavage of bovine a1(II) yields a mixture of eleven peptides. By convention, the CNBr peptides were named according to their order of elution from the ion-exchange column (Fig. 15.5.1). The peptides range in size from dipeptide to peptides of 345 residues (Table 15.5.2). For studies of CIA in mice, the peptides CB11, CB8, and CB10 are of...

Assessment Of Arthritis In The Mouse And

The presence or absence of arthritis in both the mouse and rat model is relatively easy to ascertain. In both rodent models, any or all of the four paws can be affected. The inflammation at its peak extends from the ankle all the way through the digits and is characterized by extreme swelling and erythema. Once arthritis appears, each paw is examined two to three times a week. It is not uncommon to see a normal-looking limb one day, yet find it greatly swollen the next. Although the arthritis...

Scw Pg-ps 10s Rat Arthritis Model

Gender differences, however, are less apparent when relatively high doses of cell walls are injected. Normal euthymic and athymic Lewis rats have also been studied extensively (Case et al., 1989 Sano et al., 1990, 1992, 1993 Wilder et al., 1987). Compared to euthymic Lewis rats, athymic Lewis rats develop severe, acute transient arthritis, but chronic disease is markedly blunted, indicating that acute disease is thy-mus-independent, whereas chronic disease is highly...

Streptococcal Cell Wall Arthritis

Streptococcal cell wall (SCW) arthritis in rats is an experimentally-induced inflammatory model with many features that resemble rheumatoid arthritis (RA) in humans. The model is used widely in studies of genetics, pathogenesis, and therapy of acute and chronic inflammation. Arthritis is induced in an appropriately susceptible inbred rat strain (Table 15.10.1) by an intraperitoneal injection of peptidoglycan-polysaccharide polymers (PG-PS), which are isolated from the sonicated cell walls of...

Induction Of Collageninduced Arthritis In Rats

Collagen-induced arthritis CIA using the rat was originally described by Trentham et al. 1977 . Subsequently, this model has gained widespread acceptance and has proven valuable in the study of autoimmune-mediated polyarthritis. Although the mouse model of CIA is often favored for study because of its well-characterized genetic system and plethora of immune reagents, the rat model possesses a number of features that make it desirable. These include the availability of many CIA-susceptible...

Determining Arthritis Severity by Caliper Measurement of Paw Swelling

Hold the mouse or rat in a position that allows examination of the paws. 2. Place constant tension-calipers on the ankle medial lateral and determine thickness a minimum of two times to ensure accuracy. Constant-tension calipers offer the advantage of speed but can yield falsely low values for inflamed joints if the tension is too great. 3. Repeat procedure to measure the tarsal thickness dorsal ventral of the same paw. 4. Repeat steps 2 and 3 for each paw.