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

Yang, Hyung In; Kim, Wan Uk; Min, Do Jun; Park, Sung Hwan; Hong, Yeon Sik; Lee, Sang Heon; Cho, Chul Soo; Kim, Ho Youn

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.