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Diagnostic method for detecting a GABA(A) related autoimmune disease and related subject-matter

A technology for autoimmune diseases and autoantibodies, applied in disease diagnosis, biological testing, measuring devices, etc.

Active Publication Date: 2016-05-11
INST DINVESTIGACIONES BIOMEDIQUES AUGUST PI I SUNYER +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Today, approximately 70% of cases of encephalitis of unknown etiology remain undiagnosed after adequate evaluation of the infectious etiology

Method used

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  • Diagnostic method for detecting a GABA(A) related autoimmune disease and related subject-matter
  • Diagnostic method for detecting a GABA(A) related autoimmune disease and related subject-matter
  • Diagnostic method for detecting a GABA(A) related autoimmune disease and related subject-matter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0118] Example 1: Patients

[0119] From August 2012 to February 2013, prospectively identified two patients with encephalitis, refractory seizures and the following CSF, which showed similar cell surface proteins to the nerve blanket of the rat brain Reactive patterns (index patients 1 and 2). The severity of the clinical phenomenon and the unknown antigen identity prompted us to isolate and characterize it and to retrospectively review the clinical and immune information of patients with similar characteristics.

[0120] From April 2006 to April 2013, 1134 patients with suspected diseases were studied at the Neurology Service Center of the Department of Neurology of the University of Pennsylvania Hospital or the Outpatient Department of the University of Barcelona Hospital (now the Center for Neuroimmunology, Institutd'lnvestigacionsBiomediquesAugustPiiSunyer[IDIBAPS]) Serum and CSF of patients with autoimmune encephalitis and seizures.

[0121] Of the 1134 patients, 356 (44%) ha...

Embodiment 2

[0138] Example 2: Immunohistochemistry of rat brain

[0139] The adult female Wistar rats were sacrificed without perfusion. The brain was taken out and fixed by immersing in 4% paraformaldehyde at 4°C for 1 hour, cryoprotecting in 40% sucrose for 48 hours, and embedding in a freezing composite medium. Frozen in isopentane cooled with liquid nitrogen. The 7-micron-thick tissue sections were then sequentially combined with 0.3% H 2 O 2 Incubate for 15 minutes, incubate with 5% goat serum for 1 hour, and incubate with patient serum or control serum (1:200), CSF (1:5) at 4°C overnight.

[0140] After using a suitable biotinylated secondary antibody (goat anti-human BA-3000, dilution 1:2000), as reported, the reactivity was developed by the avidin-biotin-peroxidase method. 1

Embodiment 3

[0141] Example 3: Immunocytochemistry on neuronal culture

[0142] The rat hippocampal neuron culture was prepared as reported. 5 The live neurons cultured on the coverslip were incubated with patient serum or control serum (final dilution 1:200) or CSF (1:10) at 37°C for 1 hour. After removing the medium and washing thoroughly with phosphate buffered saline (PBS), the neurons were fixed with 4% paraformaldehyde, permeabilized with 0.1% TritonX-100, and with AlexaFluor488 goat anti-human IgG (dilution 1:1000) , Invitrogen, A11013) immunolabeling. Zeiss Axiovision software (Zeiss, Thomwood, NY) was used under a fluorescence microscope, and the results were photographed.

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PUM

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Abstract

The present invention provides a use of a GABA(A)R, GABA(A)R fragment, or homolog thereof or a cell expressing the GABA(A)R, GABA(A)R fragment, or homolog thereof for the prognosis, diagnosis or treatment of an autoimmune disease in a subject, methods of prognosticating, diagnosing or treating an autoimmune disease, an autoantibody binding to a GABA(A)R, GABA(A)R fragment, or homolog thereof, a method for isolating an antibody binding to a GABA(A)R, GABA(A)R fragment, or homolog thereof, and a test kit, pharmaceutical composition and medical or diagnostic device comprising a GABA(A)R, GABA(A)R fragment, or homolog thereof.

Description

Invention field [0001] The present invention relates to predicting, diagnosing and treating newly identified autoimmune diseases, providing new cell surface autoantigens, and related means and methods for detecting and treating the autoimmune diseases. Background technique [0002] There is evidence that seizures and status pilepticus can be produced by immune responses to excitatory or inhibitory synaptic receptors or proteins that bind to these receptors. 6 These include N-methyl-D-aspartate receptor (NMDAR), α-amino-3-hydroxy-5-methyl-4-iso Azolpropionic acid receptor (AMPAR), γ-aminobutyric acid-B receptor (GABA(B)R), leucine-rich glioma inactivating protein 1 (LGI1), contact protein-related protein-like 2 ( Caspr2) and DPPX (the regulatory subunit of the Kv4.2 potassium channel). 21 Unless the immune mechanism is identified and treated, the seizures accompanying any of these conditions often resist anti-epileptic therapy. 2,15,22 In some patients, seizures or status epilepti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/94C07K14/47
CPCC07K14/4713G01N33/564G01N33/9426G01N2800/2857A61K38/00G01N2333/70571
Inventor J·达尔毛
Owner INST DINVESTIGACIONES BIOMEDIQUES AUGUST PI I SUNYER
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