Salivary gland proteome during adult development and after blood feeding of female anopheles dissidens mosquitoes (Diptera Culicidae)

Understanding changes in mosquito salivary proteins during the time that sporozoite maturation occurs and after blood feeding may give information regarding the roles of salivary proteins during the malarial transmission. Anopheles dissidens (formerly Anopheles barbirostris species A1) is a potential vector of Plasmodium vivax in Thailand. In this study, analyses of the proteomic profiles of female An. dissidens salivary glands during adult development and after blood feeding were carried out using two-dimensional gel electrophoresis coupled with nano-liquid chromatography-mass spectrometry. Results showed at least 17 major salivary gland proteins present from day one to day 21 post emergence at 8 different time points sampled. Although there was variation observed, the patterns of protein expression could be placed into one of four groups. Fifteen protein spots showed significant depletion after blood feeding with the percentages of the amount of depletion ranging from 8.5% to 68.11%. The overall results identified various proteins, including a putative mucin-like protein, an anti-platelet protein, a long form D7 salivary protein, a putative gVAG protein precursor, a D7-related 3.2 protein, gSG7 salivary proteins, and a gSG6 protein. These results allow better understanding of the changes of the salivary proteins during the adult mosquito development. They also provide candidate proteins to investigate any possible link or not between sporozoite maturation, or survival of skin stage sporozoites, and salivary proteins

Proteomic Analysis of Chikungunya Virus Infected Microgial Cells

Chikungunya virus (CHIKV) is a recently re-emerged public health problem in many countries bordering the Indian Ocean and elsewhere. Chikungunya fever is a relatively self limiting febrile disease, but the consequences of chikungunya fever can include a long lasting, debilitating arthralgia, and occasional neurological involvement has been reported. Macrophages have been implicated as an important cell target of CHIKV with regards to both their role as an immune mediator, as well evidence pointing to long term viral persistence in these cells. Microglial cells are the resident brain macrophages, and so this study sought to define the proteomic changes in a human microglial cell line (CHME-5) in response to CHIKV infection. GeLC-MS/MS analysis of CHIKV infected and mock infected cells identified some 1455 individual proteins, of which 90 proteins, belonging to diverse cellular pathways, were significantly down regulated at a significance level of p<0.01. Analysis of the protein profile in response to infection did not support a global inhibition of either normal or IRES-mediated translation, but was consistent with the targeting of specific cellular pathways including those regulating innate antiviral mechanisms

Comparative Proteomics of Activated THP-1 Cells Infected with Mycobacterium tuberculosis Identifies Putative Clearance Biomarkers for Tuberculosis Treatment.

CAPRISA, 2015.Abstract available in pdf

Identification of salivary gland proteins depleted after blood feeding in the malaria vector anopheles campestris-like mosquitoes (Diptera: Culicidae)

Malaria sporozoites must invade the salivary glands of mosquitoes for maturation before transmission to vertebrate hosts. The duration of the sporogonic cycle within the mosquitoes ranges from 10 to 21 days depending on the parasite species and temperature. During blood feeding salivary gland proteins are injected into the vertebrate host, along with malaria sporozoites in the case of an infected mosquito. To identify salivary gland proteins depleted after blood feeding of female Anopheles campestris-like, a potential malaria vector of Plasmodium vivax in Thailand, two-dimensional gel electrophoresis and nano-liquid chromatography-mass spectrometry techniques were used. Results showed that 19 major proteins were significantly depleted in three to four day-old mosquitoes fed on a first blood meal. For the mosquitoes fed the second blood meal on day 14 after the first blood meal, 14 major proteins were significantly decreased in amount. The significantly depleted proteins in both groups included apyrase, 5'-nucleotidase/apyrase, D7, D7-related 1, short form D7r1, gSG6, anti-platelet protein, serine/threonine-protein kinase rio3, putative sil1, cyclophilin A, hypothetical protein Phum_PHUM512530, AGAP007618-PA, and two non-significant hit proteins. To our knowledge, this study presents for the first time the salivary gland proteins that are involved in the second blood feeding on the day corresponding to the transmission period of the sporozoites to new mammalian hosts. This information serves as a basis for future work concerning the possible role of these proteins in the parasite transmission and the physiological processes that occur during the blood feeding

Amounts of depletion of major salivary gland proteins from unfed mosquitoes and blood fed to repletion on mice.

a<p>Spot number refers to those shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0090809#pone-0090809-g002" target="_blank">Fig. 2a</a>.</p>b<p>ASD ± SD = Average spot density ± Standard deviation.</p>c<p>Student’s <i>t</i>-test, p≤0.05.</p>d<p>Protein spot was absent.</p

Details of 17 major protein spots identified by nano-liquid chromatography-mass spectrometry in the female salivary glands of <i>An</i>. <i>dissidens</i>.

<p>Details of 17 major protein spots identified by nano-liquid chromatography-mass spectrometry in the female salivary glands of <i>An</i>. <i>dissidens</i>.</p

Expression levels of the 17 major protein spots determined at different ages in adult developmental time points.

<p>The y-axis represents relative expression level normalized with heat shock cognate (HSC) 70, and the x-axis represents different ages on days 0, 1, 2, 3, 8, 12, 16, and 21, accordingly. Different letters (a, b, c, d, e, f) indicate significantly different levels of protein expression (<i>p</i> < 0.05), i.e. groups labelled with one letter (e.g. “a”) are not significantly different from each other, but are different to groups labelled with a different letter (e.g. b, c, d, e, or f), and so on. Any given letter is only relevant within that protein (i.e. “a” in SN1 has nothing to do with “a” in SN2). Error bars are plotted for all points, but are too small to be visualised in some cases.</p

Amounts of depletion of the major salivary gland proteins after blood feeding of <i>An</i>. <i>dissidens</i> mosquitoes.

<p>Amounts of depletion of the major salivary gland proteins after blood feeding of <i>An</i>. <i>dissidens</i> mosquitoes.</p

Representative 2-DE gels of salivary gland proteins extracted from 80 female mosquitoes at different ages.

<p>(A) 0 day, (B) 1 days, (C) 3 days, (D) 12 days, (E) 16 days, and (F) 21 days old. Molecular mass markers are indicated on the left in kDa. Isoelectric points (pI) are indicated at the top. Arrows indicate major salivary gland proteins. Arrowhead indicates an internal control protein. Numbers are corresponding to the major salivary gland proteins in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0163810#pone.0163810.t001" target="_blank">Table 1</a>. The large spot at the base of the right side gels of Fig 1A and B is an artifact of separation. It was seen on the three gels shown with these samples only.</p