Epstein-barr virus replication studies and their application to vector design

Vectors containing elements of the Epstein-Barr virus genome are used primarily to maintain cloned DNA inserts as plasmids in mammalian cells. However, Epstein-Bar-virus-based vectors have also been valuable tools in the hands of those studying the life cycle of Epstein-Barr virus. In this article, we discuss those characteristics of Epstein-Barr virus and its life cycle that have been used in vector construction and describe methods that are particularly applicable to the use of Epstein-Barr-virus-based vectors.     

Malaria in macaques

Seven species of malaria naturally infect eight species of macaques in southwestern India, Sri Lanka, and Southeast Asia. Within malarious areas, the frequency of infections in infected species of macaques varies from 9 to 61%. Natural malarial infections in macaques are relatively benign. The proved or probable vectors of macaque malaria are seven species of mosquitoes that belong to the Leucosphyrus Group of the genus Anopheles. The geographic distribution of macaque malaria apparently is determined by the distribution of the Leucosphyrus Group of mosquitoes, which in turn apparently is determined by the distribution of tropical evergreen rain forest. Experimental infections with three species of macaque malaria frequently are lethal to populations or species of macaques that inhabit areas outside the geographic ranges of the parasites. In populations or species of macaques that are sympatric with experimentally virulent species of malaria, partial resistance probably evolved as a consequence of natural selection acting on favorable mutations.     

Human adenovirus cloning vectors based on infectious bacterial plasmids

By making use of the fact that human adenovirus DNA circularizes in infected cells, and that circular forms of the viral genome are infectious, we have developed an improved adenovirus-based cloning system. A deletion mutant of adenovirus type 5 (Ad5) with deletions in early regions 1 (E1) and 3 (E3) was converted to a bacterial plasmid which can regenerate infectious virus following transfection into human 293 cells. A single XbaI recognition site in the deleted E3 region serves as a site for the insertion of foreign DNA. We have used this system to clone a number of genes into the Ad5 genome and describe the insertion of the neomycin/G418 resistance marker into Ad5 as an example.     

Location of genes on chromosome arms in the Anopheles gambiae group of species and their correlation to linkage data for other anopheline mosquitoes

The use of paracentric inversions as genetic markers in the Anopheles gambiae group of mosquitoes is described. The gene for dieldrin resistance is assigned to chromosome 2 which in turn is correlated to the previous assignment of the gene to linkage group II. The locus of the enzyme phosphoglucomutase 2 (Pgm 2) is similarly assigned to chromosome 2 and evidence is presented for possible linkage between Pgm 2 and dieldrin resistance. There was no linkage or correlation of chromosome 2 and loci of the enzymes superoxide dismutase (Sod) and octanol dehydrogenase (Odh). These genes are therefore assumed to be on chromosome 3 (linkage group III). Evidence that such gene linkage group/chromosome correlations may extend to other species for which chromosome maps and homologies have been worked out is discussed.     

Inhibiting gene expression in vivo by virus-mediated small interfering RNA

Inhibiting gene expression in specific tissues and organs through intravenous injection would be the ultimately preferred method of disease therapy. Here, we report the successful delivery of lentivirus-mediated small interfering RNA (siRNA) to suppress GFP gene expression in living mice. First, a lentiviral vector with siRNA (len-siRNA) driven by H1 promoter was constructed to effectively suppress GFP expression in Mel cells. When the len-siRNA virus was injected into transgenic mice, the GFP expression was significantly suppressed (over 15% reduction) in the recipient mice compared to the control mice and the suppressing effect lasted more than 1 week after injection. Our results demonstrate a new effective approach to inhibit gene expression by siRNA and lentiviral vectors. Further development of this drug for suppression of gene expression siRNA should result in applications not only for cancers but also for infectious and immune diseases. Published in Russian in Molekulyarnaya Biologiya, 2008, Vol. 42, No. 6, pp. 990–996. The text was submitted by the authors in English.     

Mosquito seasonality and arboviral disease incidence in Murray Valley, southeast Australia

Abstract. Adult female mosquito populations were monitored at weekly intervals during spring-autumn (November-March) for 4 years (1991-95) using dry-ice-baited light traps at forty sites in the Murray Valley of Victoria, Australia. Among twenty species of mosquitoes collected, Culex annulirostriswas the most abundant (66.6 ± 9.3%) followed by Cx australicus(15.3 ± 7.7%). From a total of 476, 682 mosquitoes collected, nearly all were females and only 1295 (0.27%) were males. Mosquito population densities were generally higher in 1992-93 and 1993-94 seasons than in 1991-92 and 1994-95 seasons. Greatest densities of Cx annulirostrisand Cx australicusoccurred in 1992-93, coinciding with outbreaks of Ross River (RR) and Barman Forest (BF) arboviruses causing human polyarthritis. In the majority of shires, Cx australicuswas the predominant species from spring to early summer (November and December), then was replaced by Cx annulirostrisfrom mid-summer to autumn (January-April). In three shires, Aedes bancroftianusand Ae.sagaxpredominated during the early part of the season. Densities of both Cx annulirostrisand Cx australicuswere related to temperature. Cx australicusadults were found to be trapped when the mean ambient temperature exceeded 6d?C, with peak population recorded at 20d?C. Cx annulirostrisadult density increased when the mean temperature rose above 12d?C, reaching a peak during February and March when temperature exceeded 25d?C. Cx annulirostrisdeclined rapidly from April onwards, with no adult activity evident from May to November. Population densities of Aedesspp. were generally less than reported from earlier studies, possibly due to lower rainfall in spring and summer as well as reduced flood irrigation practices. In each year, a significant correlation was detected between Cx annulirostrisdensity and RR virus incidence in humans. As Cx annulirostrisis the predominant local mosquito species and feeds on a wide spectrum of hosts including man, it seems likely that Cx annulirostrisis the major vector of RR in the inland Murray Valley region.     

Spatial and seasonal distribution of Culicoides imicola in Iberia in relation to the transmission of African horse sickness virus

Abstract. Collections of biting midges were made over 24 months from sixty sites spread across Iberia. Information on the distribution of the vector of African horse sickness virus, Culicoides imicola , from these 3119 samples showed that this species was annually present across south-western Spain as far as 3^o53'W and throughout most of Portugal, up to 41^o5'N. C.imicola was found in all areas where African horse sickness epizootics had occurred in 1987-90 and also in areas outside the epizootic zones.
Seasonal patterns of capture success of C.imicola , from seventeen frequently sampled sites where the vector was present, usually showed a late summer-early autumn peak. At the sites furthest south there was a discrete peak, mostly in September or October, before and after which the numbers captured increased or decreased steadily. At higher latitudes peak abundances occurred as early as May or as late as November, population build up was less uniform and numbers often declined rapidly after the peak was reached. Both the distribution and seasonal abundance patterns closely matched transmission patterns of African horse sickness virus, which rose during late summer and caused most cases during the autumn months.