Helminth infections predispose mice to pneumococcal pneumonia but not to other pneumonic pathogens.

Pneumonia is the leading killer of children worldwide. Here, we report that helminth-infected mice develop fatal pneumonia when challenged with Streptococcus pneumoniae. Mice were chronically infected with either the flatworm Taenia crassiceps or the roundworm Heligmosomoides polygyrus. Upon challenge with a pneumonic type 3 strain of S. pneumoniae (A66.1), the worm-infected mice developed pneumonia at a rate and to a degree higher than age-matched control mice as measured by bioluminescent imaging and lung titers. This predisposition to pneumonia appears to be specific to S. pneumoniae, as worm-infected mice did not show evidence of increased morbidity when challenged with a lethal dose of influenza virus or sublethal doses of Staphylococcus aureus or Listeria monocytogenes. The defect was also present when worm-infected mice were challenged with a type 2 sepsis-causing strain (D39); an increased rate of pneumonia, decreased survival, and increased lung and blood titers were found. Pneumococcal colonization and immunity against acute otitis media were unaffected. Anti-helminthic treatment in the H. polygyrus model reversed this susceptibility. We conclude that helminth coinfection predisposes mice to fatal pneumococcal pneumonia by promoting increased outgrowth of bacteria in the lungs and blood. These data have broad implications for the prevention and treatment for pneumonia in the developing world, where helminth infections are endemic and pneumococcal pneumonia is common.

Fluorescent microscopy of viable Batrachochytrium dendrobatidis.

Batrachochytrium dendrobatidis ( Bd ), a chytrid fungus, is a causative agent of chytridiomycosis and amphibian population declines worldwide. The sequenced genome of Bd provides information necessary for studying the fungus and its molecular biology. Fluorescent microscopy is a technique used to image targeted molecules in live or fixed organisms to understand cellular trafficking and localization, but the use of fluorescent microscopy with Bd has not yet been demonstrated. Two fluorescent stains were tested for their use in live-cell imaging of Bd , i.e., the cell wall-specific fluorophore Solophenyl Flavine 7GFE and the DNA-specific fluorophore DRAQ5. These specific staining patterns were observed in live cultures of Bd when visualized with laser-scanning confocal microscopy.

Maternal transfer of antibodies to eggs in Xenopus laevis.

The immune system of the African clawed frog, Xenopus laevis, includes nearly the full repertoire of lymphoid organs and immune cell types found in mammals. In contrast to the mammalian immune system, the development of the amphibian immune system occurs in the open environment. Oviparity necessitates a rapid ontogeny of the immune system. X. laevis larvae become immunocompetent about 2 weeks after fertilization of the egg. During this 2-week window, larvae cannot mount an adaptive immune response to potential pathogens and presumably must depend on innate responses. In the present study, the possibility of maternal transfer of antibodies to eggs was examined. Adult female X. laevis were injected three times at weekly intervals with the hapten-carrier complex, trinitrophenylated bovine serum albumin (TNP-BSA). The sera of immunized frogs demonstrated antibody activity to BSA, TNP-BSA, and, importantly, trinitrophenylated ovalbumin (TNP-OVA) when examined by enzyme-linked immunosorbent assay (ELISA). Reactivity to TNP-OVA confirmed that antibodies were produced against TNP. The adult female frogs were induced to lay eggs by injection of human chorionic gonadotropin. Next, membrane-free extracts of the eggs were treated with protease inhibitors in order to prevent proteolysis of proteins found in the eggs. On analysis by ELISA, it was found that TNP-specific antibodies were present in the egg extracts. This demonstrated the transfer of antigen-specific antibodies from adult females to eggs in X. laevis.

Antibody response of bluegill sunfish during development of acquired resistance against the larvae of the freshwater mussel Utterbackia imbecillis.

The larvae of freshwater mussels in the order Unionoida are obligate parasites on fishes, on which they metamorphose into juveniles. Bluegill sunfish (Lepomis macrochirus) acquire resistance against glochidia of the freshwater mussel Utterbackia imbecillis after 2 infections. In order to study the systemic and mucosal antibody response associated with acquired resistance, sera from experimentally infected fish were collected at 10-d intervals during 4 sequential infection periods and from naïve fish. Enzyme-linked immunosorbant assays (ELISA) revealed that fish exhibited a humoral and mucosal antibody response around day 20 after the 1st infection which was followed by second antibody response beginning at day 60 (day 20, 3rd infection) that persisted until the end of the collection period. Western blots of glochidial proteins probed with the sera revealed that the profile of proteins recognized by antibodies produced by fish changed over the course of multiple infections. Serum collected from fish at day 20 (peak of primary response) contained antibodies against approximately 39 and 91 kDa proteins. Immunohistochemical studies on whole-mount glochidia probed with serum from these fish demonstrated that the antibodies recognize granular structures located between the larval mantle and shell. Serum collected from fish during the secondary antibody response (days 60-80) bound additional protein bands in Western blots. Those antibodies recognized other cells of the larval mantle, most prominently in a ciliated region that contains the primordia of the gills and organs of the juvenile and adult mussel.

Uptake and secretion of host proteins by Taenia crassiceps metacestodes.

Although the presence of intact host proteins in the cyst fluid of cyclophyllidean metacestodes has been well documented, the underlying reason for protein uptake is poorly understood. To investigate this discrepancy, both the cyst fluid (CF) and excreted/secreted (E/S) proteins were collected in vitro from Taenia crassiceps metacestodes 16 wk postinfection in Balb/cJ female mice. The CF and E/S were subsequently immunoblotted using rabbit anti-mouse whole serum antibodies as a probe. The results show that whole host proteins were not only internalized by metacestodes but also secreted as well. The predominant secreted host protein was 66 kDa and was confirmed to be mouse serum albumin. The amount of secreted albumin decreased daily, whereas the concentration of albumin in the cyst fluid remained consistent. This suggests that the secretion of albumin is a coordinated function rather than a random event. It is probable that albumin cycling may be an evolved mechanism providing multiple benefits for the larvae, including osmoregulation and protection from innate immune responses.

Ultrastructure of spermiogenesis and the spermatozoon in Taenia crassiceps strobilae WFU strain (Cestoda, Cyclophyllidea, Taeniidae) from golden hamsters.

Strobilae from Taenia crassiceps (WFU strain) were obtained from outbred hamsters (Mesocricetus auratus) by feeding them viable metacestodes maintained by intraperitoneal passage in female Balb/c mice. Mature and gravid proglottids from strobilae were recovered from hamster intestines and fixed for light and electron microscopy. By light microscopy, the expected structure of taeniid proglottids was observed. Ultrastructural analysis of ten proglottids showed that testicular follicles and vas deferens contained filiform spermatids, with a single axoneme, and an elongated helicoidal nucleus inserted between the axoneme and the spiraled cortical microtubules. At the apical cone, a single crest-like body was found and mature spermatids also exhibited transverse intracytoplasmic walls. The morphology and characters of the spermatids in T. crassiceps conform to type III spermiogenesis, which has been described in other taeniids.

Infrapopulation dynamics of a wild strain of Taenia crassiceps (WFU) (Cestoda: Taeniidae) in BALB/cJ mice.

Taenia crassiceps cysticerci form large infrapopulations that persist in the tissues of their rodent hosts. Early infrapopulation growth appears inhibited and is followed by rapid increases that appear not to be controlled by the host immune response. This investigation was undertaken to examine the infrapopulation growth dynamics of a normally developing strain (WFU) of T. crassiceps during a 60-day primary intraperitoneal (i.p.) infection. Three, 6, 9, 14, 28, and 60 days after i.p. inoculation of 5 cysticerci, mice were killed, and the numbers of larvae, developmental stage, and buds per larva were recorded. Larval infrapopulation abundance increased exponentially beginning on day 6 postinoculation (PI), indicating an initial lag in reproduction. A stage-structured exponential growth model, assuming no mortality, fits the larval infrapopulation dynamics in terms of the numbers of larvae in reproductive and nonreproductive stages, indicating that cysticerci evade or suppress (or both) host immune mechanisms that are parasite restrictive after the first week of infection.