Hemolysis-associated phosphatidylserine exposure promotes polyclonal plasmablast differentiation.

Antimalarial antibody responses are essential for mediating the clearance of Plasmodium parasite-infected RBCs from infected hosts. However, the rapid appearance of large numbers of plasmablasts in Plasmodium-infected hosts can suppress the development and function of durable humoral immunity. Here, we identify that the formation of plasmablast populations in Plasmodium-infected mice is mechanistically linked to both hemolysis-induced exposure of phosphatidylserine on damaged RBCs and inflammatory cues. We also show that virus and Trypanosoma infections known to trigger hemolytic anemia and high-grade inflammation also induce exuberant plasmablast responses. The induction of hemolysis or administration of RBC membrane ghosts increases plasmablast differentiation. The phosphatidylserine receptor Axl is critical for optimal plasmablast formation, and blocking phosphatidylserine limits plasmablast expansions and reduces Plasmodium parasite burden in vivo. Our findings support that strategies aimed at modulating polyclonal B cell activation and phosphatidylserine exposure may improve immune responses against Plasmodium parasites and potentially other infectious diseases that are associated with anemia.

First record of the leafhopper genus Scaphomonus Viraktamath from Korea, with description of one new species (Hemiptera: Cicadellidae: Deltocephalinae).

The leafhopper genus Scaphomonus Viraktamath, 2009 is reported from the Korean peninsula for the first time, based on the discovery of a new species: Scaphomonus naejangsanus sp. nov. A checklist and keys to world Scaphomonus species are given.

Review of the leafhopper genus Maiestas Distant (Hemiptera: Cicadellidae: Deltocephalinae) from Korea.

The Korean fauna of the leafhopper genus Maiestas Distant, 1917 is reviewed taxonomically. A total of 12 species are recognized, including three new species: M. borealis sp. nov., M. maritima sp. nov., and M. peninsularis sp. nov. The following three species are newly recorded from Korea: M. horvathi (Then), M. irisa Zhang Duan, M. yangae Zhang Duan. A check-list and key to Korean species of the genus are provided.

Rapid Detection of Urinary Tract Infections via Bacterial Nuclease Activity.

Rapid and accurate bacterial detection methods are needed for clinical diagnostic, water, and food testing applications. The wide diversity of bacterial nucleases provides a rich source of enzymes that could be exploited as signal amplifying biomarkers to enable rapid, selective detection of bacterial species. With the exception of the use of micrococcal nuclease activity to detect Staphylococcus aureus, rapid methods that detect bacterial pathogens via their nuclease activities have not been developed. Here, we identify endonuclease I as a robust biomarker for E. coli and develop a rapid ultrasensitive assay that detects its activity. Comparison of nuclease activities of wild-type and nuclease-knockout E. coli clones revealed that endonuclease I is the predominant DNase in E. coli lysates. Endonuclease I is detectable by immunoblot and activity assays in uropathogenic E. coli strains. A rapid assay that detects endonuclease I activity in patient urine with an oligonucleotide probe exhibited substantially higher sensitivity for urinary tract infections than that reported for rapid urinalysis methods. The 3 hr turnaround time is much shorter than that of culture-based methods, thereby providing a means for expedited administration of appropriate antimicrobial therapy. We suggest this approach could address various unmet needs for rapid detection of E. coli.

RPGR, a prenylated retinal ciliopathy protein, is targeted to cilia in a prenylation- and PDE6D-dependent manner.

RPGR (retinitis pigmentosa GTPase regulator) is a ciliary protein associated with several forms of inherited retinal degenerative diseases. PDE6D is a ubiquitously expressed prenyl-binding protein and involved in ciliary targeting of prenylated proteins. The current working model for the RPGR function depicts that RPGR acts as a scaffold protein to recruit cargo-loaded PDE6D to primary cilia. Here, we present evidence demonstrating an alternative relationship between RPGR and PDE6D, in which RPGR is a cargo of PDE6D for ciliary targeting. We found that the constitutive isoform of RPGR, which is prenylated, requires prenylation for its ciliary localization. We also found that there are at least two independent ciliary targeting signals in RPGR: one within the N-terminal region that contains the RCC1-like domain and the other near the prenylation site at the C-terminus. Ablation of PDE6D blocked ciliary targeting of RPGR. Our study indicates that prenylated RPGR is one of the cargos of PDE6D for ciliary trafficking and provides insight into the mechanisms by which RPGR is targeted to cilia.

The human cytomegalovirus UL133-138 gene locus attenuates the lytic viral cycle in fibroblasts.

The genomes of HCMV clinical strains (e.g. FIX, TR, PH, etc) contain a 15 kb region that encodes 20 putative ORFs. The region, termed ULb', is lost after serial passage of virus in human foreskin fibroblast (HFF) cell culture. Compared to clinical strains, laboratory strains replicate faster and to higher titers of infectious virus. We made recombinant viruses with 22, 14, or 7 ORFs deleted from the ULb' region using FIX and TR as model clinical strains. We also introduced a stop codon into single ORFs between UL133 and UL138 to prevent protein expression. All deletions within ULb' and all stop codon mutants within the UL133 to UL138 region increased to varying degrees, viral major immediate early RNA and protein, DNA, and cell-free infectious virus compared to the wild type viruses. The wild type viral proteins slowed down the viral replication process along with cell-free infectious virus release from human fibroblast cells.

Alternative splicing of the human cytomegalovirus major immediate-early genes affects infectious-virus replication and control of cellular cyclin-dependent kinase.

The major immediate-early (MIE) gene locus of human cytomegalovirus (HCMV) is the master switch that determines the outcomes of both lytic and latent infections. Here, we provide evidence that alteration in the splicing of HCMV (Towne strain) MIE genes affects infectious-virus replication, movement through the cell cycle, and cyclin-dependent kinase activity. Mutation of a conserved 24-nucleotide region in MIE exon 4 increased the abundance of IE1-p38 mRNA and decreased the abundance of IE1-p72 and IE2-p86 mRNAs. An increase in IE1-p38 protein was accompanied by a slight decrease in IE1-p72 protein and a significant decrease in IE2-p86 protein. The mutant virus had growth defects, which could not be complemented by wild-type IE1-p72 protein in trans. The phenotype of the mutant virus could not be explained by an increase in IE1-p38 protein, but prevention of the alternate splice returned the recombinant virus to the wild-type phenotype. The lower levels of IE1-p72 and IE2-p86 proteins correlated with a delay in early and late viral gene expression and movement into the S phase of the cell cycle. Mutant virus-infected cells had significantly higher levels of cdk-1 expression and enzymatic activity than cells infected with wild-type virus. The mutant virus induced a round-cell phenotype that accumulated in the G(2)/M compartment of the cell cycle with condensation and fragmentation of the chromatin. An inhibitor of viral DNA synthesis increased the round-cell phenotype. The round cells were characteristic of an abortive viral infection.

Down-regulation of locus-specific human lymphocyte antigen class I expression in Epstein-Barr virus-associated gastric cancer: implication for viral-induced immune evasion.

BACKGROUND: To understand whether the association between Epstein-Barr Virus (EBV) and gastric cancer (GC) has any role in loss of surface expression of human lymphocyte antigen (HLA) class I, the authors analyzed locus-specific transcriptional expression of HLA-A, HLA-B, HLA-C, and HLA-E along with other HLA-associated molecules (beta2-microglobulin [beta2M], cellular latent membrane protein [LMP], and transporter associated with antigen presentation [TAP]) in EBV-associated, primary GC (EBVaGC) and EBV-negative GC (EBVnGC) tissues. METHODS: Approximately 20 EBVaGC tissues and 40 EBVnGC tissues and their corresponding normal tissues were used in the study. The presence of EBV in GC was established by EBV-encoded small RNA in situ hybridization analysis and BamHI W polymerase chain reaction (PCR) analysis. Transcriptional expression of viral LMP2A and several HLA class I genes were analyzed by reverse transcriptase (RT)-PCR. Surface expression levels of HLA class I proteins in cancer samples along with their normal counterparts also were quantified by flow cytometry. RESULTS: The RT-PCR data suggested selective down-regulation of the HLA-A/HLA-B locus along with over-expression of HLA-E transcripts in EBVaGC (P < .05). This was confirmed further by the flow-cytometric studies using antibodies to HLA-ABC and HLA-E. Among the accessory molecules, LMP7 transcript was down-regulated in a number of EBVaGC tissues compared with EBVnGC. CONCLUSIONS: The current results suggested that the establishment of EBV latent infection in gastric tissues allows malignant cells to avoid the immune surveillance of both cytotoxic T-lymphocytes and natural killer cells by regulating the differential expression of HLA class I molecules.

Analysis of human lymphocyte antigen class I expression in gastric cancer by reverse transcriptase-polymerase chain reaction.

Malignant cells have been reported to escape immune surveillance by modulation of human lymphocyte antigen (HLA) class Ia molecule and/or other accessory molecules like TAP (transporter associated with antigen processing) and beta2-M expression. Most of these reports, however, are based on immunohistochemistry techniques with polymorphic- or isotype-specific antibodies. In the present study, we have instead used a locus-specific reverse transcriptase-polymerase chain reaction-based approach to detect the transcriptional expression of HLA class Ia as well as accessory molecules in gastric cancer. Our results indicate that HLA class Ia transcript is totally absent in only approximately 9% of cancer cases. Locus-specific expression of HLA-A and -B could, however, be detected in approximately 54% cases, whereas HLA-C was expressed in most of the cancer tissues. Interestingly, in some cases where HLA class Ia expression was observed, TAP1 expression could not be detected. Furthermore, we also investigated the frequency of nonclassical or HLA class Ib expression for molecules such as HLA-E and -G. HLA-G transcript was absent in gastric tissues both in cancerous and autologous normal region, whereas HLA-E was observed in a number of gastric cancers. Altogether these selective locus-specific losses of HLA class I along with impaired expression of accessory molecules may explain the complex phenomena by which gastric tumors escape both cytotoxic T-lymphocyte- as well as natural killer cell-mediated immune defense.

Simultaneous vaccination against cholera and yellow fever

A study of serum-samples from immunised individuals showed that administration of yellow-fever and cholera vaccine, simultaneously or one to three weeks apart, reduced the vibriocidal and yellow-fever-virus-neutralising antibody titres (AU)