Киберпространство и виртуальная реальность: попытка историко-философского анализа

Материалы IV Республик. науч. конф. студентов, магистрантов и аспирантов, Гомель, 12 мая 2011 г

Th2-skewed T cells correlate with B cell response to α-Gal and tick antigens in α-Gal syndrome

Tick bites have been shown to transmit a novel form of severe food allergy, the galactose-α-1,3-galactose (α-Gal) syndrome (AGS). Cellular responses to α-Gal in patients with AGS have, to date, not been thoroughly scrutinized. Therefore, we investigated T and B cell proliferation, activation, and cytokine profiles in response to tick protein extract (TE) and α-Gal-free TE in patients with AGS and in healthy controls. T and B cells from both patients and controls proliferated in response to TE, but significantly more in patients with AGS. B cell proliferation, but not T cell proliferation, in patients with AGS was reduced by removing α-Gal from the TE. In addition, TE induced a clear Th2 cytokine profile in patients with AGS. Expression of CD23 by B cells correlated only to T cell proliferation. However, both B cell proliferation and CD23 expression were reduced when CD40L and IL-4 were blocked. A large portion of the IgG1 and IgE antibodies binding TE in patients with AGS were directed against the α-Gal epitope. We have, for what we believe to be the first time, investigated T and B cell responses to α-Gal carrying tick proteins in patients with AGS, which will be essential for the understanding of the immune response against an allergenic carbohydrate transmitted by ticks

Anti-apoD antibodies capture ApoB in a detergent-free dual specific ELISA.

<p>The following capture antibodies functional under detergent-free conditions were used: A) anti-apoD (D544), B) anti-apoB (LDL17), C) anti-apoE (E981), D) anti-apoH (H219), E) anti-apoB (LDL20) and F) anti-apoJ (J29). Human EDTA finger blood plasma, prepared as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0115180#s2" target="_blank">methods</a>, was diluted with PBS containing 0.1% BSA. Detecting antibodies in A–D were LDL20-biotin (anti-apoB), D263-biotin (anti-apoD), H464-biotin (anti-apoH), 7B6-biotin (anti-IFNγ) and E981 (anti-apoE), and in E and F were D201-biotin (anti-apoD), D263-biotin (anti-apoD), LDL17-biotin (anti-apoB) and J84-biotin (anti-apoD). Note that E981 was also used for capture in C). The means ± SD of four replicates are shown. Results within each panel are from the same donor. Experiments A–D were repeated three times using the same donor, and experiments E and F were repeated four times using four different donors.</p

Intensive weight gain therapy in patients with anorexia nervosa results in improved serum tartrate-resistant acid phosphatase (TRAP) 5a and 5b isoform protein levels

Aim Tartrate-resistant acid phosphatase (TRAP) exists as isoforms 5a and 5b. TRAP 5a is a biomarker of chronic inflammation and influences adipose tissue and 5b associates with bone metabolism/pathologies. The aim was to investigate the association of serum TRAP 5a/5b isoforms with fat and bone markers and anthropometric parameters in patients with anorexia nervosa (AN) during weight gain therapy. Methods Twenty-five Swedish female AN patients, age 16-24 years, were treated for 12 weeks with a high-energy diet with six meals daily. Serum TRAP 5a/5b, markers of fat/glucose metabolism, markers of bone resorption and formation were measured. Parameters of bone and body composition were assessed by dual-energy X-ray absorptiometry and peripheral quantitative computed tomography. Results BMI increased from median 15.4 kg/m(2)to 19.0 kg/m(2),p &amp;lt; 0.0001. TRAP 5a and 5a/5b ratio increased but TRAP 5b decreased during the study. TRAP Delta 5a and Delta 5b correlated with Delta insulin and Delta adiponectin, respectively. TRAP 5b correlated with trabecular density at start but not at week 12. At 12 weeks, TRAP 5b correlated with CTX, and Delta decrease in TRAP 5b correlated to Delta increase in bone-specific alkaline phosphatase. Conclusions This clinical interventional study resulted in increased BMI in patients with AN. The decreased TRAP 5b protein levels confirm a role for TRAP 5b as a marker of bone resorption, whereas increased TRAP 5a seemed to derive from systemic changes in bone as well as metabolic changes. The combined detection of TRAP 5a and TRAP 5b in serum could be an indicator of improved bone metabolism.Funding Agencies|Swedish Research Council (VR)Swedish Research Council [K2015-99x-10363-23-4] Funding Source: Medline; ALF Grants Region Ostergotland [ALFGBG-716831] Funding Source: Medline</p

ApoD Mediates Binding of HDL to LDL and to Growing T24 Carcinoma

<div><p>Apolipoprotein (Apo) D is an important protein produced in many parts of the body. It is necessary for the development and repair of the brain and protection from oxidative stress. The purpose of this study was to investigate the extent to which apoD interacts with lipoproteins in human plasma. By using detergent-free ELISA, we show that immobilized monoclonal antibodies against apoD very efficiently bind to low density lipoprotein (LDL) from plasma; this binding is as equally efficient as binding to an anti-apoB monoclonal antibody. Adding detergent to the plasma inhibited the binding, suggesting that the binding is dependent on the presence of intact lipoprotein particles. Reversing the system by using immobilized anti-apoB revealed that the affinity of apoD for LDL is rather low, suggesting that multiple bindings are needed for a durable connection. Biosensor experiments using purified lipoproteins also showed that purified apoD and high density lipoprotein 3 (HDL3), a lipoprotein fraction rich in apoD, were both able to bind LDL very efficiently, indicating that the HDL3-LDL interaction may be a physiological consequence of the affinity of apoD for LDL. Furthermore, we found that apoD increases the binding of HDL to actively growing T24 bladder carcinoma cells but not to quiescent, contact-inhibited, confluent T24 cells. This result is especially intriguing given that the T24 supernatant only contained detectable levels of apoD after growth inhibition, raising the possibility that alternating the expression of apoD and a putative apoD-receptor could give direction to the flow of lipids. In the current paper, we conclude that apoD mediates binding of HDL to LDL and to growing T24 carcinomas, thereby highlighting the importance of apoD in lipid metabolism.</p></div

The interaction between apoD and apoB is a general mechanism.

<p>Finger blood from ten donors was assayed for A) apoD/apoB interaction in a detergent free dual-specific ELISA using D544 for capture and biotinylated LDL20 for detection B) ELISA for apoB levels and C) ELISA for apoD levels. Note that finger blood plasma, in figures B and C, was arbitrary calculated as half the finger blood volume, with the remaining volume assumed to be cells. Of the ten donors, No. 1–5 were women, and No. 6–10 were men, all between 30–62 years old. Means ± SD of four replicates are shown.</p

Biosensor monitoring of apoD-mediated binding of LDL.

<p>In the Blitz biosensor analysis, the signal corresponds to the mass collected on the biosensor surface. Here we used Streptavidin-coated sensor surfaces that were loaded with biotinylated antibody (10 µg/ml for 120 seconds) followed by apoD (2 µg/ml for 300 seconds) or buffer-control (300 seconds). The surfaces were subsequently loaded with the LDL, HDL2 or HDL3 lipoprotein particles (100 µg/ml for 300 seconds). Between loading reagents, the sensors were rinsed for 30 seconds. The buffer used for all steps and dilutions was PBS, azide, 0.1% BSA. <b>A</b>) The green, black, grey and red lines represent experiments preloaded with anti-apoD D544. The blue line indicates the control preloaded with isotype control antibody 7B6 (anti-IFNγ). The green and blue lines indicate that ApoD was preloaded, while the black, grey, and red lines indicate the buffer controls. The panel shows the final step in which the lipoprotein particles bind to the antibody loaded with or without apoD. The green, black and blue lines show binding of LDL, the grey line shows binding of HDL3 and the red line shows binding of HDL2. <b>B</b>) The binding step of the biotinylated antibody was omitted. Green, black, red and grey lines indicate experiments that used D544-biotin, and the blue line indicates that E981-biotin (anti-apoE) was used. Subsequent reactions show the addition of HDL2 in red, HDL3 in grey, apoD in green and blue, or buffer in black to the preloaded antibodies. In the last step, LDL was loaded in all the conditions.</p

VLDL is immobilized by D544.

<p>The effect of adding extra apoD (100 ng/ml) to VLDL (1000 ng/ml) was analyzed using a dual-specific ELISA with or without detergent present. The capture antibody was anti-apoD (D544), and the detection antibody was anti-apoB (LDL20-biotin). The means ± SD of four replicates are shown. Experiments were repeated three times. * p<0.05; p-values were calculated using the Mann-Whitney test with GraphPad prism 6.</p

ApoD facilitates binding of HDL to growing T24 cells.

<p>T24 cells grown in 24-well plates were washed with protein-free RPMI two times, and 200 µl/well RPMI supplemented with 0.1%BSA was added with or without HDL (total HDL, 200 ng/well) and with or without apoD (20 ng/well) or apoA1 (20 ng/well). After incubation for 1 hour at 21°C, the plate was washed three times with protein-free RPMI, and bound apoA1 was released using detergent and assayed by apoA1 ELISA. In <b>A</b>) the cells were non-confluent and in <b>B</b>) the cells were confluent. Means ± SD of four experiments are shown. ** p<0.01, ****p<0.0001; p-values were calculated using two-way ANOVA with Holm-Sidal multiple comparison test, GraphPad Prism 6.</p