Murine cerebral malaria development is independent of toll-like receptor signaling

Malaria pigment hemozoin was reported to activate the innate immunity by Toll-like receptor (TLR)-9 engagement. However, the role of TLR activation for the development of cerebral malaria (CM), a lethal complication of malaria infection in humans, is unknown. Using Plasmodium berghei ANKA (PbA) infection in mice as a model of CM, we report here that TLR9-deficient mice are not protected from CM. To exclude the role of other members of the TLR family in PbA recognition, we infected mice deficient for single TLR1, -2, -3, -4, -6, -7, or -9 and their adapter proteins MyD88, TIRAP, and TRIF. In contrast to lymphotoxin alpha-deficient mice, which are resistant to CM, all TLR-deficient mice were as sensitive to fatal CM development as wild-type control mice and developed typical microvascular damage with vascular leak and hemorrhage in the brain and lung, together with comparable parasitemia, thrombocytopenia, neutrophilia, and lymphopenia. In conclusion, the present data do not exclude the possibility that malarial molecular motifs may activate the innate immune system. However, TLR-dependent activation of innate immunity is unlikely to contribute significantly to the proinflammatory response to PbA infection and the development of fatal CM.     

Accuracy of transmission CT and FDG-PET in the detection of small pulmonary nodules with integrated PET/CT

Purpose The purpose of this study was to determine the accuracy of detection of small pulmonary nodules on quiet breathing attenuation correction CT (CTAC) and FDG-PET when performing integrated PET/CT, as compared with a diagnostic inspiratory CT scan acquired in the same imaging session.Methods PET/CT scans of 107 patients with a history of carcinoma (54 male and 53 female, mean age 57.3 years) were analyzed. All patients received an integrated PET/CT scan including a CTAC acquired during quiet respiration and a contrast-enhanced CT acquired during inspiration in the same session. Breathing CTAC scans were reviewed by two thoracic radiologists for the presence of pulmonary nodules. FDG-PET scans were reviewed to determine accuracy of nodule detection. Diagnostic CT was used as the gold standard to confirm or refute the presence of nodules.Results On the CTAC scans 200 nodules were detected, of which 183 were true positive (TP) and 17, false positive. There were 109 false negatives (FN). Overall, 51 (48%) patients had a false interpretation, including 19 in whom CT was interpreted as normal for lung nodules. The average size of the nodules missed was 3.8±2 mm (range 2–12 mm). None of the nodules missed on the CTAC scans were detected by PET. In the right lung there were 20 TP, 42 true negative (TN), 11 FP, and 34 FN interpretations with a sensitivity in nodule detection of 37% (CI 24–51%) and a specificity of 79% (CI 66–89%). In the left lungs there were 16 TP, 65 TN, 3 FP, and 23 FN interpretations, with a sensitivity of 41% (CI 26–58%) and a specificity of 96% (CI 88–99%).Conclusion The detection of small pulmonary nodules by breathing CTAC and FDG-PET is relatively poor. Therefore an additional diagnostic thoracic CT scan obtained during suspended inspiration is recommended for thorough evaluation of those patients in whom detection of pulmonary metastases is necessary for management.     

The TLR-mediated response of plasmacytoid dendritic cells is positively regulated by estradiol in vivo through cell-intrinsic estrogen receptor α signaling

Plasmacytoid dendritic cells (pDCs) produce large amounts of type I interferons (IFN-α/β) in response to viral or endogenous nucleic acids through activation of their endosomal Toll-like receptors (TLR-7 and TLR-9). Enhanced TLR-7-mediated IFN-α production by pDCs in women, compared with men, has been reported, but whether sex hormones, such as estrogens, are involved in this sex-based difference is unknown. Here we show, in humanized mice, that the TLR-7-mediated response of human pDCs is increased in female host mice relative to male. In a clinical trial, we establish that treatment of postmenopausal women with 17β-estradiol markedly enhances TLR-7- and TLR-9-dependent production of IFN-α by pDCs stimulated by synthetic ligands or by nucleic acid-containing immune complexes. In mice, we found exogenous and endogenous estrogens to promote the TLR-mediated cytokine secretion by pDCs through hematopoietic expression of estrogen receptor (ER) α. Genetic ablation of ERα gene in the DC lineage abrogated the enhancing effect of 17β-estradiol on their TLR-mediated production of IFN-α, showing that estrogens directly target pDCs in vivo. Our results uncover a previously unappreciated role for estrogens in regulating the innate functions of pDCs, which may account for sex-based differences in autoimmune and infectious diseases.     

Treatment of murine hepatocellular carcinoma using genetically modified cells to express interleukin-12

BACKGROUND AND AIM: The majority of patients cannot benefit from the conventional curative treatments that are currently used for hepatocellular carcinoma (HCC), which remains a world health problem. Interleukin (IL)-12 is one of the most potent anti-tumor cytokines. The aim of the present study was to examine the anti-tumor effect and toxicity of intrahepatic delivery of IL-12 using an ex vivo gene therapy approach in a murine model of HCC. METHODS: Syngenic fibroblasts or MM45T-Li HCC tumor cells were genetically modified in vitro to express IL-12 using a polycistronic TFG murine IL-12 retroviral vector (TFGmIL-12) coding for both p35 and p40 murine IL-12 subunits. Hepatocellular carcinoma was generated using direct intrahepatic inoculation of the tumor cell line into the left liver lobe of BALB/c mice. RESULTS: Direct liver expression of IL-12 by the injected genetically modified tumor cells induced a marked inhibition of tumor growth. This effect was associated with an early infiltration of macrophages, and lymphocytes forming numerous intralobular foci. There was no significant liver toxicity, as shown by normal biochemical liver tests. At a later time, the intralobular foci were rare and consisted mainly of CD4+ T cells, while CD8+ T cells were present in the lobule. Intrahepatic expression of IL-12 did not modify circulating or splenic B lymphocytes or natural killer (NK) cells. The inhibition of tumor growth was maintained in nude mice even when depleted in NK cells. Importantly, in a second model, treatment of established day 7 liver tumors in BALB/c mice using direct intra-tumor injection of syngenic fibroblasts that were genetically modified to express IL-12 significantly reduced tumor size. CONCLUSION: In conclusion, these data provide evidence that experimental HCC can be efficiently and safely treated using ex vivo IL-12 gene therapy, which seems promising for future clinical studies.