Autoantibodies in neurological diseases - EUROIMMUN in neurological diseases ... Golgi apparatus, cytoplasmic membrane) ... moleculare of the hippocampus (neuropil staining)

EUROIMMUN AG · Seekamp 31 · 23560 Lübeck (Germany) · Tel +49 451/5855-0 · Fax 5855-591 · [email protected] · www.euroimmun.com

Autoantibodies in neurological diseases

Cerebellum Intestine

Hippocampus HEp-2 cells


Hippocampus HEp-2 cells

Hippocampus Control transfection

Cerebellum

Cerebellum

Optic nerve

Control transfection

Anti-Hu positive Anti-NMDA-receptor positive

Anti-Yo positive Anti-aquaporin-4 positive

NMDAR (transf. cells)

AQP-4 (transf. cells)

Examples of relevant

target antigens

EUROLINEIndirect immunofl uorescence

GM3G

GM1G

GM2G

GQ1bG

GT1bG

GD1aG

GD1bG

ControlC

PNMA2 PMa2/Ta)(M

AmphiphysinA

CV2C

HuH

RiR

YoY

ControlC

PNMA2 PMa2/Ta)(M

AmphiphysinA

CV2C

HuH

RiR

YoY

ControlC

Tr (DNER)T

RecoverinR

SOX1S

TitinT

Zic4Z

GAD65GAD65GG

Hu, Ri, Yo, Tr

CV2

Ma/Ta

Amphiphysin

SOX1

Zic4

ITPR1

CARP

GAD

Recoverin

Titin

Gangliosides

MAG

Myelin

Aquaporin-4

MOG

VGKC (LGI1 + CASPR2)

NMDA receptors

AMPA receptors

GABAB receptors

DPPX

IgLON5

Glycine receptors

AChR

2

3

Autoantibodies IIFT pattern Test systems

Anti-Hu (ANNA-1*)

Autoantibodies against basic, RNA-binding proteins of the neuronal cell nuclei of the central and peripheral nervous system

Associated diseases: encephalomyelitis, subacute sensory neuronopathy (Denny-Brown syndrome), autonomous neuropathy

Associated tumours: small-cell lung carcinoma, neuroblastoma

IIFT: Granular fl uorescence of almost all neuronal nuclei on the substrates cerebellum and hippocampus. The cell nuclei of the plexus myentericus (intestinal tissue) are also positive.

EUROLINE: Positive reaction of the recombinant Hu antigen (HuD).

Anti-Ri (ANNA-2*)

Autoantibodies against neuronal cell nuclei of the central nervous system (CNS)

Associated disease: opsoclonus myoclonus syn-drome

Associated tumours: small-cell lung carcinoma, breast carcinoma

IIFT: Granular fl uorescence of almost all neuronal nuclei on the substrates cerebellum and hippocampus. The substrate intestine (plexus myentericus) shows no reac-tion.

EUROLINE: Positive reaction of the recombinant Ri anti-gen (NOVA1***).

Anti-Yo (PCA-1**)

Autoantibodies against antigens in the cytoplasm (rough ER, Golgi apparatus, cytoplasmic membrane) of the cerebellar Purkinje cells

Associated disease: cerebellar degeneration

Associated tumours: ovarian carcinoma, breast carcinoma, uterine carcinoma

IIFT: Cytoplasmic fl uorescence of the Purkinje cells on the substrate cerebellum and of individual cells in the hilus of the hippocampus. The substrate intestine (plexus myentericus) shows no specifi c reaction.

Monospecifi c test using Yo/CDR2-transfected HEK**** cells

EUROLINE: Positive reaction of the recombinant Yo antigen (CDR62*****).

*ANNA = anti-neuronal nuclear antibodies **PCA = Purkinje cell autoantibodies ***NOVA = neuronal onconeural ventral nervous system antigen ****HEK = human embryonic kidney *****CDR = cerebellar degeneration-related antigen

Intestine

Intestine


Cerebellum

Cerebellum Yo (transf. cells)

4


PCA-2*

Autoantibodies against a 280 kDa protein of the cerebellar Purkinje cells

Associated diseases: limbic/brain stem encephalitis, cerebellar ataxia, Lambert-Eaton myasthenic syndrome (LEMS), autonomous neuropathy, motor neuropathy

Associated tumours: gynaecological tumours, small-cell lung carcinoma

IIFT: Fluorescence of the cytoplasm of the cerebellar Purkinje cells, which extends into the dendrites.

Anti-Tr

Autoantibodies against DNER** protein in the Purkinje cells of the cerebellum


Associated tumour: Hodgkin‘s lymphoma

Coarse granular fl uorescence of the cytoplasm of the cerebellar Purkinje cells, dot-like staining of the molecular layer.

Monospecifi c test using DNER-transfected HEK cells.

EUROLINE: Positive reaction of the specifi c targetantigen Tr (DNER).

Anti-CARP (anti-carbonic anhydrase-related protein VIII)

Autoantibodies against CARP VIII in the Purkinje cell layer of the cerebellum

Associated diseases: paraneoplastic cerebellar degeneration, cerebellar ataxia

Associated tumours: melanoma, ovarian carcinoma

IIFT: The molecular layer and the cytoplasm of the Purkinje cells in the cerebellum show a dotted to fi ne-speckled pattern.

Monospecifi c test using CARP VIII-transfected HEK cells.

*PCA = Purkinje cell autoantibodies **DNER = delta/notch-like epidermal growth factor-related receptor

Cerebellum

Cerebellum

Cerebellum DNER (transf. cells)

CARP (transf. cells)

5


Anti-ITPR1 (Anti-Sj)

(anti-Sj / inositol 1,4,5-trisphosphate receptor)

Autoantibodies against ITPR1 in Purkinje cells of the cerebellum

Associated disease: cerebellar ataxia

Associated tumour: NSCLC in an unpublished case

IIFT: The molecular layer and the cytoplasm of the Purkinje cells in the cerebellum show a dotted to fi ne-speckled pattern.

Monospecifi c test using ITPR1-transfected HEK cells.

Anti-glia nuclear antibodies (AGNA)

Possible antigen: SOX1

Associated diseases: paraneoplastic Lambert-Eaton myasthenic syndrome (LEMS), cerebellar degeneration, sensory neuropathy

Associated tumour: small-cell lung carcinoma

IIFT: Fluorescence of the cell nuclei of the Bergmann glia in the Purkinje cell layer of the cerebellum.

Anti-Ma (Ma1, Ma2/Ta)

Autoantibodies against Ma1 (PNMA1*; 37 kDa) and Ma2/Ta (PNMA2*; 40 kDa), proteins in the nucleoli of the neuronal cell nuclei

Associated diseases: brain stem encephalitis, limbic encephalitis

Associated tumours: lung carcinoma, testicular carcinoma (esp. Ma2/Ta), breast carcinoma

IIFT: Reaction of the nerve cell nucleoli on the substrates cerebellum, cerebrum and hippocampus.

EUROLINE: Anti-Ma (Ma1 and Ma2/Ta) antibodies produce a positive reaction with the recombinant Ma2/Ta (PNMA2*) antigen.

Cerebellum

Cerebellum

Cerebellum

ITPR1 (transf. cells)

Cerebrum

PNMA1/2 = paraneoplastic antigen Ma1/2

6


Anti-CV2

Autoantibodies against CV2, a 66-kDa protein

Associated diseases: sensory and autonomous neuropathy, encephalitis, extrapyramidal motor syndrome, cerebellar degeneration

Associated tumours: small-cell lung carcinoma, thymoma

IIFT: Sand-like fl uorescence on the substrate cerebellum, most visible in the stratum moleculare.

EUROLINE: Positive reaction of the recombinant CV2.

Anti-amphiphysin

Autoantibodies against amphiphysin, a synaptic protein with a molecular weight of 128 kDa

Associated disease: paraneoplastic stiff-person syndrome

Associated tumours: breast carcinoma, small-cell lung carcinoma

IIFT: Fluorescence of the presynaptic nerve ends in the cerebellum. The nerve dendrites of the stratum moleculare show a more intense fl uorescence than the stratum granulosum. The fl uorescence pattern of the stratum granulosum resembles the pattern found with antibodies against GAD.

EUROLINE: Positive reaction of the specifi c target antigen amphiphysin.

Anti-GAD

Autoantibodies against the enzyme glutamic acid decarboxylase

Associated diseases: stiff-person syndrome, limbic encephalitis

IIFT: Patchy fl uorescence of the stratum granulosum on the substrates cerebellum and hippocampus. Fluorescence of the pancreas islets on the substrate pancreas.

Monospecifi c test using GAD-transfected HEK cells.

EUROLINE: Monospecifi c test using purifi ed GAD.

Cerebellum

Cerebellum

Cerebellum Pancreas

GAD (transf. cells)

7


Anti-ZIC4

Autoantibodies against zinc fi nger proteins (probably ZIC1, 2 and 4)



IIFT: ANA-like fl uorescence of the neuronal nuclei of the cerebellar stratum granulosum. The cell nuclei of the Purkinje cells are negative in the presence of anti-ZIC4 antibodies, whereas with ANA* all cells are stained. Antibodies against ZIC4 should be assessed not only on cerebellar sections, but also on transfected cells in parallel.

EUROLINE: Positive reaction of the specifi c targetantigen ZIC4.

Anti-MAG

Autoantibodies against myelin-associated glycoprotein, a cell membrane protein of the myelin sheaths

Associated diseases: paraproteinaemic neuropathy, particularly in the context of monoclonal IgM gammopathy, e.g. monoclonal gammopathy of undetermined signifi cance (MGUS) and Waldenström‘s syndrome

IIFT: Fluorescence of the lamina alba in the cerebellum. Annular fl uorescence on the substrate peripheral nerve.

Clinically relevant antibodies are mainly those of class IgM.

Anti-myelin

Autoantibodies against myelin and myelin proteins (e.g. MBP**) in the myelin sheath of axons

Associated diseases: diagnostic signifi cance is controversial

IIFT: Fluorescence of the lamina alba in the cerebellum. Fluo rescing hyaline cylinders on the substrate peripheral nerve.

Cerebellum Nerve (section)

Cerebellum ZIC4 (transf. cells)

Cerebellum Nerve (section)

*ANA = anti-nuclear antibodies **MBP = myelin basic protein

8


Anti-AQP-4 (NMO-IgG)

Autoantibodies against aquaporin-4 (AQP-4), a water channel of the dystroglycan protein complex

Associated diseases: neuromyelitis optica (NMO, opticospinal encephalomyelitis, Devic‘s syndrome), longitudinally extensive transverse myelitis (LETM) and recurrent optic neuritis (rec. ON). NMO, LETM and rec. ON can also occur with systemic lupus erythematosus or Sjögren‘s syndrome

IIFT: Perivascular fl uorescence with linear staining along the Virchow-Robin spaces and the microvessels in the grey and white matter on the substrate cerebellum, as well as a reticular fl uorescence on the substrate optic nerve.

Monospecifi c test using AQP4-transfected HEK cells.

Anti-MOG (anti-myelin oligodendrocyte glycoprotein)

Autoantibodies against myelin oligo-dendrocyte glycoprotein, a cell mem-brane protein of the myelin sheaths

Associated diseases: neuromyelitis optica (NMO), NMO spectrum disorders (NMOSD), acute dissemi-nated encephalomyelitis (ADEM), clinically isolated syndrome (CIS), multiple sclerosis

Associated tumour: –

IIFT: The white matter of the cerebellum shows a granular fl uorescence.

Monospecifi c test using MOG-transfected HEK cells.

Anti-NMDA receptors

Autoantibodies against glutamate receptors (type NMDA*)

Associated disease: anti-glutamate receptor (type NMDA) encephalitis (~ 60 % as paraneoplastic syndrome) Associated tumours: ovarian teratoma, testicular teratoma

IIFT: Fluorescence of the stratum moleculare of the hippocampus (neuropil staining) and the stratum granulosum of the cerebellum.

Monospecifi c test using NMDA receptor (NR1)-transfected HEK cells.

Optic nerve AQP-4 (transf. cells)

Cerebellum

MOG (transf. cells)Cerebellum

Hippocampus NMDAR (transf. cells)

Cerebellum

*NMDA = N-methyl-D-aspartate

9


Anti-AMPA receptors

Autoantibodies against glutamate receptors (type AMPA*)

Associated disease: limbic encephalitis (~ 70 % as paraneoplastic syndrome)

Associated tumours: lung carcinoma, breast carcinoma, malignant thymoma

IIFT: Fluorescence of the stratum moleculare of the hippocampus (neuropil staining), the stratum moleculare and stratum granulosum of the cerebellum and the Purkinje cells.

Monospecifi c test using AMPA1/2-transfected HEK cells.

Anti-GABAB receptors

Autoantibodies againstGABAB

** receptors

Associated disease: limbic encephalitis (~ 50 % as paraneoplastic syndrome)


IIFT: Coarse granular fl uorescence of the stratum moleculare of the hippocampus (neuropil staining) and the cerebellum. Also patchy fl uorescence of the cerebellar stratum granulosum.

Monospecifi c test using GABAB receptor-transfected HEK cells.

Anti-LGI1

Autoantibodies against LGI1***, a protein associated with voltage-gated potassium channels (VGKC)

Associated disease: limbic encephalitis

Associated tumours: thyroid carcinoma, small-cell lung carcinoma, kidney cell carcinoma, ovarian teratoma, thymoma

IIFT: Fine granular fl uorescence of the stratum moleculare of the hippocampus (neuropil staining) and the cerebellum. Also patchy fl uorescence of the cerebellar stratum granulosum. The outer molecular layer of the hippocampus shows a stronger reaction than the inner layer.

Monospecifi c test using LGI1-transfected HEK cells.

*AMPA =α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid **GABAB = γ-aminobutyric acid ***LGI1 = leucine-rich glioma-inactivated protein 1

Hippocampus

AMPAR1/R2

(transf. cells)

Cerebellum

Hippocampus GABABR (transf. cells)

Cerebellum

Hippocampus LGI1 (transf. cells)

Cerebellum

10


Anti-CASPR2

Autoantibodies against CASPR2*, a protein associated with voltage-gated potassium channels (VGKC)

Associated diseases: limbic encephalitis, neuromyotonia, Morvan‘s syndrome

Associated tumours: thymoma, uterine carcinoma

IIFT: Fluorescence of the stratum moleculare of the hippocampus (neuropil staining) and the cerebellum. Also patchy fl uorescence of the cerebellar stratum granulosum.

Monospecifi c test using CASPR2-transfected HEK cells.

Anti-DPPX

Autoantibodies against the regulatory subunit of the voltage-gated potassium channel Kv4.2

Associated disease: encephalomyelitis

IIFT: Positive reaction of the stratum granulosum and stratum moleculare of the cerebellum as well as the stratum lucidum of the hippocampus.

Monospecifi c test using DPPX**-transfected HEK cells.

Anti-IgLON5

Autoantibodies against a neuronal cell adhesion protein of the Ig superfamily

Associated diseases: parasomnia, tauopathy

IIFT: Fluorescence of the stratum moleculare of the hippocampus (neuropil staining) and the cerebellum. In addition, patchy fl uorescence of the cerebellar stratum granulosum.

Monospecifi c test using IgLON5-transfected HEK cells.

*CASPR2 = contactin-associated protein 2 **DPPX = dipeptidyl aminopeptidase-like protein 6

CASPR2 (transf. cells)

Cerebellum

Hippocampus

DPPX (transf. cells)Hippocampus

Cerebellum

Cerebellum

Hippocampus

IgLON5 (transf. cells)

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Antibodies (synonym) Antigen

(molecular weight)

Sample

material

Syndrome Most frequent tumours

Antibodies in diseases of the central nervous system

Anti-Hu (ANNA-1) Hu protein (38 kDa) Serum Encephalomyelitis,sensory neuropathy

SCLC, neuroblastoma

Anti-Ri (ANNA-2) NOVA (55 kDa and 80 kDa) Serum Opsoclonus myoclonussyndrome

Breast carcinoma, SCLC

Anti-Yo (PCA-1) CDR2, CDR62 (34 kDa and 62 kDa)

Serum Cerebellar degeneration Ovarian, breast and uterine carcinoma

PCA-2 Purkinje cell protein (280 kDa) Serum Encephalitis,neuropathy

SCLC

Anti-PNMA1 (Ma1) Ma protein (37 kDa) Serum Rhombencephalitis (brainstem), limbic encephalitis

Breast carcinoma, varioustumours

Anti-PNMA2 (Ma2/Ta) Ma protein (40 kDa) Serum Rhombencephalitis (brainstem), limbic encephalitis

Testicular carcinoma

Anti-Tr (DNER) Purkinje cell protein Serum Cerebellar degeneration Hodgkin‘s lymphoma

Anti-ITPR1 (Anti-Sj) Inositol 1,4,5 trisphosphatereceptor 1 (300 kDa)

Serum Cerebellar ataxia NSCLC in an unpublished case

Anti-CARP CARP VIII; carbonic anhydrase-related protein VIII (29 kDa)

Serum Paraneoplastic cerebellardegeneration, cerebellar ataxia

Melanoma, ovarian carcinoma

Anti-amphiphysin Amphiphysin (128 kDa) Serum Stiff-person syndrome Breast carcinoma, SCLC

Antibodies and clinical signifi cance

SCLC: small-cell lung carcinoma

12


(molecular weight)

Sample

material


Anti-CV2 66 kDa protein Serum Limbic encephalitis SCLC, thymoma

Anti-SOX1 Sry-like high mobility group box protein 1 (39 kDa)

Serum LEMS, cerebellar degeneration, sensory neuropathy

SCLC

Anti-ZIC4 Zinc fi nger protein ZIC4 (36 kDa)

Serum Cerebellar degeneration SCLC

Anti-recoverin Recoverin (23 kDa) Serum Retinopathy SCLC

Anti-GAD Glutamic acid decarboxylase (65 kDa and 67 kDa)

SerumCSF

Stiff-person syndrome Breast carcinoma, SCLC, colon carcinoma

Anti-glia nuclear antibodies (AGNA)

SOX1 protein Serum LEMS, cerebellar degeneration SCLC

Anti-AQP-4 (NMO IgG) Aquaporin-4 protein Serum Neuromyelitis optica (NMO),LETM, rec. ON

–

Anti-MOG Myelin oligodendrocyteglycoprotein (28 kDa)

Serum NMO / NMOSD, ADEM, CIS, MS –

Anti-NMDA receptors Extracellular domain of the NR1 subunit of the receptor

SerumCSF

Anti-glutamate receptor (typeNMDA) encephalitis

Teratoma (ovary, testis)

Anti-AMPA receptors GluR1 and GluR2 subunits of the receptor (each approx. 100 kDa)

SerumCSF

Limbic encephalitis Breast carcinoma, thymoma, lung carcinoma

LEMS: paraneoplastic Lambert-Eaton myasthenic syndrome LETM: longitudinally extensive transverse myelitis rec. ON: recurrent optic neuritis ADEM: acute disseminated encephalomyelitis

13


(molecular weight)

Sample

material


Anti-mGluR1 Metabotropic glutamate receptor (approx. 140 kDa)

SerumCSF

Cerebellar degeneration Hodgkin‘s lymphoma

Anti-mGluR5 Metabotropic glutamate receptor (132 kDa)

SerumCSF

Ophelia syndrome Hodgkin‘s lymphoma

Anti-GABAB receptors Extracellular domain of the GABAB subunit of the receptor

SerumCSF

Limbic encephalitis SCLC

Anti-LGI1 Protein (60 kDa) associated with voltage-gated potassium channels (VGKC)

SerumCSF

Limbic encephalitis SCLC, ovarian teratoma,thy moma, various tumours

Anti-CASPR2 Protein associated with voltage-gated potassium channels (VGKC)

SerumCSF

Limbic encephalitis,neuromyotonia, Morvan‘ssyndrome

Thymoma, uterine carcinoma

Anti-DPPX Dipeptidyl aminopeptidase-like protein 6 (98 kDa)

SerumCSF

Encephaltis, encephalomyelitis –

Anti-IgLON5 Cell adhesion protein of the Ig superfamily (37 kDa)

SerumCSF

Parasomnia, tauopathy –

Anti-GlyR Glycine receptor alpha 1 (53 kDa)

SerumCSF

PERM, stiff-person syndrome, hyperekplexia

Thymoma, Hodgkin‘s lymphoma

PERM: progressive encephalomyelitis with rigidity and myoclonus

14


(molecular weight)

Sample

material


Antibodies in diseases of the peripheral nervous system

Anti-GQ1b Tetrasialoganglioside GQ1b Serum Miller-Fisher syndrome –

Anti-GM1 Monosialoganglioside GM1 Serum Multifocal motor neuro pathy,Guillain-Barré syndrome

–

Anti-myelin Myelin Serum Diagnostic value controversial –

Anti-MAG Myelin-associated glycoprotein (100 kDa)

Serum Guillain-Barré syndrome –

Antibodies in neuromuscular diseases

Anti-AChR Acetylcholine receptor of the muscular end plate (α1:55 KDa)

Serum Myasthenia gravis Thymoma

Anti-titin Titin (3816 kDa) Serum Myasthenia gravis Thymoma

Anti-MusK Receptor tyrosine kinase, skeletal muscle specifi c (97 kDa)

Serum Myasthenia gravis Thymoma

15

EUROIMMUN AG · Seekamp 31 · 23560 Lübeck (Germany) · Tel +49 451/5855-0 · Fax 5855-591 · [email protected] · www.euroimmun.com

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19, 11/2016

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Autoantibodies in neurological diseases - EUROIMMUN in neurological diseases ... Golgi apparatus, cytoplasmic membrane) ... moleculare of the hippocampus (neuropil staining)