Subject(s)
Bone Marrow/pathology , Epstein-Barr Virus Infections/etiology , Lymph Nodes/pathology , Lymphoproliferative Disorders/etiology , Primary Myelofibrosis/drug therapy , Pyrazoles/adverse effects , Reed-Sternberg Cells/pathology , Antigens, CD/analysis , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Bone Marrow/virology , DNA, Viral/blood , Deoxycytidine/administration & dosage , Deoxycytidine/analogs & derivatives , Doxorubicin/administration & dosage , Epstein-Barr Virus Infections/drug therapy , Epstein-Barr Virus Infections/pathology , Humans , Immunocompromised Host , Lymph Nodes/virology , Lymphoproliferative Disorders/drug therapy , Lymphoproliferative Disorders/pathology , Male , Middle Aged , Nitriles , PAX5 Transcription Factor/analysis , Prednisone/administration & dosage , Primary Myelofibrosis/etiology , Pyrimidines , RNA, Viral/analysis , Reed-Sternberg Cells/chemistry , Reed-Sternberg Cells/virology , Thrombocythemia, Essential/complications , Thrombocythemia, Essential/genetics , Vinblastine/administration & dosage , GemcitabineABSTRACT
PAX5-KIDINS220 (PAX5-K220) is a novel chimeric fusion gene identified in a pediatric Philadelphia chromosome (Ph)-like acute lymphoblastic leukemia (ALL) patient, but the function of the encoded fusion protein has not yet been analyzed. Here, we report the functional analysis of PAX5-K220 in vitro. We successfully generated PAX5-K220 expressing cells and demonstrate that PAX5-K220 is a nuclear protein. Luciferase reporter assay reveals that PAX5-K220 inhibits wild-type PAX5 transcriptional activity in a dominant-negative fashion like other PAX5-related fusion proteins, and may contribute to lymphocyte differentiation block. However, although identified in Ph-like ALL, PAX5-K220 does not induce IL-3-independent proliferation when transduced in the IL-3-dependent Ba/F3 murine leukemia cells, but rather attenuates growth. These results reveal that PAX5-K220 certainly shares the character with other PAX5-related fusion proteins rather than other fusion proteins with tyrosine kinase activity identified in Ph-like ALL, and did not contribute to proliferation activity. Precise functional analysis of each differently partnered PAX5 fusion protein is warranted in the future for better understanding of PAX5-related translocations and their effects.
Subject(s)
Gene Fusion , Membrane Proteins/analysis , Membrane Proteins/genetics , Nerve Tissue Proteins/analysis , Nerve Tissue Proteins/genetics , PAX5 Transcription Factor/analysis , PAX5 Transcription Factor/genetics , Philadelphia Chromosome , Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics , Precursor Cell Lymphoblastic Leukemia-Lymphoma/pathology , Animals , B-Lymphocytes/pathology , Cell Differentiation/genetics , Cells, Cultured , Child , HEK293 Cells , Humans , Interleukin-3 , Mice , Protein-Tyrosine Kinases/metabolism , Translocation, Genetic/geneticsABSTRACT
As checkpoint molecules' inhibition may represent a therapeutic option in relapsing cases, we assessed programmed death ligands' (PD-L1/PD-L2) expression in a series of 29 primary cutaneous diffuse large B-cell lymphoma, leg-type (PCDLBCL-LT) cases. Double immunostaining for either PD-L1 or PD-L2 was associated either with PAX5 staining to evaluate tumor cells or with CD68 or CD163 staining for macrophages. The microenvironment of PCDLBCL-LT was characterized by immunostainings for CD3 (tumor-infiltrating lymphocytes), FOXP3 (regulatory T cells), programmed cell death-1, and CD33 (myeloid-derived suppressor cells). The 9p24.1 locus encoding for PD-L1/PD-L2 was evaluated by fluorescence in situ hybridization. A PD-L1 expression was observed in all cases. However, double staining with PD-L1/PAX5 identified only 1 case harboring PD-L1 expression by tumor cells. All cases displayed PD-L1 expression by numerous immune cells, characterized as CD68 CD163 M2 macrophages. A normal fluorescence in situ hybridization pattern was observed in 21 of 26 cases. Three cases (11.5%) harbored a low polysomy status including the case with PD-L1 expression by tumor cells. Interestingly, 2 cases (7.7%) exhibited a PD-L1/PD-L2 locus break-apart pattern, and PD-L2 expression by tumor cells was observed. PD-L2 expression by tumor cells was not observed in the 24 cases without 9p24.1 rearrangement. Treating patients with relapsing PCDLBCL-LT by using immune checkpoint inhibitors may have an indirect effect through immune cells, except in rare cases with 9p24.1 rearrangement leading to PD-L2 expression by tumor cells. Reprogramming tumor-associated macrophages with anticancer therapies is appealing in such lymphoma subtypes wherein M2 macrophages represent the majority of immune cells.
Subject(s)
B7-H1 Antigen/analysis , Biomarkers, Tumor/analysis , Lymphoma, Large B-Cell, Diffuse/immunology , Macrophages/immunology , Programmed Cell Death 1 Ligand 2 Protein/analysis , Skin Neoplasms/immunology , Aged , Aged, 80 and over , Antigens, CD/analysis , Antigens, Differentiation, Myelomonocytic/analysis , B7-H1 Antigen/genetics , Biomarkers, Tumor/genetics , Female , Gene Rearrangement , Genetic Loci , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Leg , Lymphocytes, Tumor-Infiltrating/immunology , Lymphocytes, Tumor-Infiltrating/pathology , Lymphoma, Large B-Cell, Diffuse/genetics , Lymphoma, Large B-Cell, Diffuse/pathology , Macrophages/pathology , Male , Middle Aged , PAX5 Transcription Factor/analysis , Programmed Cell Death 1 Ligand 2 Protein/genetics , Receptors, Cell Surface/analysis , Skin Neoplasms/genetics , Skin Neoplasms/pathology , Tumor MicroenvironmentABSTRACT
CONTEXT: - In hematopathology practice, abnormal expression of CD103 on B cells is detected by flow cytometry in hairy cell leukemia (HCL) and, in combination with other phenotypic and morphologic findings, provides diagnostic specificity and sensitivity. Immunostaining on paraffin sections makes it possible to perform immunophenotyping in situ. However, normal bone marrow contains CD103-positive cells, which are not B cells, making it difficult to be certain about low-level involvement by HCL. OBJECTIVE: - To develop dual immunostaining for confirmation that CD103 is expressed in B cells, which may be highly desirable in assessing low-level involvement by HCL. DESIGN: - We developed a dual immunostaining approach using a B-cell marker, Pax5, expressed in the nucleus, in combination with a membrane marker, CD103. RESULTS: - We analyzed 6 HCLs, 7 marginal zone lymphomas, 12 lymphoplasmacytic lymphomas, 7 follicular lymphomas, 5 chronic lymphocytic leukemias, 5 mantle cell lymphomas, 1 multiple myeloma (lymphocytic variant), and 3 bone marrows not involved by any B-cell neoplasm. Our dual staining approach confirmed that only the neoplastic cells of HCL were positive for both CD103 and Pax5. CONCLUSIONS: - This dual-staining method is particularly helpful in cases with low-level involvement by HCL and can be used for determining minimal residual disease after treatment.
Subject(s)
Antigens, CD/analysis , Biomarkers, Tumor/analysis , Integrin alpha Chains/analysis , Leukemia, Hairy Cell/diagnosis , PAX5 Transcription Factor/analysis , Bone Marrow/metabolism , Bone Marrow/pathology , Humans , Immunohistochemistry/methods , Immunophenotyping/methods , Leukemia, Hairy Cell/metabolism , Leukemia, Hairy Cell/pathology , Leukemia, Lymphocytic, Chronic, B-Cell/diagnosis , Leukemia, Lymphocytic, Chronic, B-Cell/metabolism , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Lymphoma, Non-Hodgkin/diagnosis , Lymphoma, Non-Hodgkin/metabolism , Lymphoma, Non-Hodgkin/pathology , Multiple Myeloma/diagnosis , Multiple Myeloma/metabolism , Multiple Myeloma/pathology , Neoplasm, Residual , Paraffin Embedding , Sensitivity and Specificity , Staining and Labeling/methods , Waldenstrom Macroglobulinemia/diagnosis , Waldenstrom Macroglobulinemia/metabolism , Waldenstrom Macroglobulinemia/pathologyABSTRACT
Anaplastic lymphoma kinase (ALK)-negative anaplastic large cell lymphoma (ALCL) is a subtype of T-cell lymphomas that may mimic several other malignancies morphologically. With the help of immunohistochemistry, most cases of ALCL can be diagnosed on the basis of expression of T-cell lineage associated antigens. However, aberrations in the expression of immunohistochemical markers pose diagnostic challenges. The morphological and immunophenotypic features of ALCL show considerable overlap with classical Hodgkin lymphoma (CHL), which is a B-cell lymphoma. The 2008 World Health Organization Classification of Tumours of Haematopoietic and Lymphoid Tissues suggests that staining for the B-cell transcription factor, paired box 5 (PAX-5), is helpful in differentiating between them, as it is weakly positive in most CHL and should be negative in ALCL. We report a rare case of ALK-negative ALCL, which was positive for PAX-5 and CD15, mimicking CHL by immunohistochemistry, resulting in a diagnostic dilemma.
Subject(s)
Biomarkers, Tumor/analysis , Lymphoma, Large-Cell, Anaplastic/diagnosis , Lymphoma, Large-Cell, Anaplastic/pathology , PAX5 Transcription Factor/biosynthesis , Aged , Anaplastic Lymphoma Kinase , Diagnosis, Differential , Female , Hodgkin Disease/diagnosis , Hodgkin Disease/pathology , Humans , PAX5 Transcription Factor/analysis , Receptor Protein-Tyrosine KinasesABSTRACT
The aim of our study was to evaluate the immunohistochemical expression of SOX11, PAX5, TTF-1 and ISL-1 in medulloblastoma (MB) to investigate their diagnostic usefulness. METHODS: Immunohistochemical expression of PAX5 (two antibodies: Dako, DAK-Pax5; and BD, clone 24), TTF-1 (Dako, 8G7G3/1), SOX11 (CL0142; Abcam) and ISL-1 (1 H9, Abcam) was analyzed using the h-score and Remmele score in 25 cases of MB. RESULTS: There were 18 MBs of classic and 7 of desmoplastic type. SOX11 was strongly expressed in all tumors. The expression of PAX5 was higher and more frequent in a case of DAK-Pax5 clone (25/25) than clone 24 (6/25). ISL-1 was positive in 11 (44%) and TTF-1 in 3 (12%) cases. ISL-1 expression correlated positively (p<0.001), while TTF-1 correlated negatively with the age of patients (p=0.039). PAX5 expression correlated with ISL-1 (p=0.039) and showed a trend toward higher expression in the desmoplastic subtype (p=0.069). CONCLUSIONS: SOX11 is strongly and robustly expressed in MBs. PAX5 expression pattern differs substantially among two antibody clones. TTF-1 and ISL-1 is associated with the age of patients.
Subject(s)
Cerebellar Neoplasms/diagnosis , DNA-Binding Proteins/biosynthesis , LIM-Homeodomain Proteins/biosynthesis , Medulloblastoma/diagnosis , PAX5 Transcription Factor/biosynthesis , SOXC Transcription Factors/biosynthesis , Transcription Factors/biosynthesis , Adolescent , Adult , Biomarkers, Tumor/analysis , Cerebellar Neoplasms/metabolism , Child , Child, Preschool , DNA-Binding Proteins/analysis , Female , Humans , Immunohistochemistry , Infant , LIM-Homeodomain Proteins/analysis , Male , Medulloblastoma/metabolism , PAX5 Transcription Factor/analysis , SOXC Transcription Factors/analysis , Transcription Factors/analysis , Young AdultABSTRACT
Olfactory neuroblastoma (ONB) is a rare neoplasm of the sinonasal area with neuroendocrine differentiation. ISL-1, TTF-1 and PAX5 are transcription factors that are frequently upregulated in tumors showing neuroendocrine differentiation. The aim of our study was to evaluate these markers in a group of ONBs. We included 11 ONBs from 4 large university hospitals. Immunohistochemical expression of TTF-1, PAX5 and ISL-1 was evaluated. TTF-1, ISL-1 and PAX5 were expressed in 3/11 cases (27.27%, h-score: 3-45), 7/11 cases (63.64%, h-score: 23-200), and in 3/11 cases (27.77%, h-score 3-85), respectively. The patient with the strongest PAX5 reactivity exhibited an aggressive clinical course with rapid dissemination to the spine and death shortly after the diagnosis. No significant correlation in the expression of PAX5 and TTF-1 (ï² = 0.43; p = 0.18) was observed. ISL-1 is widely expressed in tumors with neuroendocrine differentiation and therefore of limited value in their differential diagnosis. TTF-1 positivity does not exclude the diagnosis of primary ONB, although usually only a small percentage of cells are positive. PAX5 expression is infrequent (27.27%) in ONB; however, if present it can be associated with a very aggressive clinical course.
Subject(s)
DNA-Binding Proteins/biosynthesis , Esthesioneuroblastoma, Olfactory/metabolism , LIM-Homeodomain Proteins/biosynthesis , Nose Neoplasms/metabolism , PAX5 Transcription Factor/biosynthesis , Transcription Factors/biosynthesis , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/analysis , DNA-Binding Proteins/analysis , Esthesioneuroblastoma, Olfactory/pathology , Female , Humans , Immunohistochemistry , LIM-Homeodomain Proteins/analysis , Male , Middle Aged , Nasal Cavity/pathology , Nose Neoplasms/pathology , PAX5 Transcription Factor/analysis , Transcription Factors/analysis , Young AdultABSTRACT
Thyroid transcription factor-1 (TTF-1) is a marker of tumors of pulmonary and thyroid origin, and its expression practically excludes diagnosis of Merkel cell carcinoma (MCC). However, TTF-1 expression in combined MCC was recently reported. PAX5 is a marker of B-cell origin that is also expressed in most classical MCC cases; however, its expression was not described in combined MCC. The authors decided to evaluate the expression of both these markers in a group of 5 combined MCCs (2 with invasive squamous cell carcinoma, 2 with squamous cell carcinoma in situ, and 1 with basal cell carcinoma). Expression of TTF-1 was found in 4 of 5 cases; in 3 cases, the marker was shown in the MCC component (weakly in 2 cases and strongly in 1 case), whereas the non-MCC component presented TTF-1 expression in 2 cases. A weak-to-moderate immunoreactivity for PAX5 was identified in all cases of the MCC component but in none of the non-MCC component. The results show that the expression of TTF-1 is a frequent finding in combined MCC and can be present in the neuroendocrine component, which differs from the conventional MCC. In contrast, PAX5 expression pattern is similar to that of the classical MCC.
Subject(s)
Biomarkers, Tumor/analysis , Carcinoma in Situ/chemistry , Carcinoma, Basal Cell/chemistry , Carcinoma, Merkel Cell/chemistry , Carcinoma, Squamous Cell/chemistry , Neoplasms, Complex and Mixed/chemistry , Nuclear Proteins/analysis , PAX5 Transcription Factor/analysis , Skin Neoplasms/chemistry , Transcription Factors/analysis , Aged , Aged, 80 and over , Carcinoma in Situ/pathology , Carcinoma, Basal Cell/pathology , Carcinoma, Merkel Cell/pathology , Carcinoma, Squamous Cell/pathology , Cell Differentiation , Europe , Female , Humans , Immunohistochemistry , Male , Neoplasms, Complex and Mixed/pathology , Predictive Value of Tests , Skin Neoplasms/pathology , Thyroid Nuclear Factor 1ABSTRACT
Mature B cell neoplasms cover a spectrum of diseases involving lymphoid tissues (lymphoma) or blood (leukemia), with an overlap between these two presentations. Previous studies describing equine lymphoid neoplasias have not included analyses of clonality using molecular techniques. The objective of this study was to use molecular techniques to advance the classification of B cell lymphoproliferative diseases in five adult equine patients with a rare condition of monoclonal gammopathy, B cell leukemia, and concurrent lymphadenopathy (lymphoma/leukemia). The B cell neoplasms were phenotypically characterized by gene and cell surface molecule expression, secreted immunoglobulin (Ig) isotype concentrations, Ig heavy-chain variable (IGHV) region domain sequencing, and spectratyping. All five patients had hyperglobulinemia due to IgG1 or IgG4/7 monoclonal gammopathy. Peripheral blood leukocyte immunophenotyping revealed high proportions of IgG1- or IgG4/7-positive cells and relative T cell lymphopenia. Most leukemic cells lacked the surface B cell markers CD19 and CD21. IGHG1 or IGHG4/7 gene expression was consistent with surface protein expression, and secreted isotype and Ig spectratyping revealed one dominant monoclonal peak. The mRNA expression of the B cell-associated developmental genes EBF1, PAX5, and CD19 was high compared to that of the plasma cell-associated marker CD38. Sequence analysis of the IGHV domain of leukemic cells revealed mutated Igs. In conclusion, the protein and molecular techniques used in this study identified neoplastic cells compatible with a developmental transition between B cell and plasma cell stages, and they can be used for the classification of equine B cell lymphoproliferative disease.
Subject(s)
B-Lymphocytes , Horse Diseases/genetics , Leukemia, B-Cell/veterinary , Lymphatic Diseases/veterinary , Lymphopenia/veterinary , Lymphoproliferative Disorders/veterinary , Paraproteinemias/veterinary , Animals , Antigens, CD19/analysis , B-Lymphocytes/immunology , B-Lymphocytes/metabolism , B-Lymphocytes/pathology , Horses , Immunoglobulin Heavy Chains/genetics , Immunophenotyping , Leukemia, B-Cell/genetics , Leukemia, B-Cell/immunology , Lymphatic Diseases/genetics , Lymphatic Diseases/immunology , Lymphopenia/genetics , Lymphopenia/immunology , Lymphoproliferative Disorders/classification , Lymphoproliferative Disorders/genetics , Lymphoproliferative Disorders/immunology , PAX5 Transcription Factor/analysis , Paraproteinemias/genetics , Paraproteinemias/immunology , Plasma Cells , Receptors, Complement 3d/analysis , T-LymphocytesABSTRACT
We describe herein the case of a 64-year-old man with a diagnosis of plasma cell myeloma (PCM). A chromosome analysis based on G-banding and spectral karyotyping revealed the following complex karyotype: 46,XY,del(3)(p?), t(4;15)(q31;q24),t(9;14;11)(p13;q32;q13),add(15)(q24),add(18)(q21). Fluorescence in situ hybridization (FISH) detected one signal each for the immunoglobulin heavy chain (IGH) and cyclin D1 (CCND1) genes, and three fusion signals of IGH and CCND1. FISH analysis of metaphase spreads revealed fusion signals on the derivative chromosomes 9, 11, and 14. Immunohistochemical analysis identified abnormal expression of CCND1 and PAX5. PAX5-positive PCM is rare because the down-regulation of PAX5 is essential for the terminal differentiation of B cells into plasma cells. To the best of our knowledge, this is the first reported case of a novel complex variant translocation of t(11;14)(q13;q32) and t(9;14)(p13;q32).
Subject(s)
Multiple Myeloma/genetics , Multiple Myeloma/pathology , PAX5 Transcription Factor/analysis , Plasma Cells/pathology , Translocation, Genetic , Chromosome Banding , Chromosomes, Human, Pair 11/genetics , Chromosomes, Human, Pair 14/genetics , Chromosomes, Human, Pair 9/genetics , Cyclin D1/genetics , Gene Expression Regulation, Neoplastic , Humans , Immunoglobulin Heavy Chains/genetics , In Situ Hybridization, Fluorescence , Karyotyping , Male , Middle Aged , PAX5 Transcription Factor/genetics , Plasma Cells/metabolismABSTRACT
Lung cancer is the leading cause of cancer-related death in both men and women and consists of different histological types. Histopathological examination and accurate subtype diagnosis has become increasingly important in guiding patient management and, as such, is the most important currently available lung cancer "biomarker". In this study, we examined the expression of PAX2 and PAX5 by immunohistochemistry in 47 cases of lung cancer and 13 cases of pneumonia. The results demonstrated that PAX2 were detected in 82.8% (24/29) of NSCLC, 0% (0/18) of SCLC and 7.7% (1/13) of pneumonia, respectively; However, PAX5 were detected in 15/18 cases (83.3%) of SCLC, 6.8% (2/29) of NSCLC and 7.7% (1/13) of pneumonia. Further, the samples with lymphatic metastasis had remarkable higher positive PAX2 or PAX5 than that without metastases. Overall, our data indicated that PAX2 and PAX5 differentially expressed in NSCLC and SCLC. Thus, PAX2 and PAX5 are useful biomarker in the differential diagnosis of lung cancer.
Subject(s)
Biomarkers, Tumor/analysis , Carcinoma, Non-Small-Cell Lung/diagnosis , Lung Neoplasms/diagnosis , PAX2 Transcription Factor/biosynthesis , PAX5 Transcription Factor/biosynthesis , Small Cell Lung Carcinoma/diagnosis , Adult , Aged , Diagnosis, Differential , Female , Humans , Immunohistochemistry , Male , Middle Aged , PAX2 Transcription Factor/analysis , PAX5 Transcription Factor/analysisABSTRACT
AIMS: The purpose of this study was to compare the immunohistochemical staining profiles of PAX8-polyclonal, PAX8-monoclonal, PAX5-monoclonal and PAX6-monoclonal antibodies in several histological types of primary thoracic and thyroid tumours. In addition, we analysed PAX8 mRNA expression by using in-situ hybridization. METHODS AND RESULTS: We compared polyclonal PAX8 and monoclonal PAX8, PAX5 and PAX6 antibodies in 962 samples (687 lung carcinomas, 40 malignant pleural mesotheliomas, 138 thymic tumours and 97 thyroid tumours) using the tissue microarray technique. Among thyroid tumours, the monoclonal and polyclonal PAX8 antibodies showed a high positive rate (98.0%). Of 167 polyclonal PAX8 antibody-positive tumours, except for thyroid tumours, 54 cases tested positive for PAX5 and/or PAX6 (31 lung carcinomas and 23 thymic tumours). No PAX8 mRNA expression was detected using RNAscope (in-situ hybridization technique) other than in thyroid tumours. A portion of polyclonal PAX8 antibody-positive tumours showed cross-reactivity for PAX5 or PAX6 protein. CONCLUSIONS: Monoclonal PAX8 antibody showed high specificity to thyroid tumours and was superior to the polyclonal antibody.
Subject(s)
Antibodies, Monoclonal/immunology , Paired Box Transcription Factors/analysis , Paired Box Transcription Factors/immunology , Thoracic Neoplasms/diagnosis , Thyroid Neoplasms/diagnosis , Animals , Biomarkers/analysis , Eye Proteins/analysis , Eye Proteins/immunology , Homeodomain Proteins/analysis , Homeodomain Proteins/immunology , Humans , In Situ Hybridization , Lung Neoplasms/diagnosis , Mice , PAX5 Transcription Factor/analysis , PAX5 Transcription Factor/immunology , PAX6 Transcription Factor , PAX8 Transcription Factor , Pleural Neoplasms/diagnosis , Repressor Proteins/analysis , Repressor Proteins/immunology , Thymus Neoplasms/diagnosis , Tissue Array AnalysisSubject(s)
Carcinoma, Merkel Cell/diagnosis , Leukemia, Lymphocytic, Chronic, B-Cell/diagnosis , Aged , Carcinoma, Merkel Cell/pathology , Cell Transformation, Neoplastic , Female , Flow Cytometry , Humans , Leukemia, Lymphocytic, Chronic, B-Cell/complications , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Leukocytosis/pathology , Lymphatic Diseases/pathology , Neoplasm Metastasis , PAX5 Transcription Factor/analysisABSTRACT
AIMS: The purpose of this study was to evaluate the expression patterns of B-cell specific activator protein (BSAP)/PAX5 and PAX8 in a wide variety of B-cell and T-cell neoplasms. METHODS AND RESULTS: A wide range of B-cell and T-cell neoplasms were subjected to immunohistochemical staining with antibodies against BSAP/PAX5 and PAX8 (polyclonal, pPAX8; monoclonal, mPAX8). Ten non-neoplastic lymph node specimens were examined with the same panel. All of the tested neoplastic and non-neoplastic B-cells reacted with the BSAP/PAX5 and pPAX8 antibodies, but did not show reactivity with the mPAX8 antibody. All tested T-cell neoplasms were negative using the BSAP/PAX5, pPAX8 and mPAX8 antibodies. CONCLUSIONS: This is the first study to show the absence of reactivity to an mPAX8 antibody in an expanded panel of B-cell lymphomas as well as in a variety of T-cell neoplasms. In contrast to the mPAX8 antibody, the pPAX8 antibody shows nuclear positivity in non-neoplastic B cells and mature B-cell neoplasms; however, this expression is probably a result of cross-reactivity with PAX5. Given that many laboratories use the pPAX8 antibody, a clear understanding of the differential staining patterns is necessary. The differential diagnosis of a B-cell lymphoma should be entertained when a pPAX8-positive, epithelial marker-negative neoplasm of uncertain primary origin is encountered.
Subject(s)
Biomarkers, Tumor/analysis , Lymphoma, B-Cell/metabolism , Lymphoma, T-Cell/metabolism , PAX5 Transcription Factor/biosynthesis , Paired Box Transcription Factors/biosynthesis , Humans , Immunohistochemistry , Lymphoma, B-Cell/diagnosis , Lymphoma, T-Cell/diagnosis , PAX5 Transcription Factor/analysis , PAX8 Transcription Factor , Paired Box Transcription Factors/analysisABSTRACT
Recently, reports using immunohistochemistry and a polyclonal antibody directed against the N-terminal region of PAX8 describe PAX8 expression in malignant lymphomas. As the N-terminal regions of PAX family members, including the B-cell transcription factor PAX5, have high sequence homology, we investigated PAX8 positivity in malignant lymphomas. Comparative sequence analysis between the N- and C-terminal regions of PAX8 and PAX5 proteins confirmed homologies of 70% and 39%, respectively. We then compared the results using N-terminal (high homology) and C-terminal (lower homology) anti-PAX8 antibodies to assess PAX8 expression in reactive tissues, diffuse large B-cell lymphoma and classical Hodgkin lymphoma, using routine immunohistochemical methods. Expression of PAX8 was also assessed in diffuse large B-cell lymphoma and classical Hodgkin lymphoma cell lines using real-time qRT-PCR methods. Our results show that reactive and neoplastic B-cells are positive for PAX8 using the N-terminal antibody, but negative for PAX8 when the C-terminal antibody was used. PAX8 mRNA levels were not detected in any of the B-cell lymphoma cell lines studied. These results indicate that benign and malignant B-cells do not express PAX8. We conclude that positivity for PAX8 reported by others in B-cell lymphomas is likely due to cross-reactivity between the N-terminal regions of PAX8 and PAX5, due to the high sequence homology of these two regions.
Subject(s)
Biomarkers, Tumor/analysis , Lymphoma, B-Cell/metabolism , PAX5 Transcription Factor/analysis , Paired Box Transcription Factors/analysis , Amino Acid Sequence , Antibody Specificity , Cross Reactions , Humans , Immunohistochemistry , Molecular Sequence Data , PAX5 Transcription Factor/chemistry , PAX5 Transcription Factor/metabolism , PAX8 Transcription Factor , Paired Box Transcription Factors/chemistry , Paired Box Transcription Factors/metabolism , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino AcidSubject(s)
Antigens, CD20/analysis , Ki-1 Antigen/analysis , Lewis X Antigen/analysis , Lymphoma, T-Cell, Peripheral/immunology , Aged , Humans , Lymphoma, T-Cell, Peripheral/diagnosis , Lymphoma, T-Cell, Peripheral/drug therapy , Lymphoma, T-Cell, Peripheral/pathology , Male , PAX5 Transcription Factor/analysisABSTRACT
Histiocytic and interdigitating dendritic cell sarcomas are rare tumors originating from bone marrow-derived myeloid stem cells. Recent studies have shown evidence of cross-lineage transdifferentiation of B cells in follicular lymphoma to histiocytic and dendritic cell sarcomas. In this study, we report the morphologic, molecular and cytogenetic analysis of seven cases of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) associated with histiocytic and dendritic cell sarcomas. All seven patients were elderly males (median age 71 years). The B-cell neoplasms preceded the development of the histiocytic and dendritic cell sarcomas in six of seven patients, and one patient had both tumors diagnosed at the same time. The tumors included four interdigitating dendritic cell sarcomas: one Langerhans cell sarcoma, one histiocytic sarcoma and one immature neoplasm with evidence of histiocytic origin. Laser-capture microdissection and PCR analysis showed identical clonal immunoglobulin gene rearrangements in the two phenotypically distinct components in all cases. There was a preferential usage of IGHV4-39 by the V-D-J gene rearrangement. By fluorescence in situ hybridization (FISH) analysis, two cases showed deletion 17p in both components, whereas four cases had normal cytogenetic findings by FISH in the CLL/SLL cells, but acquired cytogenetic abnormalities in the corresponding histiocytic and dendritic tumors. Chromosome 17p abnormalities were the most common cytogenetic abnormality detected in the sarcomas, seen in five of six cases studied. Compared with the CLL/SLL cells, the histiocytic/dendritic cells were largely negative for PAX5, but showed strong expression of PU.1 and variable and weak expression of CEBPß. Our study provides evidence for transdifferentiation of CLL/SLL B cells to tumors of dendritic and less often histiocytic lineage, and suggests that secondary genetic events may play a role in this phenomenon.
Subject(s)
Cell Transdifferentiation , Dendritic Cell Sarcoma, Interdigitating , Histiocytic Sarcoma , Langerhans Cell Sarcoma , Leukemia, Lymphocytic, Chronic, B-Cell , Aged , Aged, 80 and over , Base Sequence , Biomarkers, Tumor/analysis , CCAAT-Enhancer-Binding Protein-beta/analysis , Chromosome Aberrations , Chromosomes, Human, Pair 17 , Dendritic Cell Sarcoma, Interdigitating/genetics , Dendritic Cell Sarcoma, Interdigitating/immunology , Dendritic Cell Sarcoma, Interdigitating/pathology , Gene Rearrangement, B-Lymphocyte , Histiocytic Sarcoma/genetics , Histiocytic Sarcoma/immunology , Histiocytic Sarcoma/pathology , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Langerhans Cell Sarcoma/genetics , Langerhans Cell Sarcoma/immunology , Langerhans Cell Sarcoma/pathology , Laser Capture Microdissection , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/immunology , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Male , Maryland , Middle Aged , Molecular Sequence Data , PAX5 Transcription Factor/analysis , Polymerase Chain Reaction , Proto-Oncogene Proteins/analysis , Trans-Activators/analysis , V(D)J RecombinationABSTRACT
This study aimed to identify the frequency and distribution of developing B cell populations in the kidney of the rainbow trout, using four molecular B cell markers that are highly conserved between species, including two transcription factors, Pax5 and EBF1, recombination-activating gene RAG1, and the immunoglobulin heavy chain mu. Three distinct B cell stages were defined: early developing B cells (CLP, pro-B, and early pre-B cells), late developing B cell (late pre-B, immature B, and mature B cells), and IgM-secreting cells. Developmental stage-specific, combinatorial expression of Pax5, EBF1, RAG1 and immunoglobulin mu was determined in trout anterior kidney cells by flow cytometry. Trout staining patterns were compared to a well-defined primary immune tissue, mouse bone marrow, and using mouse surface markers B220 and CD43. A remarkable level of similarity was uncovered between the primary immune tissues of both species. Subsequent analysis of the entire trout kidney, divided into five contiguous segments K1-K5, revealed a complex pattern of early developing, late developing, and IgM-secreting B cells. Patterns in anterior kidney segment K1 were most similar to those of mouse bone marrow, while the most posterior part of the kidney, K5, had many IgM-secreting cells, but lacked early developing B cells. A potential second B lymphopoiesis site was uncovered in segment K4 of the kidney. The B cell patterns, or "B cell signatures" described here provide information on the relative abundance of distinct developing B cell populations in the trout kidney, and can be used in future studies on B cell development in other vertebrate species.
Subject(s)
B-Lymphocytes/immunology , Bone Marrow/immunology , Kidney/immunology , Lymphopoiesis , Trout/immunology , Animals , B-Lymphocytes/metabolism , Biomarkers/analysis , Blotting, Western , Cell Differentiation , Flow Cytometry , Gene Expression , Homeodomain Proteins/analysis , Immunoglobulin Heavy Chains/analysis , Immunoglobulin mu-Chains/analysis , Leukocyte Common Antigens/analysis , Leukosialin/analysis , Mice , PAX5 Transcription Factor/analysis , Precursor Cells, B-Lymphoid , Trans-Activators/analysis , Transcription Factors/genetics , Trout/geneticsABSTRACT
OBJECTIVE: Undifferentiated tumors and hematolymphoid neoplasms can be diagnostically challenging due to potential overlap of morphologic features and variant antigen expression. PAX-5, a transcription factor expressed throughout B-cell maturation, is detected in most B-cell neoplasms including those that lack expression of mature B-cell markers, such as classical Hodgkin lymphoma (cHL), B-lymphoblastic leukemia and B-cell lymphomas following rituximab therapy. The lack of PAX-5 expression in most CD30-positive non-hematopoietic malignancies (embryonal carcinoma and seminoma) and T-cell lymphomas, such as anaplastic large cell lymphoma (ALCL), suggests that the absence of PAX-5 may be used to confirm non-B-cell lineage. The goal of this study was to retrospectively assess PAX-5 immunoreactivity in diagnostic samples of hematolymphoid and other non-hematopoietic malignancies. DESIGN: Diagnostic lymph node, decalcified core bone marrow biopsies and tissue sections from 111 archived paraffin-embedded tissue blocks and a tissue lymphoma microarray were immunostained using a monoclonal antibody to PAX-5. The corresponding hematoxylin and eosin stained tissue sections and additional immunostains were simultaneously evaluated. PAX-5 immunoreactivity in neoplastic cells was scored as positive or negative. This study was exempted by the Institutional Review Board for Human Research. RESULTS: Nuclear PAX-5 immunoreactivity was detected in 88% (36/41) of Hodgkin lymphoma, all cases of diffuse large B-cell lymphoma (n=72), small B-cell lymphomas (n=5), B-lymphoblastic leukemia/lymphoma and mixed phenotype acute leukemia with B-cell lineage (n=5). PAX-5 was not detected in ALCL (n=22), T-cell lymphoblastic leukemia/lymphoma, mixed phenotype acute leukemia with T-cell lineage (n=5), acute myeloid leukemia (n=4), carcinoid tumors with typical morphology (n=5), melanoma (n=3), and undifferentiated/metastatic tumors (n=8). Non-neoplastic bone marrow sections showed scattered nuclear staining in small B-cell lymphocytes/hematogones. The detection of PAX-5 immunoreactivity resulted in the reclassification of two cases of ALCL to cHL. CONCLUSION: Overall, our results demonstrate that including PAX-5 in a panel with other immunomarkers helps establish B-cell lineage and increases diagnostic yield.
Subject(s)
Biomarkers, Tumor/analysis , Lymphoma/diagnosis , PAX5 Transcription Factor/analysis , Diagnosis, Differential , Hodgkin Disease/diagnosis , Humans , Immunohistochemistry , Lymphoma, Large B-Cell, Diffuse/diagnosis , Lymphoma, Large-Cell, Anaplastic/diagnosisABSTRACT
BACKGROUND: The hierarchical organization of hematopoiesis with unidirectional lineage determination has become a questionable tenet in view of the experimental evidence of reprogramming and transdifferentiation of lineage-determined cells. Clinical examples of hematopoietic lineage plasticity are rare. Here we report on a patient who presented with an acute B-lymphoblastic leukemia and developed a Langerhans' cell sarcoma 9 years later. We provide evidence that the second neoplasm is the result of transdifferentiation. DESIGN AND METHODS: B-cell acute lymphoblastic leukemia was diagnosed in an 11-year old boy in 1996. Treatment according to the ALL-BFM-1995 protocol resulted in a complete remission. Nine years later, in 2005, Langerhans' cell sarcoma was diagnosed in a supraclavicular lymph node. Despite treatment with different chemotherapy protocols the patient had progressive disease. Finally, he received an allogeneic peripheral blood stem cell transplant and achieved a continuous remission. Molecular studies of IGH- and TCRG-gene rearrangements were performed with DNA from the Langerhans' cell sarcoma and the cryopreserved cells from the acute B-lymphoblastic leukemia. The expression of PAX5 and ID2 was analyzed with real-time reverse transcriptase polymerase chain reaction. RESULTS: Identical IGH-rearrangements were demonstrated in the acute B-lymphoblastic leukemia and the Langerhans' cell sarcoma. The key factors required for B-cell and dendritic cell development, PAX5 and ID2, were differentially expressed, with a strong PAX5 signal in the acute B-lymphoblastic leukemia and only a weak expression in the Langerhans' cell sarcoma, whereas ID2 showed an opposite pattern. CONCLUSIONS: The identical IGH-rearrangement in both neoplasms indicates transdifferentiation of the acute B-lymphoblastic leukemia into a Langerhans' cell sarcoma. Loss of PAX5 and the acquisition of ID2 suggest that these key factors are involved in the transdifferentiation from a B-cell phenotype into a Langerhans'/dendritic cell phenotype. (Clinical trial registration at: Deutsches KrebsStudienRegister, http://www.studien.de, study-ID:8).