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Article ; Online: Somatic mutations in FAS pathway increase hemophagocytic lymphohistiocytosis risk in T- and/or NK-cell lymphoma patients.

Liu, Ying / Sardana, Rohan / Nemirovsky, David / Frosina, Denise / Jungbluth, Achim / Johnson, William T / Vardhana, Santosha A / Arcila, Maria E / Horwitz, Steven M / Derkach, Andriy / Dogan, Ahmet / Xiao, Wenbin

Blood advances

2024

Abstract: While significant progress has been made in understanding the genetic basis of primary hemophagocytic lymphohistiocytosis (HLH), the pathogenesis of secondary HLH, the more prevalent form, remains unclear. Among the various conditions giving rise to ... ...

Abstract	While significant progress has been made in understanding the genetic basis of primary hemophagocytic lymphohistiocytosis (HLH), the pathogenesis of secondary HLH, the more prevalent form, remains unclear. Among the various conditions giving rise to secondary HLH, HLH in lymphoma patients (HLH-L) accounts for a substantial proportion. In this study, we investigated the role of somatic mutations in the pathogenesis of HLH-L in a cohort of patients with T- and/or NK-cell lymphoma. We identified a 3-time higher frequency of mutations in FAS pathway in patients with HLH-L. Patients harbouring these mutations had a 5-time increased HLH-L risk. These mutations were independently associated with inferior outcome. Hence, our study demonstrates the association between somatic mutations in FAS pathway and HLH-L. Further studies are warranted on the mechanistic role of these mutations in HLH-L.
Language	English
Publishing date	2024-04-09
Publishing country	United States
Document type	Journal Article
ZDB-ID	2915908-8
ISSN	2473-9537 ; 2473-9529
ISSN (online)	2473-9537
ISSN	2473-9529
DOI	10.1182/bloodadvances.2023011733
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Zs.A 7153: Show issues

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Article ; Online: A Novel Technique for the Generation of Multitissue Blocks Using a Carrier.

Frosina, Denise / Jungbluth, Achim A

Applied immunohistochemistry & molecular morphology : AIMM

2016 Volume 24, Issue 9, Page(s) 668–672

Abstract: Paraffin blocks containing several tissues have become a major tool in surgical pathology. As multitissue blocks, they usually consist of few rather large samples and as tissue microarrays (TMAs) they may contain up to several hundred small-sized tissue ... ...

Abstract	Paraffin blocks containing several tissues have become a major tool in surgical pathology. As multitissue blocks, they usually consist of few rather large samples and as tissue microarrays (TMAs) they may contain up to several hundred small-sized tissue cores. We developed a novel approach to generate multitissue blocks using a carrier in which tissue samples are inserted. Normal tissues with homogenous consistency such as liver, spleen, and lung seem to be ideal carriers. Carrier tissue size can be freely chosen to accommodate number and size of sample tissues as desired. As the carrier tissue serves as a scaffold for the inserted tissue, even small-sized tissues will stay exactly as placed in the carrier. This makes carrier-based multitissue blocks (CBMTBs) an ideal approach when exact orientation of layers is important, for example in normal GIT tissues. The carrier tissue approach can also be used for few large-sized sample tissues or to generate classical TMAs with sample cores of <1 mm. As the newly generated CBMTB or carrier-based tissue microarray is completely reembedded after assembling, sectioning of CBMTBs is comparable with sectioning a normal solid tissue block producing virtually no sample loss and requiring minimal trimming and consequently vastly increasing yield.
MeSH term(s)	Humans ; Immunohistochemistry ; Paraffin Embedding/methods
Language	English
Publishing date	2016-10
Publishing country	United States
Document type	Journal Article
ZDB-ID	1473273-7
ISSN	1533-4058 ; 1062-3345 ; 1541-2016
ISSN (online)	1533-4058
ISSN	1062-3345 ; 1541-2016
DOI	10.1097/PAI.0000000000000239
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Zs.A 3783: Show issues

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Article ; Online: Immunohistochemical Detection of Cancer-Testis Antigen PRAME.

Lezcano, Cecilia / Müller, Annette M / Frosina, Denise / Hernandez, Enmily / Geronimo, Jerica A / Busam, Klaus J / Jungbluth, Achim A

International journal of surgical pathology

2021 Volume 29, Issue 8, Page(s) 826–835

Abstract: Cancer-testis (CT) antigens were identified by their ability to elicit T- or B-cell immune responses in the autologous host. They are typically expressed in a wide variety of neoplasms and in normal adult tissues are restricted to testicular germ cells. ... ...

Abstract	Cancer-testis (CT) antigens were identified by their ability to elicit T- or B-cell immune responses in the autologous host. They are typically expressed in a wide variety of neoplasms and in normal adult tissues are restricted to testicular germ cells. PReferentially expressed Antigen of Melanoma (PRAME) is a member of the family of nonclassical CT antigens being expressed in a few other normal tissues besides testis. Interestingly, knowledge about the protein expression of many CT antigens is still incomplete due to the limited availability of reagents for their immunohistochemical detection. Here, we tested several commercially available serological reagents and identified a monoclonal antibody suitable for the immunohistochemical detection of PRAME in formalin-fixed paraffin-embedded specimens. We also tested a wide array of normal and neoplastic tissues. PRAME protein expression in normal tissues is congruent with original molecular data being present in the testis, and at low levels in the endometrium, adrenal cortex, and adult as well as fetal ovary. In tumors, there is diffuse PRAME immunoreactivity in most metastatic melanomas, myxoid liposarcomas, and synovial sarcomas. Other neoplasms such as seminomas and carcinomas of various origins including endometrial, serous ovarian, mammary ductal, lung, and renal showed an intermediate proportion of cases and variable extent of tumor cells positive for PRAME protein expression. As seen with other CT antigens, hepatocellular and colorectal carcinoma, Leydig cell tumors, mesothelioma, and leiomyosarcoma are poor expressers of PRAME.
MeSH term(s)	Antibodies, Monoclonal/immunology ; Antibodies, Monoclonal/metabolism ; Antigens, Neoplasm/immunology ; Antigens, Neoplasm/isolation & purification ; Antigens, Neoplasm/metabolism ; Biomarkers, Tumor/analysis ; Cell Line, Tumor ; Female ; Humans ; Immunohistochemistry/methods ; Male ; Neoplasms/diagnosis ; Neoplasms/pathology
Chemical Substances	Antibodies, Monoclonal ; Antigens, Neoplasm ; Biomarkers, Tumor ; PRAME protein, human
Language	English
Publishing date	2021-04-23
Publishing country	United States
Document type	Journal Article
ZDB-ID	1336393-1
ISSN	1940-2465 ; 1066-8969
ISSN (online)	1940-2465
ISSN	1066-8969
DOI	10.1177/10668969211012085
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Zs.A 4500: Show issues

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Article ; Online: Expression Analysis of GD2 by Immunohistochemistry in Invasive Breast Carcinoma: Clinical and Pathologic Correlation.

Zhong, Elaine / Brogi, Edi / D'Alfonso, Timothy M / Wen, Hannah / Frosina, Denise / Cheung, Nai-Kong / Jungbluth, Achim A / Ross, Dara S

Applied immunohistochemistry & molecular morphology : AIMM

2021 Volume 30, Issue 2, Page(s) 113–118

Abstract: The glycosphingolipid disialoganglioside GD2 is a cell surface-associated antigen expressed on tumors of neuroectodermal origin that serves as a target of immunotherapy in select cancer types. Information about the expression of GD2 in breast cancer is ... ...

Abstract	The glycosphingolipid disialoganglioside GD2 is a cell surface-associated antigen expressed on tumors of neuroectodermal origin that serves as a target of immunotherapy in select cancer types. Information about the expression of GD2 in breast cancer is limited. In the present study, we investigate the utility of GD2 as a potential biomarker for targeted treatment. The study cohort consists of 386 breast carcinomas of several histologic types. GD2 expression was assessed in both whole tumor sections and tissue microarrays with anti-GD2 3F8 monoclonal antibody immunohistochemistry and correlated with clinicopathologic features and survival outcomes. A total of 134 (35%) breast carcinomas were positive for GD2, with a median H-score of 100. 3F8 staining displayed granular and predominantly cytoplasmic or perinuclear patterns, which was confined to the neoplastic tissue in nearly all cases. GD2 positivity was significantly associated with tumor histologic type (P=0.0015), low grade (P<0.0001), estrogen receptor positivity (P<0.0001), low stage (P=0.0014), and multifocality (P=0.022). Event-free survival and overall survival of patients with GD2-positive and GD2-negative tumors were not significantly different. Our results support further assessment of GD2 using the 3F8 antibody as a predictive and prognostic biomarker in breast cancer.
MeSH term(s)	Antineoplastic Agents/therapeutic use ; Breast Neoplasms/diagnosis ; Breast Neoplasms/drug therapy ; Female ; Gangliosides/metabolism ; Humans ; Immunohistochemistry ; Immunotherapy
Chemical Substances	Antineoplastic Agents ; Gangliosides
Language	English
Publishing date	2021-09-16
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1473273-7
ISSN	1533-4058 ; 1062-3345 ; 1541-2016
ISSN (online)	1533-4058
ISSN	1062-3345 ; 1541-2016
DOI	10.1097/PAI.0000000000000974
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Zs.A 3783: Show issues

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Article ; Online: mTOR Pathway Activation Assessed by Immunohistochemistry in Cervical Biopsies of HPV-associated Endocervical Adenocarcinomas (HPVA): Correlation With Silva Invasion Patterns.

Segura, Sheila / Stolnicu, Simona / Boros, Monica / Park, Kay / Ramirez, Pedro / Salvo, Gloria / Frosina, Denise / Jungbluth, Achim / Soslow, Robert A

Applied immunohistochemistry & molecular morphology : AIMM

2021 Volume 29, Issue 7, Page(s) 527–533

Abstract: The Silva pattern of invasion, recently introduced to stratify patients at risk for lymph node metastases in human papillomavirus-associated endocervical adenocarcinomas (HPVAs), can only be assessed in cone and loop electrosurgical excision procedure ... ...

Abstract	The Silva pattern of invasion, recently introduced to stratify patients at risk for lymph node metastases in human papillomavirus-associated endocervical adenocarcinomas (HPVAs), can only be assessed in cone and loop electrosurgical excision procedure excisions with negative margins or in a hysterectomy specimen. Previous studies found associations between destructive stromal invasion patterns (Silva patterns B and C) and mutations in genes involved in the MEK/PI3K pathways that activate the mammalian target of rapamycin (mTOR) pathway. The primary aim of this study was to use cervical biopsies to determine whether markers of mTOR pathway activation associate with aggressive invasion patterns in matched excision specimens. The status of the markers in small biopsy specimens should allow us to predict the final and biologically relevant pattern of invasion in a resection specimen. Being able to predict the final pattern of invasion is important, since prediction as Silva A, for example, might encourage conservative clinical management. If the pattern in the resection specimen is B with lymphovascular invasion or C, further surgery can be performed 34 HPVA biopsies were evaluated for expression of pS6, pERK, and HIF1α. Immunohistochemical stains were scored semiquantitatively, ranging from 0 to 4+ with scores 2 to 4+ considered positive, and Silva pattern was determined in follow-up excisional specimens. Silva patterns recognized in excisional specimens were distributed as follows: pattern A (n=8), pattern B (n=4), and pattern C (n=22). Statistically significant associations were found comparing pS6 and pERK immunohistochemistry with Silva pattern (P=0.034 and 0.05, respectively). Of the 3 markers tested, pERK was the most powerful for distinguishing between pattern A and patterns B and C (P=0.026; odds ratio: 6.75, 95% confidence interval: 1.111-41.001). Although the negative predictive values were disappointing, the positive predictive values were encouraging: 90% for pERK, 88% for pS6 and 100% for HIF1α. mTOR pathway activation assessed by immunohistochemistry in cervical biopsies of HPVA correlate with Silva invasion patterns.
MeSH term(s)	Adenocarcinoma/enzymology ; Adenocarcinoma/pathology ; Adenocarcinoma/virology ; Adult ; Biopsy ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Papillomaviridae/metabolism ; Papillomavirus Infections/enzymology ; Papillomavirus Infections/pathology ; TOR Serine-Threonine Kinases/metabolism ; Uterine Cervical Neoplasms/enzymology ; Uterine Cervical Neoplasms/pathology ; Uterine Cervical Neoplasms/virology
Chemical Substances	MTOR protein, human (EC 2.7.1.1) ; TOR Serine-Threonine Kinases (EC 2.7.11.1)
Language	English
Publishing date	2021-01-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1473273-7
ISSN	1533-4058 ; 1062-3345 ; 1541-2016
ISSN (online)	1533-4058
ISSN	1062-3345 ; 1541-2016
DOI	10.1097/PAI.0000000000000915
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Zs.A 3783: Show issues

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Article ; Online: Performance of DAXX Immunohistochemistry as a Screen for DAXX Mutations in Pancreatic Neuroendocrine Tumors.

Hechtman, Jaclyn F / Klimstra, David S / Nanjangud, Gouri / Frosina, Denise / Shia, Jinru / Jungbluth, Achim A

Pancreas

2019 Volume 48, Issue 3, Page(s) 396–399

Abstract: Objectives: DAXX immunohistochemistry (IHC) is often used as a surrogate for sequencing. We aimed to elucidate the sensitivity of IHC for DAXX mutation.: Methods: All pancreatic neuroendocrine tumors (PanNETs) with DAXX mutations detected by ... ...

Abstract	Objectives: DAXX immunohistochemistry (IHC) is often used as a surrogate for sequencing. We aimed to elucidate the sensitivity of IHC for DAXX mutation. Methods: All pancreatic neuroendocrine tumors (PanNETs) with DAXX mutations detected by sequencing and a subset of DAXX wild-type PanNETs were analyzed for DAXX expression by IHC. Results: Of 154 PanNETs with MSK-IMPACT testing, 36 (30%) harbored DAXX mutations. DAXX mutations were associated with TSC2 mutations (46% vs 10%, P < 0.0001), tended to co-occur with MEN1 mutations (63% vs 49%, P = 0.11), and tended to be mutually exclusive with ATRX mutations (11% vs 25%, P = 0.053). Of 27 available DAXX mutant PanNETs, 23 lost DAXX expression (85.2%). All 4 DAXX mutants with retained expression harbored DAXX mutations within the SUMO-interacting motif of the last exon. Telomere-specific fluorescence in situ hybridization demonstrated alternative lengthening of telomeres in all 4 cases. Of 20 PanNETs with wild-type DAXX, 19 retained DAXX IHC expression (95%). Conclusions: The sensitivity and specificity of IHC for DAXX mutation are 85% and 95%, respectively. Last exon DAXX mutant PanNETs often show alternative lengthening of telomeres despite retained DAXX expression, likely due to escape of nonmediated decay.
MeSH term(s)	Adaptor Proteins, Signal Transducing/biosynthesis ; Adaptor Proteins, Signal Transducing/genetics ; Biomarkers, Tumor/biosynthesis ; Biomarkers, Tumor/genetics ; DNA Mutational Analysis ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Mutation ; Neuroendocrine Tumors/diagnosis ; Neuroendocrine Tumors/genetics ; Neuroendocrine Tumors/metabolism ; Nuclear Proteins/biosynthesis ; Nuclear Proteins/genetics ; Pancreatic Neoplasms/diagnosis ; Pancreatic Neoplasms/genetics ; Pancreatic Neoplasms/metabolism ; Retrospective Studies ; Telomere Homeostasis/genetics ; X-linked Nuclear Protein/biosynthesis ; X-linked Nuclear Protein/genetics
Chemical Substances	Adaptor Proteins, Signal Transducing ; Biomarkers, Tumor ; DAXX protein, human ; Nuclear Proteins ; ATRX protein, human (EC 3.6.4.12) ; X-linked Nuclear Protein (EC 3.6.4.12)
Language	English
Publishing date	2019-02-07
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	632831-3
ISSN	1536-4828 ; 0885-3177
ISSN (online)	1536-4828
ISSN	0885-3177
DOI	10.1097/MPA.0000000000001256
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Zs.A 2160: Show issues

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Article ; Online: Comparison of Immunohistochemistry, Next-generation Sequencing and Fluorescence In Situ Hybridization for Detection of MTAP Loss in Pleural Mesothelioma.

Febres-Aldana, Christopher A / Chang, Jason C / Jungbluth, Achim A / Adusumilli, Prasad S / Bodd, Francis M / Frosina, Denise / Geronimo, Jerica A / Hernandez, Enmily / Irawan, Helen / Offin, Michael D / Rekhtman, Natasha / Travis, William D / Vanderbilt, Chad / Zauderer, Marjorie G / Zhang, Yanming / Ladanyi, Marc / Yang, Soo-Ryum / Sauter, Jennifer L

Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

2024 Volume 37, Issue 3, Page(s) 100420

Abstract: 9p21 deletions involving MTAP/CDKN2A genes are detected in diffuse pleural mesotheliomas (DPM) but are absent in benign mesothelial proliferations. Loss of MTAP expression by immunohistochemistry (IHC) is well accepted as a surrogate for 9p21 deletion to ...

Abstract	9p21 deletions involving MTAP/CDKN2A genes are detected in diffuse pleural mesotheliomas (DPM) but are absent in benign mesothelial proliferations. Loss of MTAP expression by immunohistochemistry (IHC) is well accepted as a surrogate for 9p21 deletion to support a diagnosis of DPM. Accurate interpretation can be critical in the diagnosis of DPM, but variations in antibody performance may impact interpretation. The objectives of this study were to compare the performance of MTAP monoclonal antibodies (mAbs) EPR6893 and 1813 and to compare MTAP expression by IHC with 9p21 copy number status in DPM. Cytoplasmic expression of MTAP IHC with mAbs EPR6893 (ab126770; Abcam) and 1813 (NBP2-75730, Novus Biologicals) was evaluated in 56 DPM (47 epithelioid, 7 biphasic, and 2 sarcomatoid) profiled by targeted next-generation sequencing. 9p21 Copy number status was assessed by Fraction and Allele-Specific Copy Number Estimates from Tumor Sequencing (FACETS) analysis and also by CDKN2A fluorescence in situ hybridization in discrepant cases when material was available. MTAP mAb 1813 showed stronger immunoreactivity, more specific staining, and no equivocal interpretations compared to mAb EPR6893 which showed equivocal staining in 19 (34%) of cases due to weak or heterogenous immunoreactivity, lack of definitive internal positive control, and/or nonspecific background staining. MTAP expression with mAb 1813 showed near perfect agreement with 9p21 copy number by combined FACETS/fluorescence in situ hybridization calls (κ = 0.85; 95% CI, 0.71-0.99; P < .001). MTAP IHC with mAb 1813 was 96% sensitive, 86% specific, and 93% accurate for 9p21 homozygous deletion. The findings of this study suggest that interpretation of MTAP IHC is improved with mAb 1813 because mAb EPR6893 was often limited by equivocal interpretations. We show that MTAP IHC and molecular assays are complementary in detecting 9p21 homozygous deletion. MTAP IHC may be particularly useful for low tumor purity samples and in low-resource settings.
MeSH term(s)	Humans ; Biomarkers, Tumor/analysis ; Cyclin-Dependent Kinase Inhibitor p16/genetics ; High-Throughput Nucleotide Sequencing ; Homozygote ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Mesothelioma/diagnosis ; Mesothelioma/genetics ; Mesothelioma/pathology ; Mesothelioma, Malignant/genetics ; Pleural Neoplasms/diagnosis ; Pleural Neoplasms/genetics ; Pleural Neoplasms/pathology ; Sequence Deletion ; Ubiquitin Thiolesterase/genetics
Chemical Substances	Biomarkers, Tumor ; Cyclin-Dependent Kinase Inhibitor p16 ; Ubiquitin Thiolesterase (EC 3.4.19.12)
Language	English
Publishing date	2024-01-05
Publishing country	United States
Document type	Comparative Study ; Journal Article
ZDB-ID	645073-8
ISSN	1530-0285 ; 0893-3952
ISSN (online)	1530-0285
ISSN	0893-3952
DOI	10.1016/j.modpat.2023.100420
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Zs.A 2450: Show issues

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Article ; Online: Pan-Trk immunohistochemistry is a sensitive and specific ancillary tool for diagnosing secretory carcinoma of the salivary gland and detecting ETV6-NTRK3 fusion.

Xu, Bin / Haroon Al Rasheed, Mohamed R / Antonescu, Cristina R / Alex, Deepu / Frosina, Denise / Ghossein, Ronald / Jungbluth, Achim A / Katabi, Nora

Histopathology

2019 Volume 76, Issue 3, Page(s) 375–382

Abstract: Aims: Secretory carcinoma (SC) of the salivary gland typically harbours ETV6-NTRK3 fusion, which can be utilised clinically to assist with diagnosis. Pan-Trk inhibitor therapy has demonstrated drastic responses in patients with NTRK-translocated tumours, ...

Abstract	Aims: Secretory carcinoma (SC) of the salivary gland typically harbours ETV6-NTRK3 fusion, which can be utilised clinically to assist with diagnosis. Pan-Trk inhibitor therapy has demonstrated drastic responses in patients with NTRK-translocated tumours, including SC. Pan-Trk immunohistochemistry (IHC) is emerging as a sensitive and specific tool for detecting NTRK1, NTRK2 and NTRK3 fusions in various cancers. We aimed to establish the specificity and sensitivity of pan-Trk IHC in diagnosing SC and detecting ETV6-NTRK3 fusion. A literature review on the utility of pan-Trk IHC was conducted. Methods and results: Pan-Trk IHC was performed on 83 salivary gland neoplasms (29 SCs and 54 non-SCs). ETV6-NTRK3 fusion status was established in 25 cases. With any staining (nuclear or cytoplasmic) as a positive threshold, the sensitivity and specificity of pan-Trk IHC were 90% and 70% in diagnosing SC, and 100% and 0% in detecting NTRK3 fusion. When only pan-Trk nuclear staining was considered as positive, the sensitivity and specificity were 69% and 100% in diagnosing SC, and 92% and 100% in detecting NTRK3 fusion. Conclusions: Nuclear pan-Trk IHC is highly specific for SC diagnosis, with a specificity approaching 100%, making it a useful and precise diagnostic tool for differentiating SC from its histological mimics. On the other hand, any pan-Trk staining (nuclear or cytoplasmic) is highly sensitive for SC, and can serve as an attractive, cheap, fast and accessible screening tool for selecting patients to undergo confirmative molecular testing for clinical trials using TRK inhibitors.
MeSH term(s)	Carcinoma/diagnosis ; Carcinoma/genetics ; Carcinoma/metabolism ; Carcinoma/pathology ; Cohort Studies ; Humans ; Immunohistochemistry ; Oncogene Proteins, Fusion/genetics ; Oncogene Proteins, Fusion/metabolism ; Retrospective Studies ; Salivary Gland Neoplasms/diagnosis ; Salivary Gland Neoplasms/genetics ; Salivary Gland Neoplasms/metabolism ; Salivary Gland Neoplasms/pathology ; Salivary Glands/metabolism ; Salivary Glands/pathology ; Sensitivity and Specificity
Chemical Substances	ETV6-NTRK3 fusion protein, human ; Oncogene Proteins, Fusion
Language	English
Publishing date	2019-12-11
Publishing country	England
Document type	Journal Article
ZDB-ID	131914-0
ISSN	1365-2559 ; 0309-0167
ISSN (online)	1365-2559
ISSN	0309-0167
DOI	10.1111/his.13981
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Zs.A 1328: Show issues

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Article ; Online: CytoLyt fixation significantly inhibits MIB1 immunoreactivity whereas alternative Ki-67 clone 30-9 is not susceptible to the inhibition: Critical diagnostic implications.

Buonocore, Darren J / Konno, Fumiko / Jungbluth, Achim A / Frosina, Denise / Fayad, Mariam / Edelweiss, Marcia / Lin, Oscar / Rekhtman, Natasha

Cancer cytopathology

2019 Volume 127, Issue 10, Page(s) 643–649

Abstract: Background: The Ki-67 proliferation marker has multiple diagnostic and prognostic applications. Although several clones to the Ki-67 antigen are commercially available, the MIB1 clone is widely recommended in the surgical pathology literature for ... ...

Abstract	Background: The Ki-67 proliferation marker has multiple diagnostic and prognostic applications. Although several clones to the Ki-67 antigen are commercially available, the MIB1 clone is widely recommended in the surgical pathology literature for neuroendocrine tumors. In our cytopathology practice, we have encountered unexpectedly low MIB1 immunoreactivity in CytoLyt-fixed cell blocks (CBs). The current study evaluated the impact of fixatives, CB processing, and immunocytochemical (ICC) procedures on Ki-67 immunoreactivity. Methods: Test CBs were prepared from freshly resected tumors, and multiple variables in the MIB1 ICC procedure were tested, including CytoLyt versus formalin collection media, MIB1 versus other Ki-67 clones including 30-9, and other variables. MIB1 versus Ki-67 30-9 clones were tested in parallel on CytoLyt-fixed CBs from clinical samples of small cell lung carcinoma (SCLC). Results: In the test CBs (n = 10), the mean MIB1 labeling index was 10% in CytoLyt versus 47% in formalin (P = .0116), with a mean loss of reactivity in matched CBs of 37% (up to 70%). None of the procedure modifications tested in 223 individual ICC reactions recovered MIB1 reactivity in CytoLyt except for switching to the Ki-67 30-9 antibody. In CytoLyt-fixed SCLC samples (n = 14), the Ki-67 30-9 antibody demonstrated expected ranges of reactivity (mean, 83%; range, 60%-100%), whereas MIB1 demonstrated markedly inhibited labeling (mean, 60%; range, 10%-95%) (P = .0058). Conclusions: CytoLyt fixation substantially inhibits MIB1 immunoreactivity, whereas the Ki-67 30-9 clone is not susceptible to inhibition. Markedly discrepant MIB1 reactivity may present a pitfall in the diagnosis of SCLC and may lead to the incorrect prognostic stratification of other tumor types. For laboratories using CytoLyt, we recommend using the Ki-67 30-9 antibody rather than the MIB1 antibody.
MeSH term(s)	Antibodies, Antinuclear/chemistry ; Antibodies, Antinuclear/immunology ; Antibodies, Monoclonal/chemistry ; Antibodies, Monoclonal/immunology ; Diagnosis, Differential ; Formaldehyde/adverse effects ; Humans ; Immunohistochemistry ; Ki-67 Antigen/chemistry ; Ki-67 Antigen/immunology ; Ki-67 Antigen/metabolism ; Lung Neoplasms/immunology ; Lung Neoplasms/metabolism ; Lung Neoplasms/pathology ; Small Cell Lung Carcinoma/immunology ; Small Cell Lung Carcinoma/metabolism ; Small Cell Lung Carcinoma/pathology ; Tissue Fixation/methods
Chemical Substances	Antibodies, Antinuclear ; Antibodies, Monoclonal ; Ki-67 Antigen ; MIB-1 antibody ; Formaldehyde (1HG84L3525)
Language	English
Publishing date	2019-08-09
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2594979-2
ISSN	1934-6638 ; 1934-662X
ISSN (online)	1934-6638
ISSN	1934-662X
DOI	10.1002/cncy.22170
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Zs.A 6893: Show issues

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Article ; Online: Immunohistochemical Detection of γ/δ T Lymphocytes in Formalin-fixed Paraffin-embedded Tissues.

Jungbluth, Achim A / Frosina, Denise / Fayad, Miriam / Pulitzer, Melissa P / Dogan, Ahmet / Busam, Klaus J / Imai, Naoko / Gnjatic, Sacha

Applied immunohistochemistry & molecular morphology : AIMM

2018 Volume 27, Issue 8, Page(s) 581–583

Abstract: T lymphocytes can be distinguished based on the composition of the T-cell receptor (TCR) chain in α/β T cells and γ/δ T cells. Correspondingly, α/β lymphomas can be distinguished from γ/δ lymphomas. The latter are rare neoplasms, which are usually ... ...

Abstract	T lymphocytes can be distinguished based on the composition of the T-cell receptor (TCR) chain in α/β T cells and γ/δ T cells. Correspondingly, α/β lymphomas can be distinguished from γ/δ lymphomas. The latter are rare neoplasms, which are usually confined to particular organs and tissues and carry a dismal prognosis. Until recently, monoclonal antibody (mAb) clone g3.20 to the TCR γ-chain was the reagent of choice for the immunohistochemical detection of γ/δ T cells and lymphomas in standard formalin-fixed paraffin-embedded tissues. However, due to technical problems, mAb g3.20 became recently unavailable. Our attempts to identify another commercially available clone to the TCR γ-chain were unsuccessful. However, we were able to identify a mAb (clone H-41, SC-100289; Santa Cruz, Dallas, TX) to the TCR δ-chain. H-41 works well in immunohistochemistry on paraffin-embedded tissue and comparison with previously stained cases, shows superior immunolabeling to mAb g3.20. H-41 to the TCR δ-chain appears to be a suitable reagent for the replacement of mAb g3.20.
MeSH term(s)	Antibodies, Monoclonal/immunology ; Formaldehyde ; Humans ; Immunohistochemistry ; Lymphoma/diagnosis ; Lymphoma/immunology ; Paraffin Embedding ; Receptors, Antigen, T-Cell, gamma-delta/metabolism ; T-Lymphocytes/immunology
Chemical Substances	Antibodies, Monoclonal ; Receptors, Antigen, T-Cell, gamma-delta ; Formaldehyde (1HG84L3525)
Language	English
Publishing date	2018-02-28
Publishing country	United States
Document type	Journal Article
ZDB-ID	1473273-7
ISSN	1533-4058 ; 1062-3345 ; 1541-2016
ISSN (online)	1533-4058
ISSN	1062-3345 ; 1541-2016
DOI	10.1097/PAI.0000000000000650
Database	MEDical Literature Analysis and Retrieval System OnLINE

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