Objective: Calreticulin is a multicompartmental protein which regulates many important cellular responses. The aim of this study was to elucidate whether the intensity and location of calreticulin overexpression in tumor cells are related to the elevated humoral immunity to calreticulin in patients with benign or malignant breast disease. Methods: This study involved 27 patients with benign and 58 patients with malignant breast tumors before surgical resection and 38 healthy volunteers. Cytoplasmatic or membranous calreticulin overexpression in malignant or benign cells in paraffin-embedded tissues was determined using immunohistochemistry. Levels of the serum anti-calreticulin autoantibodies were detected by ELISA. Results: Statistically significant differences between serum levels of IgA of anti-calreticulin antibodies in controls and patients with breast tumors, and between controls and patients with nonmalignant breast diseases were found, but no statistically significant differences were found between levels of serum IgG anti-calreticulin antibodies. Humoral immunity to calreticulin developed against cytoplasmatic and co-localized membranous calreticulin was not correlated to the intensity of its overexpression and was present even in the absence of its membranous localization. Conclusions: The degree of calreticulin overexpression in lobular breast carcinoma is lower than in ductal breast carcinoma. Elevated concentrations of anti-calreticulin IgA antibodies were present more frequently in patients with metastasis in locoregional lymph nodes in comparison to anti-calreticulin IgG antibodies.

1.
Michalak M, Robert Parker JM, Opas M: Ca2+ signaling and calcium binding chaperones of the endoplasmic reticulum. Cell Calcium 2002;32:269–278.
2.
Qiu Y, Michalak M: Transcriptional control of the calreticulin gene in health and disease. Int J Biochem Cell Biol 2009;41:531–538.
3.
Erić A, Juranić Z, Milovanović Z, Marković I, Inić M, Stanojević-Bakić N, Vojinović-Golubović V: Effects of humoral immunity and calreticulin overexpression on postoperative course in breast cancer. Pathol Oncol Res 2009;15:89–90.
4.
Bini L, Magi B, Marzocchi B, Arcuri F, Tripodi S, Cintorino M, Sanchez JC, Frutiger S, Hughes G, Pallini V, Hochstrasser DF, Tosi P: Protein expression profiles in human breast ductal carcinoma and histologically normal tissue. Electrophoresis 1997;18:2832–2841.
5.
Lwin ZM, Guo C, Salim A, Yip GW, Chew FT, Nan J, Thike AA, Tan PH, Bay BH: Clinicopathological significance of calreticulin in breast invasive ductal carcinoma. Mod Pathol 2010;23:1559–1566.
6.
Liang S, Singh M, Gam LH: The differential expression of aqueous soluble proteins in breast normal and cancerous tissues in relation to ethnicity of the patients; Chinese, Malay and Indian. Dis Markers 2010;28:149–65.
7.
Baltatzis GE, Gaitanarou H, Arnogianaki N, Misitzis J, Voloudakis-Baltatzis IE: Biochemical and ultrastructural correlations of calreticulin and thioredoxin expression in breast mucinous carcinoma and infiltrating ductal carcinoma non-special type. Ultrastruct Pathol 2011;35:7–13.
8.
Gromov P, Gromova I, Bunkenborg J, Cabezon T, Moreira JM, Timmermans-Wielenga V, Roepstorff P, Rank F, Celis JE: Up-regulated proteins in the fluid bathing the tumour cell microenvironment as potential serological markers for early detection of cancer of the breast. Mol Oncol 2010;4:65–89.
9.
Zhu N, Wang Z: Calreticulin expression is associated with androgen regulation of the sensitivity to calcium ionophore-induced apoptosis in LNCaP prostate cancer cells. Cancer Res 1999;59:1896–1902.
10.
Kageyama S, Isono T, Iwaki H, Wakabayashi Y, Okada Y, Kontani K, Yoshimura K, Terai A, Arai Y, Yoshiki T: Identification by proteomic analysis of calreticulin as a marker for bladder cancer and evaluation of the diagnostic accuracy of its detection in urine. Clin Chem 2004;50:857–866.
11.
Roy L, Laboissière S, Abdou E, Thibault G, Hamel N, Taheri M, Boismenu D, Lanoix J, Kearney RE, Paiement J: Proteomic analysis of the transitional endoplasmic reticulum in hepatocellular carcinoma: an organelle perspective on cancer. Biochim Biophys Acta 2010;1804:1869–1881.
12.
Chen CN, Chang CC, Su TE, Hsu WM, Jeng YM, Ho MC, Hsieh FJ, Lee PH, Kuo L, Lee H, Chang KJ: Identification of calreticulin as a prognosis marker and angiogenic regulator in human gastric cancer. Ann Surg Oncol 2009;16:524–533.
13.
Fraser SA, Karimi R, Michalak M, Hudig D: Perforin lytic activity is controlled by calreticulin. J Immunol 2000;164:4150–4155.
14.
Dupuis M, Schaerer E, Krause KH, Tschopp J: The calcium-binding protein calreticulin is a major constituent of lytic granules in cytolytic T lymphocytes. J Exp Med 1993;177:1–7.
15.
Andrin C, Pinkoski MJ, Burns K, Atkinson EA, Krahenbuhl O, Hudig D, Fraser SA, Winkler U, Tschopp J, Opas M, Bleackley RC, Michalak M: Interaction between a Ca2+-binding protein calreticulin and perforin, a component of the cytotoxic T-cell granules. Biochemistry 1998;37:10386–10394.
16.
Sipione S, Ewen C, Shostak I, Michalak M, Bleackley RC: Impaired cytolytic activity in calreticulin-deficient CTLs. J Immunol 2005;174:3212–3219.
17.
Valck C, Ramírez G, López N, Ribeiro CH, Maldonado I, Sánchez G, Ferreira VP, Schwaeble W, Ferreira A: Molecular mechanisms involved in the inactivation of the first component of human complement by Trypanosoma cruzi calreticulin. Mol Immunol 2010;47:1516–1521.
18.
Schroeder H, Skelly PJ, Zipfel PF, Losson B, Vanderplasschen A: Subversion of complement by hematophagous parasites. Dev Comp Immunol 2009;33:5–13.
19.
Gold LI, Eggleton P, Sweetwyne MT, Van Duyn LB, Greives MR, Naylor SM, Michalak M, Murphy-Ullrich JE: Calreticulin: non-endoplasmic reticulum functions in physiology and disease. FASEB J 2010;24:665–683.
20.
Obeid M, Tesniere A, Ghiringhelli F, Fimia GM, Apetoh L, Perfettini JL, Castedo M, Mignot G, Panaretakis T, Casares N, Métivier D, Larochette N, Van Endert P, Ciccosanti F, Piacentini M, Zitvogel L, Kroemer G: Calreticulin exposure dictates the immunogenicity of cancer cell death. Nat Med 2007;13:54–61.
21.
Chaput N, De Botton S, Obeid M, Apetoh L, Ghiringhelli F, Panaretakis T, Flament C, Zitvogel L, Kroemer G: Molecular determinants of immunogenic cell death: surface exposure of calreticulin makes the difference. J Mol Med 2007;85:1069–1076.
22.
Obeid M: ERP57 membrane translocation dictates the immunogenicity of tumor cell death by controlling the membrane translocation of calreticulin. J Immunol 2008;181:2533–2543.
23.
Panaretakis T, Joza N, Modjtahedi N, Tesniere A, Vitale I, Durchschlag M, Fimia GM, Kepp O, Piacentini M, Froehlich KU, Van Endert P, Zitvogel L, Madeo F, Kroemer G: The co-translocation of ERp57 and calreticulin determines the immunogenicity of cell death. Cell Death Differ 2008;15:1499–1509.
24.
Hong SH, Misek DE, Wang H, Puravs E, Giordano TJ, Greenson JK, Brenner DE, Simeone DM, Logsdon CD, Hanash SM: An autoantibody-mediated immune response to calreticulin isoforms in pancreatic cancer. Cancer Res 2004;64:5504– 5510.
25.
Orth T, Dorner T, Meyer Zum Buschenfelde KH, Mayet WJ: Complete congenital heart block is associated with increased autoantibody titers against calreticulin. Eur J Clin Invest 1996;26:205–215.
26.
Pekáriková A, Sánchez D, Palová-Jelínková L, Simsová M, Benes Z, Hoffmanová I, Drastich P, Janatková I, Mothes T, Tlaskalová-Hogenová H, Tucková L: Calreticulin is a B cell molecular target in some gastrointestinal malignancies. Clin Exp Immunol 2010;160:215–222.
27.
Sánchez D, Palová-Jelínková L, Felsberg J, Šimšová M, Pekáriková A, Pecharová B, Swoboda I, Mothes T, Mulder CJ, Beneš Z, Tlaskalová-Hogenová H, Tučková L: Anti-calreticulin immunoglobulin A (IgA) antibodies in refractory coeliac disease. Clin Exp Immunol 2008;153:351–359.
28.
Sánchez D, Tučková L, Mothes T, Kreisel W, Beneš Z, Tlaskalová-Hogenová H: Epitopes of calreticulin recognised by IgA autoantibodies from patients with hepatic and coeliac disease. J Autoimmun 2003;21:383–392.
29.
Arranz E, Ferguson A: Intestinal antibody pattern of celiac disease: occurrence in patients with normal jejunal biopsy histology. Gastroenterol 1993;104:1263–1272.
30.
Stinton LM, Swain M, Myers MP, Shaheen AA, Fritzler MJ: Autoantibodies to GW bodies and other autoantigens in primary biliary cirrhosis. Clin Exp Immunol 2010;163:147–156.
31.
Routsias JG, Tzioufas AG: B-cell epitopes of the cytoplasmatic autoantigens Ro/SSA and La/SSB: tools to study the regulation of the autoimmune response. J Autoimmun 2010;35:256–264.
32.
Juranic ZD, Neskovic-Konstantinovic Z, Stanojkovic TP, Zizak Z, Srdic T, Stanojevic-Bakic N, Milosevic D, Jovanovic D: The antitumor immune response in HER-2 positive, metastatic breast cancer patients. J Transl Med 2005;3:13.
33.
Zappasodi R, Pupa SM, Ghedini GC, Bongarzone I, Magni M, Cabras AD, Colombo MP, Carlo-Stella C, Gianni AM, Di Nicola M: Improved clinical outcome in indolent B-cell lymphoma patients vaccinated with autologous tumor cells experiencing immunogenic death. Cancer Res 2010;70:9062–9072.
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.