HER2/neu Testing In 432 Consecutive Breast Cancer Cases using FISH and IHC - A Comparative Study.

INTRODUCTION: The development of trastuzumab, a HER-2/neu targeted monoclonal antibody resulted in significant improvements in clinical response and survival in HER-2/neu gene amplified group of patients. Thus, accurate assessment of HER-2/neu status becomes critical. Fluorescence In Situ Hybridization (FISH) and Immunohistochemistry (IHC) are the most commonly used methods for this purpose and specific recommendations exist with regard to the concordance to be observed between the two tests. AIM: Here, we report and evaluate the concordance rate between FISH and IHC for HER-2/neu status in breast cancer specimens. MATERIALS AND METHODS: Archival paraffin blocks of tumour tissue from 450 patients of breast cancer were analyzed for Her-2/neu status using FISH and IHC. RESULTS: There was a highly significant concordance between the results of FISH and IHC assays in HER-2/neu status assessment in invasive breast cancer cases. There were inverse associations between the expression of Oestrogen Receptors/Progesterone Receptors (ER/PR) and HER-2/neu amplification. CONCLUSION: Although, IHC gave significant concordant results with FISH in determining HER-2/neu status, its subjective grading system precludes its use as a gold standard. FISH should always be used to determine true gene amplification when the IHC results are equivocal.

Chronic lymphoproliferative disorders at an Indian tertiary cancer centre - the panel sufficiency in the diagnosis of chronic lymphocytic leukaemia.

BACKGROUND: Flow cytometry has come to occupy the vanguard of the high through put diagnostic techniques that have been used to differentiate between various chronic lymphoproliferative disorders (CLPD). However, economic considerations have created the need for minimal consensus panels that can yield maximum information at reasonable costs. AIMS: To collect, analyse and correlate the morphologic, immunophenotypic, and the cytogenetic data from the cases of chronic lymphoproliferative disorders, which were diagnosed at an Indian speciality cancer centre. METHODS AND MATERIAL: The morphology was recorded after staining the samples with the Leishman or the MGG stains. The lineage assignment was done by using three colour flow cytometry with a primary panel of antibodies. For the cytogenetic studies, the short term culture of the sample cells were arrested by using colcemid and they were G-banded by using trypsin and Giemsa stain. FISH studies were conducted by using a CLL-specific diagnostic kit. RESULTS AND CONCLUSIONS: A total of 66 cases were evaluated, which had a median age of 64.5 years and a sex ratio of 2.3:1. Of these 66 cases, 40 cases were of CLL and 9 cases were of atypical CLL. 17 cases were classified as CLPD and these included 13 cases of Non-Hodgkin's Lymphoma, two cases of Hairy Cell Leukaemia, one case of Follicular Lymphoma and one case of Prolymphocytic Leukaemia. In immunophenotyping, the lack of expression of CD22 had the highest correlation with a definitive diagnosis of CLL. Cytogenetics demonstrated a classical follicular lymphoma abnormality, t (14; 18) (q32; q21), in one case. A basic minimal panel is sufficient for the routine diagnosis of CLL. However, the stratification of CLPD requires the use of more extensive panels.

CD34 positive-microgranular variant of acute promyelocytic leukemia in a child.

Acute promyelocytic leukemia (APL) is a subtype of acute myeloid leukemia (AML) in which abnormal promyelocytes predominate. APL is rare in children (approximately 10% of childhood AML) and is characterized by a higher incidence of hyperleukocytosis, an increased incidence of microgranular morphology, the presence of balanced t(15;17)(q22;q11.2-12) translocation, and more frequent occurrence of the PML-RARα isoforms bcr 2 and bcr 3 compared to adults. The cytomorphology of microgranular variant blasts is obviously different from AML M3 blasts; these cells have a nongranular or hypogranular cytoplasm or contain fine dust-like cytoplasmic azurophil granules that may not be apparent by light microscopy. This case report emphasizes the importance of a high index of suspicion for the diagnosis of APL, the hypogranular variant in particular. They are responsive to differentiation therapy with all trans-retinoic acid and complete remission in seen in >80% cases.

An uncommon case of an adult with del(5)(q) in acute lymphoblastic leukemia.

Del(5)(q) is a common chromosomal abnormality with favourable prognosis in Myelodysplastic Syndrome (MDS) and Acute myeloid leukemia (AML). However, del(5)(q) is also seen rarely in Acute lymphoblastic leukemia (ALL) and its significance remains poorly understood. We present here, a case report of diagnosis of an adult 75 year old patient of ALL with a cytogenetic abnormality of del(5)(q32). His clinical features, morphology and immunophenotyping findings were suggestive of T-ALL. Relevant literature has been reviewed and discussed.

Her-2/neu overexpression due to polysomy 17 in breast cancer: molecular testing to guide therapeutic options.

BACKGROUND: Fluorescent in situ hybridization (FISH) equivocal results for Her-2/neu still pose a diagnostic dilemma in oncology practice. In this study, we evaluate if Her-2/neu mRNA expression is an alternative to FISH for detecting Her-2/neu positivity. PATIENTS AND METHODS: Archival paraffin blocks of 54 breast cancer patients were analyzed for Her-2/neu status using immunohistochemistry (IHC), FISH, and Her-2/neu gene expression using mRNA. RESULTS: There was a 100% positive agreement and 64.7% negative agreement of Her-2/neu mRNA expression with respect to the reference standard (FISH), with the kappa value for agreement being 0.36. mRNA levels correlated positively and strongly with FISH ratio and IHC positivity. For Her-2/neu mRNA expression, Her-2/neu copy number was a significant predictor indicating that mRNA expression is independent of polysomy status. CONCLUSIONS: Her-2/neu mRNA expression may help tide over ambiguity posed by polysomy and FISH equivocal samples.

Accelerated phase of chediak higashi syndrome mimicking lymphoma--a case report.

SUMMARY: Chediak Higashi Syndrome (CHS) is a rare autosomal recessive disease, characterized by partial oculocutaneous albinism, frequent pyogenic infections, and the presence of abnormal large granules in leukocytes and other granule containing cells. The abnormal granules are readily seen in blood and marrow granulocytes. About 50% to 85% of patients eventually enter an accelerated phase, manifested by fever, lymphadenopathy, anemia, jaundice, neutropenia, thrombocytopenia, and widespread lymphohistiocytic organ infiltrates. The first accelerated phase of CHS may occur shortly after birth or several years later. Most patients undergo a variable period of recurrent infections before going into the accelerated phase. Therefore, primary presentation in the accelerated phase is unusual. This case was referred to our institution that is a tertiary care cancer centre, with a clinical diagnosis of lymphoma/leukemia. Hence this interesting case of CHS in accelerated phase at presentation is described. The child had 1-month history of fever, bilateral neck swellings, and loss of appetite. On the basis of the clinical presentation, hematologic, and histopathologic findings, a diagnosis of accelerated phase of CHS was made. The child was treated with antipyretics, antibiotics, and stem cell transplantation was suggested to him. When the child presents to a hospital with oculocutaneous albinism and recurrent infections, careful examination of the peripheral blood smear by an experienced morphologist cannot be overemphasized. A high degree of awareness and early recognition of the syndrome, could lead to the institution of the only possible curative treatment, bone marrow transplant, before the accelerated phase supervenes.