Maintenance of long-lived plasma cells and serological memory despite mature and memory B cell depletion during CD20 immunotherapy in mice

CD20 mAb-mediated B cell depletion is an effective treatment for B cell malignancies and some autoimmune diseases. However, the full effects of B cell depletion on natural, primary, and secondary Ab responses and the maintenance of Ag-specific serum Ig levels are largely unknown. The relationship between memory B cells, long-lived plasma cells, and long-lived humoral immunity also remains controversial. To address the roles of B cell subsets in the longevity of humoral responses, mature B cells were depleted in mice using CD20 mAb. Peritoneal B cell depletion reduced natural and Ag-induced IgM responses. Otherwise, CD20+ B cell depletion prevented humoral immune responses and class switching and depleted existing and adoptively transferred B cell memory. Nonetheless, B cell depletion did not affect serum Ig levels, Ag-specific Ab titers, or bone marrow Ab-secreting plasma cell numbers. Coblockade of LFA-1 and VLA-4 adhesion molecules temporarily depleted long-lived plasma cells from the bone marrow. CD20+ B cell depletion plus LFA-1/VLA-4 mAb treatment significantly prolonged Ag-specific plasma cell depletion from the bone marrow, with a significant decrease in Ag-specific serum IgG. Collectively, these results support previous claims that bone marrow plasma cells are intrinsically long-lived. Furthermore, these studies now demonstrate that mature and memory B cells are not required for maintaining bone marrow plasma cell numbers, but are required for repopulation of plasma cell-deficient bone marrow. Thereby, depleting mature and memory B cells does not have a dramatic negative effect on preexisting Ab levels.     

Comparison of temporal changes in psychological distress after hematopoietic stem cell transplantation among the underlying diseases of Japanese adult patients

Background  

The onset and course of irritable bowel syndrome (IBS) are strongly influenced by psychological factors, and treatment often includes cognitive-behavioral therapy. We conducted a study of the relationships between cognitive appraisal of IBS symptoms and negative mood for the subtypes of IBS.     

Membrane topology and essential amino acid residues of Phs1, a 3-hydroxyacyl-CoA dehydratase involved in very long-chain fatty acid elongation

Yeast Phs1 is the 3-hydroxyacyl-CoA dehydratase that catalyzes the third reaction of the four-step cycle in the elongation of very long-chain fatty acids (VLCFAs). In yeast, the hydrophobic backbone of sphingolipids, ceramide, consists of a long-chain base and an amide-linked C26 VLCFA. Therefore, defects in VLCFA synthesis would be expected to greatly affect sphingolipid synthesis. In fact, in this study we found that reduced Phs1 levels result in significant impairment of the conversion of ceramide to inositol phosphorylceramide. Phs1 proteins are conserved among eukaryotes, constituting a novel protein family. Phs1 family members exhibit no sequence similarity to other dehydratase families, so their active site sequence and catalytic mechanism have been completely unknown. Here, by mutating 22 residues conserved among Phs1 family members, we identified six amino acid residues important in Phs1 function, two of which (Tyr-149 and Glu-156) are indispensable. We also examined the membrane topology of Phs1 using an N-glycosylation reporter assay. Our results suggest that Phs1 is a membrane-spanning protein that traverses the membrane six times and has an N terminus and C terminus facing the cytosol. The important amino acids are concentrated in or near two of the six proposed transmembrane regions. Thus, we also propose a catalytic mechanism for Phs1 that is not unlike mechanisms used by other hydratases active in lipid synthesis.     

Pedoecological effects of a sand-fixing poplar (Populus simonii Carr.) forest in a desertified sandy land of Inner Mongolia, China

To assess the pedoecological effects of a 23-year old poplar ( Populus simonii Carr.) forest on soil amelioration and vegetation restoration via soil erosion reduction and atmospheric dust retention in a desertified sandy land ecosystem, daily dynamics of wind speed, sand transport and dust deposition rates were monitored over an erosive period from April through June in 2001, using fixed observation sites located at different positions within and around the forest. Soil and vegetation characteristics at these sites were also measured. The observation sites were placed at distances of 15H (as control), 6H and 3H (H is mean tree height) from the forest edge of the windward side (abbreviated CK, 6H-W and 3H-W respectively), forest center (FC), and at distances of 0H, 6H and 8H from the forest edge of the leeward side (FE-L, 6H-L and 8H-L respectively). Daily mean wind speed was significantly lower in different observation sites than CK, with FC having the greatest reduction of wind speed and 6H-W the least reduction. Daily transport rate of sand by wind was also significantly lower in different observation sites than CK, with FE-L having the greatest reduction of wind erosion and 6H-W the least reduction. The fact that the poplar forest will lose its functions against wind at a distance of about 12-fold tree height from the forest edge of the leeward side suggests that the effective wind-preventing range of the poplar forest is about 150 m. There was marked spatial and seasonal distribution of dust-fall rate. Over space, the rate of dust-fall was much greater within the forest than outside the forest. Over time, the daily dust deposition rate was greatest in April, followed in decreasing order by May, June, July, September and August, closely linked to the seasonal distribution pattern of dust storm. Significant positive changes in soil and vegetation parameters of the different observation sites during the 23 years that the poplar forest was established suggest the perceptible pedoecological effects of the poplar forest on soil development and restorative succession of plant community within its immediate vicinity through windbreak, soil erosion reduction and atmospheric dust retention. Understanding these pedoecological effects may aid in the design of protective forest systems in arid and semi-arid areas.     

Oxidized forms of peroxiredoxins and DJ-1 on two-dimensional gels increased in response to sublethal levels of paraquat

We previously found hydroperoxide-responsive proteins (HPRPs), which are comprised of peroxiredoxin I(Prx I), Prx II, Prx III, Prx VI, HSP27, G3PDH and two unidentified proteins (HPRP-2' and HPRP-5'), in human umbilical vein endothelial cells. It was demonstrated by two-dimensional polyacrylamide gel electrophoresis (2D PAGE) that most HPRPs are converted into variants with lower pI upon exposure to hydroperoxides. In this study, we examined the HPRP response on 2D gels upon exposure of human endothelial cells (ECV304) to paraquat (PQ²⁺), which generates reactive oxygen species (ROS) within cells. PQ²⁺ exerted cytotoxic effects in a dose- (10μM-10mM) and time- (24-168h) dependent manner. Two-dimensional PAGE analysis revealed that HPRP-2', and oxidized forms of Prx I, Prx II and Prx III were clearly increased upon exposure of cells to sublethal levels of PQ²⁺. Microsequence analysis revealed that both HPRP-2 and -2' were identical with human DJ-1. Moreover immunoblot analysis confirmed the increase of oxidized forms of Prx II, Prx III and DJ-1 in response to sublethal levels of PQ²⁺. PQ²⁺ treatment failed to increase fluorescence intensity derived from DCF, which is believed to be an indicator for intracellular levels of hydroperoxide. Although pentachlorophenol (PCP), an uncoupler of the mitochondrial respiratory chain, clearly elevated the fluorescence, PCP had no effect on HPRP response. These observations indicated that DCF-derived fluorescence is not correlated with HPRP response. We consider that the response of Prxs and DJ-1 on 2D gels could reflect endogenous production of ROS in PQ²⁺-treated cells, and might be a sensitive indicator of oxidative stress status.     

Changes in Lysozyme due to Interaction with Vaporized Hexanal

In order to elucidate the mechanism of the alteration of proteins induced by vaporized aldehydes, unmodified and chemically-modified lysozymes were exposed in the solid state to vaporized hexanal at 50°C and 5.8 or 75% relative humidity (RH). On exposure at 75%RH, the unmodified lysozyme exhibited polymerization, browning, loss of solubility, fluorescence production and impairment of lysine, tryptophan and methionine residues. Methionine residues seemed to be oxidized to methionine sulfoxide residues. The polymerization did not proceed at 5.8RH. All the above alterations were almost completely prevented by the removal of oxygen from the reaction cells. Acetylation of lysozyme retarded these alterations fairly well except that the impairment of tryptophan residues was unaffected.

On the basis of all the results it is suggested that at the first step the concerned reaction essentially requires hexanal derivatives such as peroxyhexanoic acid and/or related radicals induced through the reaction with oxygen. The second step seems to consist at least of two routes which are independent of each other and require water. One route is assumed to be an amino-carbonyl reaction involving lysine residues. The other route seems responsible for the attack on tryptophan and methionine residues through oxidation involving the radicals.