Autoantibodies in neurological diseases

Hu, Ri, Yo, Tr

CV2
A
Amphiphysin A
Amphiphysin G
GM1
Ma/Ta
C
CV2 Amphiphysin
Cerebellum Intestine Hippocampus Control transfection C
CV2 G
GM2 SOX1
PNMA2
P
(M
Ma2/Ta) Zic4
PNMA2
P
R
Ri G
GM3 ITPR1
(M
Ma2/Ta)
CARP
R
Ri Y
Yo G
GD1a GAD
Hippocampus HEp-2 cells Cerebellum NMDAR (transf. cells) Recoverin
H
Hu
Titin
Anti-Hu positive Anti-NMDA-receptor positive Y
Yo G
GD1b
R
Recoverin Gangliosides
MAG
H
Hu S
SOX1 G
GT1b
Myelin
T
Titin
Aquaporin-4
G
GQ1b MOG
Z
Zic4 VGKC (LGI1 + CASPR2)
Cerebellum Intestine Cerebellum Control transfection
NMDA receptors
G
GAD65
AMPA receptors
Tr (DNER)
T GABAB receptors
DPPX
C
Control C
Control C
Control IgLON5
Hippocampus HEp-2 cells Optic nerve AQP-4 (transf. cells) Glycine receptors
Anti-Yo positive Anti-aquaporin-4 positive AChR

Indirect immunofluorescence EUROLINE Examples of relevant

target antigens

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 Autoantibodies IIFT pattern Test systems

Anti-Hu (ANNA-1*) IIFT: Granular fluorescence of almost all neuronal nuclei

on the substrates cerebellum and hippocampus. The
Autoantibodies against basic, RNA- cell nuclei of the plexus myentericus (intestinal tissue)
binding proteins of the neuronal cell are also positive.
nuclei of the central and peripheral
nervous system EUROLINE: Positive reaction of the recombinant
Hu antigen (HuD).
Associated diseases: encephalomyelitis, subacute
sensory neuronopathy (Denny-Brown syndrome),
autonomous neuropathy

Associated tumours: small-cell lung carcinoma, Cerebellum Intestine

neuroblastoma

Anti-Ri (ANNA-2*) IIFT: Granular fluorescence of almost all neuronal nuclei

on the substrates cerebellum and hippocampus. The
Autoantibodies against neuronal cell substrate intestine (plexus myentericus) shows no reac-
nuclei of the central nervous system tion.
(CNS)
EUROLINE: Positive reaction of the recombinant Ri anti-
Associated disease: opsoclonus myoclonus syn- gen (NOVA1***).
drome

Associated tumours: small-cell lung carcinoma,

breast carcinoma

Cerebellum Intestine

Anti-Yo (PCA-1**) IIFT: Cytoplasmic fluorescence of the Purkinje cells

on the substrate cerebellum and of individual cells in
Autoantibodies against antigens the hilus of the hippocampus. The substrate intestine
in the cytoplasm (rough ER, Golgi (plexus myentericus) shows no specific reaction.
apparatus, cytoplasmic membrane) of Intestine
the cerebellar Purkinje cells Monospecific test using Yo/CDR2-transfected HEK****
cells
Associated disease: cerebellar degeneration

Associated tumours: ovarian carcinoma, breast EUROLINE: Positive reaction of the recombinant Yo
carcinoma, uterine carcinoma antigen (CDR62*****).
Cerebellum Yo (transf. cells)

*
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
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 Autoantibodies IIFT pattern Test systems

PCA-2* IIFT: Fluorescence of the cytoplasm of the cerebellar

Purkinje cells, which extends into the dendrites.
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
Cerebellum

Anti-Tr Coarse granular fluorescence of the cytoplasm

of the cerebellar Purkinje cells, dot-like staining of the
Autoantibodies against DNER** protein molecular layer.
in the Purkinje cells of the cerebellum
Monospecific test using DNER-transfected HEK cells.
Associated disease: cerebellar degeneration

Associated tumour: Hodgkin‘s lymphoma EUROLINE: Positive reaction of the specific target
antigen Tr (DNER).

Cerebellum DNER (transf. cells)

Anti-CARP (anti-carbonic anhydrase- IIFT: The molecular layer and the cytoplasm of the
related protein VIII) Purkinje cells in the cerebellum show a dotted to fine-
speckled pattern.
Autoantibodies against CARP VIII
in the Purkinje cell layer of the Monospecific test using CARP VIII-transfected HEK cells.
cerebellum

Associated diseases: paraneoplastic cerebellar

degeneration, cerebellar ataxia

Associated tumours: melanoma, ovarian carcinoma

Cerebellum CARP (transf. cells)

* **
PCA = Purkinje cell autoantibodies DNER = delta/notch-like epidermal growth factor-related receptor
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Autoantibodies IIFT pattern Test systems

Anti-ITPR1 (Anti-Sj) IIFT: The molecular layer and the cytoplasm of the
(anti-Sj / inositol 1,4,5-trisphosphate Purkinje cells in the cerebellum show a dotted to fine-
receptor) speckled pattern.

Autoantibodies against ITPR1 in Monospecific test using ITPR1-transfected HEK cells.

Purkinje cells of the cerebellum

Associated disease: cerebellar ataxia

Associated tumour: NSCLC in an unpublished case

Cerebellum ITPR1 (transf. cells)

Anti-glia nuclear antibodies (AGNA) IIFT: Fluorescence of the cell nuclei of the Bergmann
glia in the Purkinje cell layer of the cerebellum.
Possible antigen: SOX1

Associated diseases: paraneoplastic Lambert-

Eaton myasthenic syndrome (LEMS), cerebellar
degeneration, sensory neuropathy

Associated tumour: small-cell lung carcinoma

Cerebellum

Anti-Ma (Ma1, Ma2/Ta) IIFT: Reaction of the nerve cell nucleoli on the
substrates cerebellum, cerebrum and hippocampus.
Autoantibodies against Ma1 (PNMA1*;
37 kDa) and Ma2/Ta (PNMA2*; 40 kDa), EUROLINE: Anti-Ma (Ma1 and Ma2/Ta) antibodies
proteins in the nucleoli of the neuronal produce a positive reaction with the recombinant Ma2/
cell nuclei Ta (PNMA2*) antigen.

Associated diseases: brain stem encephalitis, limbic

encephalitis

Associated tumours: lung carcinoma, testicular

carcinoma (esp. Ma2/Ta), breast carcinoma Cerebellum Cerebrum

PNMA1/2 = paraneoplastic antigen Ma1/2

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Autoantibodies IIFT pattern Test systems

Anti-CV2 IIFT: Sand-like fluorescence on the substrate

cerebellum, most visible in the stratum moleculare.
Autoantibodies against CV2, a 66-kDa
protein EUROLINE: Positive reaction of the recombinant CV2.

Associated diseases: sensory and autonomous

neuropathy, encephalitis, extrapyramidal motor
syndrome, cerebellar degeneration

Associated tumours: small-cell lung carcinoma,

thymoma

Cerebellum

Anti-amphiphysin IIFT: Fluorescence of the presynaptic nerve ends in

the cerebellum. The nerve dendrites of the stratum
Autoantibodies against amphiphysin, moleculare show a more intense fluorescence than the
a synaptic protein with a molecular stratum granulosum. The fluorescence pattern of the
weight of 128 kDa stratum granulosum resembles the pattern found with
antibodies against GAD.
Associated disease: paraneoplastic stiff-person
syndrome
EUROLINE: Positive reaction of the specific target
Associated tumours: breast carcinoma, small-cell antigen amphiphysin.
lung carcinoma

Cerebellum

Anti-GAD IIFT: Patchy fluorescence of the stratum granulosum

on the substrates cerebellum and hippocampus.
Autoantibodies against the enzyme Fluorescence of the pancreas islets on the substrate
glutamic acid decarboxylase pancreas.
GAD (transf. cells)
Associated diseases: stiff-person syndrome, limbic Monospecific test using GAD-transfected HEK cells.
encephalitis

EUROLINE: Monospecific test using purified GAD.

Cerebellum Pancreas

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 Autoantibodies IIFT pattern Test systems

Anti-ZIC4 IIFT: ANA-like fluorescence of the neuronal nuclei of the

cerebellar stratum granulosum. The cell nuclei of the
Autoantibodies against zinc finger Purkinje cells are negative in the presence of anti-ZIC4
proteins antibodies, whereas with ANA* all cells are stained.
(probably ZIC1, 2 and 4) Antibodies against ZIC4 should be assessed not only
on cerebellar sections, but also on transfected cells in
Associated disease: cerebellar degeneration parallel.
Associated tumour: small-cell lung carcinoma
EUROLINE: Positive reaction of the specific target
antigen ZIC4.
Cerebellum ZIC4 (transf. cells)

Anti-MAG IIFT: Fluorescence of the lamina alba in the cerebellum.

Annular fluorescence on the substrate peripheral nerve.
Autoantibodies against myelin-
associated glycoprotein, a cell Clinically relevant antibodies are mainly those of class
membrane protein of the myelin IgM.
sheaths

Associated diseases: paraproteinaemic neuropathy,

particularly in the context of monoclonal IgM
gammopathy, e.g. monoclonal gammopathy
of undetermined significance (MGUS) and
Waldenström‘s syndrome Cerebellum Nerve (section)

Anti-myelin IIFT: Fluorescence of the lamina alba in the cerebellum.

Fluorescing hyaline cylinders on the substrate
Autoantibodies against myelin and peripheral nerve.
myelin proteins (e.g. MBP**) in the
myelin sheath of axons

Associated diseases: diagnostic significance is

controversial

Cerebellum Nerve (section)

* **
ANA = anti-nuclear antibodies MBP = myelin basic protein
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 Autoantibodies IIFT pattern Test systems

Anti-AQP-4 (NMO-IgG) IIFT: Perivascular fluorescence with linear staining

along the Virchow-Robin spaces and the microvessels
Autoantibodies against aquaporin-4 in the grey and white matter on the substrate
(AQP-4), a water channel of the cerebellum, as well as a reticular fluorescence on the
dystroglycan protein complex Cerebellum substrate optic nerve.

Associated diseases: neuromyelitis optica (NMO, Monospecific test using AQP4-transfected HEK cells.
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
Optic nerve AQP-4 (transf. cells)

Anti-MOG (anti-myelin IIFT: The white matter of the cerebellum shows a

oligodendrocyte glycoprotein) granular fluorescence.

Autoantibodies against myelin oligo- Monospecific test using MOG-transfected HEK cells.
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: – Cerebellum MOG (transf. cells)

Anti-NMDA receptors IIFT: Fluorescence of the stratum moleculare of the

hippocampus (neuropil staining) and the stratum
Autoantibodies against glutamate granulosum of the cerebellum.
receptors (type NMDA*)
Cerebellum Monospecific test using NMDA receptor (NR1)-
Associated disease: anti-glutamate receptor (type
NMDA) encephalitis (~ 60 % as paraneoplastic
transfected HEK cells.
syndrome)

Associated tumours: ovarian teratoma, testicular

teratoma

Hippocampus NMDAR (transf. cells)

*
NMDA = N-methyl-D-aspartate
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 Autoantibodies IIFT pattern Test systems

Anti-AMPA receptors IIFT: Fluorescence of the stratum moleculare of

the hippocampus (neuropil staining), the stratum
Autoantibodies against glutamate moleculare and stratum granulosum of the cerebellum
receptors (type AMPA*) and the Purkinje cells.
Cerebellum
Associated disease: limbic encephalitis Monospecific test using AMPA1/2-transfected HEK cells.
(~ 70 % as paraneoplastic syndrome)

Associated tumours: lung carcinoma, breast

carcinoma, malignant thymoma

AMPAR1/R2
Hippocampus (transf. cells)

Anti-GABAB receptors IIFT: Coarse granular fluorescence of the stratum

moleculare of the hippocampus (neuropil staining)
Autoantibodies against and the cerebellum. Also patchy fluorescence of the
GABAB** receptors cerebellar stratum granulosum.
Cerebellum
Associated disease: limbic encephalitis Monospecific test using GABAB receptor-transfected
(~ 50 % as paraneoplastic syndrome)
HEK cells.
Associated tumour: small-cell lung carcinoma

Hippocampus GABABR (transf. cells)

Anti-LGI1 IIFT: Fine granular fluorescence of the stratum

moleculare of the hippocampus (neuropil staining)
Autoantibodies against LGI1***, a and the cerebellum. Also patchy fluorescence of the
protein associated with voltage-gated cerebellar stratum granulosum. The outer molecular
potassium channels (VGKC) Cerebellum layer of the hippocampus shows a stronger reaction
than the inner layer.
Associated disease: limbic encephalitis

Associated tumours: thyroid carcinoma, small-cell Monospecific test using LGI1-transfected HEK cells.
lung carcinoma, kidney cell carcinoma, ovarian
teratoma, thymoma

Hippocampus LGI1 (transf. cells)

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

Anti-CASPR2 IIFT: Fluorescence of the stratum moleculare of the

hippocampus (neuropil staining) and the cerebellum.
Autoantibodies against CASPR2*, a Also patchy fluorescence of the cerebellar stratum
protein associated with voltage-gated granulosum.
potassium channels (VGKC) Cerebellum
Monospecific test using CASPR2-transfected HEK cells.
Associated diseases: limbic encephalitis,
neuromyotonia, Morvan‘s syndrome

Associated tumours: thymoma, uterine carcinoma

Hippocampus CASPR2 (transf. cells)

Anti-DPPX IIFT: Positive reaction of the stratum granulosum and

stratum moleculare of the cerebellum as well as the
Autoantibodies against the regulatory stratum lucidum of the hippocampus.
subunit of the voltage-gated
potassium channel Kv4.2 Monospecific test using DPPX**-transfected HEK cells.
Cerebellum
Associated disease: encephalomyelitis

Hippocampus DPPX (transf. cells)

Anti-IgLON5 IIFT: Fluorescence of the stratum moleculare of the

hippocampus (neuropil staining) and the cerebellum. In
Autoantibodies against a neuronal cell addition, patchy fluorescence of the cerebellar stratum
adhesion protein of the Ig superfamily granulosum.
Hippocampus
Associated diseases: parasomnia, tauopathy Monospecific test using IgLON5-transfected HEK cells.

Cerebellum IgLON5 (transf. cells)

* **
CASPR2 = contactin-associated protein 2 DPPX = dipeptidyl aminopeptidase-like protein 6
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Antibodies and clinical significance
Antibodies (synonym) Antigen Sample Syndrome Most frequent tumours
(molecular weight) material

Antibodies in diseases of the central nervous system

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

Anti-Ri (ANNA-2) NOVA (55 kDa and 80 kDa) Serum Opsoclonus myoclonus Breast carcinoma, SCLC
syndrome

Anti-Yo (PCA-1) CDR2, CDR62 (34 kDa and Serum Cerebellar degeneration Ovarian, breast and uterine
62 kDa) carcinoma

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

neuropathy

Anti-PNMA1 (Ma1) Ma protein (37 kDa) Serum Rhombencephalitis (brain Breast carcinoma, various
stem), limbic encephalitis tumours

Anti-PNMA2 (Ma2/Ta) Ma protein (40 kDa) Serum Rhombencephalitis (brain Testicular carcinoma
stem), limbic encephalitis

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

Anti-ITPR1 (Anti-Sj) Inositol 1,4,5 trisphosphate Serum Cerebellar ataxia NSCLC in an unpublished case
receptor 1 (300 kDa)

Anti-CARP CARP VIII; carbonic anhydrase- Serum Paraneoplastic cerebellar Melanoma, ovarian carcinoma
related protein VIII (29 kDa) degeneration, cerebellar ataxia

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

SCLC: small-cell lung carcinoma

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Antibodies (synonym) Antigen Sample Syndrome Most frequent tumours
(molecular weight) material

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

Anti-SOX1 Sry-like high mobility group Serum LEMS, cerebellar degeneration, SCLC
box protein 1 (39 kDa) sensory neuropathy

Anti-ZIC4 Zinc finger protein ZIC4 Serum Cerebellar degeneration SCLC

(36 kDa)

Anti-recoverin Recoverin (23 kDa) Serum Retinopathy SCLC

Anti-GAD Glutamic acid decarboxylase Serum Stiff-person syndrome Breast carcinoma, SCLC, colon
(65 kDa and 67 kDa) CSF carcinoma

Anti-glia nuclear SOX1 protein Serum LEMS, cerebellar degeneration SCLC

antibodies (AGNA)

Anti-AQP-4 (NMO IgG) Aquaporin-4 protein Serum Neuromyelitis optica (NMO), –

LETM, rec. ON

Anti-MOG Myelin oligodendrocyte Serum NMO / NMOSD, ADEM, CIS, MS –

glycoprotein (28 kDa)

Anti-NMDA receptors Extracellular domain of the Serum Anti-glutamate receptor (type Teratoma (ovary, testis)
NR1 subunit of the receptor CSF NMDA) encephalitis

Anti-AMPA receptors GluR1 and GluR2 subunits of Serum Limbic encephalitis Breast carcinoma, thymoma,
the receptor (each approx. CSF lung carcinoma
100 kDa)
LEMS: paraneoplastic Lambert-Eaton myasthenic syndrome LETM: longitudinally extensive transverse myelitis rec. ON: recurrent optic neuritis ADEM: acute disseminated encephalomyelitis

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Antibodies (synonym) Antigen Sample Syndrome Most frequent tumours
(molecular weight) material

Anti-mGluR1 Metabotropic glutamate Serum Cerebellar degeneration Hodgkin‘s lymphoma

receptor (approx. 140 kDa) CSF

Anti-mGluR5 Metabotropic glutamate Serum Ophelia syndrome Hodgkin‘s lymphoma

receptor (132 kDa) CSF

Anti-GABAB receptors Extracellular domain of Serum Limbic encephalitis SCLC

the GABAB subunit of the CSF
receptor
Anti-LGI1 Protein (60 kDa) associated Serum Limbic encephalitis SCLC, ovarian teratoma,
with voltage-gated potassium CSF thymoma, various tumours
channels (VGKC)
Anti-CASPR2 Protein associated with Serum Limbic encephalitis, Thymoma, uterine carcinoma
voltage-gated potassium CSF neuromyotonia, Morvan‘s
channels (VGKC) syndrome
Anti-DPPX Dipeptidyl aminopeptidase- Serum Encephaltis, encephalomyelitis –
like protein 6 (98 kDa) CSF

Anti-IgLON5 Cell adhesion protein of the Serum Parasomnia, tauopathy –

Ig superfamily (37 kDa) CSF

Anti-GlyR Glycine receptor alpha 1 Serum PERM, stiff-person syndrome, Thymoma, Hodgkin‘s
(53 kDa) CSF hyperekplexia lymphoma

PERM: progressive encephalomyelitis with rigidity and myoclonus

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Antibodies (synonym) Antigen Sample Syndrome Most frequent tumours
(molecular weight) material

Antibodies in diseases of the peripheral nervous system

Anti-GQ1b Tetrasialoganglioside GQ1b Serum Miller-Fisher syndrome –

Anti-GM1 Monosialoganglioside GM1 Serum Multifocal motor neuropathy, –

Guillain-Barré syndrome

Anti-myelin Myelin Serum Diagnostic value controversial –

Anti-MAG Myelin-associated Serum Guillain-Barré syndrome –

glycoprotein (100 kDa)

Antibodies in neuromuscular diseases

Anti-AChR Acetylcholine receptor of Serum Myasthenia gravis Thymoma
the muscular end plate (α1:
55 KDa)
Anti-titin Titin (3816 kDa) Serum Myasthenia gravis Thymoma

Anti-MusK Receptor tyrosine kinase, Serum Myasthenia gravis Thymoma

skeletal muscle specific
(97 kDa)

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EUROIMMUN AG · Seekamp 31 · 23560 Lübeck (Germany) · Tel +49 451/ 58 55-0 · Fax 58 55-591 · info@euroimmun.de · www.euroimmun.com