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Article ; Online: The status of anti-ovarian antibody immunoassays: Valid or invalid, in or out?

Pires, Eusebio S / Parikh, Firuza R

American journal of reproductive immunology (New York, N.Y. : 1989)

2022 Volume 89, Issue 2, Page(s) e13617

Abstract: There has been a paradign shift in the status of immunoassays. There used to be a time where immunoassays had a very narrow role in clinical medicine, but that is not the case in today's world. Immunoassays have taken a central role in helping us better ... ...

Abstract	There has been a paradign shift in the status of immunoassays. There used to be a time where immunoassays had a very narrow role in clinical medicine, but that is not the case in today's world. Immunoassays have taken a central role in helping us better understand and treat human diseases. The literature around anti-ovarian antibodies (AOA) immunoassay testings have been conflicting. Researchers challenged the specificity of the reported assays, but a systematic study was never elucidated on what/who the trouble maker was in rendering these tests so nonspecific. Attempts were made by our group in Mumbai, India, to throw light on the culprit behind the nonspecificity casative factor in the immunoassays and a method to overcome this was reported and published. This review highlights the stories back five and a half decades to date, to demonstrate where the status of AOA testing was, is and will be.
MeSH term(s)	Female ; Humans ; Ovary ; Autoantibodies ; Immunoassay/methods ; India
Chemical Substances	Autoantibodies
Language	English
Publishing date	2022-09-18
Publishing country	Denmark
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	604542-x
ISSN	1600-0897 ; 0271-7352 ; 8755-8920 ; 1046-7408
ISSN (online)	1600-0897
ISSN	0271-7352 ; 8755-8920 ; 1046-7408
DOI	10.1111/aji.13617
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Article: The Unmysterious Roles of HSP90: Ovarian Pathology and Autoantibodies.

Pires, Eusebio S

Advances in anatomy, embryology, and cell biology

2017 Volume 222, Page(s) 29–44

Abstract: The heat shock proteins (HSPs) are a group of evolutionarily conserved proteins with important physiological functions, whose synthesis is enhanced by elevated temperature or other stresses. HSPs show high sequence homology between different species, ... ...

Abstract	The heat shock proteins (HSPs) are a group of evolutionarily conserved proteins with important physiological functions, whose synthesis is enhanced by elevated temperature or other stresses. HSPs show high sequence homology between different species, from bacteria to humans. Despite the significant degree of evolutionary conservation, HSPs are highly immunogenic. Of the several HSPs, HSP90 is an abundant, constitutively expressed chaperone constituting around 1-2% of total cellular protein under non-stress conditions. This protein from even the most distantly related eukaryotes has 50% amino acid identity, and all have more than 40% identity with the Escherichia coli protein. They are immunodominant antigens for many common microbes, and thus their epitopes are recognized by the immune system. As HSPs are overexpressed at sites of acute and chronic inflammation, individuals are likely to be sensitized during the course of a microbial infection encountered during life. This chapter considers the evidence of a role for HSP90 in autoimmune ovarian failure, where autoantibodies to it have been observed in patients, and has been correlated to infertility.
MeSH term(s)	Animals ; Antibody Formation/immunology ; Autoantibodies/immunology ; Female ; HSP90 Heat-Shock Proteins/immunology ; Humans ; Immunodominant Epitopes/immunology ; Ovary/immunology ; Ovary/pathology
Chemical Substances	Autoantibodies ; HSP90 Heat-Shock Proteins ; Immunodominant Epitopes
Language	English
Publishing date	2017
Publishing country	Germany
Document type	Journal Article ; Review
ISSN	0301-5556
ISSN	0301-5556
DOI	10.1007/978-3-319-51409-3_2
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Article ; Online: Cancer-oocyte SAS1B protein is expressed at the cell surface of multiple solid tumors and targeted with antibody-drug conjugates.

Mandal, Arabinda / Shetty, Jagathpala / Tran, Christine A / Olson, Walter C / Mandal, Mriganka / Ban, Bhupal / Pires, Eusebio S / Adair, Sara J / Bauer, Todd W / Slingluff, Craig L / Herr, John C

Journal for immunotherapy of cancer

2024 Volume 12, Issue 3

Abstract: Background: S: Methods: SAS1B expression in human normal and cancer tissues was determined ...

Abstract	Background: S Methods: SAS1B expression in human normal and cancer tissues was determined by immunohistochemistry, and complementary DNA (cDNA) libraries were employed to PCR amplify human SAS1B and its transcripts. Monoclonal antibodies (mAbs) to human SAS1B were generated using mouse hybridomas. SAS1B deletion constructs were developed to map SAS1B's epitope, enabling the creation of a blocking peptide. Indirect immunofluorescence (IIF) of human transfected normal and cancer cells was performed to assess SAS1B expression. SAS1B intracellular versus surface expression in normal and tumor tissues was evaluated by flow cytometry after staining with anti-SAS1B mAb, with specificity confirmed with the blocking peptide. Human cancer lines were treated with increasing mAb and ADC concentrations. ATP was quantitated as a measure of cell viability. Results: SAS1B expression was identified in a subset of human cancers and the cytoplasm of pancreatic islet cells. Two new SAS1B splice variants were deduced. Monoclonal antibodies were generated to SAS1B splice variant A. The epitope for mAbs SB2 and SB5 is between SAS1B amino acids 32-39. IIF demonstrated intracellular SAS1B expression in transfected kidney cells and on the cell surface of squamous cell lung carcinoma. Flow cytometry demonstrated intracellular SAS1B expression in all tumors and some normal cells. However, surface expression of SAS1B was identified only on cancer cells. SB2 ADC mediated dose-dependent cytotoxic killing of multiple human cancer lines. Conclusion: SAS1B is a novel cancer-oocyte antigen with cell surface expression restricted to cancer cells. In vitro, it is an effective target for antibody-mediated cancer cell lysis. These findings support further exploration of SAS1B as a potential therapeutic cancer target in multiple human cancers, either with ADC or as a chimeric antigen receptor-T (CAR-T) cell target.
MeSH term(s)	Male ; Humans ; Mice ; Animals ; Immunoconjugates/pharmacology ; Immunoconjugates/therapeutic use ; Semen ; Oocytes/metabolism ; Neoplasms/drug therapy ; Neoplasms/genetics ; Neoplasms/metabolism ; Antibodies, Monoclonal/pharmacology ; Antibodies, Monoclonal/therapeutic use ; Epitopes ; Peptides/metabolism
Chemical Substances	Immunoconjugates ; Antibodies, Monoclonal ; Epitopes ; Peptides
Language	English
Publishing date	2024-03-13
Publishing country	England
Document type	Journal Article
ZDB-ID	2719863-7
ISSN	2051-1426 ; 2051-1426
ISSN (online)	2051-1426
ISSN	2051-1426
DOI	10.1136/jitc-2023-008430
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Article ; Online: Multiplicity of molecular and cellular targets in human ovarian autoimmunity: an update.

Pires, Eusebio S

Journal of assisted reproduction and genetics

2010 Volume 27, Issue 9-10, Page(s) 519–524

Abstract: Purpose: To provide an update of putative auto-antigens identified and proposed to be involved in human ovarian autoimmunity.: Methods: Review of literature pertaining to ovarian auto-antigens / proteins identified with various immunological tools ... ...

Abstract	Purpose: To provide an update of putative auto-antigens identified and proposed to be involved in human ovarian autoimmunity. Methods: Review of literature pertaining to ovarian auto-antigens / proteins identified with various immunological tools using sera of infertile women as a probe for investigation. Results: An overview of autoimmune targets known till date in the study of human ovarian autoimmunity. Conclusions: Anti-ovarian antibodies (AOA) to multiple components and compartments of the ovary are present in the sera of infertile women. Researchers propose that these AOA may be responsible for ovarian failures and therefore render women to be infertile. Evaluation of AOA can be effective as a prognostic factor in the treatment of infertile patients and for the IVF-ET program.
MeSH term(s)	Autoantibodies/immunology ; Autoantigens/immunology ; Autoimmune Diseases/immunology ; Autoimmunity/immunology ; Female ; Humans ; Ovarian Diseases/immunology
Chemical Substances	Autoantibodies ; Autoantigens
Language	English
Publishing date	2010-06-03
Publishing country	Netherlands
Document type	Journal Article ; Review
ZDB-ID	1112577-9
ISSN	1573-7330 ; 1058-0468
ISSN (online)	1573-7330
ISSN	1058-0468
DOI	10.1007/s10815-010-9440-5
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Article ; Online: Evaluation of SAS1B as a target for antibody-drug conjugate therapy in the treatment of pancreatic cancer.

Knapp, Kiley A / Pires, Eusebio S / Adair, Sara J / Mandal, Arabinda / Mills, Anne M / Olson, Walter C / Slingluff, Craig L / Parsons, J Thomas / Bauer, Todd W / Bullock, Timothy N / Herr, John C

Oncotarget

2018 Volume 9, Issue 10, Page(s) 8972–8984

Abstract: Successful therapeutic options remain elusive for pancreatic cancer. The exquisite sensitivity and specificity of humoral and cellular immunity may provide therapeutic approaches if antigens specific for pancreatic cancer cells can be identified. Here we ...

Abstract	Successful therapeutic options remain elusive for pancreatic cancer. The exquisite sensitivity and specificity of humoral and cellular immunity may provide therapeutic approaches if antigens specific for pancreatic cancer cells can be identified. Here we characterize SAS1B (ovastacin,
Language	English
Publishing date	2018-01-04
Publishing country	United States
Document type	Journal Article
ZDB-ID	2560162-3
ISSN	1949-2553 ; 1949-2553
ISSN (online)	1949-2553
ISSN	1949-2553
DOI	10.18632/oncotarget.23944
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Article ; Online: A block in the road to fertility: autoantibodies to heat-shock protein 90-beta in human ovarian autoimmunity.

Pires, Eusebio S / Khole, Vrinda V

Fertility and sterility

2008 Volume 92, Issue 4, Page(s) 1395–1409

Abstract: ... infertility referral center and research institution.: Patient(s): Fifty women ... normally menstruating fertile women as controls.: Intervention(s): None.: Main outcome measure(s ... Identification and complete characterization of a 90-kd protein, the most immunodominant autoantigen.: Result(s ...

Abstract	Objective: To report autoantibodies to human heat-shock protein 90-beta (HSP90 beta) in sera of women with infertility. Design: Prospective, controlled observations. Setting: Major urban infertility referral center and research institution. Patient(s): Fifty women with premature ovarian failure, 65 infertile women enrolled in the in vitro fertilization-embryo transfer program, and 60 normally menstruating fertile women as controls. Intervention(s): None. Main outcome measure(s): Identification and complete characterization of a 90-kd protein, the most immunodominant autoantigen. Result(s): Our previous studies employing a novel blocking demonstrated several cellular and molecular ovarian antigenic targets using patient's serum. Of all these antigens, the 90-kd protein designated as EP90 was found to be conserved across species, was serine-threonine phosphorylated, and was expressed from the primordial stage to the graafian-stage ooplasm of the oocytes during follicular development. Using high-throughput proteomic technologies like liquid chromatography/mass spectrometry, matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF), and tandem mass spectrometry analysis revealed the identity of this protein to be HSP90 beta. Commercially available recombinant protein immunoreacted with the sera from patients with antiovarian antibodies against the 90-kd antigen. In parallel, using monoclonal antibody to human HSP90, we found that it reacts with the eluted protein from a crude ovarian extract. Conclusion(s): This is the first report to show the presence of ovarian autoantibodies to human HSP90 in sera of women with infertility. This protein could be involved in human ovarian autoimmunity and thereby be a causative factor in early ovarian failure.
MeSH term(s)	Animals ; Autoantibodies/blood ; Autoantibodies/isolation & purification ; Autoantibodies/metabolism ; Autoimmune Diseases/complications ; Autoimmune Diseases/immunology ; Autoimmunity/immunology ; Autoimmunity/physiology ; Case-Control Studies ; Female ; Fertility/immunology ; HSP90 Heat-Shock Proteins/immunology ; Humans ; Infertility, Female/etiology ; Infertility, Female/immunology ; Mice ; Ovary/immunology ; Ovary/metabolism ; Primary Ovarian Insufficiency/blood ; Primary Ovarian Insufficiency/complications ; Primary Ovarian Insufficiency/immunology ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Swine
Chemical Substances	Autoantibodies ; HSP90 Heat-Shock Proteins ; HSP90AB1 protein, human
Language	English
Publishing date	2008-11-19
Publishing country	United States
Document type	Journal Article
ZDB-ID	80133-1
ISSN	1556-5653 ; 0015-0282
ISSN (online)	1556-5653
ISSN	0015-0282
DOI	10.1016/j.fertnstert.2008.08.068
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Article ; Online: Correction: Membrane associated cancer-oocyte neoantigen SAS1B/ovastacin is a candidate immunotherapeutic target for uterine tumors.

Pires, Eusebio S / D'Souza, Ryan S / Needham, Marisa A / Herr, Austin K / Jazaeri, Amir A / Li, Hui / Stoler, Mark H / Anderson-Knapp, Kiley L / Thomas, Theodore / Mandal, Arabinda / Gougeon, Alain / Flickinger, Charles J / Bruns, David E / Pollok, Brian A / Herr, John C

Oncotarget

2017 Volume 8, Issue 9, Page(s) 16099

Language	English
Publishing date	2017-06-19
Publishing country	United States
Document type	Journal Article ; Published Erratum
ZDB-ID	2560162-3
ISSN	1949-2553 ; 1949-2553
ISSN (online)	1949-2553
ISSN	1949-2553
DOI	10.18632/oncotarget.15755
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Article ; Online: Anti-HSP90 autoantibodies in sera of infertile women identify a dominant, conserved epitope EP6 (380-389) of HSP90 beta protein.

Pires, Eusebio S / Choudhury, Asmita K / Idicula-Thomas, Susan / Khole, Vrinda V

Reproductive biology and endocrinology : RB&E

2011 Volume 9, Page(s) 16

Abstract: ... on prediction and validation of the immunodominant epitope/s of this protein using sera from infertile women ...

Abstract	Background: We earlier reported a simple specific test for detection of anti-ovarian antibodies in infertile women and identified number of specific molecular and cellular targets of which human heat shock protein 90-beta (HSP90 beta) was found to be the most immunodominant. The present study focuses on prediction and validation of the immunodominant epitope/s of this protein using sera from infertile women having anti-HSP90 autoantibodies. Methods: Delineation of the immunodominant epitopes of HSP90 beta was done by using epitope prediction algorithms and 10 peptides (EP1-EP10) were custom synthesized. Their immunoreactivity was measured by ELISA using sera from patients and controls. To determine the most immunodominant epitope, the results were subjected to statistical analysis. The immunoreactivity of the immunodominant peptides were confirmed by dot blots using sera from patients. A rabbit polyclonal antibody against the immunodominant epitope was generated and its immunoreactivity to the parent protein in ovarian extracts as well in oocytes and embryos was investigated. Results: Experimentally and statistically, peptide EP6 (380-389) seems to be the major antigenic epitope for the serum antibody binding followed by EP1 (1-12) and EP8 (488-498). Predicted 3D structures of these peptides demonstrated that they exist in the loop conformation which is the most mobile part of the protein. Also, analysis of the sequences of HSP90 beta across several species reveals that EP6 peptide forms a part of a well conserved motif. The polyclonal antibody generated to the immunodominant epitope- EP6 confirms similar biochemical and cellular immunoreactivity as seen with the patients' sera having anti-HSP90 autoantibodies. Conclusions: The decapeptide EP6 is a major immunogenic epitope of HSP90 followed by EP1 and EP8. Knowledge of binding epitopes on the autoantigen is necessary to understand the subsequent pathologic events. The study might generate new tools for the detection of disease-inducing epitopes and a possible therapeutic intervention.
MeSH term(s)	Animals ; Autoantibodies/blood ; Autoantigens/immunology ; Female ; HSP90 Heat-Shock Proteins/immunology ; Humans ; Immunodominant Epitopes/immunology ; Infertility, Female/immunology ; Oligopeptides/immunology ; Peptide Fragments/immunology ; Rabbits
Chemical Substances	Autoantibodies ; Autoantigens ; HSP90 Heat-Shock Proteins ; Immunodominant Epitopes ; Oligopeptides ; Peptide Fragments
Language	English
Publishing date	2011-01-27
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1477-7827
ISSN (online)	1477-7827
DOI	10.1186/1477-7827-9-16
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Article ; Online: Anti-HSP90 autoantibodies in sera of infertile women identify a dominant, conserved epitope EP6 (380-389) of HSP90 beta protein

Idicula-Thomas Susan / Choudhury Asmita K / Pires Eusebio S / Khole Vrinda V

Reproductive Biology and Endocrinology, Vol 9, Iss 1, p

2011 Volume 16

Abstract: ... on prediction and validation of the immunodominant epitope/s of this protein using sera from infertile women ...

Abstract	Abstract Background We earlier reported a simple specific test for detection of anti-ovarian antibodies in infertile women and identified number of specific molecular and cellular targets of which human heat shock protein 90-beta (HSP90 beta) was found to be the most immunodominant. The present study focuses on prediction and validation of the immunodominant epitope/s of this protein using sera from infertile women having anti-HSP90 autoantibodies. Methods Delineation of the immunodominant epitopes of HSP90 beta was done by using epitope prediction algorithms and 10 peptides (EP1-EP10) were custom synthesized. Their immunoreactivity was measured by ELISA using sera from patients and controls. To determine the most immunodominant epitope, the results were subjected to statistical analysis. The immunoreactivity of the immunodominant peptides were confirmed by dot blots using sera from patients. A rabbit polyclonal antibody against the immunodominant epitope was generated and its immunoreactivity to the parent protein in ovarian extracts as well in oocytes and embryos was investigated. Results Experimentally and statistically, peptide EP6 (380-389) seems to be the major antigenic epitope for the serum antibody binding followed by EP1 (1-12) and EP8 (488-498). Predicted 3D structures of these peptides demonstrated that they exist in the loop conformation which is the most mobile part of the protein. Also, analysis of the sequences of HSP90 beta across several species reveals that EP6 peptide forms a part of a well conserved motif. The polyclonal antibody generated to the immunodominant epitope- EP6 confirms similar biochemical and cellular immunoreactivity as seen with the patients' sera having anti-HSP90 autoantibodies. Conclusions The decapeptide EP6 is a major immunogenic epitope of HSP90 followed by EP1 and EP8. Knowledge of binding epitopes on the autoantigen is necessary to understand the subsequent pathologic events. The study might generate new tools for the detection of disease-inducing epitopes and a possible therapeutic intervention.
Keywords	Physiology ; QP1-981 ; Science ; Q ; DOAJ:Physiology ; DOAJ:Biology ; DOAJ:Biology and Life Sciences
Subject code	570
Language	English
Publishing date	2011-01-01T00:00:00Z
Publisher	BioMed Central
Document type	Article ; Online
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Article ; Online: Can anti-ovarian antibody testing be useful in an IVF-ET clinic?

Pires, Eusebio S / Parikh, Firuza R / Mande, Purvi V / Uttamchandani, Shonali A / Savkar, Sujata / Khole, Vrinda V

Journal of assisted reproduction and genetics

2010 Volume 28, Issue 1, Page(s) 55–64

Abstract: ... infertility reference centre and National research institute.: Patient(s): Five hundred seventy infertile ... women enrolled for IVF-ET.: Intervention(s): AOA testing, corticosteroid therapy and IVF-ET/ICSI ... Main outcome measure(s): Comparable clinical outcome and significance of AOA testing established ...

Abstract	Objective: To establish importance of anti-ovarian antibodies (AOA) testing in infertile women. Design: A clinical reproductive outcome comparative study between two groups of women undergoing IVF-ET. Group 1 consists of women tested positive for AOA, put on corticosteroid therapy, reverted to AOA negative and then taken up for IVF-ET. Group 2 were seronegative for AOA. Setting: Major urban infertility reference centre and National research institute. Patient(s): Five hundred seventy infertile women enrolled for IVF-ET. Intervention(s): AOA testing, corticosteroid therapy and IVF-ET/ICSI. Main outcome measure(s): Comparable clinical outcome and significance of AOA testing established. Results: AOA positive serum samples were sent periodically to re-investigate presence of AOA after corticosteroid therapy and women turned AOA negative were taken up for IVF-ET. Of the 70/138 women in group 1 who were treated with corticosteroids and turned seronegative for AOA, 22/70 were poor responders and needed donor oocyte-recipient cycles. Results demonstrated that fertilization and clinical pregnancy rates between both groups are comparable. Nevertheless, it is also observed that there is poor response to stimulation protocol, smaller number of oocytes retrieved and more spontaneous abortions in group 1 women. Hence not all outcomes following the treatment are comparable between the two groups. Usefulness of the test was established in two case studies. Conclusions: AOA testing could be included in the battery of tests investigating and treating infertility.
MeSH term(s)	Adrenal Cortex Hormones/therapeutic use ; Adult ; Antibodies, Anti-Idiotypic/blood ; Antibodies, Anti-Idiotypic/immunology ; Autoantibodies/blood ; Autoantibodies/immunology ; Female ; Humans ; Infertility, Female/drug therapy ; Infertility, Female/immunology ; Oocytes/immunology ; Ovary/immunology ; Pregnancy ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic/methods
Chemical Substances	Adrenal Cortex Hormones ; Antibodies, Anti-Idiotypic ; Autoantibodies
Language	English
Publishing date	2010-10-12
Publishing country	Netherlands
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1112577-9
ISSN	1573-7330 ; 1058-0468
ISSN (online)	1573-7330
ISSN	1058-0468
DOI	10.1007/s10815-010-9488-2
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