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Volume 118, Issue 3, Pages 573-581 (March 2000) Autoimmune-related pancreatitis is associated with autoantibodies and a Th1/Th2-type cellular immune response Kazuichi Okazaki*, Kazushige Uchida*, Masaya Ohana*, Hiroshi Nakase*, Suguru Uose*, Maki Inai*, Yumi Matsushima*, Kenji Katamura‡, Katsuyuki Ohmori§, Tsutomu Chiba* Gastroenterology Volume 118, Issue 3, Pages 573-581 (March 2000) DOI: 10.1016/S0016-5085(00)70264-2 Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 1 Serum levels of anti-LF antibody using ELISA. The specific absorbance levels of serum ALF in patients with AIP (n = 17; 1.276 ± 0.276) were significantly higher than those in patients with gallstone-related pancreatitis (0.513 ± 0.165), patients with alcoholic chronic pancreatitis (0.628 ± 0.113), and normal subjects (0.369 ± 0.122). The dotted line indicates mean absorbance + 3 SD of normal controls (0.735). There were no significant differences in the autoantibodies between primary (●) and secondary (○) AIP. Gastroenterology 2000 118, 573-581DOI: (10.1016/S0016-5085(00)70264-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 2 Serum levels of ACA-II antibody using ELISA. Serum levels of ACA-II in patients with AIP (1.032 ± 0.445) were significantly higher than those in patients with gallstone-related pancreatitis (0.513 ± 0.127), patients with alcoholic chronic pancreatitis (0.613 ± 0.268), and normal subjects (0.541 ± 0.122). The dotted line indicates mean absorbance + 3 SD of normal controls (0.907). There were no significant differences in the autoantibodies between primary (●) and secondary (○) AIP. Gastroenterology 2000 118, 573-581DOI: (10.1016/S0016-5085(00)70264-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 3 Relationship between the serum levels of ACA-II and ALF in AIP patients. There was no significant relationship between ACA-II and ALF (y = 0.112x + 0.876) with heterogeneous prevalence. ●, Primary AIP; ○, secondary AIP. Gastroenterology 2000 118, 573-581DOI: (10.1016/S0016-5085(00)70264-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 4 Ratios of activated CD3+ cells by flow cytometry. The subset of PBLs was examined in 13 of 17 patients with AIP, patients with control pancreatitis (n = 17; 9 gallstone-related and 8 alcoholic pancreatitis), and normal subjects (n = 17) by flow cytometry. The ratios of HLA-DR+CD3+ (30.2% ± 10.7%) in patients with AIP were significantly higher than those in control pancreatitis patients (15.3% ± 3.0%) or normal controls (13.2% ± 9.2%). However, activated CD3+ cells for early antigens (CD69+, CD25+, and CD71+) were not significantly changed. Gastroenterology 2000 118, 573-581DOI: (10.1016/S0016-5085(00)70264-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 5 Ratios of activated CD4+ and CD8+ cells by flow cytometry. The subset of PBLs was examined in 13 of 17 patients with AIP, patients with control pancreatitis (n = 17; 9 gallstone-related and 8 alcoholic pancreatitis), and normal subjects (n = 17) by flow cytometry. The ratios of HLA-DR+CD4+ (17.0% ± 4.5%) and HLA-DR+CD8+ (25.6% ± 4.8%) PBLs in AIP patients were significantly higher than those in control pancreatitis patients (HLA-DR+CD4+, 8.5% ± 1.5%; HLA-DR+CD8+, 10.2% ± 3.6%) or normal controls (HLA-DR+CD4+, 8.0% ± 3.3%; HLA-DR+CD8+, 13.2% ± 5.6%). CD4+CD45RO+ (33.2% ± 3.3%) and CD8+CD45RO+ (22.2% ± 4.9%) memory cells in patients with AIP tended to be increased compared with control pancreatitis patients (CD4+CD45RO+, 24.4% ± 3.7%; CD8+CD45RO+, 15.8% ± 2.7%) or normal controls (CD4+CD45RO+, 18.3% ± 3.3%; CD8+CD45RO+, 14.0% ± 3.4%), although they were not significantly different. Gastroenterology 2000 118, 573-581DOI: (10.1016/S0016-5085(00)70264-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 6 Intracellular cytokines of PBLs were studied in 11 of 17 patients with AIP, patients with control pancreatitis (n = 13; 7 gallstone-related and 6 alcoholic pancreatitis), and normal subjects (n = 13). Flow-cytometric analysis of intracellular cytokines showed that the ratios of CD4+ cells producing IFN-γ in AIP (16.4% ± 5.6%) were significantly higher than those in control pancreatitis patients (7.5% ± 2.7%) and normal subjects (7.2% ± 2.8%). However, the ratios of CD4+ cells producing IL-4 were not different between AIP (2.7% ± 0.8%) and control patients (2.2% ± 0.9%) or normal subjects (1.9% ± 0.7%). Gastroenterology 2000 118, 573-581DOI: (10.1016/S0016-5085(00)70264-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 7 Medium levels of cytokines using ELISA. IFN-γ and IL-4 cytokines in the culture medium secreted from PBLs in 11 of 17 patients with AIP, patients with control pancreatitis (n = 13; 7 gallstone-related and 6 alcoholic pancreatitis), and normal subjects (n = 13) were measured by the ELISA system. The medium levels of IFN-γ secreted by the PMA/ionomycin-stimulated lymphocytes of AIP (n = 11; 196 ± 56 pg/mL) were significantly higher than those in controls (control patients: n = 13, 35 ± 23 pg/mL; normal subjects: n = 13, 30 ± 19 pg/mL). However, IL-4 levels in both groups were below detectable levels (10 pg/mL). Gastroenterology 2000 118, 573-581DOI: (10.1016/S0016-5085(00)70264-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions