Autoantibodies as biomarker of paraneoplastic encephalitis associated with testicular cancer
This disclosure describes methods and compositions for detecting the presence of a Kelch-like protein 11 autoantibody in a biological sample. Also provided are methods of treating subjects with testicular cancer or a premalignant condition and paraneoplastic encephalomyelitis.
1. A method of detecting the presence of a Kelch-like protein 11 (KLHL11) autoantibody in a biological sample from a male subject presenting with encephalitis, comprising the steps of:
(a) contacting the biological sample with a KLHL11 polypeptide or antigenic fragment thereof, the polypeptide or fragment thereof having at least 95% identity to one or more sequences of SEQ ID NOs: 1-30; and
(b) detecting the presence of binding of the KLHL11 polypeptide or antigenic fragment thereof to the KLHL11 autoantibody in the biological sample.
2. The method of claim 1 , wherein the KLHL11 polypeptide or antigenic fragment is heterologously-expressed on the surface of a cell, a phage, or a virus.
3. The method of claim 1 , wherein the KLHL11 polypeptide or antigenic fragment thereof is expressed in a phage display or eukaryotic cell display library.
4. The method of claim 1 , wherein the KLHL11 polypeptide or antigenic fragment thereof is an isolated, purified KLHL11 polypeptide or fragment thereof.
5. The method of claim 1 , wherein the KLHL11 polypeptide or antigenic fragment is an isolated, purified KLHL11 polypeptide or fragment thereof that is immobilized on a solid carrier.
6. The method of claim 1 , wherein the KLHL11 polypeptide or antigenic fragment comprises the sequence of SEQ ID NO:1.
7. The method of claim 1 , wherein the KLHL11 polypeptide or antigenic fragment thereof is in brain tissue or other nervous tissue.
8. The method of claim 1 , wherein the biological sample is serum, plasma, cerebrospinal fluid, blood, or urine.
9. The method of claim 1 , wherein the male subject has one or more of ataxia, cognitive decline, mood disorder, hearing loss, tinnitus, trigeminal neuralgia, vertigo, diplopia, dysarthria, or other neurological deficit.
10. The method of claim 1 , wherein the male subject has one or more of ataxia, hearing loss, tinnitus, or vertigo.
11. The method of claim 1 , wherein the male subject has at least one of:
(i) a cerebrospinal fluid protein count of greater than 30 mg per deciliter;
(ii) a cerebrospinal fluid blood cell count of at least 1 per cubic millimeter;
(iii) two or more cerebrospinal oligoclonal bands; or
(iv) a CSF index greater than 0.6.
12. The method of claim 1 , wherein the male subject has at least one risk factor for testicular cancer.
13. The method of claim 1 , wherein the male subject has not been previously diagnosed with testicular cancer.
14. The method of claim 1 , wherein the male subject has been previously treated for testicular cancer or a premalignant condition or is being treated for testicular cancer or a premalignant condition.
15. The method of claim 1 , wherein the presence of the binding of the KLHL11 polypeptide or antigenic fragment thereof to said KLHL11 autoantibody in the biological sample indicates that the subject has or is at risk of having testicular cancer or a premalignant testicular condition.
16. The method of claim 15 , wherein the testicular cancer is seminoma.
17. The method of claim 1 , wherein the presence of the binding of the KLHL11 polypeptide or antigenic fragment thereof to said KLHL11 autoantibody in the biological sample indicates that the subject has paraneoplastic encephalitis.
18. The method of claim 15 , wherein the premalignant testicular condition is at least one of testicular microlithiasis, testicular atrophy, or testicular fibrosis.
19. The method of claim 1 , wherein step (b) of detecting is performed by at least one of immunohistochemical staining, immunoprecipitation, microarray analysis, enzyme-linked immunosorbent assay (ELISA), or Western blot analysis.
20. The method of claim 1 , wherein the KLHL11 polypeptide or fragment thereof is in a tissue section,
wherein step (a) comprises contacting the tissue section with the biological sample and a detectably-labeled secondary antibody under conditions in which a complex is formed between KLHL11 polypeptides in the tissue section, a corresponding KLHL11 autoantibody in the biological sample, if present, and the detectably-labeled secondary antibody, and
wherein step (b) comprises identifying a pattern of complex formation in the tissue sample by detecting the detectably-labeled secondary antibody, wherein the presence of a pattern of complex formation is indicative of the presence of a KLHL11 autoantibody in the biological sample, and wherein the absence of a pattern of complex formation is indicative of the absence of a KLHL11 autoantibody in the biological sample.
21. The method of claim 20 , wherein the pattern of complex formation in the brain tissue is sparse yet widespread throughout the brain, including the hippocampus, cerebral cortex, cerebellum, midbrain, and brain stem, and is enriched in perivascular tissue, perimeningeal tissue, and white matter tracts.
22. The method of claim 1 , wherein the KLHL11 polypeptide or fragment thereof is heterologously-expressed on the surface of a cell,
wherein step (a) comprises contacting the cell that heterologously-expresses the KLHL11 polypeptide or antigenic fragment thereof on its surface with the biological sample and a detectably-labeled secondary antibody under conditions in which a complex is formed between KLHL11 polypeptides in the tissue section, a corresponding KLHL11 autoantibody in the biological sample, if present, and the detectably-labeled secondary antibody, and
wherein step (b) comprises detecting the detectably-labeled secondary antibody, wherein detecting the presence of the detectably-labeled secondary antibody is indicative of the presence of a KLHL11 autoantibody in the biological sample, and wherein not detecting the presence of the detectably-labeled secondary antibody is indicative of the absence of a KLHL11 autoantibody in the biological sample.
23. The method of claim 1 comprising
(a) contacting the biological sample with a KLHL11 polypeptide having at least 95% identity to SEQ ID NO:1.
24. The method of claim 1 comprising
(a) contacting the biological sample with a KLHL11 polypeptide having at least 95% identity to one or more sequences of SEQ ID NOs: 1-30.
25. The method of claim 1 comprising
(a) contacting the biological sample with a KLHL11 polypeptide having at least 97% identity to one or more sequences of SEQ ID NOs: 1-30 or fragment thereof having at least 97% identity to one or more sequences of SEQ ID NOs: 1-30.
26. The method of claim 1 , wherein the KLHL11 polypeptide or antigenic fragment thereof, is truncated at one or both termini by one or more amino acids.