Figure 2.

Effects of pretransfer treatment with A213 in murine autoimmune sialadenitis (MIS). (a) Saliva was collected from recombination‐activating gene 1 (Rag‐1)^–/– mice after adoptive transfer. The saliva volume was measured on days 0 and 45 after transfer, adjusted for body weight and calculated relative to the volume measured at baseline [A213‐treated mice (n = 11) and phosphate‐buffered saline (PBS)‐treated mice (n = 8)]. Statistical significance was determined using Student's t‐test. *P < 0·05 (day 45 after adoptive transfer versus day 0), not significant (n.s.) between A213‐ and PBS‐treated mice at days 0 and 45. (b) Comparison of haematoxylin and eosin (H&E)‐stained salivary gland sections of muscarinic acetylcholine receptor (M3R)^–/–→Rag‐1^–/– mice pretransfer treated with A213 and PBS on day 45. Representative images obtained from 13 to 16 mice per group. (c) Histological focus score of inflammatory lesions in salivary glands between M3R^–/–→Rag‐1^–/– mice pretransfer treated with A213 and PBS on day 45. The focus score was assessed in a blinded manner. Data are mean ± standard deviation (s.d.) of 13–16 mice per group. Statistical significance was determined using Student's t‐test; n.s. between A213‐ and PBS‐treated mice. (d,e) Splenocytes (left panel) and cervical lymph node (cLN) cells (right panel) obtained from M3R^–/–→Rag‐1^–/– mice on day 45 pretransfer treated with A213 or PBS were cultured with or without M3R peptide mixture. Interferon (IFN)‐γ (d) and interleukin (IL)‐17 (e) levels in culture supernatants were analysed by enzyme‐linked immunosorbent assay (ELISA). Data are mean ± standard error of the mean (s.e.m.) of six mice per group; n.s. (Student's t‐test); not detected (n.d.).