ABSTRACT
Background: Lymphoid malignancies are a heterogeneous group of disorders which may be difficult to differentiate from reactive proliferations even after immunohistochemistry. Polymerase chain reaction (PCR) is believed to be a good adjunct tool for diagnosis. Materials and Methods: We examined 24 cases of neoplastic and non-neoplastic lymphoproliferative lesions in this study and evaluated the PCR as an additional tool in the confirmation of the diagnosis. Two different PCR methodologies were evaluated. Results: In the evaluation of the T-cell PCR, it was seen that the correlation using both the commercial kits and the custom-synthesized primers was highly significant at a P value of <0.05. In the evaluation of the B-cell PCR, it was seen that the correlation using both the commercial kits and the custom-synthesized primers was not significant using either method (P > 0.05). Conclusions: Both the methods showed an excellent concordance for T-cell γ gene rearrangements, However, the same was not seen in the B-cell receptor rearrangements. This may be because of the small sample size or the inability of consensus V primers to recognize complementary DNA sequences in all of the V segments.
Subject(s)
Clone Cells , DNA Primers/genetics , Humans , Lymphoproliferative Disorders/diagnosis , Lymphoproliferative Disorders/genetics , Lymphoproliferative Disorders/pathology , Pathology, Molecular/methods , Polymerase Chain Reaction/methods , Reagent Kits, Diagnostic , T-Lymphocytes/cytologyABSTRACT
La papulosis linfomatoide (PL) es considerada en la actualidad una forma indolente de linfoma cutáneo CD30+. Su presentación es más frecuente entre la 4º y 5º décadas de la vida, con un discreto predominio en el sexo masculino (1,5/1). Su mecanismo etiopatogénico es complejo y se ha vinculado principalmente con factores genéticos e inmunitarios. Aunque exhibe características clínicas de benignidad, se manifiesta histológicamente con rasgos de malignidad. Las técnicas inmunohistoquímicas resultan de utilidad a los fines diagnósticos y recientemente han permitido la identificación de un nuevo tipo de PL que remeda un linfoma cutáneo primario agresivo a células T epidermotrópico CD8+ (propuesto como PL tipo D). Puede hallarse asociada a otros trastornos linfoproliferativos y a entidades inflamatorias con perfil de citocinas Th2, entre otros. El diagnóstico diferencial con otros linfomas cutáneos, y especialmente con los casos que presentan el antígeno CD30, puede a veces resultar muy dificultoso. Si bien la PL tiene un curso clínico benigno, quienes la padecen tienen un mayor riesgo de desarrollar una segunda neoplasia. Las opciones terapéuticas disponibles son múltiples; sin embargo, ninguna de ellas ha resultado hasta ahora completamente eficaz.
Subject(s)
Humans , Lymphoma, T-Cell/classification , Lymphoma, T-Cell/pathology , Lymphoma, T-Cell/therapy , Lymphomatoid Papulosis/genetics , Lymphomatoid Papulosis/pathology , /analysis , Diagnosis, Differential , Immunohistochemistry , Skin Neoplasms/genetics , Prognosis , Lymphoproliferative Disorders/genetics , Lymphoproliferative Disorders/pathologyABSTRACT
La pleiotropía de la leucemia linfocítica crónica (LLC) indica la aparición de otros tipos de enfermedades linfoproliferativas en ancestros del probando. En 35 pares de padres-descendientes con LLC en los padres, 27 pares (77%) tenían también LLC. en los descendientes, igual en herencia materna y paterna. En 8 descendientes (23%) sin LLC, hubo 6 casos con otra enfermedad linfoproliferativa y dos casos con plicitemia vera. Por ende, la pleitropía de la LLC comprende tanto trastornos linfoproliferativos como mieloproliferativos. En relación con la anticipación, no se observó ninguna diferencia en la edad al inicio de la enfermedad entre padres e hijos. Se debate que estos hallazgos puedan estar de acuerdo con un modo no mendeliano de transmisión de la LLC con muchos alelos de bajo riesgo que confieren un pequeño riesgo fenotípico de LLC. Además, la transmisión de los genes no alélicos refleja la impronta (silenciamiento) materna de los genes paternos como consecuencia del microquimerismo relacionado con el embarazo.
Subject(s)
Humans , Leukemia, Lymphocytic, Chronic, B-Cell , Lymphoma, Non-Hodgkin , Lymphoproliferative Disorders , Lymphoproliferative Disorders/geneticsABSTRACT
OBJETIVO: Determinar a utilidade, na prática rotineira, da análise da clonalidade dos linfócitos T e B nos tecidos pulmonares por reação em cadeia da polimerase no diagnóstico das doenças linfoproliferativas pulmonares. MÉTODOS: Avaliaram-se, mediante análise imunohistoquímica e rearranjo molecular dos genes, 8 casos de pneumonia intersticial linfocítica (PIL) e 7 casos de doenças linfoproliferativas pulmonares. RESULTADOS: Todos os 8 casos de PIL expressaram imunocoloração moderada a forte para CD3, em contraste com apenas 2 casos de linfoma e 1 caso de pseudolinfoma. Rearranjo gênico foi detectado em 4 de 8 casos de PIL, o que mudou o diagnóstico de PIL para linfoma, indicando, assim, a importância da detecção de rearranjo gênico em casos de PIL. Nesta situação, rearranjo gênico usando-se os pares de primers VH/JH e Vgama11/Jgama12 foi detectado em 3 e 1 casos de PIL, respectivamente, e não foram detectadas anormalidades gênicas usando-se as pares Dbeta1/Jbeta2 e Vgama101/Jgama12. Uma associação positiva foi detectada entre a intensidade de imunoexpressão CD20 e CD68 e rearranjo gênico usando-se o par de primers VH/JH. Antes do rearranjo gênico, 4 pacientes com PIL morreram rapidamente, enquanto que, após o rearranjo gênico, apenas 1 paciente com PIL morreu. CONCLUSÕES: A detecção de células B e T monoclonais por imunofenotipagem e reação em cadeia da polimerase mostrou impacto no diagnóstico de linfomas pulmonares em pacientes previamente diagnosticados com PIL. Portanto, imunofenotipagem e reação em cadeia da polimerase devem ser incluídas como métodos de 'padrão ouro' na rotina diagnóstica.
OBJECTIVE: To determine the usefulness, in routine practice, of using polymerase chain reaction to analyze B and T lymphocyte clonality in pulmonary tissue as a tool for the diagnosis of pulmonary lymphoproliferative disorders. METHODS: Immunohistochemistry and molecular gene rearrangement analysis were performed in order to assess 8 cases of lymphoid interstitial pneumonia (LIP) and 7 cases of pulmonary lymphoproliferative disorders. RESULTS: All 8 cases of LIP presented moderate to strong immunostaining for CD3, compared with only 2 cases of lymphoma and 1 case of pseudolymphoma (p = 0.02). Gene rearrangement was detected in 4 of the 8 cases, which changed the diagnosis from LIP to lymphoma, showing the importance of gene rearrangement detection in cases of LIP. In this situation, gene rearrangement using the VH/JH and Vgamma11/Jgamma12 primer pairs was detected in 3 cases and 1 case, respectively, and no gene abnormalities were found using the Dbeta1/Jbeta2 and Vgamma101/Jgamma12 primer pairs in any of the cases. A significant positive association was found between the intensity of CD20 and CD68 expression and gene rearrangement using the VH/JH primer pair. Prior to the gene rearrangement, 4 patients with LIP died quickly, whereas only one patient with LIP died after the gene rearrangement. CONCLUSIONS: Detection of monoclonal B and T cells by immunophenotyping and polymerase chain reaction had an impact on the diagnosis of pulmonary lymphomas in patients previously diagnosed with LIP. Therefore, immunophenotyping and polymerase chain reaction should be used as 'gold standard' techniques in routine practice.
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child, Preschool , Female , Humans , Male , Middle Aged , Gene Rearrangement , Immunophenotyping , Lung Diseases, Interstitial/immunology , Lung Neoplasms/immunology , Lymphoma/immunology , Antigens, CD/analysis , Case-Control Studies , Diagnosis, Differential , DNA Primers , Feasibility Studies , Gene Rearrangement, B-Lymphocyte, Heavy Chain/genetics , Gene Rearrangement, B-Lymphocyte, Heavy Chain/immunology , Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor/genetics , Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor/immunology , Lung Diseases, Interstitial/diagnosis , Lung Diseases, Interstitial/genetics , Lung Neoplasms/diagnosis , Lung Neoplasms/genetics , Lymphoid Tissue/pathology , Lymphoma/diagnosis , Lymphoma/genetics , Lymphoproliferative Disorders/diagnosis , Lymphoproliferative Disorders/genetics , Lymphoproliferative Disorders/immunology , Polymerase Chain Reaction , Pseudolymphoma/diagnosis , Pseudolymphoma/genetics , Pseudolymphoma/immunology , Retrospective StudiesABSTRACT
BACKGROUND AND OBJECTIVES: Molecular events that precede transformations from lymphomatoid palulosis (LyP) to mycosis fungoides (MF) or to cutaneous anaplastic large cell lymphoma (ALCL) in the CD 30(+) cutaneous lymphoproliferative diseases (LPDs) are not known. Altered p(53) gene may be responsible since overexpression of the p(53) gene product has been reported in higher, but not in lower grades of cutaneous lymphomas. Expression of the anaplastic lymphoma kinase (ALK) gene product has also been described as an important prognostic indicator in ALCL. ALK positive systemic nodal ALCL are associated with a good prognosis. However, primary cutaneous ALCL that are ALK negative have a better overall survival. The current study was done to see if mutated p(53) gene or ALK reactivity were poor prognostic indicators in those patients with CD 30(+) cutaneous LPD who showed progression of the disease. METHODS: Mutations of the p(53) gene and expression of the ALK gene product were analysed in 36 patients (23 of LyP and 13 of CD30(+) cutaneous ALCL). Follow up data were available up till 5 yr in all patients. RESULTS: Clinical progression or histological transformation in sequential biopsy specimens was found in 9 of 36 patients. Transformation occurred in 5 patients (4 from LyP to ALCL and 1 from MF to ALCL) and clinical progression in 4 patients with ALCL. Mutations of the p(53) gene were found in two biopsy specimens of LyP. ALK gene products were not detected in any of the biopsy specimens of LyP and primary cutaneous ALCL. INTERPRETATION AND CONCLUSION: Although 9 of 36 patients with cutaneous CD30(+) LPDs had progression of their disease, neither mutations of the p(53) gene nor ALK immunoreactivity were found in any of these biopsies. The two cases of LyP that had mutated p(53) gene in their biopsy specimens showed no progression of their disease in the 5 yr follow up period. It appears that these molecular events may not play any significant role in the pathogenesis, progression or transformation of cutaneous CD30(+) LPD.