We constructed miniaturized autoantigen arrays to perform large-scale multiplex characterization of autoantibody responses directed against structurally diverse autoantigens, using submicroliter quantities of clinical samples. Autoantigen microarrays were produced by attaching hundreds of proteins, peptides and other biomolecules to the surface of derivatized glass slides using a robotic arrayer. Arrays were incubated with patient serum, and spectrally resolvable fluorescent labels were used to detect autoantibody binding to specific autoantigens on the array. We describe and characterize arrays containing the major autoantigens in eight distinct human autoimmune diseases, including systemic lupus erythematosus and rheumatoid arthritis. This represents the first report of application of such technology to multiple human disease sera, and will enable validated detection of antibodies recognizing autoantigens including proteins, peptides, enzyme complexes, ribonucleoprotein complexes, DNA and post-translationally modified antigens. Autoantigen microarrays represent a powerful tool to study the specificity and pathogenesis of autoantibody responses, and to identify and define relevant autoantigens in human autoimmune diseases.

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Abstract. The response of a surface-trapped river plume to an upwelling favorable wind is studied using a three-dimensional model in a simple, rectangular domain. Model simulations demonstrate that the plume thins and is advected offshore by the cross-shore Ekman transport. The thinned plume is susceptible to significant mixing because of the vertically sheared horizontal currents. The Ekman dynamics and shear-induced mixing cause the plume to evolve to a quasi-steady uniform thickness, which can be estimated by a critical Richardson number criterion. Although the mixing rate decreases slowly in time, mixing continues under a sustained upwelling wind until the plume is destroyed. Mixing persists at the seaward plume front because of an Ekman straining mechanism in which there is a balance between the advection of cross-shore salinity gradients and vertical mixing. The plume mixing rate observed is similar to the mixing law obtained by previous studies of one-dimensional mixing, although the river plume mixing is essentially twodimensional. IntroductionIt has long been recognized that local winds play an important role in the dynamics of river plumes. A theoretical study by Csanady [1978] Although the basic tendency for the plume to spread offshore during upwelling winds has been observed in the aforementioned studies, none of these studies quantifies the plume motions in response to upwelling winds nor determines whether or not the Ekman physics are the only important part of the dynamical balance. Fong et al. [1997] provide one of the first quantitative tests of the plume response to alongshore winds based on observations of the western Gulf of Maine plume. They find that the motions at the seaward front of the plume are approximately described by an Ekman-dominated alongshore momentum balance. It is likely that the Ekman physics is important for the entire plume behavior, and one might expect the Ekman response to place strong constraints on how the structure of the plume is modified during an up- welling favorable wind event. The previous studies suggest that one consequence of upwelling winds is to thin the plume. However, the details of this thinning process have not been quantified or described in any detail.The wind-induced mixing of a river plume has received little attention in previous studies. Masse and Murthy [1992] observe the spreading of the thermally driven Niagara River plume to behave qualitatively consistently with the Ekman response. They suggest that the secondary effect of winds is to mix the plume and ambient waters. They argue that strong upwelling winds will enhance plume mixing by blowing the plume offshore and weakening the vertical density gradients. The enhanced shears induced by the thinning plume may make the plume more susceptible to shear-induced turbulent mixing. Souza and Simpson [1997] also note that winds may be important in driving mixing in a plume but do not describe the mechanism by which it would be accomplished. There remains the questions of how the thinning and sp...

The diversity of autoimmune responses poses a formidable challenge to the development of antigen-specific tolerizing therapy. We developed 'myelin proteome' microarrays to profile the evolution of autoantibody responses in experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis (MS). Increased diversity of autoantibody responses in acute EAE predicted a more severe clinical course. Chronic EAE was associated with previously undescribed extensive intra- and intermolecular epitope spreading of autoreactive B-cell responses. Array analysis of autoantigens targeted in acute EAE was used to guide the choice of autoantigen cDNAs to be incorporated into expression plasmids so as to generate tolerizing vaccines. Tolerizing DNA vaccines encoding a greater number of array-determined myelin targets proved superior in treating established EAE and reduced epitope spreading of autoreactive B-cell responses. Proteomic monitoring of autoantibody responses provides a useful approach to monitor autoimmune disease and to develop and tailor disease- and patient-specific tolerizing DNA vaccines.

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