Tolerogenic Effect of Orally Administrated Type II Collagen in CIA Animal (DBA/1 mice) / 대한류마티스학회지

OBJECTIVE: To investigate the dosage of bovine type II collagen (BnCII) for the induction of oral tolerance in CIA animals, and to verify the changes of immune response and TGF-beta production of mesenteric lymph node cells in tolerized CIA animals. METHODS: Oral tolerance was induced by feeding of variable doses (5 microgram, 10 microgram, 20 microgram and 40 microgram) of BnCII to DBA/1 mice 4 times per week during 2 weeks, and control mice were given ovalbumin (1000 microgram), before immunization. We examed clinical assessment ; incidence of arthritis, severity of arthritis, arthritic limb by visual analysis. IgG antibodies to BnCII were measured by ELISA, T cell responses to BnCII and PHA were quantified by antigen (CII)-induced 3H-thymidine incorporation into lymphocytes of mesenteric lymph node, draining lymph node, and spleen. TGF-beta in supernatants obtained from lymph node culture medium was measured by ELISA. RESULTS: Arthritis limbs were observed in 100% of control at 5 weeks after subcutaneous BnCII injection. The incidences of CIA in all tolerized group were significantly lower than that in control 5 weeks after immunization (control 100 % vs. 5 microgram feeding group: 50%, 10 microgram feeding group: 50%, 20 microgram feeding group: 50%, 40 microgram feeding group: 55.5%, P<0.01). In comparison to control, mean articular indices were lower in all tolerized groups (control 5.13 : 5 microgram feeding group 3.50, 10 microgram feeding group 2.75, 20 microgram feeding group 2.87, 40 microgram feeding group 2.63, P<0.05). Arthritic limbs were also significantly lower in tolerized groups (control 58.3 : 5 microgram feeding group 20.8, 10 microgram feeding group 16.7, 20 microgram feeding group 20.8, 40 microgram feeding group 20.8, P<0.05). The titers of IgG antibody to CII were lower in tolerized group than that in control [tolerized group ; median 10 (min. 0, max. 48), control ; median 33 (min. 8.6, max. 101), P<0.05]. The proliferative responses to BnCII were significantly suppressed in tolerized (control 8010+/-2319cpm, tolerized group 4500+/-2060cpm, P<0.01). High TGF-beta production was noted in tolerized group (control ; 28pg/ml, BnCII feeding group ; 73pg/ml). CONCLUSION: Oral tolerance in DBA/1 mice was successfully induced from low doses of BnCII (5 microgram) and suppressed T and B cell function in conjunction with increased TGF-beta production may play an important role for the induction of CII induced oral tolerance in DBA/1 mice.

Tolerogenic Effect of Orally Administrated Type II Collagen in CIA Animal (DBA/1 mice) / 대한류마티스학회지

OBJECTIVE: To investigate the dosage of bovine type II collagen (BnCII) for the induction of oral tolerance in CIA animals, and to verify the changes of immune response and TGF-beta production of mesenteric lymph node cells in tolerized CIA animals. METHODS: Oral tolerance was induced by feeding of variable doses (5 microgram, 10 microgram, 20 microgram and 40 microgram) of BnCII to DBA/1 mice 4 times per week during 2 weeks, and control mice were given ovalbumin (1000 microgram), before immunization. We examed clinical assessment ; incidence of arthritis, severity of arthritis, arthritic limb by visual analysis. IgG antibodies to BnCII were measured by ELISA, T cell responses to BnCII and PHA were quantified by antigen (CII)-induced 3H-thymidine incorporation into lymphocytes of mesenteric lymph node, draining lymph node, and spleen. TGF-beta in supernatants obtained from lymph node culture medium was measured by ELISA. RESULTS: Arthritis limbs were observed in 100% of control at 5 weeks after subcutaneous BnCII injection. The incidences of CIA in all tolerized group were significantly lower than that in control 5 weeks after immunization (control 100 % vs. 5 microgram feeding group: 50%, 10 microgram feeding group: 50%, 20 microgram feeding group: 50%, 40 microgram feeding group: 55.5%, P<0.01). In comparison to control, mean articular indices were lower in all tolerized groups (control 5.13 : 5 microgram feeding group 3.50, 10 microgram feeding group 2.75, 20 microgram feeding group 2.87, 40 microgram feeding group 2.63, P<0.05). Arthritic limbs were also significantly lower in tolerized groups (control 58.3 : 5 microgram feeding group 20.8, 10 microgram feeding group 16.7, 20 microgram feeding group 20.8, 40 microgram feeding group 20.8, P<0.05). The titers of IgG antibody to CII were lower in tolerized group than that in control [tolerized group ; median 10 (min. 0, max. 48), control ; median 33 (min. 8.6, max. 101), P<0.05]. The proliferative responses to BnCII were significantly suppressed in tolerized (control 8010+/-2319cpm, tolerized group 4500+/-2060cpm, P<0.01). High TGF-beta production was noted in tolerized group (control ; 28pg/ml, BnCII feeding group ; 73pg/ml). CONCLUSION: Oral tolerance in DBA/1 mice was successfully induced from low doses of BnCII (5 microgram) and suppressed T and B cell function in conjunction with increased TGF-beta production may play an important role for the induction of CII induced oral tolerance in DBA/1 mice.

Change of Anti-type II Collagen Antibody in Rheumatoid Arthritis and Degenerative Arthritis / 대한정형외과학회잡지

PURPOSE: To evaluate the chronological change of serum anti-type II collagen antibody level according to the type of immunoglobulin (i.e. IgG, IgM) in chronic arthritic patients. MATERIALS AND METHODS: The serum levels of anti-type II collagen antibody were determined in three groups (control, degenerative arthritis and rheumatoid arthritis) with ELISA method. In each person, the serum levels of antibody IgG, IgM against human, bovine and chicken type II collagens were determined separately. RESULTS: Since correlation coefficients of ELISA showed high values except in the case of denatured human collagen, ELISA test is regarded as reliable. In terms of denatured human collagen, no constant denaturation pattern induced variable results. With regards to chronological changes, there was no change of IgG titer in all three groups. IgM titer of control group was not varied according to time change. But IgM titer of arthritis group changed from time to time. Chronological changes of antibody titers depended on the types of antigen. In case of human collagen, there were changes of antibody titer in the degenerative arthritis and rheumatoid arthritis groups. And in the case of bovine collagen, change of antibody titer was observed only in the rheumatoid arthritis group. CONCLUSION: Chronological change of IgG titer was not observed in the arthritis group but IgM titer changes are observed in that group. Thus, it might be a topic of future research to evaluate the relationship between the chronological change of IgM antibody and disease aggravation.

Differences in anti-type II Collegen antibody titers Among Degenerative Arthritis, Rheumatoid Arthritis and Control Groups / 대한정형외과학회잡지

Collagen is the major structural protein in the human body, especially in connective tissues. There are more than 13 types of collagen. Among them, type II collagen is a main component of articular cartilage structure. Altered immunological conditions against type II collagen may be closely related to the pathologic conditions of joint, especially arthritis. Since 1977, animal model for collageninduced arthritis(CIA) has been developed and used in the investigation of arthritis. In those animals, high titers of anti-type II collagen antibody were noticed. Pathologic findings were similar to rheumatoid arthritis of human, which suggested that rheumatoid arthritis might be one of the autoimmune diseases. There had been many reports about elevation of serum and synovial level of anti-type II collagen antibody in rheumatoid arthritis patients. But majority of them did not discriminate the antibody titers according to the type of immunoglobulin(i.e. IgG, IgM). And the question whether the elevated antibody titers are cause or effect of the arthritis is still in controversy. In this study, the serum levels of anti-type II collagen antibody were determined in 82 persons(35 degenerative arthritis patients, 24 rheumatoid arthritis patients and 22 normal controls without any joint problem) via ELISA method. In each person the serum IgG, IgM and IgG+M+A antibody levels against bovine type IIcollagen and chicken typeII collagen were determined individually. Statistical evaluation of these data among degenerative arthritis group, rheumatoid arthritis group and normal control group was performed. The results were as follows; 1. Degenerative arthritis group revealed significant elevation of anti-type II collagen antibody(IgG, IgG+M+A) compared to normal control(p < 0.05). 2. Rheumatoid arthritis group showed significant elvation of IgM and IgG+M+A compared to normal control. 3. Between degenerative arthritis and rheumatoid arthritis group, no sigificant difference was noticed. 4. Rheumatoid arthritis group showed significant increase of IgM antibody level compared to normal control. 5. Female rheumatoid arthritis group showed significant increase of IgM level compared to female degenerative arthritis group. These findings suggested that the elevation of anti-type II collagen antibody titer is not specific for rheumatoid arthritis and related with general pathologies destroying articular cartilage. And it is suggested that anti-type II collagen antibody associated with rheumatoid arthritis is mainly IgM proportion, especially in female patients. So further investigation of anti-type II collagen antibody associated with rheumatoid arthritis is needed to target IgM antibody.