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
Cerebellum Intestine
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 Intestine
Cerebellum
Cerebellum Yo (transf. cells)
4
Autoantibodies IIFT pattern Test systems
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 disease: cerebellar degeneration
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
Autoantibodies IIFT pattern Test systems
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
Autoantibodies IIFT pattern Test systems
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
Autoantibodies IIFT pattern Test systems
Anti-ZIC4
Autoantibodies against zinc fi nger proteins (probably ZIC1, 2 and 4)
Associated disease: cerebellar degeneration
Associated tumour: small-cell lung carcinoma
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
Autoantibodies IIFT pattern Test systems
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
Autoantibodies IIFT pattern Test systems
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)
Associated tumour: small-cell lung carcinoma
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
Autoantibodies IIFT pattern Test systems
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)
11
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
Antibodies (synonym) Antigen
(molecular weight)
Sample
material
Syndrome Most frequent tumours
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
Antibodies (synonym) Antigen
(molecular weight)
Sample
material
Syndrome Most frequent tumours
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
Antibodies (synonym) Antigen
(molecular weight)
Sample
material
Syndrome Most frequent tumours
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
FA_1111_I_U
K_A
19, 11/2016