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

ABSTRACT

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.