Single-cell suspensions of BALF or lung cells were incubated on ice for 20 min with supernatant from hybridoma 2.4.G2 to block Fc receptors and then stained with the indicated Abs. Female C57BL/6 mice at 6 to 8 weeks of age were purchased from the Animal Institute of Chinese Medical Academy (Beijing, China). Control mice were inoculated with the same volume of PBS containing the same concentration of sucrose as the corresponding viral inoculum. A wide array of both viral vectors such as retrovirus, lentivirus, herpes simplex virus, recombinant adeno-associated virus (rAAV), and adenovirus (Ad), in addition to nonviral gene delivery systems such as naked plasmid DNA and lipid vesicles, are currently being tested for various applications. Reactivation of productive cytomegalovirus replication can occur in vitro in experimental conditions, which suggests that cell differentiation pathways act as determinants of reactivation . Mice were killed after the injection of 1 × 105 NG4TL4-TK cells on 7, 14, and 21 d (n = 3). Many studies have provided evidence that RONS may have antimicrobial roles in host defense during both the innate and adaptive phases of the immune response (reviewed in Ref.
CL2MDP-lip were administered to mice 2 days before infection with HSV-1 (following day 4 of exercise), as well as on day 4 postinfection, to deplete lung macrophages. Induction of TNF-α expression in SJPL cells.We first used reverse transcription-PCR to identify the proinflammatory cytokines whose expression was induced in SJPL cells infected with A/Sydney/5/97 (H3N2) influenza virus (MOI, 2). The open triangle represents the mean value for each group. Therefore, in the present study we aimed to determine the role of neutrophils in a relevant host-pneumovirus interaction by using antibody-mediated neutrophil depletion in PVM infected mice. Cytosol and nuclear protein extracts were prepared from lung tissue using Nuclear/Cytosol Fractionation kit (K266–100, Biovision) following protocols provided by the manufacturer. The fraction of the airway surface area that was inflamed (defined by the presence of extravasated leukocytes) or denuded (defined by exposed basement membrane) was calculated using standard stereological approaches . Mice were sacrificed at days 0, 7, 10, 14, 21, 28, 56, and 77 days postinfection (p.i.) for virus titer determination, immunohistochemistry (IHC), and RT-PCR.
The primers were designed based on a highly conserved region of the ICP4 gene to amplify a 1500 bp fragment. Most of the viruses inducing asthma exacerbation are aerocontaminant ones. Bioluminescence of mice were analysed using Xenogen IVIS200 imaging system (Xenogen Corp.) at the indicated time points. The lungs were finely minced and then suspended in RPMI 1640 (Life Sciences Technologies) containing collagenase-dispase (1 μg/mL) (Roche, Indianapolis, IN, USA). Pulmonary function tests. The samples with a cycle threshold value less than or equal to 45 were considered as positive. Th1 cytokines (interferon gamma, TNF-α) produce the pro-inflammatory responses to fight viruses and other intracellular parasites, and to eliminate cancer cells.
Proliferation was measured by adding 50 μl [3H]thymidine to 100 μl of cell culture and harvesting 12 h later for detection of incorporated 3H by scintillation counting. Interestingly, like pandemic H1N1 influenza virus, SwIV also infects the lower respiratory tract of pigs. The mice were anesthetized with Zoletil (Virbac, Carros, France) and infected with 5 LD50 in 50 μl intranasally. (San Jose, CA), and confirmed to be >95% pure by analytical high-performance liquid chromatography at the National Institute of Allergy and Infections Diseases peptide core facility (Bethesda, MD). Compared to CPA treatment alone, infection with GLV-1h68 or combination treatment leads to an up-regulation of several pro-inflammatory cytokines and chemokines (Table 3C and D), like seen in the virus-treated versus untreated tumors, affirming that virus infection leads to a pro-inflammatory status within the tumor. UV inactivation of Ad gene expression was confirmed by infecting follicular skin DCs with UV-irradiated Ad vector encoding enhanced green fluorescent protein (AdEGFP) or untreated AdEGFP and visualizing under a fluorescent microscope (data not shown). Cathelicidins are other cationic peptides with LL-37 as the only human member and cathelicidin-related antimicrobial peptide (CRAMP) being its murine counterpart (reviewed in Refs.
Erickson et al. The ability of VLPs to package exogenous DNA makes them promising vectors for gene therapy . Furthermore, lack of Fas-FasL interaction in ECTV-infected cocultures can lead to disturbances in proinflammatory chemokines production. In this respect, further research is required into the possible involvement of this concomitance, through the use of live BVDV and BHV-1 vaccines, in the triggering of an impaired pulmonary immune response. The receptor for advanced glycation end products (RAGE) was the first receptor demonstrated to bind HMGB1 , and HMGB1 signaling through RAGE was found to promote chemotaxis and the production of cytokines in a process that involves the activation of the transcription factor nuclear factor-κB (NF-κB) [13, 14]. Useful information can be obtained on the fine structure of the lung, using samples taken soon after death. There are, however, a few studies that have not supported a correlation between metastatic calcification and the level of the calcium–phosphate ion product (5, 36–38).