The genomic sequence of Mycoplasma pneumoniae establish this cell-wall-less prokaryote as among the smallest known microorganisms capable of self-replication. However, this genomic simplicity and corresponding biosynthetic austerity are sharply contrasted by the complex terminal structure found in this species. This tip structure (attachment organelle) directs colonization of the human respiratory mucosa, leading to bronchitis and atypical pneumonia. Furthermore, formation of a second tip structure appears to precede cell division, implying temporal regulation. However, the organization, regulation, and assembly of the attachment organelle in M. pneumoniae are poorly understood, and no counterparts have been identified among the walled bacteria. M. pneumoniae possesses a cytoskeleton-like structure required to localize adhesin proteins to the attachment organelle. The cytadherence-associated proteins HMW1, HMW2, and HMW3 are components of the mycoplasma cytoskeleton, with HMW1 localizing strictly along the filamentous extensions from the cell body and HMW3 being a key structural element of the terminal organelle. Disruptions in hmw2 result in the loss of HMW1 and HMW3. However, the hmw1 and hmw3 genes were transcribed and translated at wild-type levels in hmw2 mutants. HMW1 and HMW3 were relatively stable in the wild-type background over 8 h but disappeared in the mutants over this time period. Evaluation of recombinant HMW1 levels in mycoplasma mutants suggested a requirement for the C-terminal domain of HMW1 for turnover. Finally, an apparent defect in the processing of the precursor for the adhesin protein P1 was noted in the HMW ؊ mutants.Members of the genus Mycoplasma are distinguished by their lack of a cell wall or cell wall precursors, the incorporation of cholesterol in their membranes, the use of UGA as a codon for tryptophan rather than termination, and their extremely small genomes (Ն580 kbp). In addition, several mycoplasma species possess a complex terminal structure that directs the colonization of vertebrate host cells and is thought to function in cell division and gliding motility. The presence of this complex organelle stands in marked contrast to the perception of mycoplasmas as otherwise simple wall-less prokaryotes (1). In Mycoplasma pneumoniae, a major cause of primary atypical community-acquired pneumonia, the terminal (attachment) organelle is seen by transmission electron microscopy of thin sections as a membrane-bound extension of the cell body, with an electron-dense core associated with a cytoskeleton-like network of mycoplasma proteins (2). The organization, regulation, and assembly of the terminal organelle of M. pneumoniae is poorly understood, largely due to the stringent nutritional requirements and poor growth yields of mycoplasmas, as well as a general lack of genetic tools for use in these prokaryotes. Finally, no counterparts to the terminal organelle have been identified among the walled bacteria.Adherence of M. pneumoniae to host respiratory epithelium (cytadheren...

Two experiments were conducted to evaluate the efficacy of beta-glucan on growth performance, nutrient digestibility, and immunity in weanling pigs. In Exp. 1, 210 weanling pigs (6.38 +/- 0.92 kg of BW) were fed dietary beta-glucan (0, 0.01, 0.02, 0.03, or 0.04%) for 5 wk. In Exp. 2, 168 pigs (6.18 +/- 1.31 kg of BW) were fed no beta-glucan or antibiotics (T1), 0.02% beta-glucan (T2), only antibiotics (T3), or 0.02% beta-glucan with antibiotics (T4) for 8 wk. In Exp. 2, the antibiotics fed were apramycin and carbadox in phase I (0 to 2 wk) and carbadox and chlortetracycline in phase II (3 to 8 wk). During Exp. 2, the performance study was conducted for 5 wk, and the immune response was tested until 8 wk. In Exp. 1, there was a trend for a linear increase (P = 0.068) in ADG as the dietary beta-glucan concentration increased in the diet. The digestibilities of DM, GE, CP, ether extract, Ca, and P increased linearly (P < 0.05) in the beta-glucan-supplemented pigs. In Exp. 2, the overall ADG was greater (P < 0.05) in treatment T4 compared with the control group (T1). Also, except for P, this group showed greater (P < 0.05) nutrient digestibilities than the control group. In Exp. 2, at d 15, 24, and 46 antibody titers were measured by ELISA against Pasteurella multocida type A and D after vaccination with atrophic rhinitis, and they differed significantly (P < 0.05) with no particular trend. Flow cytometry was used to determine porcine lymphocyte subpopulations at 4 and 8 wk of Exp. 2. There was an increase in CD4 cells (P < 0.05) and a trend for an increase in CD8 cells (P < 0.10) at 8 wk in pigs fed the T2 diet compared with the other groups. Overall, increasing the dietary concentrations of beta-glucan did not improve ADG without antibiotic, and in weanling pigs antibiotics seem to be more effective in improving nutrient digestibilities and growth performance than beta-glucan.

No abstract

Toll-like receptor (TLR) ligands are strongly considered immune-adjuvants for cancer immunotherapy and have been shown to exert direct anti-cancer effects. This study was performed to evaluate the synergistic anti-cancer and anti-metastatic effects of the TLR7 agonist imiquimod (IMQ) during radiotherapy for melanoma. The pretreatment of B16F10 or B16F1 cells with IMQ combined with γ-ionizing radiation (IR) led to enhanced cell death via autophagy, as demonstrated by increased expression levels of autophagy-related genes, and an increased number of autophagosomes in both cell lines. The results also confirmed that the autophagy process was accelerated via the reactive oxygen species (ROS)-mediated MAPK and NF-κB signaling pathway in the cells pretreated with IMQ combined with IR. Mice subcutaneously injected with melanoma cells showed a reduced tumor growth rate after treatment with IMQ and IR. Treatment with 3-methyladenine (3-MA), ameliorated the anti-cancer effect of IMQ combined with IR. Additionally, the combination therapy enhanced anti-cancer immunity, as demonstrated by an increased number of CD8+ T cells and decreased numbers of regulatory T cells (Treg) and myeloid-derived suppressor cells (MDSCs) in the tumor lesions. Moreover, the combination therapy decreased the number of metastatic nodules in the lungs of mice that were injected with B16F10 cells via the tail vein. In addition, the combination therapy enhanced systemic anti-cancer immunity by increasing the abundances of T cell populations expressing IFN-γ and TNF-α. Therefore, these findings suggest that IMQ could serve as a radiosensitizer and immune booster during radiotherapy for melanoma patients.

Phagocytosis is an essential mechanism in innate immune defense, and in maintaining homeostasis to eliminate apoptotic cells or microbes, such as Mycobacterium tuberculosis, Salmonella enterica, Streptococcus pyogenes and Legionella pneumophila. After internalizing microbial pathogens via phagocytosis, phagosomes undergo a series of ‘maturation’ steps, to form an increasingly acidified compartment and subsequently fuse with the lysosome to develop into phagolysosomes and effectively eliminate the invading pathogens. Through this mechanism, phagocytes, including macrophages, neutrophils and dendritic cells, are involved in the processing of microbial pathogens and antigen presentation to T cells to initiate adaptive immune responses. Therefore, phagocytosis plays a role in the bridge between innate and adaptive immunity. However, intracellular bacteria have evolved diverse strategies to survive and replicate within hosts. In this review, we describe the sequential stages in the phagocytosis process. We also discuss the immune evasion strategies used by pathogens to regulate phagosome maturation during intracellular bacterial infection, and indicate that these might be used for the development of potential therapeutic strategies for infectious diseases.