8A). We also evaluated in vitro AMs cytokine production in the presence or absence click here of MP extracts.

AMs were obtained from BALF at 6 day after two IP immunizations with MP extracts plus alum or with alum alone. When we compared the concentration of cytokines in the culture supernatants, the production of IL-1β, IL-6, MIP-2, and RANTES after 8 h incubation, was strikingly greater in the former than the latter ( Fig. 8B), indicating that pre-immunization with MP extracts augmented the potential reactivity of AMs to the extracts. Taken together, it is likely that AMs primed with two IP immunizations with MP extracts constituted the major effector cells in facilitating the initial neutrophilic/lymphocytice infiltration after IT. Previous studies have demonstrated that mycoplasma cell wall antigens activate MAPK-NF-κB signaling in AMs through TLR-2. Thus, we

evaluated the expression of TLR-2 on AMs at 6 day after GW786034 in vivo two IP immunizations with MP extracts plus alum or alum alone. The number of TLR-2 positive AMs in the alveolar spaces 4.6 times greater in model E (86.9%) than in model D (18.5%) (Fig. 9A and B), whereas the number of TLR-2 positive epithelial cells was even between model D and E. The expression of mRNA in BALF cells obtained from model E were 3.0 times higher than in model D (Fig. 9C). To elucidate that TLR-2 signaling is involved in cytokine production, we performed an inhibition assay using a MAPK inhibitor, SP600125 for JNK (c-Jun N-terminal kinase) enzymes. The inhibitor completely abrogated the production of IL-6 by AM stimulated with MP extracts in vitro ( Fig. 9D). The aim of this study was to reproduce the inflammatory processes seen in human MP pneumonia using a novel mouse MP pneumonia model. None of the previously reported murine models [26], [31], [33], [36], [37], [38], [39], [40], [41], [42] and [43] or other animal models [44], [45], [46], [47] and [48] described persistent plasma

cell infiltration in the PBVA. Thus, this is the first report that describes a murine model with longstanding plasma cell infiltration Thymidine kinase in the PBVA. This study also described the chronological sequence of inflammatory events in MP pneumonia both quantitatively and qualitatively. We confirmed that neutrophil infiltration precedes lymphocyte infiltration in both lung sections and BALF, which is consistent with previous studies [33], [36] and [40]. The peaked cell density was 4-fold higher in model E than D, suggesting that the effect of pre-immunization with MP extracts was a potent promoter of neutrophil infiltration. Similarly, in model E, lymphocyte infiltration peaked at 48 h, lasting up to 336 h, an effect that was not observed in model D.