Considering the B-cell-mediated pivotal role in the adaptive immune system, we next aimed to check the serum level of immunoglobulins in those two strains of mice. To achieve this, we assessed the serum level of immunoglobulin in age-matched and sex-matched AKR/J and SAMP1/Yit mice strains, and observed that the serum contents of immunoglobulin were almost similar, except that a minor decrease was noted in IgG3 of AKR/J mice compared with that of SAMP1/Yit mice (Fig. 7b). The SAMP1/Yit mice exhibit serious B-cell defects, so they

may generate a differential pattern of adaptive immune functions by producing less serum immunoglobulin compared with AKR/J strain. A decreased production of regulatory cytokines was observed in TLR-stimulated MLN B cells from SAMP1/Yit mice; for this reason, we speculated that these B cells may fail to inhibit mTOR inhibitor inflammation. To confirm our speculation of whether B cells from SAMP1/Yit mice can modulate inflammatory consequences, peritoneal macrophages were isolated C59 wnt datasheet from AKR/J mice, and co-cultured

with purified MLN B cells from SAMP1/Yit or AKR/J mice, then stimulated with LPS and CpG-DNA. The IL-1β contents in culture supernatants were examined by EIA. As shown in Fig. 8(a), LPS and CpG-DNA did not stimulate IL-1β production by MLN B cells without peritoneal macrophages. Following the co-culture with peritoneal macrophages, significant amounts of IL-1β were observed in the supernatant of TLR ligand-stimulated cells. Moreover, we also noticed that the SAMP1/Yit B cells co-cultured with macrophages did not regulate/inhibit but rather enhanced IL-1β secretion by macrophages, which implies with Olson’s findings that the SAMP1/Yit B cells might be pathogenic.43 On the other hand, in the case of AKR/J B cells when co-cultured with LPS or CpG-DNA-treated macrophages, they neither induced nor reduced but instead maintained a steady state of IL-1β content as produced by the macrophages treated with the

respective ligands and without co-culture. We therefore conclude that the B cells from SAMP1/Yit Interleukin-2 receptor mice were found to be solely pathogenic whereas those from AKR/J groups were non-pathogenic. Apart from AKR/J macrophages, using the SAMP1/Yit or AKR/J B-cell co-culturing system we also tested our hypothesis in SAMP/Yit mouse macrophages co-cultured with B cells from both mice. With this co-culture system including the peritoneal macrophages from SAMP1/Yit and the B cells from both mice, the effects of LPS or CpG-DNA for IL-1β production by SAMP1/Yit macrophages was lower and the B cells from both the mouse strains were found to increase IL-1β production (Fig. 8b), which implies that the later system employing the diseased model of mouse peritoneal macrophages did not represent any conclusive data towards our proposed hypothesis. Interferon-γ is a Th1-type cytokine produced mainly by T cells upon inflammation.