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Diagnostic methods and related topics for the detection of GABA(a)-associated autoimmune diseases

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

Active Publication Date: 2020-11-06
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 methods and related topics for the detection of GABA(a)-associated autoimmune diseases
  • Diagnostic methods and related topics for the detection of GABA(a)-associated autoimmune diseases
  • Diagnostic methods and related topics for the detection of GABA(a)-associated autoimmune diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0122] Embodiment 1: patient

[0123] From August 2012 to February 2013, two patients were prospectively identified with encephalitis, refractory seizures, and CSF that showed a similarity to the cell surface protein of the neural blanket of the rat brain Reactive pattern (index patients 1 and 2). The severity of the clinical picture and the unknown antigenic identity prompted us to isolate and characterize it and to review retrospectively the clinical and immunological information of patients with similar characteristics.

[0124] From April 2006 to April 2013, 1134 patients were studied at the Neurology Department of the Hospital of the University of Pennsylvania or the Neurological Service of the Outpatient Department of the University Hospital of Barcelona (now, Center of Neuroimmunology, Institut d'lnvestigacions Biomediques August Pi i Sunyer [IDIBAPS]). Serum and CSF of a patient with suspected autoimmune encephalitis and seizures.

[0125] Of 1134 patients, 356 (44%)...

Embodiment 2

[0142] Example 2: Immunohistochemistry of Rat Brain

[0143] Adult female Wistar rats were sacrificed without perfusion, brains were removed and fixed by immersion in 4% paraformaldehyde at 4°C for 1 hour, cryoprotected in 40% sucrose for 48 hours, embedded in cryocomplex medium and Snap freeze in isopentane chilled with liquid nitrogen. 7 µm thick tissue sections were subsequently sequentially treated with 0.3% HO 2 o 2 Incubate for 15 minutes, 1 hour with 5% goat serum, and overnight at 4°C with patient serum or control serum (1:200), CSF (1:5).

[0144] After using an appropriate biotinylated secondary antibody (goat anti-human BA-3000, dilution 1:2000), reactivity was developed with the avidin-biotin-peroxidase method as reported. 1

Embodiment 3

[0145] Example 3: Immunocytochemistry on Neuronal Cultures

[0146] Rat hippocampal neuronal cultures were prepared as reported. 5 Live neurons grown on coverslips were incubated with patient serum or control serum (final dilution 1:200) or CSF (1:10) at 37°C for 1 hr. After removing the medium and washing extensively with phosphate-buffered saline (PBS), neurons were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, and stained with Alexa Fluor 488 goat anti-human IgG (dilution 1:1000, Invitrogen, A11013) immunolabeling. Results were photographed under a fluorescence microscope using Zeiss Axiovision software (Zeiss, Thomwood, NY).

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PUM

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Abstract

The present invention provides GABA(A)R, GABA(A)R fragments or homologues thereof or cells expressing GABA(A)R, GABA(A)R fragments or homologues thereof for prognostic, diagnostic or therapeutic testing Uses for autoimmune diseases in patients, methods for predicting, diagnosing or treating autoimmune diseases, autoantibodies binding to GABA(A)R, GABA(A)R fragments or homologues thereof, for isolating and binding to GABA(A) Methods for antibodies binding to R, GABA(A)R fragments or homologues thereof, and test kits, pharmaceutical compositions and medical or diagnostic applications comprising GABA(A)R, GABA(A)R fragments or homologues thereof device.

Description

[0001] field of invention [0002] The present invention relates to prediction, diagnosis and treatment of newly identified autoimmune diseases, provision of new cell surface autoantigens and related means and methods for detection and treatment of said autoimmune diseases. Background technique [0003] There is evidence that seizures and status epilepticus can result from immune responses directed against excitatory or inhibitory synaptic receptors or proteins that bind these receptors. 6 These include the N-methyl-D-aspartate receptor (NMDAR), α-amino-3-hydroxy-5-methyl-4-iso azole propionic acid receptor (AMPAR), gamma-aminobutyric acid-B receptor (GABA(B)R), leucine-rich glioma inactivating protein 1 (LGI1), contactin-associated protein-like 2 ( Caspr2) and DPPX (regulatory subunit of the Kv4.2 potassium channel). 21 Seizures associated with any of these disorders are often resistant to antiepileptic therapy unless the immune mechanism is identified and treated. 2,15,2...

Claims

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Application Information

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