title: Molecular mimicry between HSP 65 of Mycobacterium leprae and cytokeratin 10 of the host keratin; role in pathogenesis of leprosy.
abstract: Mycobacteria are known to induce autoimmune response in the host. Anti-host keratrin antibodies (AkAbs) might be responsible for the autoimmune phenomena in leprosy patients as majority of leprosy lesions are manifested in the skin and occurrence of keratosis is not an uncommon feature. The aim of this study was to find out the level of AkAbs in leprosy patients across the spectrum and to explore its correlation with the clinical manifestation of the disease. Further, mimicking epitopes of keratin and Mycobacterium leprae components were characterized. We screened 140 leprosy patients (27 BT, 28 BL, 41 LL, 25 T1R, 19 ENL), 74 healthy controls (HC) and 3 psoriasis patients as positive control. Highest AkAbs level was observed in the psoriasis patients followed by T1R, LL, BL, ENL, TT/BT. AkAbs level was significantly (p<0.05) higher in all the groups of leprosy patients except TT/BT in comparison to HC. Significant positive correlation was found between number of lesions and level of AkAbs in leprosy patients. Highest lympho-proliferation for keratin protein was observed in T1R, followed by BL/LL, TT/BT, ENL. Lympho-proliferation was significantly (p<0.05) higher in all groups of leprosy patients except ENL in comparison to HC. Interestingly, it was noted that hyperimmunization of inbred strains of female BALB/c mice and rabbit with M. leprae soluble antigen (MLSA) induce higher level of AkAbs. The percentage of FoxP3(+) expressing Treg cells (total CD4(+)CD25(+)FoxP3(+) andCD4(+)CD25(+hi)FoxP3(+)) in splenocytes and lymph nodes of hyperimmunized mice were declined in comparison to control mice. Further, it was found that this autoimmune response can be adoptively transferred in naïve mice by splenocytes and lymph node cells as well as T cells. Comparative molecular characterization between keratin and MLSA noted a cross-reactivity/similarity between these two antigens. The cross-reactive protein of keratin was found to be in molecular weight range ≈74-51kDa and at pI 4.5 while the cross-reactive protein of MLSA was found to be in molecular weight ≈65kDa and at pI 4-4.5. Cross-reactive protein of keratin and MLSA was identified and characterized by MALDI-TOF/TOF analysis and Mascot software. It was found that the keratin (host protein) which reacted with anti-M. leprae sera is cytokeratin-10 and MLSA which reacted with anti-keratin sera is heat shock protein 65 (HSP 65). Seven B-cell epitopes of cytokeratin-10 and HSP 65 was found to be similar by multiple sequence alignment using ClustalW server and out of which 6 B-cell epitopes were found to be on the surface of HSP 65. In conclusion, our study provides evidence for the existence of molecular mimicry between cytokeratin-10 of keratin (host protein) and 65kDa HSP (groEL2) of M. leprae. Presence of heightened CMI response of leprosy patients to keratin and positive correlation of AkAbs level with number of lesions of leprosy patients showed the clinical evidence for its role in the pathogenesis in leprosy.
authors.affiliation: Department of Immunology, National JALMA Institute for Leprosy and Other Mycobacterial Diseases, (ICMR) Tajganj, Agra, India.
authors.forename: Itu
authors.initials: I
authors.lastname: Singh
authors.forename: Asha Ram
authors.initials: AR
authors.lastname: Yadav
authors.forename: Keshar Kunja
authors.initials: KK
authors.lastname: Mohanty
authors.forename: Kiran
authors.initials: K
authors.lastname: Katoch
authors.forename: Deepa
authors.initials: D
authors.lastname: Bisht
authors.forename: Prashant
authors.initials: P
authors.lastname: Sharma
authors.forename: Bhawna
authors.initials: B
authors.lastname: Sharma
authors.forename: U D
authors.initials: UD
authors.lastname: Gupta
authors.forename: Utpal
authors.initials: U
authors.lastname: Sengupta
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D000942:Antigens, Bacterial / Q000008:administration & dosage / Q000276:immunology; D001323:Autoantibodies / Q000096:biosynthesis / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry* / Q000276:immunology; D016022:Case-Control Studies; D018834:Chaperonin 60 / Q000737:chemistry* / Q000276:immunology; D003429:Cross Reactions; D005260:Female; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D006801:Humans; D007114:Immunization; D053550:Keratin-10 / Q000737:chemistry* / Q000276:immunology; D007918:Leprosy / Q000276:immunology* / Q000382:microbiology / Q000517:prevention & control*; D008198:Lymph Nodes / Q000166:cytology / Q000276:immunology; D051379:Mice; D018716:Molecular Mimicry; D009166:Mycobacterium leprae / Q000276:immunology*; D011817:Rabbits; D012720:Severity of Illness Index; D012867:Skin / Q000276:immunology / Q000382:microbiology; D013154:Spleen / Q000166:cytology / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology / Q000382:microbiology
chemical_list: D000942:Antigens, Bacterial; D001323:Autoantibodies; D001426:Bacterial Proteins; D018834:Chaperonin 60; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; C507493:KRT10 protein, human; C074119:heat-shock protein 65, Mycobacterium; D053550:Keratin-10