Biomedical subjects

Brian A Cobb

Publications and source records attributed to Brian A Cobb.

7 recordsLinked to original sources

PubMed2006

A bacterial carbohydrate links innate and adaptive responses through Toll-like receptor 2.

Commensalism is critical to a healthy Th1/Th2 cell balance. Polysaccharide A (PSA), which is produced by the intestinal commensal Bacteroides fragilis, activates CD4+ T cells, resulting in a Th1 response correcting the Th2 cell skew of germ-free mice. We identify Toll-like receptors as crucial to the convergence of innate and adaptive responses stimulated by PSA. Optimization of the Th1 cytokine interferon-gamma in PSA-stimulated dendritic cell-CD4+ T cell co-cultures depends on both Toll-like receptor (TLR) 2 and antigen presentation. Synergy between the innate and adaptive responses was also shown when TLR2-/- mice exhibited impaired intraabdominal abscess formation in response to B. fragilis. Commensal bacteria, using molecules like PSA, potentially modulate the Th1/Th2 cell balance and the response to infection by coordinating both the innate and adaptive pathways.

Animals↗

PubMed2006

Structure and function relations with a T-cell-activating polysaccharide antigen using circular dichroism.

Studies centered on understanding how molecular structure affects biological function have historically focused on proteins. Circular dichroism (CD) is commonly used to analyze protein secondary structure, yet its application to other molecules is far less explored. In fact, little is known about how glycan conformation might affect function, likely because of a lack of tools for measuring dynamic structural changes of carbohydrates. In the present study, we developed a method based on CD to monitor conformational changes in the zwitterionic T-cell-activating glycoantigen polysaccharide A1 (PSA). We found that PSA helical structure produces a CD spectrum that is strikingly similar to proteins rich in alpha-helical content and is equally sensitive to nonpolar solvents. Like conventional T-cell-dependent proteins, PSA requires processing before major histocompatibility complex class II (MHCII) binding. CD spectra of PSA fragments of varying sizes indicated that fragments smaller than three repeating units lack helical content and are incapable of MHCII binding. Likewise, neutralization of charged groups in the repeating unit resulted in major conformational changes as measured by CD, which correlated with a lack of MHCII presentation. These data represent two significant findings: CD can be used to measure conformational changes in carbohydrates and the functional epitope from PSA is dependent on a specific conformation that is stabilized by adjacent repeating units and a zwitterionic charge motif. As a result, this work demonstrates that CD is a valuable tool for use in functional glycomics efforts that seek to align chemical and conformational structure with biological activity.

Circular Dichroism↗

PubMed2005

Coming of age: carbohydrates and immunity.

Adaptive immune responses have long been considered the "territory" of antigenic proteins, whereas carbohydrates are characterized as T-cell-independent antigens that are not typically recognized by the complete adaptive machinery. The current modus operandi when searching for dominant epitopes is the use of synthetic peptides designed from the primary structure of interesting target proteins; however, there is growing evidence that sugars can also play a critical role in immune recognition. Findings reported in this issue of the European Journal of Immunology begin to shed light on the differences in protein glycosylation that can occur in association with disorders like rheumatoid arthritis and the effect these changes have on collagen recognition by the immune system. Other recent studies have shown that immunodominant glycopeptide "remnant" epitopes as well as glycosylation changes on self-proteins can generate autoimmunity. Finally, some types of carbohydrates are now known to be processed and presented to T cells by class II MHC. Taken together, these advances illustrate a clear importance for carbohydrate recognition in foreign and self antigens by the adaptive immune system. With the common presence of carbohydrate molecules on eukaryotic, prokaryotic, and viral surfaces, the impact of carbohydrates on adaptive immunity is now indisputable.

Animals↗

PubMed2005

Zwitterionic capsular polysaccharides: the new MHCII-dependent antigens.

The immune system has evolved the ability for T cells to recognize nearly any biological polymer, including peptides, protein superantigens, and glycolipids through presentation by the major histocompatibility complex (MHC) proteins such as MHC class I (MHCI), MHC class II (MHCII), and CD1. A recent and unexpected addition to this list is the zwitterionic capsular polysaccharide (ZPS). These bacterial molecules utilize MHCII presentation to activate T cells via recognition by alphabeta T cell receptor (alphabetaTCR) proteins. In this review, we explore what is currently known about ZPS processing and presentation within antigen-presenting cells (APCs) and the immune response that follows.

Abscess↗

PubMed2004

Polysaccharide processing and presentation by the MHCII pathway.

The adaptive immune system functions through the combined action of antigen-presenting cells (APCs) and T cells. Specifically, class I major histocompatibility complex antigen presentation to CD8(+) T cells is limited to proteosome-generated peptides from intracellular pathogens while the class II (MHCII) endocytic pathway presents only proteolytic peptides from extracellular pathogens to CD4(+) T cells. Carbohydrates have been thought to stimulate immune responses independently of T cells; however, zwitterionic polysaccharides (ZPSs) from the capsules of some bacteria can activate CD4(+) T cells. Here we show that ZPSs are processed to low molecular weight carbohydrates by a nitric oxide-mediated mechanism and presented to T cells through the MHCII endocytic pathway. Furthermore, these carbohydrates bind to MHCII inside APCs for presentation to T cells. Our observations begin to elucidate the mechanisms by which some carbohydrates induce important immunologic responses through T cell activation, suggesting a fundamental shift in the MHCII presentation paradigm.

Animals↗

PubMed2002

Factors influencing alpha-crystallin association with phospholipid vesicles.

PURPOSE: Lens lipids undergo a number of changes with age, including an overall increase in phospholipid acyl chain saturation and a decrease in length. In addition, the amount of membrane bound alpha-crystallin increases dramatically with age and with the onset of cataract. The aim of this study was to determine if a link exists between age and cataract associated changes in lens lipids and the changes in alpha-crystallin membrane association. METHODS: Protein-free lipid vesicles composed of a wide variety of synthetic and lens-derived lipid vesicles were formed by sonication. These vesicles were used with fluorescent native and recombinant alpha-crystallin conjugates in vesicle binding assays. Vesicles were collected by centrifugation and bound alpha-crystallin was quantified with fluorescence intensity measurements. RESULTS: alpha-Crystallin complexes showed remarkably similar binding profiles for all lipid vesicles tested, regardless of lipid origin, phospholipid head group, acyl chain length or saturation, and inclusion of cholesterol. In addition, recombinant alpha-crystallin complexes bind to these vesicles in a manner that is essentially indistinguishable from that of native human and bovine alpha-crystallins. Unlike alpha-crystallin binding to lens membranes containing intrinsic proteins, binding of alpha-crystallin to protein-free vesicles was very high capacity and unsaturable. CONCLUSIONS: We conclude from these data that the binding of alpha-crystallin to lens membranes is not lipid-specific. Furthermore, protein post-translational changes, such as phosphorylation, do not appear to alter alpha-crystallin binding to these vesicles. Given the linearity of the binding curves, we propose that the only limiting factor for normal alpha-crystallin membrane binding is available surface area on the bilayer. Finally, the present data suggests that increased in vivo membrane association of alpha-crystallin is not a result of lipid changes, but more likely a result of non-lipid factors such as the accumulation of high molecular weight forms of alpha-crystallin.

Animals↗

PubMed2002

alpha-Crystallin chaperone-like activity and membrane binding in age-related cataracts.

alpha-Crystallin, the major protein component of the vertebrate lens, is thought to play a critical role in the maintenance of transparency through its ability to inhibit stress-induced protein aggregation. However, during aging and cataract formation the amount of membrane-bound alpha-crystallin increases significantly while high molecular weight complexes (HMWCs) comprised of alpha-crystallin and other lens crystallins accumulate. These and other recent data suggest a possible link between cataract formation, the formation of high molecular weight alpha-crystallin aggregates, and the progressive increase in membrane association of alpha-crystallin. To better understand these processes, we characterized the chaperone-like activity (CLA) and subunit exchange of membrane bound alpha-crystallin. In addition, we measured the membrane binding properties of in vitro constituted HMWCs to understand the mechanism by which increased alpha-crystallin is bound to the membrane of old and cataractous lens cells in vivo. Membrane-associated alpha-crystallin complexes have measurably reduced CLA compared to complexes in solution; however, membrane binding does not alter the time required for alpha-crystallin complexes to reach subunit exchange equilibrium. In addition, HMWCs prepared in vitro have a profoundly increased membrane binding capacity as compared to native alpha-crystallin. These results are consistent with a model in which increased membrane binding of alpha-crystallin is an integral step in the pathogenesis of many forms of cataracts.

1,2-Dipalmitoylphosphatidylcholine↗