[Detection of specific T cells for 264-272 and 149-157 peptides of p53 protein in peripheral blood of patients with breast cancer]. / Detección de linfocitos T citotóxicos contra los epítopos 264-272 y 149-157 de la proteína p53 en sangre periférica de pacientes con cáncer de mama.

BACKGROUND AND OBJECTIVE: p53 protein is overexpressed in nearly half of all human tumours. An HLA-A2.1-restricted immunological response mediated by anti-p53 CD8+ T cells directed against the wild type p53 264-272 epitope has been demonstrated in patients with head and neck squamous carcinomas. The existence of such a response in patients with other cancer types could be determinant for the development of specific antitumour vaccines targeting the p53 protein. We aimed to determine the presence of anti-p53 specific CD8+ T cells in peripheral blood of breast cancer patients in vivo. PATIENTS AND METHOD: p53 264-272-specific CD8+ T cells were directly enumerated in the peripheral circulation of patients with breast cancer using tetrameric p53 264-272/HLA-A2.1 complexes by multicolor flow cytometry. The same procedure was used to enumerate T cells specific for another HLA-A2.1 restricted wild type p53 epitope, p53 (149-157). RESULTS: The 99th percentile of the concentration of anti-p53 cells in 6 HLA A2- breast cancer patients was 1/5634 (cut-off point). The median counts of anti-p53264-272 and anti-p53149-157 lymphocytes in 14 HLA A2.1+ patients were 1/2383 and 1/2335 respectively. All of the HLA A2+ patients had concentrations of anti-p53 lymphocytes above the cut-off point for at least one of the epitopes: 13/14 (93%) for p53(264-272) and 11/12 (92%) for p53(149-157). CONCLUSIONS: A specific immunological response mediated by anti-p53 CD8+ T cells has been detected in patients with breast carcinoma. More studies are needed to confirm these results and to determine its usefulness for the development of p53-based vaccines.

Detección de linfocitos T citotóxicos contra los epítopos 264-272 y 149-157 de la proteína p53 en sangre periférica de pacientes con cáncer de mama / Detection of specific T cells for 264-272 and 149-157 peptides of p53 protein in peripheral blood of patients with breast cancer

FUNDAMENTO Y OBJETIVO: La proteína p53 está sobreexpresada en la mitad de los tumores humanos.Se ha descrito que se produce una respuesta inmunológica específica mediada por linfocitosT citotóxicos dirigidos contra el epítopo 264-272 de p53 en pacientes con cáncer de cabezay cuello. Demostrar que se produce ese tipo de respuesta en otros tumores podría serdeterminante para el desarrollo de vacunas antitumorales específicas anti-p53. El objetivo deeste estudio fue demostrar in vivo la existencia de linfocitos T citotóxicos específicos anti-p53en sangre periférica de pacientes con cáncer de mama.PACIENTES Y MÉTODO: Se realizó la determinación mediante citometría de flujo del recuento delinfocitos T citotóxicos específicos dirigidos contra los epítopos HLA A2 restringidos 264-272 y149-147 de la proteína p53 en sangre periférica de pacientes con cáncer de mama.RESULTADOS: El percentil 99 de la concentración de células T anti-p53 en los pacientes conHLA A2.1 negativo fue 1/5.634 (punto de corte). En los pacientes con HLA A2 positivo, la medianade linfocitos anti-p53264-272 fue de 1/2.383 y la de linfocitos anti-p53149-157, de 1/2.335.Todos los pacientes con HLA A2 positivo presentaron recuentos de linfocitos anti-p53 por encimadel punto de corte para al menos uno de los 2 epítopos: 13/14 (93%) para p53264-272 y11/12 (92%) para p53149-157.CONCLUSIONES: Ha sido posible demostrar in vivo que hay una respuesta inmunológica específicamediada por linfocitos T citotóxicos anti-p53 en pacientes con cáncer de mama. Son necesariosfuturos estudios para confirmar estos resultados y determinar su utilidad para el diseño y eldesarrollo de vacunas dirigidas contra la proteína p53

BACKGROUND AND OBJECTIVE: p53 protein is overexpresed in nearly half of all human tumours. AnHLA-A2.1-restricted immunological response mediated by anti-p53 CD8+ T cells directedagainst the wild type p53 264-272 epitope has been demonstrated in patients with head andneck squamous carcinomas. The existence of such a response in patients with other cancer typescould be determinant for the development of specific antitumour vaccines targeting thep53 protein. We aimed to determine the presence of anti-p53 specific CD8+ T cells in peripheralblood of breast cancer patients in vivo.PATIENTS AND METHOD: p53 264-272-specific CD8+ T cells were directly enumerated in the peripheralcirculation of patients with breast cancer using tetrameric p53 264-272/HLA-A2.1complexes by multicolor flow cytometry. The same procedure was used to enumerate T cellsspecific for another HLA-A2.1 restricted wild type p53 epitope, p53 (149-157).RESULTS: The 99th percentile of the concentration of anti-p53 cells in 6 HLA A2– breast cancerpatients was 1/5634 (cut-off point). The median counts of anti-p53264-272 and anti-p53149-157lymphocytes in 14 HLA A2.1+ patients were 1/2383 and 1/2335 respectively. All of the HLAA2+ patients had concentrations of anti-p53 lymphocytes above the cut-off point for at leastone of the epitopes: 13/14 (93%) for p53264-272 and 11/12 (92%) for p53149-157.CONCLUSIONS: A specific immunological response mediated by anti-p53 CD8+ T cells has beendetected in patients with breast carcinoma. More studies are needed to confirm these resultsand to determine its usefulness for the development of p53-based vaccines

Common variants on chromosomes 2q35 and 16q12 confer susceptibility to estrogen receptor-positive breast cancer.

Familial clustering studies indicate that breast cancer risk has a substantial genetic component. To identify new breast cancer risk variants, we genotyped approximately 300,000 SNPs in 1,600 Icelandic individuals with breast cancer and 11,563 controls using the Illumina Hap300 platform. We then tested selected SNPs in five replication sample sets. Overall, we studied 4,554 affected individuals and 17,577 controls. Two SNPs consistently associated with breast cancer: approximately 25% of individuals of European descent are homozygous for allele A of rs13387042 on chromosome 2q35 and have an estimated 1.44-fold greater risk than noncarriers, and for allele T of rs3803662 on 16q12, about 7% are homozygous and have a 1.64-fold greater risk. Risk from both alleles was confined to estrogen receptor-positive tumors. At present, no genes have been identified in the linkage disequilibrium block containing rs13387042. rs3803662 is near the 5' end of TNRC9 , a high mobility group chromatin-associated protein whose expression is implicated in breast cancer metastasis to bone.

Results of a pilot trial of immunotherapy with dendritic cells pulsed with autologous tumor lysates in patients with advanced cancer.

AIMS AND BACKGROUND: The purpose of the study was to test the immunological and clinical effects of infusions of dendritic cells pulsed with autologous tumor lysate in patients with advanced cancer. PATIENTS AND METHODS: Peripheral blood mononuclear cells from 15 patients with metastatic cancer (melanoma in 10, lung cancer in 2, renal cell carcinoma in 1, sarcoma in 1, breast cancer in 1) were harvested by leukapheresis after mobilization with GM-CSF (5 microg/kg/day s.c. for 4 days). Mononuclear cells were separated and cultured in GM-CSF (1000 U/ml) and interleukin-4 (1000 U/ml) for 7 days. Phenotype was assessed by 2-color flow cytometry and immunocytochemistry. On day 6, dendritic cells were pulsed with 1 g of fresh autologous tumor lysate for 24 h and infused intravenously. Interleukin-2 (6 million IU), interferon a (4 million IU) and GM-CSF (400 microg) were injected s.c. daily for 10 days beginning on the day of dendritic cell infusion. Treatment was repeated every 21 days for 3 courses. RESULTS: The morphology, immunocytochemistry and phenotype of cultured cells was consistent with dendritic cells: intense positivity for HLA-DR and CD86, with negativity for markers of other lineages, including CD3, CD4, CD8 and CD14. More than 5 x 10(7) dendritic cells were injected in all patients. Nine patients developed >5 mm delayed type cutaneous hypersensitivity reactions to tumor lysate+/-GM-CSF after the first immunization (larger than GM-CSF in all cases). Median delayed type cutaneous hypersensitivity to lysate +/- GM-CSF was 3 cm after the third immunization. One melanoma patient with skin, liver, lung and bone metastases had a partial response lasting 8 months (followed by progression in the brain). Seven patients had stable disease for >3 months, and 7 had progression. CONCLUSIONS: Infusion of tumor lysate-pulsed dendritic cells induces a strong cell-mediated antitumor immune reaction in patients with advanced cancer and has some clinical activity.

Clear cell carcinoma of the ovary diagnosed in ascitic fluid. A case report.

BACKGROUND: Clear cell carcinoma of the ovary (CCC) is a rare variety of ovarian cancer. CASE: A case of CCC in a 49-year-old woman was diagnosed in asciticfluid on thin-layer preparations. Peritoneal fluid cytology revealed papillary clusters of cells with clear cytoplasm and extracellular hyaline material generally without neoplastic cells. The tumor was excised, and the histologic sections confirmed the cytologic diagnosis. CONCLUSION: CCC has a distinctive cytomorphologic appearance, and the entity may be diagnosed on ascitic fluid cytology.

Extraovarian primary peritoneal carcinoma. A case report

Se presenta un caso de carcinoma peritoneal primario extraovárico. Este raro e interesante tumor se caracteriza por una carcinomatosis peritoneal y por un patrón histológico similar al del carcinoma papilar seroso de ovario, pero sin comprometer o sólo afectando microscópicamente la superficie de los ovarios. La paciente presentaba hinchazón abdominal, ascitis con citología positiva y el CA-125 elevado en el suero. En el momento de la cirugía se encontró una afectación peritoneal difusa o multifocal, existiendo afectación microscópica de la superficie serosa ovárica. Las células tumorales fueron positivas para citoqueratina (AE1/AE3), antígeno de membrana epitelial, B72.3, Ber- EP4, Leu-M1, p53, antígeno carcinoembrionario, fosfatasa alcalina placentaria, proteína S-100, CA-125 y receptores estrogénicos

One case of extraovarian primary peritoneal carcinoma is reported. This rare interesting tumor is characterized by a peritoneal carcinomatosis with ascites and by a histological pattern similar to ovarian serous papillary carcinoma, but it either spares the ovaries or involves the surface of the ovaries only microscopically. The patient had abdominal swelling, ascites with positive cytology, and elevated serum CA-125. At surgery multifocal or diffuse peritoneal involvement was found. Microscopic ovarian serosal surface involvement was demonstrated. Tumor cells stained positive for cytokeratin (AE1/AE3), epithelial membrane antigen, B72.3, Ber-EP4, Leu-M1, p53, carcinoembryonic antigen, placental alkaline phosphatase, S-100, CA-125, and estrogen receptors

Primary pulmonary botryomycosis diagnosed by fine needle aspiration cytology: a case report.

BACKGROUND: Primary pulmonary botryomycosis is an uncommon suppurative bacterial infection of the lung. Cytologic findings by conventional methods and thin-layer preparations in1 case are presented. CASE: A 45-year-old man had primary pulmonary botryomycosis diagnosed by fine needle aspiration cytology (FNAC). Aspiration smears were characterized by densely packed microorganisms surrounded by polymorphonuclear leukocytes. Microbiologic analysis of cytologic material revealed colonies of Streptococcus constellatus. CONCLUSION: FNAC diagnosis of pulmonary botryomycosis offers the opportunity to distinguish this bacterial infection from lung cancer and avoid unnecessary surgery.

Linfoma primario cutáneo anaplásico de células grandes CD30 (Ki-1) positivo simulando un sarcoma epitelioide / CD30 (Ki-1) primary cutaneous anaplastic large-cell lymphoma mimicking an epithelioid sarcoma

Presentamos un caso de linfoma primario cutáneo de células grandes CD30 (Ki-1) positivo con características histológicas que recuerdan a un sarcoma epitelioide. El linfoma primario cutáneo anaplásico de células grandes CD30 positivo es una entidad clinicopatológica recientemente descrita, reconocida por criterios inmunohistoquímicos. Describimos el caso de una mujer de 74 años de edad con un tumor ulcerado en el tronco, de 3 meses de evolución. El diagnóstico se hizo en base a los hallazgos inmunohistoquímicos. No se detectó afectación sistémica. La paciente está bien, sin enfermedad extracutánea concurrente, tras 6 años de seguimiento. Los estudios clínicos indican un mejor pronóstico en los linfomas cutáneos anaplásicos de células grandes CD30 positivos que en los linfomas anaplásicos de células grandes CD30 positivos nodales y que en los otros linfomas cutáneos anaplásicos de células grandes. Este es pues, un caso de regresión espontánea y seguimiento favorable en este tipo de linfomas cutáneos, que tiene unos rasgos morfológicos peculiares, como es su parecido morfológico con un sarcoma epitelioide (AU)

Dermal analogue tumor of the salivary gland diagnosed by fine needle aspiration cytology. A case report.

BACKGROUND: Dermal analogue tumor of the salivary gland is a rare form of salivary gland adenoma. Cytologic and histologic findings of 1 case are presented. CASE: A 74-year-old-female had a dermal analogue tumor in the parotid gland diagnosed by fine needle aspiration cytology (FNAC). Aspiration smears were characterized by aggregates of uniform epithelial cells, and cell groups bordered on thick, basement membrane-like material. The tumor was excised, and the histologic sections confirmed the cytologic diagnosis. CONCLUSION: Dermal analogue tumor of the salivary gland has a distinctive cytomorphologic appearance, and diagnosis of this neoplasm by FNAC is possible. Preoperative FNAC of salivary gland lesions is important in planning the most appropriate type of treatment.

Extramedullary plasmacytoma presenting as a primary mass in the breast. A case report.

BACKGROUND: Extramedullary plasmacytoma (EMP) of the breast is extremely rare, especially that not associated with multiple myeloma. CASE: A case of plasmacytoma of the breast in a 73-year-old man was diagnosed by fine needle aspiration cytology (FNAC). Aspiration smears revealed a dispersed population of plasmacytoid cells at various stages of maturation. The tumor was excised, and the histologic sections confirmed the cytologic diagnosis. CONCLUSION: FNAC diagnosis of plasmacytoma of the breast offers the opportunity to distinguish these neoplasms from primary mammary tumors and avoid unnecessary surgery.

Malignant phyllodes tumor with pleomorphic liposarcomatous stroma diagnosed by fine needle aspiration cytology: a case report.

BACKGROUND: Liposarcomatous differentiation within a phyllodes tumor is extremely rare. Cytologic and histologic findings of a case of malignant phyllodes tumor with liposarcomatous stroma of the breast are presented. CASE: A 45-year-old female had a malignant phyllodes tumor with pleomorphic liposarcomatous stroma diagnosed by fine needle aspiration (FNA) cytology. The cytologic findings were representative of the histologic features. CONCLUSION: Malignant phyllodes tumor of the breast can be diagnosed by FNA. It is very important to acknowledge the morphologic variants of sarcomatous stroma and to recognize the cytologic features of such rare tumors to prevent misdiagnosis as primary sarcomas of the breast. Preoperative diagnosis is important in planning the most appropriate type of treatment. It is also important to follow patients for long periods for recurrence and metastasis after surgery for this tumor.