Expression and distribution of Toll-like receptors 11–13 in the brain during murine neurocysticercosis

The functions of Toll-like receptors (TLRs) 11–13 in central nervous system (CNS) infections are currently unknown. Using a murine model of neurocysticercosis, we investigated the expression and distribution of TLRs 11–13 by using both gene specific real-time PCR analysis and in situ immunofluoresence microscopy in both control and neurocysticercosis brains. In the mock infected brain, mRNAs of TLRs 11–13 were constitutively expressed. Parasite infection caused an increase of both mRNAs and protein levels of all three TLRs by several fold. All three TLR proteins were present in both CNS and immune cell types. Among them TLR13 was expressed the most in terms of number of positive cells and brain areas expressing it, followed by TLR11 and TLR12 respectively. Among the nervous tissue cells, TLRs 11–13 protein levels appeared highest in neurons. However, TLR13 expression was also present in ependymal cells, endothelial cells of pial blood vessels, and astrocytes. In contrast, infiltrating CD11b and CD11c positive myeloid cells predominantly produced TLR11 protein, particularly early during infection at 1 wk post infection (~50% cells). TLRs 12 and 13 proteins were present on approximately 5% of infiltrating immune cells. The infiltrating cells positive for TLRs 11–13 were mostly of myeloid origin, CD11b+ cells. This report provides a comprehensive analysis of the expression of TLRs 11–13 in normal and parasite infected mouse brains and suggests a role for them in CNS infections

Attenuated Response of Aged Mice to Respiratory Francisella novicida Is Characterized by Reduced Cell Death and Absence of Subsequent Hypercytokinemia

Pneumonia and pulmonary infections are major causes of mortality among the growing elderly population. Age associated attenuations of various immune parameters, involved with both innate and adaptive responses are collectively known as immune senescence. These changes are likely to be involved with differences in host susceptibility to disease between young and aged individuals.The objective of this study was to assess potential age related differences in the pulmonary host response in mice to the Gram-negative respiratory pathogen, Francisella novicida. We intranasally infected mice with F. novicida and compared various immune and pathological parameters of the pulmonary host response in both young and aged mice.We observed that 20% of aged mice were able to survive an intranasal challenge with F. novicida while all of their younger cohorts died consistently within 4 to 6 days post infection. Further experiments revealed that all of the aged mice tested were initially able to control bacterial replication in the lungs as well as at distal sites of replication compared with young mice. In addition, the small cohort of aged survivors did not progress to a severe sepsis syndrome with hypercytokinemia, as did all of the young adult mice. Finally, a lack of widespread cell death in potential aged survivors coupled with a difference in cell types recruited to sites of infection within the lung confirmed an altered host response to Francisella in aged mice

Rapid dissemination of Francisella tularensis and the effect of route of infection

<p>Abstract</p> <p>Background</p> <p><it>Francisella tularensis </it>subsp. <it>tularensis </it>is classified as a Category A bioweapon that is capable of establishing a lethal infection in humans upon inhalation of very few organisms. However, the virulence mechanisms of this organism are not well characterized. <it>Francisella tularensis </it>subsp. <it>novicida</it>, which is an equally virulent subspecies in mice, was used in concert with a microPET scanner to better understand its temporal dissemination in vivo upon intranasal infection and how such dissemination compares with other routes of infection. Adult mice were inoculated intranasally with <it>F. tularensis </it>subsp. <it>novicida </it>radiolabeled with ⁶⁴Cu and imaged by microPET at 0.25, 2 and 20 hours post-infection.</p> <p>Results</p> <p>⁶⁴Cu labeled <it>F. tularensis </it>subsp. <it>novicida </it>administered intranasally or intratracheally were visualized in the respiratory tract and stomach at 0.25 hours post infection. By 20 hours, there was significant tropism to the lung compared with other tissues. In contrast, the images of radiolabeled <it>F. tularensis </it>subsp. <it>novicida </it>when administered intragastrically, intradermally, intraperitoneally and intravenouslly were more generally limited to the gastrointestinal system, site of inoculation, liver and spleen respectively. MicroPET images correlated with the biodistribution of isotope and bacterial burdens in analyzed tissues.</p> <p>Conclusion</p> <p>Our findings suggest that Francisella has a differential tissue tropism depending on the route of entry and that the virulence of Francisella by the pulmonary route is associated with a rapid bacteremia and an early preferential tropism to the lung. In addition, the use of the microPET device allowed us to identify the cecum as a novel site of colonization of <it>Francisella tularensis </it>subsp. <it>novicida </it>in mice.</p

Differential Release and Phagocytosis of Tegument Glycoconjugates in Neurocysticercosis: Implications for Immune Evasion Strategies

Neurocysticercosis (NCC) is an infection of the central nervous system (CNS) by the metacestode of the helminth Taenia solium. The severity of the symptoms is associated with the intensity of the immune response. First, there is a long asymptomatic period where host immunity seems incapable of resolving the infection, followed by a chronic hypersensitivity reaction. Since little is known about the initial response to this infection, a murine model using the cestode Mesocestoides corti (syn. Mesocestoides vogae) was employed to analyze morphological changes in the parasite early in the infection. It was found that M. corti material is released from the tegument making close contact with the nervous tissue. These results were confirmed by infecting murine CNS with ex vivo–labeled parasites. Because more than 95% of NCC patients exhibit humoral responses against carbohydrate-based antigens, and the tegument is known to be rich in glycoconjugates (GCs), the expression of these types of molecules was analyzed in human, porcine, and murine NCC specimens. To determine the GCs present in the tegument, fluorochrome-labeled hydrazides as well as fluorochrome-labeled lectins with specificity to different carbohydrates were used. All the lectins utilized labeled the tegument. GCs bound by isolectinB4 were shed in the first days of infection and not resynthesized by the parasite, whereas GCs bound by wheat germ agglutinin and concavalinA were continuously released throughout the infectious process. GCs bound by these three lectins were taken up by host cells. Peanut lectin-binding GCs, in contrast, remained on the parasite and were not detected in host cells. The parasitic origin of the lectin-binding GCs found in host cells was confirmed using antibodies against T. solium and M. corti. We propose that both the rapid and persistent release of tegumental GCs plays a key role in the well-known immunomodulatory effects of helminths, including immune evasion and life-long inflammatory sequelae seen in many NCC patients

Changes in Gene Expression of Pial Vessels of the Blood Brain Barrier during Murine Neurocysticercosis

<div><p>In murine neurocysticercosis (NCC), caused by infection with the parasite <i>Mesocestoides corti</i>, the breakdown of the Blood Brain Barrier (BBB) and associated leukocyte infiltration into the CNS is dependent on the anatomical location and type of vascular bed. Prior studies of NCC show that the BBB comprised of pial vessels are most affected in comparison to the BBB associated with the vasculature of other compartments, particularly parenchymal vessels. Herein, we describe a comprehensive study to characterize infection-induced changes in the genome wide gene expression of pial vessels using laser capture microdissection microscopy (LCM) combined with microarray analyses. Of the 380 genes that were found to be affected, 285 were upregulated and 95 were downregulated. Ingenuity Pathway Analysis (IPA) software was then used to assess the biological significance of differentially expressed genes. The most significantly affected networks of genes were “inflammatory response, cell-to-cell signaling and interaction, cellular movement”, “cellular movement, hematological system development and function, immune cell trafficking, and “antimicrobial response, cell-to-cell signaling and interaction embryonic development”. RT-PCR analyses validated the pattern of gene expression obtained from microarray analysis. In addition, chemokines CCL5 and CCL9 were confirmed at the protein level by immunofluorescence (IF) microscopy. Our data show altered gene expression related to immune and physiological functions and collectively provide insight into changes in BBB disruption and associated leukocyte infiltration during murine NCC.</p> </div

Top biological functions associated with differentially expressed genes.

<p>Top biological functions associated with differentially expressed genes.</p

Hierarchical cluster analysis of differentially expressed probe sets in pial endothelial cells.

<p>First 3 columns represent 3 biological replicates of 3 wks p.i. mice whereas last 3 columns represent 3 independent mock-infected samples (A). A bar showing the color range used for denoting the upregulated and downregulated genes (B).</p

Parasite infection induced chemokines in pial endothelial cells and their known role in leukocyte trafficking.

<p>Parasite infection induced chemokines in pial endothelial cells and their known role in leukocyte trafficking.</p