Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques

Purpose The aim of our study was the characterization of anti-cytoplasmic antibodies by home-made morphological and biochemical techniques. Indeed, indirect immunofluorescence (IIF) on HEp-2 cell line is not always exhaustive in relation to the complexity of the antigens involved. Methods Nine serum...
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Gespeichert in:

Autor*in:	Alpini, Claudia [verfasserIn] Lotzniker, Milvia Valaperta, Serenella Bottone, Maria Grazia Malatesta, Manuela Montanelli, Alessandro Merlini, Giampaolo

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2012

Schlagwörter:	Anti-endoplasmatic reticulum antibodies Anti-Golgi apparatus antibodies Anti-lysosome/endosome antibodies

Anmerkung:	© Springer-Verlag 2012

Übergeordnetes Werk:	Enthalten in: Autoimmunity highlights - Mailand : Springer Milan, 2010, 3(2012), 2 vom: 09. Juni, Seite 79-85
Übergeordnetes Werk:	volume:3 ; year:2012 ; number:2 ; day:09 ; month:06 ; pages:79-85

Links:	Volltext

DOI / URN:	10.1007/s13317-012-0033-4

Katalog-ID:	SPR031317537

Internformat


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520			\|a Purpose The aim of our study was the characterization of anti-cytoplasmic antibodies by home-made morphological and biochemical techniques. Indeed, indirect immunofluorescence (IIF) on HEp-2 cell line is not always exhaustive in relation to the complexity of the antigens involved. Methods Nine serum samples with anti-cytoplasmic antibodies (2 anti-Golgi apparatus, 3 with diffuse pattern and 4 with lysosome/endosome-like pattern) were tested with fluorescent confocal microscopy, Western blot analysis and, when necessary, with electron microscopy technique. Results Confirmation of the IIF staining pattern was performed in confocal microscopy by comparison with the respective antibody marker. The anti-endoplasmatic reticulum positivity was also confirmed by electron microscopy evaluation. Both anti-lysosome/endosome and anti-endoplasmatic reticulum positivity have been definitely identified by Western blot through clear reactivity with calreticulin and LC3B, respectively. Conclusions These results do not aim at representing a standard routine laboratory procedure. Electron microscopy evaluation cannot be proposed as a routine approach, but confocal microscopy technique may be offered in centralized reference laboratories. Newer technologies, especially multiplex immunoassay, can also lead to an easier identification of these autoantibodies, without recurring to a home-made immunoblotting. Only with a complete characterization we will be able to define the clinical relevance of anti-cytoplasmic antibodies, which are still considered as “esoteric” and not as “diagnostic” antibodies.
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allfields	10.1007/s13317-012-0033-4 doi (DE-627)SPR031317537 (SPR)s13317-012-0033-4-e DE-627 ger DE-627 rakwb eng Alpini, Claudia verfasserin aut Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques 2012 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer-Verlag 2012 Purpose The aim of our study was the characterization of anti-cytoplasmic antibodies by home-made morphological and biochemical techniques. Indeed, indirect immunofluorescence (IIF) on HEp-2 cell line is not always exhaustive in relation to the complexity of the antigens involved. Methods Nine serum samples with anti-cytoplasmic antibodies (2 anti-Golgi apparatus, 3 with diffuse pattern and 4 with lysosome/endosome-like pattern) were tested with fluorescent confocal microscopy, Western blot analysis and, when necessary, with electron microscopy technique. Results Confirmation of the IIF staining pattern was performed in confocal microscopy by comparison with the respective antibody marker. The anti-endoplasmatic reticulum positivity was also confirmed by electron microscopy evaluation. Both anti-lysosome/endosome and anti-endoplasmatic reticulum positivity have been definitely identified by Western blot through clear reactivity with calreticulin and LC3B, respectively. Conclusions These results do not aim at representing a standard routine laboratory procedure. Electron microscopy evaluation cannot be proposed as a routine approach, but confocal microscopy technique may be offered in centralized reference laboratories. Newer technologies, especially multiplex immunoassay, can also lead to an easier identification of these autoantibodies, without recurring to a home-made immunoblotting. Only with a complete characterization we will be able to define the clinical relevance of anti-cytoplasmic antibodies, which are still considered as “esoteric” and not as “diagnostic” antibodies. Anti-endoplasmatic reticulum antibodies (dpeaa)DE-He213 Anti-Golgi apparatus antibodies (dpeaa)DE-He213 Anti-lysosome/endosome antibodies (dpeaa)DE-He213 Lotzniker, Milvia aut Valaperta, Serenella aut Bottone, Maria Grazia aut Malatesta, Manuela aut Montanelli, Alessandro aut Merlini, Giampaolo aut Enthalten in Autoimmunity highlights Mailand : Springer Milan, 2010 3(2012), 2 vom: 09. Juni, Seite 79-85 (DE-627)631492992 (DE-600)2563376-4 2038-3274 nnns volume:3 year:2012 number:2 day:09 month:06 pages:79-85 https://dx.doi.org/10.1007/s13317-012-0033-4 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2005 GBV_ILN_2014 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 3 2012 2 09 06 79-85
spelling	10.1007/s13317-012-0033-4 doi (DE-627)SPR031317537 (SPR)s13317-012-0033-4-e DE-627 ger DE-627 rakwb eng Alpini, Claudia verfasserin aut Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques 2012 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer-Verlag 2012 Purpose The aim of our study was the characterization of anti-cytoplasmic antibodies by home-made morphological and biochemical techniques. Indeed, indirect immunofluorescence (IIF) on HEp-2 cell line is not always exhaustive in relation to the complexity of the antigens involved. Methods Nine serum samples with anti-cytoplasmic antibodies (2 anti-Golgi apparatus, 3 with diffuse pattern and 4 with lysosome/endosome-like pattern) were tested with fluorescent confocal microscopy, Western blot analysis and, when necessary, with electron microscopy technique. Results Confirmation of the IIF staining pattern was performed in confocal microscopy by comparison with the respective antibody marker. The anti-endoplasmatic reticulum positivity was also confirmed by electron microscopy evaluation. Both anti-lysosome/endosome and anti-endoplasmatic reticulum positivity have been definitely identified by Western blot through clear reactivity with calreticulin and LC3B, respectively. Conclusions These results do not aim at representing a standard routine laboratory procedure. Electron microscopy evaluation cannot be proposed as a routine approach, but confocal microscopy technique may be offered in centralized reference laboratories. Newer technologies, especially multiplex immunoassay, can also lead to an easier identification of these autoantibodies, without recurring to a home-made immunoblotting. Only with a complete characterization we will be able to define the clinical relevance of anti-cytoplasmic antibodies, which are still considered as “esoteric” and not as “diagnostic” antibodies. Anti-endoplasmatic reticulum antibodies (dpeaa)DE-He213 Anti-Golgi apparatus antibodies (dpeaa)DE-He213 Anti-lysosome/endosome antibodies (dpeaa)DE-He213 Lotzniker, Milvia aut Valaperta, Serenella aut Bottone, Maria Grazia aut Malatesta, Manuela aut Montanelli, Alessandro aut Merlini, Giampaolo aut Enthalten in Autoimmunity highlights Mailand : Springer Milan, 2010 3(2012), 2 vom: 09. Juni, Seite 79-85 (DE-627)631492992 (DE-600)2563376-4 2038-3274 nnns volume:3 year:2012 number:2 day:09 month:06 pages:79-85 https://dx.doi.org/10.1007/s13317-012-0033-4 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2005 GBV_ILN_2014 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 3 2012 2 09 06 79-85
allfields_unstemmed	10.1007/s13317-012-0033-4 doi (DE-627)SPR031317537 (SPR)s13317-012-0033-4-e DE-627 ger DE-627 rakwb eng Alpini, Claudia verfasserin aut Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques 2012 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer-Verlag 2012 Purpose The aim of our study was the characterization of anti-cytoplasmic antibodies by home-made morphological and biochemical techniques. Indeed, indirect immunofluorescence (IIF) on HEp-2 cell line is not always exhaustive in relation to the complexity of the antigens involved. Methods Nine serum samples with anti-cytoplasmic antibodies (2 anti-Golgi apparatus, 3 with diffuse pattern and 4 with lysosome/endosome-like pattern) were tested with fluorescent confocal microscopy, Western blot analysis and, when necessary, with electron microscopy technique. Results Confirmation of the IIF staining pattern was performed in confocal microscopy by comparison with the respective antibody marker. The anti-endoplasmatic reticulum positivity was also confirmed by electron microscopy evaluation. Both anti-lysosome/endosome and anti-endoplasmatic reticulum positivity have been definitely identified by Western blot through clear reactivity with calreticulin and LC3B, respectively. Conclusions These results do not aim at representing a standard routine laboratory procedure. Electron microscopy evaluation cannot be proposed as a routine approach, but confocal microscopy technique may be offered in centralized reference laboratories. Newer technologies, especially multiplex immunoassay, can also lead to an easier identification of these autoantibodies, without recurring to a home-made immunoblotting. Only with a complete characterization we will be able to define the clinical relevance of anti-cytoplasmic antibodies, which are still considered as “esoteric” and not as “diagnostic” antibodies. Anti-endoplasmatic reticulum antibodies (dpeaa)DE-He213 Anti-Golgi apparatus antibodies (dpeaa)DE-He213 Anti-lysosome/endosome antibodies (dpeaa)DE-He213 Lotzniker, Milvia aut Valaperta, Serenella aut Bottone, Maria Grazia aut Malatesta, Manuela aut Montanelli, Alessandro aut Merlini, Giampaolo aut Enthalten in Autoimmunity highlights Mailand : Springer Milan, 2010 3(2012), 2 vom: 09. Juni, Seite 79-85 (DE-627)631492992 (DE-600)2563376-4 2038-3274 nnns volume:3 year:2012 number:2 day:09 month:06 pages:79-85 https://dx.doi.org/10.1007/s13317-012-0033-4 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2005 GBV_ILN_2014 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 3 2012 2 09 06 79-85
allfieldsGer	10.1007/s13317-012-0033-4 doi (DE-627)SPR031317537 (SPR)s13317-012-0033-4-e DE-627 ger DE-627 rakwb eng Alpini, Claudia verfasserin aut Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques 2012 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer-Verlag 2012 Purpose The aim of our study was the characterization of anti-cytoplasmic antibodies by home-made morphological and biochemical techniques. Indeed, indirect immunofluorescence (IIF) on HEp-2 cell line is not always exhaustive in relation to the complexity of the antigens involved. Methods Nine serum samples with anti-cytoplasmic antibodies (2 anti-Golgi apparatus, 3 with diffuse pattern and 4 with lysosome/endosome-like pattern) were tested with fluorescent confocal microscopy, Western blot analysis and, when necessary, with electron microscopy technique. Results Confirmation of the IIF staining pattern was performed in confocal microscopy by comparison with the respective antibody marker. The anti-endoplasmatic reticulum positivity was also confirmed by electron microscopy evaluation. Both anti-lysosome/endosome and anti-endoplasmatic reticulum positivity have been definitely identified by Western blot through clear reactivity with calreticulin and LC3B, respectively. Conclusions These results do not aim at representing a standard routine laboratory procedure. Electron microscopy evaluation cannot be proposed as a routine approach, but confocal microscopy technique may be offered in centralized reference laboratories. Newer technologies, especially multiplex immunoassay, can also lead to an easier identification of these autoantibodies, without recurring to a home-made immunoblotting. Only with a complete characterization we will be able to define the clinical relevance of anti-cytoplasmic antibodies, which are still considered as “esoteric” and not as “diagnostic” antibodies. Anti-endoplasmatic reticulum antibodies (dpeaa)DE-He213 Anti-Golgi apparatus antibodies (dpeaa)DE-He213 Anti-lysosome/endosome antibodies (dpeaa)DE-He213 Lotzniker, Milvia aut Valaperta, Serenella aut Bottone, Maria Grazia aut Malatesta, Manuela aut Montanelli, Alessandro aut Merlini, Giampaolo aut Enthalten in Autoimmunity highlights Mailand : Springer Milan, 2010 3(2012), 2 vom: 09. Juni, Seite 79-85 (DE-627)631492992 (DE-600)2563376-4 2038-3274 nnns volume:3 year:2012 number:2 day:09 month:06 pages:79-85 https://dx.doi.org/10.1007/s13317-012-0033-4 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2005 GBV_ILN_2014 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 3 2012 2 09 06 79-85
allfieldsSound	10.1007/s13317-012-0033-4 doi (DE-627)SPR031317537 (SPR)s13317-012-0033-4-e DE-627 ger DE-627 rakwb eng Alpini, Claudia verfasserin aut Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques 2012 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer-Verlag 2012 Purpose The aim of our study was the characterization of anti-cytoplasmic antibodies by home-made morphological and biochemical techniques. Indeed, indirect immunofluorescence (IIF) on HEp-2 cell line is not always exhaustive in relation to the complexity of the antigens involved. Methods Nine serum samples with anti-cytoplasmic antibodies (2 anti-Golgi apparatus, 3 with diffuse pattern and 4 with lysosome/endosome-like pattern) were tested with fluorescent confocal microscopy, Western blot analysis and, when necessary, with electron microscopy technique. Results Confirmation of the IIF staining pattern was performed in confocal microscopy by comparison with the respective antibody marker. The anti-endoplasmatic reticulum positivity was also confirmed by electron microscopy evaluation. Both anti-lysosome/endosome and anti-endoplasmatic reticulum positivity have been definitely identified by Western blot through clear reactivity with calreticulin and LC3B, respectively. Conclusions These results do not aim at representing a standard routine laboratory procedure. Electron microscopy evaluation cannot be proposed as a routine approach, but confocal microscopy technique may be offered in centralized reference laboratories. Newer technologies, especially multiplex immunoassay, can also lead to an easier identification of these autoantibodies, without recurring to a home-made immunoblotting. Only with a complete characterization we will be able to define the clinical relevance of anti-cytoplasmic antibodies, which are still considered as “esoteric” and not as “diagnostic” antibodies. Anti-endoplasmatic reticulum antibodies (dpeaa)DE-He213 Anti-Golgi apparatus antibodies (dpeaa)DE-He213 Anti-lysosome/endosome antibodies (dpeaa)DE-He213 Lotzniker, Milvia aut Valaperta, Serenella aut Bottone, Maria Grazia aut Malatesta, Manuela aut Montanelli, Alessandro aut Merlini, Giampaolo aut Enthalten in Autoimmunity highlights Mailand : Springer Milan, 2010 3(2012), 2 vom: 09. Juni, Seite 79-85 (DE-627)631492992 (DE-600)2563376-4 2038-3274 nnns volume:3 year:2012 number:2 day:09 month:06 pages:79-85 https://dx.doi.org/10.1007/s13317-012-0033-4 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2005 GBV_ILN_2014 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 3 2012 2 09 06 79-85
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source	Enthalten in Autoimmunity highlights 3(2012), 2 vom: 09. Juni, Seite 79-85 volume:3 year:2012 number:2 day:09 month:06 pages:79-85
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author	Alpini, Claudia
spellingShingle	Alpini, Claudia misc Anti-endoplasmatic reticulum antibodies misc Anti-Golgi apparatus antibodies misc Anti-lysosome/endosome antibodies Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques
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topic_title	Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques Anti-endoplasmatic reticulum antibodies (dpeaa)DE-He213 Anti-Golgi apparatus antibodies (dpeaa)DE-He213 Anti-lysosome/endosome antibodies (dpeaa)DE-He213
topic	misc Anti-endoplasmatic reticulum antibodies misc Anti-Golgi apparatus antibodies misc Anti-lysosome/endosome antibodies
topic_unstemmed	misc Anti-endoplasmatic reticulum antibodies misc Anti-Golgi apparatus antibodies misc Anti-lysosome/endosome antibodies
topic_browse	misc Anti-endoplasmatic reticulum antibodies misc Anti-Golgi apparatus antibodies misc Anti-lysosome/endosome antibodies
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title	Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques
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title_full	Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques
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author_browse	Alpini, Claudia Lotzniker, Milvia Valaperta, Serenella Bottone, Maria Grazia Malatesta, Manuela Montanelli, Alessandro Merlini, Giampaolo
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title_sort	characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques
title_auth	Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques
abstract	Purpose The aim of our study was the characterization of anti-cytoplasmic antibodies by home-made morphological and biochemical techniques. Indeed, indirect immunofluorescence (IIF) on HEp-2 cell line is not always exhaustive in relation to the complexity of the antigens involved. Methods Nine serum samples with anti-cytoplasmic antibodies (2 anti-Golgi apparatus, 3 with diffuse pattern and 4 with lysosome/endosome-like pattern) were tested with fluorescent confocal microscopy, Western blot analysis and, when necessary, with electron microscopy technique. Results Confirmation of the IIF staining pattern was performed in confocal microscopy by comparison with the respective antibody marker. The anti-endoplasmatic reticulum positivity was also confirmed by electron microscopy evaluation. Both anti-lysosome/endosome and anti-endoplasmatic reticulum positivity have been definitely identified by Western blot through clear reactivity with calreticulin and LC3B, respectively. Conclusions These results do not aim at representing a standard routine laboratory procedure. Electron microscopy evaluation cannot be proposed as a routine approach, but confocal microscopy technique may be offered in centralized reference laboratories. Newer technologies, especially multiplex immunoassay, can also lead to an easier identification of these autoantibodies, without recurring to a home-made immunoblotting. Only with a complete characterization we will be able to define the clinical relevance of anti-cytoplasmic antibodies, which are still considered as “esoteric” and not as “diagnostic” antibodies. © Springer-Verlag 2012
abstractGer	Purpose The aim of our study was the characterization of anti-cytoplasmic antibodies by home-made morphological and biochemical techniques. Indeed, indirect immunofluorescence (IIF) on HEp-2 cell line is not always exhaustive in relation to the complexity of the antigens involved. Methods Nine serum samples with anti-cytoplasmic antibodies (2 anti-Golgi apparatus, 3 with diffuse pattern and 4 with lysosome/endosome-like pattern) were tested with fluorescent confocal microscopy, Western blot analysis and, when necessary, with electron microscopy technique. Results Confirmation of the IIF staining pattern was performed in confocal microscopy by comparison with the respective antibody marker. The anti-endoplasmatic reticulum positivity was also confirmed by electron microscopy evaluation. Both anti-lysosome/endosome and anti-endoplasmatic reticulum positivity have been definitely identified by Western blot through clear reactivity with calreticulin and LC3B, respectively. Conclusions These results do not aim at representing a standard routine laboratory procedure. Electron microscopy evaluation cannot be proposed as a routine approach, but confocal microscopy technique may be offered in centralized reference laboratories. Newer technologies, especially multiplex immunoassay, can also lead to an easier identification of these autoantibodies, without recurring to a home-made immunoblotting. Only with a complete characterization we will be able to define the clinical relevance of anti-cytoplasmic antibodies, which are still considered as “esoteric” and not as “diagnostic” antibodies. © Springer-Verlag 2012
abstract_unstemmed	Purpose The aim of our study was the characterization of anti-cytoplasmic antibodies by home-made morphological and biochemical techniques. Indeed, indirect immunofluorescence (IIF) on HEp-2 cell line is not always exhaustive in relation to the complexity of the antigens involved. Methods Nine serum samples with anti-cytoplasmic antibodies (2 anti-Golgi apparatus, 3 with diffuse pattern and 4 with lysosome/endosome-like pattern) were tested with fluorescent confocal microscopy, Western blot analysis and, when necessary, with electron microscopy technique. Results Confirmation of the IIF staining pattern was performed in confocal microscopy by comparison with the respective antibody marker. The anti-endoplasmatic reticulum positivity was also confirmed by electron microscopy evaluation. Both anti-lysosome/endosome and anti-endoplasmatic reticulum positivity have been definitely identified by Western blot through clear reactivity with calreticulin and LC3B, respectively. Conclusions These results do not aim at representing a standard routine laboratory procedure. Electron microscopy evaluation cannot be proposed as a routine approach, but confocal microscopy technique may be offered in centralized reference laboratories. Newer technologies, especially multiplex immunoassay, can also lead to an easier identification of these autoantibodies, without recurring to a home-made immunoblotting. Only with a complete characterization we will be able to define the clinical relevance of anti-cytoplasmic antibodies, which are still considered as “esoteric” and not as “diagnostic” antibodies. © Springer-Verlag 2012
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title_short	Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques
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author2	Lotzniker, Milvia Valaperta, Serenella Bottone, Maria Grazia Malatesta, Manuela Montanelli, Alessandro Merlini, Giampaolo
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Electron microscopy evaluation cannot be proposed as a routine approach, but confocal microscopy technique may be offered in centralized reference laboratories. Newer technologies, especially multiplex immunoassay, can also lead to an easier identification of these autoantibodies, without recurring to a home-made immunoblotting. 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Characterization for anti-cytoplasmic antibodies specificity by morphological and molecular techniques

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