Complement receptors 1 and 2 influence the immune environment in a B cell receptor-independent manner

The CD21/35 proteins are complement receptors implicated in controlling and interpreting activation states of the innate and acquired immune responses. One defect of CD21/35(-/-) animals is depressed production of Ag-specific IgG3 which we show is evident in vivo but not in vitro. Gene expression profiles obtained from naive wild-type and CD21/35(-/-) splenocytes demonstrated enhanced expression of inflammatory mediators from CD11b(+) splenocytes in the CD21/35(-/-) animals. Splenocyte populations between wild-type and CD21/35(-/-) mice were similar except for a moderate increase in GR1(low)CD31(+) immature myeloid cells. Furthermore, depletion of neutrophils and other GR1-expressing cells alleviates elevated inflammatory gene expression in the CD21/35(-/-) spleen. Complement activation also plays a key role in the differential gene expression observed in the CD21/35-deficient mouse as depletion of C3 or inhibition of C3a receptor signaling within the animal returned inflammatory gene expression within the spleen to wild-type levels. Finally, C3 depletion before immunization allowed for the enhanced production of Ag-specific IgG3 production in the CD21/35(-/-) mouse compared with mock-depleted animals. These data suggest that the overall environment of the CD21/35(-/-) spleen is quite different from that of the wild-type animal perhaps due to altered complement convertase activity. This difference may be responsible for a number of the phenotypes ascribed to the deficiency of CD21/35 proteins on B cells and follicular dendritic cells.     

High Expression of CD244 and SAP Regulated CD8+ T Cell Responses of Patients with HTLV-I Associated Neurologic Disease

HTLV-I-specific CD8⁺ T cells have been characterized with high frequencies in peripheral blood and cerebrospinal fluid and production of proinflammatory cytokines, which contribute to central nervous system inflammation in HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). However, little is known about the differences in CD8⁺ T cell activation status between asymptomatic carrier (ACs) and patients with HAM/TSP. The expression of CD244, a signaling lymphocyte activation molecule (SLAM) family receptor, was significantly higher on CD8⁺ T cells in HTLV-I-infected patients, both ACs and patients with HAM/TSP, than those on healthy normal donors (NDs). Blockade of CD244 inhibited degranulation and IFN-γ production in CD8⁺ T cells of patients with HAM/TSP, suggesting that CD244 is associated with effector functions of CD8⁺ T cells in patients with HAM/TSP. Moreover, SLAM-associated protein (SAP) was overexpressed in patients with HAM/TSP compared to ACs and NDs. SAP expression in Tax-specific CTLs was correlated in the HTLV-I proviral DNA loads and the frequency of the cells in HTLV-I-infected patients. SAP knockdown by siRNA also inhibited IFN-γ production in CD8⁺ T cells of patients with HAM/TSP. Thus, the CD244/SAP pathway was involved in the active regulation of CD8⁺ T cells of patients with HAM/TSP, and may play roles in promoting inflammatory neurological disease.     

Looking at lipid rafts?

The notion that microdomains enriched in certain specialized lipids exist in membranes has been both attractive and controversial since it was first proposed that such domains, termed rafts, might act as apical sorting devices in epithelial cells. The observation that certain lipids are not extractable in cold nonionic detergent supports the raft concept, but the nature of the in vivo correlate of such detergent-resistant membranes remains enigmatic. In principle, microscopy should be able to determine whether the postulated rafts exist. This article focuses on recent microscopy experiments addressing this question. Several, but not all, results support the raft concept, but further definition of the structure, dynamics and function of lipid domains in various biological contexts is urgently required.     

Mechanical ventilation uncouples synthesis and assembly of elastin and increases apoptosis in lungs of newborn mice. Prelude to defective alveolar septation during lung development?

Prolonged mechanical ventilation (MV) with O2-rich gas inhibits lung growth and causes excess, disordered accumulation of lung elastin in preterm infants, often resulting in chronic lung disease (CLD). Using newborn mice, in which alveolarization occurs postnatally, we designed studies to determine how MV with either 40% O2 or air might lead to dysregulated elastin production and impaired lung septation. MV of newborn mice for 8 h with either 40% O2 or air increased lung mRNA for tropoelastin and lysyl oxidase, relative to unventilated controls, without increasing lung expression of genes that regulate elastic fiber assembly (lysyl oxidase-like-1, fibrillin-1, fibrillin-2, fibulin-5, emilin-1). Serine elastase activity in lung increased fourfold after MV with 40% O2, but not with air. We then extended MV with 40% O2 to 24 h and found that lung content of tropoelastin protein doubled, whereas lung content of elastin assembly proteins did not change (lysyl oxidases, fibrillins) or decreased (fibulin-5, emilin-1). Quantitative image analysis of lung sections showed that elastic fiber density increased by 50% after MV for 24 h, with elastin distributed throughout the walls of air spaces, rather than at septal tips, as in control lungs. Dysregulation of elastin was associated with a threefold increase in lung cell apoptosis (TUNEL and caspase-3 assays), which might account for the increased air space size previously reported in this model. Our findings of increased elastin synthesis, coupled with increased elastase activity and reduced lung abundance of proteins that regulate elastic fiber assembly, could explain altered lung elastin deposition, increased apoptosis, and defective septation, as observed in CLD.     

Anti-CD22-MCC-DM1 and MC-MMAF conjugates: impact of assay format on pharmacokinetic parameters determination

CD22 represents a promising target for antibody-drug conjugate therapy in the context of B cell malignancies since it rapidly internalizes, importing specifically bound antibodies with it. To determine the pharmacokinetic parameters of anti-CD22-MCC-DM1 and MC-MMAF conjugates, various approaches to quantifying total and conjugated antibody were investigated. Although the total antibody assay formats gave similar results for both conjugates, the mouse pharmacokinetic profile for the anti-CD22-MCC-DM1 and MC-MMAF appeared significantly different depending on the conjugated antibody assay format. Since these differences significantly impacted the PK parameters determination, we investigated the effect of the drug/antibody ratio on the total and conjugated antibody quantification using multiple assay formats. Our investigations revealed the limitations of some assay formats to quantify anti-CD22-MCC-DM1 and MC-MMAF with different drug load and in the context of a heterogeneous ADC population highlight the need to carefully plan the assay strategy for the total and conjugated antibody quantification in order to accurately determine the ADC PK parameters.     

Agrin and neuregulin, expanding roles and implications for therapeutics

Agrin and neuregulin are broadly expressed molecules that have significant developmental roles. Here we review the diverse temporal and spatial expression patterns and functions of these molecules and the impact that dysregulation may have on a number of disease states. Many know agrin as a modulator of synaptogenesis and the neuregulins for their prominent role in breast cancer; this review elaborates on many of the other proposed functions for these molecules both in the nervous system and elsewhere. In several instances we discuss the possible use of agrin, neuregulin and related molecules as therapeutic agents.     

Groundwater nutrient concentrations near an incised midwestern stream: effects of floodplain lithology and land management

It has been recognized that subsurface lithology plays an important role in controlling nutrient cycling and transport in riparian zones. In Iowa and adjacent states, the majority of alluvium preserved in small and moderate sized valleys consists of Holocene-age organic-rich, and fine-grained loam. In this paper, we describe and evaluate spatial and temporal patterns of lithology and groundwater nutrient concentrations at a riparian well transect across Walnut Creek at the Neal Smith National Wildlife Refuge in Jasper County, Iowa. Land treatment on one side of the stream reduced the grass cover to bare ground and allowed assessment of the effects of land management on nutrient concentrations. Results indicated that groundwater in Holocene alluvium is very nutrient rich with background concentrations of nitrogen, phosphorus and dissolved organic carbon that exceed many environmentally sensitive criteria. Average concentrations of ammonium exceeded 1 mg/l in several wells under grass cover whereas nitrate concentrations exceeded 20 mg/l in wells under bare ground. Phosphate concentrations ranged from 0.1 to 1.3 mg/l and DOC concentrations exceeded 5 mg/l in many wells. Denitrification, channel incision, land management and geologic age of alluvium were found to contribute to variable nutrient loading patterns at the site. Study results indicated that riparian zones of incised streams downcutting through nutrient-rich Holocene alluvium can potentially be a significant source of nutrient loadings to streams.     

Structure-activity relationships of 2-chloro-N6-substituted-4'-thioadenosine-5'-N,N-dialkyluronamides as human A3 adenosine receptor antagonists

On the basis of potent and selective A(3) adenosine receptor (AR) antagonist, 2-chloro-N(6)-(3-iodobenzyl)-4'-thioadenosine-5'-N,N-dimethyluronamide, structure-activity relationships were studied for a series of 5'-N,N-dialkyluronamide derivatives, synthesized from D-gulonic gamma-lactone. From this study, it was revealed that removal of the hydrogen bond-donating ability of the 5'-uronamide was essential for the pure A(3)AR antagonism. 5'-N,N-Dimethyluronamide derivatives exhibited higher binding affinity than larger 5'-N,N-dialkyl or 5'-N,N-cycloalkylamide derivatives, indicating that steric factors are crucial in binding to the human A(3)AR. A N(6)-(3-bromobenzyl) derivative 6c (K(i)=9.32 nM) exhibited the highest binding affinity at the human A(3)AR with very low binding affinities to other AR subtypes.