A new role for PGRP-S (Tag7) in immune defense: lymphocyte migration is induced by a chemoattractant complex of Tag7 with Mts1

PGRP-S (Tag7) is an innate immunity protein involved in the antimicrobial defense systems, both in insects and in mammals. We have previously shown that Tag7 specifically interacts with several proteins, including Hsp70 and the calcium binding protein S100A4 (Mts1), providing a number of novel cellular functions. Here we show that Tag7–Mts1 complex causes chemotactic migration of lymphocytes, with NK cells being a preferred target. Cells of either innate immunity (neutrophils and monocytes) or acquired immunity (CD4⁺ and CD8⁺ lymphocytes) can produce this complex, which confirms the close connection between components of the 2 branches of immune response.     

Effects of microclimate on daily flight activity ofSimulium arcticum Malloch (Diptera: Simuliidae)

Microclimate was found to affect the daily flight activity ofSimulium arcticum in a pasture situation in central Alberta, Canada. Vapour pressure, air temperature, light intensity, and wind gust velocity had the greatest correlation to flight activity. These variables were correlated to support the concept that they influence the energy and water balances of a black fly. Flight activity is a response of the black fly to alterations of its energy and water balances.     

A unique DR-related B cell differentiation antigen

The Ia or class II molecules in both mouse and man are the basis for the genetic control of the immune response. In addition to DR, other class II antigens have been described in man. We describe a new human Ia antigen K19, recognized by three monoclonal antibodies (HK-9, HK-19, and HK-20). This antigen has the general biochemical characteristics of an Ia antigen but is different from a DR antigen. Further, this antigen is found only on mature B lymphocytes and not on monocytes and activated T cells. Thus, this antigen may represent a new Ia-like molecule that is preferentially expressed on mature B cells.     

Plasminogen activator and collagenase production by cultured capillary endothelial cells

Cultured bovine capillary endothelial (BCE) cells produce low levels of collagenolytic activity and significant amounts of the serine protease plasminogen activator (PA). When grown in the presence of nanomolar quantities of the tumor promoter 12-O-tetradecanoyl phorbol-13-acetate (TPA), BCE cells produced 5-15 times more collagenolytic activity and 2-10 times more PA than untreated cells. The enhanced production of these enzymes was dependent on the dose of TPA used, with maximal response at 10(-7) to 10(-8) M. Phorbol didecanoate (PDD), an analog of TPA which is an active tumor promoter, also increased protease production. 4-O-methyl-TPA and 4α-PDD, two analogs of TPA which are inactive as tumor promoters, had no effect on protease production. Increased PA and collagenase activities were detected within 7.5 and 19 h, respectively, after the addition of TPA. The TPA-stimulated BCE cells synthesized a urokinase-type PA and a typical vertebrate collagenase. BCE cells were compared with bovine aortic endothelial (BAE) cells and bovine embryonic skin (BES) fibroblasts with respect to their production of protease in response to TPA. Under normal growth conditions, low levels of collagenolyic activity were detected in the culture fluids from BCE, BAE, and BES cells. BCE cells produced 5-13 times the basal levels of collagenolytic activity in response to TPA, whereas BAE cells and BES fibroblasts showed a minimal response to TPA. Both BCE and BAE cells exhibited relatively high basal levels of PA, the production of which was stimulated approximately threefold by the addition of TPA. The observation that BCE cells and not BAE cells produced high levels of both PA and collagenase activities in response to TPA demonstrates a significant difference between these two types of endothelial cells and suggests that the enhanced detectable activities are a property unique to bovine capillary and microvessel and endothelial cells.     

Separation of dissociated thyroid follicular and parfollicular cells: association of serotonin binding protein with parafollicular cells

Parafollicular cells (PC) of the sheep thyroid gland are neural crest derivatives that synthesize and release the biogenic amine serotonin (5-HT) as well as the hormone calcitonin. The thyroid also contains a highly specific serotonin-binding protein (SBP). Separation of dissociated thyroid cells was done to study the cellular localization of SBP and to develop a means of isolating PC for study. Various methods were used to obtain an enriched and purified population of PC. Minced thyroid glands were enzymatically dissociated and the cells were layered on a Ficoll linear density gradient. Fractions obtained from the gradient were examined for cell number, viability, 5-HT concentration, SBP activity, and morphology by electron microscopy. One of the fractions was found to be enriched in PC. High levels of 5-HT and SBP were also found in this fraction, whereas these levels were low where the majority of cells were found. This PC-rich fraction, however, contained numerous follicular cells (FC); therefore, additional approaches to cell separation were used. FC can be stimulated in vitro with thyroid stimulating hormone (TSH) to become intensely phagocytic. When stimulated cells were incubated in the presence of silica microspheres, the FC engulfed the microspheres, which were toxic to them. PC did not become phagocytic and were unharmed by the microspheres. Suspended cells, after incubation with microspheres, were centrifuged on a discontinuous gradient, and a PC-rich fraction was obtained. Silica, however, interfered with analysis of SBP. Another method to take advantage of the phagocytic potential of FC was therefore used. TSH-stimulated cell suspensions were passed through a column of sepharose to which thyroglobulin had been coupled. Stimulated FC apparently adhered to the beads and were retained by the columns. Fractions eluting from the columns were greatly enriched with PC. These fractions contained high levels of 5-HT and SBP, and considerably reduced FC contamination was found by quantitative electron microscopy. It is concluded that SBP is localized to PC in the sheep thyroid. The idea that these cells resemble serotonergic neurons in their mechanisms of 5-HT storage is supported.     

California encephalitis virus activity in mosquitoes and horses in southern Ontario, 1975.

A study was undertaken in 1975 to determine California encephalitis virus activity in southern Ontario. Three thousand and sixty-one mosquitoes, primarily Aedes species, were divided into 104 pools and inoculated into suckling mice. Isolates of snowshoe hare virus were obtained from one pool each of Aedes fitchii and A. triseriatus mosquitoes collected in the Guelph area. Serological testing of horse sera revealed extensive virus activity in southern Ontario and indicated that horses may serve as excellent monitors for California encephalitis virus.     

Insoluble non-collagenous cartilage glycoproteins with aggregating sub-units.

Two insoluble non-collagenous glycoprotein fractions (A and G) have been separated from puppy rib cartilage, following extraction of most of the proteoglycan and digestion of the insoluble residue with purified collagenase. After reduction, alkylation and extraction with sodium dodecylsulfate most of each protein is solubilized. Gel electrophoresis of solubilized A or G shows the presence of either one or two bands and gel chromatography shows both high and low molecular weight peaks. The production of a low molecular weight electrophoresis band from the high molecular weight Sephadex fraction indicates that there is aggregation and disaggregation of sub-units in sodium dodecylsulfate. Both A and G are high in aspartate plus glutamate and have a low hydroxyproline content. The insoluble A and G both contain hexose, uronic acid, galactosamine, glucosamine and a small amount of sialic acid, but they differ in their contents of hexose and six amino acids. They both form single bands in CsCl gradients but they differ in density. Electron microscopy shows that both insoluble glycoprotein fractions stain with lead, ruthenium red, or alcian blue plus phosphotungstate and that G contains many fine filaments. Material with the same appearance and staining properties was found to occur on the surface of collagen fibres in the undigested cartilage residue.     

Circadian rhythms of sensitivity to insecticides in Musca domestica (Diptera, Muscidae)

Circadian rhythms of LD₅₀ values to DDT, dieldrin and malathion, topically applied, were determined for houseflies reared under LD 14:10 with dawn at 06.00 hr. There was a marked increase in susceptibility at 05.00 hr in each case. With dawn at 18.00 hr., DDT LD₅₀ values were lowest at 17.00 hr indicating independence of the flies' biological clocks from clock time of day. Flies reared under LD 18:6 and 10:14 also had circadian rhythms of sensitivity to DDT. Mean daily LD₅₀ values were inversely related to photophase length. The ratios of mean daily LD₅₀ to pre-dawn values were greatest for the longer photophases. Flies reared under LD 14:10 until the pupal stage, then DD until testing showed a normal circadian rhythm. Flies reared in total darkness (DD) showed no diel variations in susceptibility. W.H.O. standard strain flies were used for all the experiments. A fully susceptible (Cooper) and a DDT resistant (DEH-DOV) strain also showed significant circadian rhythms of sensitivity to DDT.

---

Zusammenfassung Circadianrhythmen der LD 50-Werte gegenüber DDT, Dieldrin und Malathion-topical angewandt wurden bei Stubenfliegen ermittelt, die bei 14:10 h-Tag mit Tagesanbruch um 6.00 Uhr gezüchtet wurden. In allen Fällen war die Empfindlichkeit um 5.00 Uhr wesentlich erhöht. Bei Tagesanbruch um 18.00 Uhr waren die niedrigsten LD 50-Werte um 17.00 Uhr. Dies weist auf die Unabhängigkeit der biologischen Uhr der Fliegen von der Tageszeit hin. Fliegen, die bei 18:6 oder bei 10:14 LD gezüchtet wurden, zeigten ebenfalls einen Circadianrhythmus hinsichtlich der Empfindlichkeit gegenüber DDT. Die mittleren LD 50-Werte waren umgekehrt proportional zur Länge der Photophase. Das Verhältnis der mittleren täglichen LD 50-Werte zu den Vortagesanbruchwerten war am grössten bei längerer Photophase. Fliegen, die bei 14:10 LD bis zum Puppenstadium und anschliessend bei DD bis zur Testung gehalten wurden, zeigten einen normalen circadianen Rhythmus. Bei Züchtung in völliger Dunkelheit zeigten sie keine Tagesschwankungen in der Empfindlichkeit. Für alle Versuche wurde ein WHO-Standardstamm benutzt. Zwei andere Stämme, einer voll empfindlich (Cooper), der andere resistent (DEH-DOV) zeigten ebenfalls signifikante Circadianrhythmen in der DDT-Empfindlichkeir.

     

Hsps are up-regulated in melanoma tissue and correlate with patient clinical parameters

Heat shock proteins (hsps) have been studied in numerous cancer types, but a clear view of their clinical relevance in melanoma remains elusive. Therefore, the aim of this study was to investigate the expression of hsps in melanoma with respect to patient clinical parameters. Using Western immunoblotting, hsps 90, 70, 60, 40 and 32 were observed to be widely expressed in metastatic melanomas (n = 31), while immunofluorescence demonstrated that in the majority of samples these hsps, apart from hsp32, were increased in expression in melanoma cells compared with surrounding non-melanoma cells in situ (n = 8). Correlating hsp expression with patient clinical parameters indicated that greater hsp90 (P < 0.02) and hsp40 (P < 0.03) expression correlated with advanced stage (stage III Vs stage IV), while in the case of hsp40, this was additionally associated with reduced patient survival (P < 0.05). In contrast, higher hsp32 expression was associated with improved patient survival (P < 0.007). On the other hand, the expression of the other hsps did not correlate with any obtainable patient clinical parameters. This study provides further evidence for the importance of hsps in melanoma and for their use as therapeutic targets and biomarkers, but larger-scale follow-up studies are required to confirm these results.

Electronic supplementary material

The online version of this article (doi:10.1007/s12192-012-0363-1) contains supplementary material, which is available to authorized users.