Abstract
Hsp60 (also called Cpn60) is a chaperonin with essential functions for cell physiology and survival. Additionally, its involvement in the pathogenesis of a variety of diseases (e.g., some autoimmune disorders and cancer) is becoming evident with new research. For example, the distribution and levels of Hsp60 in cells and tissues have been found altered in many pathologic conditions, and the significance of these alterations is being investigated in a number of laboratories. The aim of this ongoing research is to determine the meaning of these Hsp60 alterations with regard to pathogenetic mechanisms, diagnosis, classification of lesions, and assessing prognosis and response to treatment.
Hsp60 occurs in the mitochondria, i.e., its typical residence according to classic knowledge, and also in other locales, such as the cytosol, the cell membrane, the intercellular space, and biological fluids (e.g., blood and cerebrospinal fluid). Detection and quantitative determinations in all these locations are becoming essential components of laboratory pathology in clinics and research. Consequently, immunohistochemistry targeting Hsp60 is also becoming essential for pathologists and researchers interested in disorders involving this chaperonin.
In this chapter, we summarize some recent discoveries on the participation of Hsp60 in the pathogenesis of human diseases, and describe in detail how to perform immunohistochemical reactions for detecting the chaperonin, determining its location, and measuring its quantitative levels.
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References
Levy-Rimler G, Bell RE, Ben-Tal N, Azem A (2002) Type I chaperonins: not all are created equal. FEBS Lett 529:1–5
Hansen JJ, Bross P, Westergaard M, Nielsen MN, Eiberg H, Borglum AD, Mogensen J, Kristiansen K, Bolund L, Gregersen N (2003) Genomic structure of the human mitochondrial chaperonin genes: HSP60 and HSP10 are localised head to head on chromosome 2 separated by a bidirectional promoter. Hum Genet 112:71–77
Soltys BJ, Gupta RS (1999) Mitochondrial proteins at unexpected cellular locations: export of proteins from mitochondria from an evolutionary perspective. Int Rev Cytol 194:133–196
Cappello F, Conway de Macario E, Marasà L, Zummo G, Macario AJL (2008) Hsp60 expression, new locations, functions and perspectives for cancer diagnosis and therapy. Cancer Biol Ther 7:801–809
Macario AJL, Conway de Macario E (2008/2009) The chaperoning system: physiology and pathology. In: Gerbino A, Crescimanno G, Zummo G (eds) Experimental medicine reviews, vol 2/3. Plumelia, Bagheria, Italy, pp 9–21
Cappello F, Bellafiore M, Palma A, David S, Marciano V, Bartolotta T, Sciume C, Modica G, Farina F, Zummo G, Bucchieri F (2003) 60 kDa chaperonin (HSP60) is over-expressed during colorectal carcinogenesis. Eur J Histochem 47:105–110
Rappa F, Farina F, Zummo G, David S, Campanella C, Carini F, Tomasello G, Damiani P, Cappello F, Conway de Macario E, Macario AJL (2012) HSP-molecular chaperones in cancer biogenesis and tumor therapy: an overview. Anticancer Res 32:5139–5150
Campanella C, Rappa F, Sciumè C, Marino Gammazza A, Barone R, Bucchieri F, David S, Curcurù G, Caruso Bavisotto C, Pitruzzella A, Geraci G, Modica G, Farina F, Zummo G, Fais S, Conway de Macario E, Macario AJL, Cappello F (2015) Heat shock protein 60 levels in tissue and circulating exosomes in human large bowel cancer before and after ablative surgery. Cancer 121:3230–3239
Rappa F, Unti E, Baiamonte P, Cappello F, Scibetta N (2013) Different immunohistochemical levels of Hsp60 and Hsp70 in a subset of brain tumors and putative role of Hsp60 in neuroepithelial tumorigenesis. Eur J Histochem 57:e20
Hamelin C, Cornut E, Poirier F, Pons S, Beaulieu C, Charrier JP, Haïdous H, Cotte E, Lambert C, Piard F, Ataman-Önal Y, Choquet-Kastylevsky G (2011) Identification and verification of heat shock protein 60 as a potential serum marker for colorectal cancer. FEBS J 278:4845–4859
Soltys BJ, Gupta RS (1997) Cell surface localization of the 60 kDa heat shock chaperonin protein (hsp60) in mammalian cells. Cell Biol Int 21:315–320
Lin L, Kim SC, Wang Y, Gupta S, Davis B et al (2007) HSP60 in heart failure: abnormal distribution and role in cardiac myocyte apoptosis. Am J Physiol Heart Circ Physiol 293:H2238–H2247
Piselli P, Vendetti S, Vismara D, Cicconi R, Poccia F et al (2000) Different expression of CD44, ICAM-1, and HSP60 on primary tumor and metastases of a human pancreatic carcinoma growing in scid mice. Anticancer Res 20:825–831
Shin BK, Wang H, Yim AM, Le Naour F, Brichory F et al (2003) Global profiling of the cell surface proteome of cancer cells uncovers an abundance of proteins with chaperone function. J Biol Chem 278:7607–7616
Dziewanowska K, Carson AR, Patti JM, Deobold CF, Bayles KW et al (2000) Staphylococcal fibronectin binding protein interacts with heat shock protein 60 and integrins: role in internalization by epithelial cells. Infect Immun 68:6321–6328
Merendino AM, Bucchieri F, Campanella C, Marcianò V, Ribbene A et al (2010) Hsp60 is actively secreted by human tumor cells. PLoS One 5:e9247
Campanella C, Bucchieri F, Merendino AM, Fucarino A, Burgio G et al (2012) The odyssey of Hsp60 from tumor cells to other destinations includes plasma membrane-associated stages and Golgi and exosomal protein-trafficking modalities. PLoS One e42008:7
Osterloh A, Meier-Stiegen F, Veit A, Fleischer B, von Bonin A et al (2004) Lipopolysaccharide-free heat shock protein 60 activates T cells. J Biol Chem 279:47906–47911
Chen W, Syldath U, Bellmann K, Burkart V, Kolb H (1999) Human 60- kDa heat shock protein: a danger signal to the innate immune system. J Immunol 162:3212
Marino Gammazza A, Rizzo M, Citarrella R, Rappa F, Campanella C et al (2014) Elevated blood Hsp60, its structural similarities and cross-reactivity with thyroid molecules, and its presence on the plasma membrane of oncocytes point to the chaperonin as an immunopathogenic factor in Hashimoto's thyroiditis. Cell Stress Chaperones 19:343–353
Quintana FJ, Cohen IR (2011) The HSP60 immune system network. Trends Immunol 32:89–95
Ikawa S, Weinberg RA (1992) An interaction between p21ras and heat shock protein hsp60, a chaperonin. Proc Natl Acad Sci U S A 89:2012–2016
Barazi HO, Zhou L, Templeton NS, Krutzsch HC, Roberts DD (2002) Identification of heat shock protein 60 as a molecular mediator of alpha 3 beta 1 integrin activation. Cancer Res 62:1541–1548
Ghosh JC, Dohi T, Kang BH, Altieri DC (2008) Hsp60 regulation of tumor cell apoptosis. J Biol Chem 283:5188–5194
Laad AD, Thomas ML, Fakih AR, Chiplunkar SV (1999) Human gamma delta T cells recognize heat shock protein-60 on oral tumor cells. Int J Cancer 80:709–714
Hanlon JG, Adams K, Rainbow AJ, Gupta RS, Singh G (2001) Induction of Hsp60 by Photofrin-mediated photodynamic therapy. J Photochem Photobiol B 64:55–61
Jalili A, Makowski M, Switaj T, Nowis D, Wilczynski GM et al (2004) Effective photoimmunotherapy of murine colon carcinoma induced by the combination of photodynamic therapy and dendritic cells. Clin Cancer Res 10:4498–4508
Korbelik M, Sun J, Cecic I (2005) Photodynamic therapy-induced cell surface expression and release of heat shock proteins: relevance for tumor response. Cancer Res 65:1018–1026
Pace A, Barone G, Lauria A, Martorana A, Piccionello AP et al (2013) Hsp60, a novel target for antitumor therapy: structure-function features and prospective drugs design. Curr Pharm Des 19:2757–2764
Macario AJL, Conway de Macario E (2004) The pathology of anti-stress mechanisms: a new frontier. Stress 7:243–249
Landstein D, Ulmansky R, Naparstek Y ((2015)) HSP60: a double edge sword in autoimmunity. Oncotarget 6:32299–32300
Raska M, Weigl E (2005) Heat shock proteins in autoimmune diseases. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 149:243–249
Xu Q, Schett G, Perschinka H, Mayr M, Egger G et al (2000) Serum soluble heat shock protein 60 is elevated in subjects with atherosclerosis in a general population. Circulation 102:14–20
Rizzo M, Macario AJL, Conway de Macario E, Gouni-Berthold I, Berthold HK et al (2011) Heat shock protein-60 and risk for cardiovascular disease. Curr Pharm Des 17:3662–3668
Wick G, Jakic B, Buszko M, Wick MC, Grundtman C (2014) The role of heat shock proteins in atherosclerosis. Nat Rev Cardiol 11:516–529
Almanzar G, Öllinger R, Leuenberger J, Onestingel E, Rantner B et al (2012) Autoreactive HSP60 epitope-specific T-cells in early human atherosclerotic lesions. J Autoimmun 39:441–450
Grundtman C, Kreutmayer SB, Almanzar G, Wick MC, Wick G (2011) Heat shock protein 60 and immune inflammatory responses in atherosclerosis. Arterioscler Thromb Vasc Biol 31:960–968
Matsuura E, Kobayashi K, Matsunami Y, Shen L, Quan N et al (2009) Autoimmunity, infectious immunity, and atherosclerosis. J Clin Immunol 29:714–721
Alard JE, Dueymes M, Youinou P, Jamin C (2008) HSP60 and anti-HSP60 antibodies in vasculitis: they are two of a kind. Clin Rev Allergy Immunol 35:66–71
Rauch J, Dieudé M, Subang R, Levine JS (2010) The dual role of innate immunity in the antiphospholipid syndrome. Lupus 19:347–353
Choi B, Choi M, Park C, Lee EK, Kang DH et al (2015) Cytosolic Hsp60 orchestrates the survival and inflammatory responses of vascular smooth muscle cells in injured aortic vessels. Cardiovasc Res 106:498–508
Krenn V, Vollmers HP, von Landenberg P, Schmausser B, Rupp M (1996) Immortalized B lymphocytes from rheumatoid synovial tissue show specificity for bacterial HSP60. Virchows Arch 427:511–518
Vercoulen Y, van Teijlingen NH, de Kleer IM, Kamphuis S, Albani S et al (2009) Heat shock protein 60 reactive T cells in juvenile idiopathic arthritis: what is new? Arthritis Res Ther 11:231
Huang MN, Yu H, Moudgil KD (2010) The involvement of heat-shock proteins in the pathogenesis of autoimmune arthritis: a critical appraisal. Semin Arthritis Rheum 40:164–175
Lang A, Benke D, Eitner F, Engel D, Ehrlich S et al (2005) Heat shock protein 60 is released in immune-mediated glomerulonephritis and aggravates disease: in vivo evidence for an immunologic danger signal. J Am Soc Nephrol 16:383–391
Slot MC, Theunissen R, van Paassen P, Damoiseaux JG, Cohen Tervaert JW (2006) Evaluation of antibodies against human HSP60 in patients with MPO-ANCA associated glomerulonephritis: a cohort study. J Autoimmune Dis 5:3–4
Elst EF, Klein M, de Jager W, Kamphuis S, Wedderburn LR et al (2008) Hsp60 in inflamed muscle tissue is the target of regulatory autoreactive T cells in patients with juvenile dermatomyositis. Arthritis Rheum 58:547–555
Kasperkiewicz M, Tukaj S, Gembicki AJ, Silló P, Görög A et al (2014) Evidence for a role of autoantibodies to heat shock protein 60, 70, and 90 in patients with dermatitis herpetiformis. Cell Stress Chaperones 19:837–843
Seung NR, Park EJ, Kim CW, Kim KH, Kim KJ et al (2007) Comparison of expression of heat-shock protein 60, toll-like receptors 2 and 4, and T-cell receptor gammadelta in plaque and guttate psoriasis. J Cutan Pathol 34:903–911
Marino Gammazza A, Colangeli R, Orban G, Pierucci M, Di Gennaro G, Lo Bello M, D’Aniello A, Bucchieri F, Pomara C, Valentino M, Muscat R, Benigno A, Zummo G, Conway de Macario E, Cappello F, Di Giovanni G, Macario AJL (2015) Hsp60 response in exper- imental and human temporal lobe epilepsy. Sci Rep 5:9434
Tomasello G, Rodolico V, Zerilli M, Martorana A, Bucchieri F et al (2011) Changes in immunohistochemical levels and subcellular localization after therapy and correlation and colocalization with CD68 suggest a pathogenetic role of Hsp60 in ulcerative colitis. Appl Immunohistochem Mol Morphol 19:552–561
Cappello F, Conway de Macario E, Di Felice V, Zummo G, Macario AJL (2009) Chlamydia trachomatis infection and anti-Hsp60 immunity: the two sides of the coin. PLoS Pathog 5:e1000552
Karlin S, Brocchieri L (2000) Heat shock protein 60 sequence comparisons: duplications, lateral transfer, and mitochondrial evolution. Proc Natl Acad Sci U S A 97:11348–11353
Zügel U, Kaufmann SH (1999) Role of heat shock proteins in protection from and pathogenesis of infectious diseases. Clin Microbiol Rev 12:19–39
Pockley AG (2003) Heat shock proteins as regulators of the immune response. Lancet 362:469–476
Res PC, Schaar CG, Breedveld FC, van Eden W, van Embden JD et al (1988) Synovial fluid T cell reactivity against 65kDa heat shock protein of mycobacteria in early chronic arthritis. Lancet 2:478–480
Georgopoulos C, Mc Farland H (1993) Heat shock proteins in multiple sclerosis and other autoimmune diseases. Immunol Today 14:373–375
Elfaitouri A, Herrmann B, Bölin-Wiener A, Wang Y, Gottfries CG et al (2013) Epitopes of microbial and human heat shock protein 60 and their recognition in myalgic encephalomyelitis. PLoS One 8:e8115
Child D, Smith C, Williams C (1993) Heat shock protein and the double insult theory for the development of insulin-dependent diabetes. J R Soc Med 86:217–219
Gammazza AM, Bucchieri F, Grimaldi LM, Benigno A, Conway de Macario E et al (2012) The molecular anatomy of human Hsp60 and its similarity with that of bacterial orthologs and acetylcholine receptor reveal a potential pathogenetic role of anti-chaperonin immunity in myasthenia gravis. Cell Mol Neurobiol 32:943–947
Rodolico V, Tomasello G, Zerilli M, Martorana A, Pitruzzella A et al (2010) Hsp60 and Hsp10 increase in colon mucosa of Crohn’s disease and ulcerative colitis. Cell Stress Chaperones 15:877–884
Macario AJL, Conway de Macario E (2005) Sick chaperones, cellular stress and disease. N Engl J Med 353:1489–1501
Cappello F, Marino Gammazza A, Palumbo Piccionello A, Campanella C, Pace A et al (2014) Hsp60 chaperonopathies and chaperonotherapy:targets and agents. Expert Opin Ther Targets 18:185–208
Acknowledgments
A.J.L.M. and E.C. de M. were partially supported by IMET; A.J.L.M. and F.C. were partially supported by IEMEST. This work was done under the umbrella of the agreement between the Euro-Mediterranean Institute of Science and Technology (IEMEST; Italy) and the Institute of Marine and Environmental Technology (IMET; USA) signed March 2012 (this is IMET contribution number IMET 17-195). In this work were used instruments provided by the Euro-Mediterranean Institute of Science and Technology, and funded by the Italian National Operational Programme for Research and Competitiveness 2007–2013 grant (Project code: PONa3_00210, European Regional Development Fund).
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Cappello, F., Conway de Macario, E., Rappa, F., Zummo, G., Macario, A.J.L. (2018). Immunohistochemistry of Human Hsp60 in Health and Disease: From Autoimmunity to Cancer. In: Calderwood, S., Prince, T. (eds) Chaperones. Methods in Molecular Biology, vol 1709. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7477-1_21
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DOI: https://doi.org/10.1007/978-1-4939-7477-1_21
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