Perforin and Fas cytolytic pathways coordinately shape the selection and diversity of CD8+-T-cell escape variants of influenza virus

Antigenic variation is a viral strategy exploited to promote survival in the face of the host immune response and represents a major challenge for efficient vaccine development. Influenza viruses are pathogens with high transmissibility and mutation rates, enabling viral escape from immunity induced by prior infection or vaccination. Intense selection from neutralizing antibody drives antigenic changes in the surface glycoproteins, resulting in emergence of new strains able to reinfect hosts immune to previously circulating viruses. CD8+ cytotoxic T cells (CTLs) also provide protective immunity from influenza virus infection and may contribute to the antigenic evolution of influenza viruses. Utilizing mice transgenic for an influenza virus NP366-374 peptide-specific T-cell receptor, we demonstrated that the respiratory tract is a suitable site for generation of escape variants of influenza virus selected by CTL in vivo. In this report the contributions of the perforin and Fas pathways utilized by influenza virus-specific CTLs in viral clearance and selection of CTL escape variants have been evaluated. While transgenic CTLs deficient in either perforin- or Fas-mediated pathways are efficient in initial pulmonary viral control, variant virus emergence was observed in all the mice studied, although the spectrum of viral CTL escape variants selected varied profoundly. Thus, a less-restricted repertoire of escape variants was observed in mice with an intact perforin cytotoxic pathway compared with a limited variant diversity in perforin pathway-deficient mice, although maximal variant diversity was observed in mice having both Fas and perforin pathways intact. We conclude that selection of viral CTL escape variants reflects coordinate action between the tightly controlled perforin/granzyme pathway and the more promiscuous Fas/FasL pathway.     

PC3 overexpression affects the pattern of cell division of rat cortical precursors

The PC3 gene is transiently expressed during neurogenesis in precursor cells of the telencephalic ventricular/subventricular zone, and is rapidly downregulated before cell migration and differentiation. It is thought to have a role in controlling cell proliferation, but its precise function is not known. Here we present evidence that PC3, when overexpressed in vitro by retroviral-mediated gene transfer, acts by interfering with the normal pattern of cell division. Firstly, we report evidence that PC3 overexpression reduces the rate of cell proliferation in both NIH 3T3 cells and embryonic precursor cells from the rat cerebral cortex. Secondly, when studying the pattern of BrdU dilution in clones of cortical precursors, we observe that clones transduced with PC3 show an asymmetric pattern of BrdU dilution more frequently than clones transduced with a control vector. We discuss the hypothesis that the higher number of PC3 transduced clones showing an asymmetric pattern of BrdU dilution may be due to an increase in asymmetric cell divisions.     

What have tissue culture studies told us about the development of oligodendrocytes?

One major success of studying neural cell development in tissue culture has been the discovery of the O-2A cell. This bipotential cell generates oligodendrocytes or, under certain conditions, a type of astrocyte. This essay considers the evidence that the characteristic properties demonstrated by the O-2A cells in vitro are an accurate reflection of oligodendrocyte development in vivo.     

Genome structure and apomixis in Phoenicaulis (Brassicaceae; Boechereae)

Apomixis in crucifer (Brassicaceae) species is rare, reported in just four genera (Boechera, Draba, Erysimum, and Parrya), and one intergeneric hybrid (Raphanobrassica). It is well studied only in Boechera, where it is widespread among 100+ recognized species. However, its occurrence in related genera of the tribe Boechereae has not been documented previously. Here we analyzed genome evolution, mode of reproduction, and fertility of the monospecific Boechereae genus Phoenicaulis (P. cheiranthoides), endemic to the northwestern United States. We discovered that the species encompasses diploid (2n = 2x = 14), triploid (2n = 3x = 21), and tetraploid (2n = 4x = 28) populations. Comparative chromosome painting analyses revealed that the three karyotypes are essentially structurally identical, differing only in the presence of a largely heterochromatic chromosome (Het) in the triploid and tetraploid cytotypes. The genome structure of Phoenicaulis appeared identical to that of Boechera species previously analyzed, suggesting genomic stasis despite the morphological and molecular divergence of the two genera. This genome colinearity extended to the presence and structure of the Het chromosomes, which are closely associated with apomictic reproduction in Boechera. Interestingly, all three cytotypes of Phoenicaulis proved to be apomictic, regardless of the presence or absence of a Het chromosome, and sexual populations have yet to be identified.     

Intestinal perfusion indicates high reliance on paracellular nutrient absorption in an insectivorous bat Tadarida brasiliensis

Flying vertebrates have been hypothesized to have a high capacity for paracellular absorption of nutrients. This could be due to high permeability of the intestines to nutrient-sized molecules (i.e., in the size range of amino acids and glucose, MW 75–180 Da). We performed intestinal luminal perfusions of an insectivorous bat, Tadarida brasiliensis. Using radio-labeled molecules, we measured the uptake of two nutrients absorbed by paracellular and transporter-mediated mechanisms (l-proline, MW 115 Da, and d-glucose, MW 180 Da) and two carbohydrates that have no mediated transport (l-arabinose, MW 150 Da, and lactulose, MW 342 Da). Absorption of lactulose (0.61 ± 0.06 nmol min^? 1 cm^? 1) was significantly lower than that of the smaller arabinose (1.09 ± 0.04 nmol min^? 1 cm^? 1). Glucose absorption was significantly lower than that of proline at both nutrient concentrations (10 mM and 75 mM). Using the absorption of arabinose to estimate the portion of proline absorption that is paracellular, we calculated that 25.1 ± 3.0% to 66.2 ± 7.8% of proline absorption is not transporter-mediated (varying proline from 1 mM to 75 mM). These results confirm our predictions that 1) paracellular absorption is molecule size selective, 2) absorption of proline would be greater than glucose absorption in an insectivore, and 3) paracellular absorption represents a large fraction of total nutrient absorption in bats.