Low apoptotic index & bak expression in EBV-associated childhood classical Hodgkin lymphoma.

Background & objectives: Association of Epstein-Barr virus (EBV) with Hodgkin lymphoma (HL) is particularly high in low-income countries, and resistance to apoptosis might play a role in pathogenesis and survival. Data from previous studies are not consistent, and none is available in children. Thus this study was undertaken on Indian children with classical Hodgkin lymphoma to assess the significance of bcl-2, bak and p53 expression, and apoptotic index in relation with EBV status and treatment outcome with chemotherapy alone. Methods: Children (age<15 yr) with classical HL (n=143) were included in the study. Bcl-2, bak, p53, Ki67 and latent membrane protein-1 (LMP1) were detected by immunohistochemistry in pre-treatment lymph node biopsies. Apoptotic index was assessed by TdT-dUTP nick-end labelling (TUNEL). Results: Bcl-2, bak, p53 were expressed above positivity threshold in 83.3, 94.0 and 7.1 per cent of the cases respectively. More than 10 per cent of apoptotic tumour cells were seen in 60.4 per cent of the cases. 131 (91.6%) cases were EBV associated. EBV-positive cases had a significantly lower mean bak expression (p=0.001) and a lower apoptotic index, without higher proliferation index. Advanced stage showed a borderline association with bcl-2 expression in >25 per cent of tumour cells and p53 negative tumours. In univariate analysis, p53 positive cases, which were significantly associated with B symptoms, had a poorer overall survival (P=0.03) while low proliferation index was associated with poorer failurefree survival. Neither EBV status nor any of the apoptotic parameters studied showed independent association with survival. Interpretation & conclusion: EBV detection in children with classical Hodgkin lymphoma was associated with significant lower bak expression and with lower spontaneous apoptosis of H-RS cells suggesting that EBV-LMP1 might downregulate bak pro-apoptotic protein. this needs to be substantiated further.

Diagnosis and grading of non-Hodgkin's lymphomas on fine needle aspiration cytology.

Fine needle aspiration cytology (FNAC) enjoys popularity among clinicians worldwide, as a first line of investigation in all patients with lymphadenopathy and is preferred over biopsy because of its minimally invasive nature and cost-effectiveness. Although non-Hodgkin's lymphomas (NHL) are conventionally diagnosed and graded on biopsy specimens, it may be useful to be able to not only diagnose but also grade these cases on FNAC smears. The WHO and REAL classifications forming the basis of treatment in some centres rely on clinical features, immunocytochemistry and cytogenetics, which are beyond reach of most centres in the developing countries. This study therefore is aimed at diagnosing and grading NHLs on morphological parameters. The cytologic grading accuracy is compared with the histologic grades assigned according to the International Working Formulation (IWF) system which relies solely on morphological features, most important of which is cell size. Ninety five cases were retrieved over a 3 year period (May 2000 to April 2003). These were (i) cases where a cytological diagnosis of NHL or suspicious of NHL was made and corresponding histological sections available and (ii) cases where a diagnosis of NHL was made in histology and corresponding FNAC smears were available irrespective of the cytological diagnosis. The diagnostic accuracy of FNAC for NHLs was determined using histology as the gold standard. Cases were also graded on FNAC smears using a three tier grading system based upon cell size into low, intermediate and high grades. Cytologically assigned grades were correlated with the corresponding histological grades (IWF) to determine grading accuracy. An accurate diagnosis of NHL was thus possible in 67/95 (70.5%) cases. Overall accurate grading was seen in 65/95 (68%) cases using cytological criteria. Accurate cytologic grading was possible in 14/15 (93.33%) low grade, 11/18 (61.11%) intermediate and 40/62 (64.5%) high grade non Hodgkin's lymphomas. Kappa statistics revealed a very good agreement between cytological and histological grades for low grade NHL. The kappa scores for intermediate and high grade NHLs indicated moderate agreement. Using the two-tier system grading the kappa value for high grade lymphomas improved to 0.72, indicating good concordance. This study highlights the utility of FNAC as a morphological tool for diagnosing and grading NHLs in a significant number of cases. This modality may assist clinicians in management of cases of NHLs, especially in centres working within the constraints of limited availability or non availability of ancillary techniques.

Etoposide, vinblastine, adriamycin and prednisolone (EVAP) combination chemotherapy as first-line treatment for Hodgkin's disease.

BACKGROUND: Mechlorethamine, vincristine, procarbazine, prednisolone (MOPP) and doxorubicin, bleomycin, vinblastine, dacarbazine (ABVD) are well established first-line chemotherapy protocols for the treatment of Hodgkin's disease. The aim of this study was to try a new combination of drugs that individually have a proven efficacy in Hodgkin's disease but have less incidence of severe nausea, vomiting, pulmonary toxicity and sterility. METHODS: This prospective, single-arm study enrolled 66 newly diagnosed, previously untreated patients of Hodgkin's disease with stages IA (bulky)-IVB disease. They were given 6-8 courses of etoposide, vinblastine, doxorubicin and prednisolone (EVAP) as first-line chemotherapy between January 1992 and December 1997. Radiotherapy (RT) was given to the involved fields of those patients who had bulky (> or = 10 cm) stages I or II disease at presentation. The end-points were (i) complete and overall response; (ii) disease-free and overall survival; and (iii) toxicity. RESULTS: Complete response was seen in 78.8% and partial response in 12.2% of patients; the overall response rate was 91%. The median follow up was 48 months. The 5-year overall and disease-free survivals were 72% and 62%, respectively. There were 3 episodes of grade IV neutropenia requiring hospitalization. One patient developed avascular necrosis of the femur. There were 2 deaths during treatment, one due to chemotoxicity, and another due to progressive disease. CONCLUSION: The overall and complete responses were fractionally inferior to the recently published hybrid MOPP/ ABV combination and that of ABVD chemotherapy. The advantages of the EVAP combination are absence of pulmonary toxicity, markedly lower incidence of sterility and nausea and vomiting. EVAP is an attractive option and a randomized trial is warranted to assess its efficacy against established protocols.