Defining the expression hierarchy of latent T-cell epitopes in Epstein-Barr virus infection with TCR-like antibodies

Epstein-Barr virus (EBV) is a gamma herpesvirus that causes a life-long latent infection in human hosts. The latent gene products LMP1, LMP2A and EBNA1 are expressed by EBV-associated tumors and peptide epitopes derived from these can be targeted by CD8 Cytotoxic T-Lymphocyte (CTL) lines. Whilst CTL-based methodologies can be utilized to infer the presence of specific latent epitopes, they do not allow a direct visualization or quantitation of these epitopes. Here, we describe the characterization of three TCR-like monoclonal antibodies (mAbs) targeting the latent epitopes LMP1[subscript 125–133], LMP2A[subscript 426–434] or EBNA1[subscript 562–570] in association with HLA-A0201. These are employed to map the expression hierarchy of endogenously generated EBV epitopes. The dominance of EBNA1[subscript 562–570] in association with HLA-A0201 was consistently observed in cell lines and EBV-associated tumor biopsies. These data highlight the discordance between MHC-epitope density and frequencies of associated CTL with implications for cell-based immunotherapies and/or vaccines for EBV-associated disease

S-100 protein positive cells in nasopharyngeal carcinoma (NPC): absence of prognostic significance. A clinicopathological and immunohistochemical study of 40 cases

An immunohistochemical study of S-100 protein in 43 nasopharyngeal carcinomas (NPC) of known clinical evolution (33 primary and 10 metastatic) is presented. Sixty per cent of primary site cases as well as all metastatic forms showed S-100 protein positive cells intermingled with tumour cells. These S-100 positive elements were identified as Langerhans cells. No significant differences were found when correlating S-100 protein positivity and histological NPC variants, neither in age nor in sex of patients. Statistical analysis failed to demonstrate any positive correlation between S-100 protein reactivity and clinical survival

Immunohistochemical estimation of cell cycle phase in laryngeal neoplasia

We previously developed an immunohistochemical method for estimating cell cycle state and phase in tissue samples, including biopsies that are too small for flow cytometry. We have used our technique to examine whether primary abnormalities of the cell cycle exist in laryngeal neoplasia. Antibodies against the markers of cell cycle entry, minichromosome maintenance protein-2 (Mcm-2) and Ki67, and putative markers of cell cycle phase, cyclin D1 (G1-phase), cyclin A (S-phase), cyclin B1 (G2-phase) and phosphohistone H3 (Mitosis) were applied to paraffin-embedded sections of normal larynx (n=8), laryngeal dysplasia (n=10) and laryngeal squamous cell carcinoma (n=10). Cells expressing each marker were determined as a percentage of total cells, termed the labelling index (LI), and as a percentage of Mcm-2-positive cells, termed the labelling fraction (LF). The frequency of coexpression of each putative phase marker was investigated by confocal microscopy. There was a correlation between Mcm-2 and Ki67 LIs (ρ=0.93) but Mcm-2 LIs were consistently higher. All cells expressing a phase marker coexpressed Mcm-2, whereas Ki67 was not expressed in a proportion of these cells. The putative phase markers showed little coexpression. Labelling index values increased on progression from normal larynx through laryngeal dysplasia to squamous cell carcinoma for Mcm-2 (P=0.001), Ki67 (P=0.0002), cyclin D1 (P=0.015), cyclin A (P=0.0001) and cyclin B1 (P=0.0004). There was no evidence of an increase in the LF for any phase marker. Immunohistochemistry can be used to estimate cell cycle state and phase in laryngeal biopsies. Our data argues against primary cell cycle phase abnormalities in laryngeal neoplasia

Towards a strategy for rehabilitation and development of irrigation systems in the Batticaloa district, Sri Lanka : a concept paper

Fra NORAGRIC. Report to NORAD. Sri Lanka, Batticaloa.Agriculture and fisheries are the mainstay of the rural household economies of Batticaloa district. Agriculture in the district is synonymous with rice cultivation as the latter remains the most important agricultural activity for the vast majority of its farm households. However, a wide range of subsidiary food crops and tree crops like coconut and cashew are also cultivated in the highlands. While the availability of a variety of soils offers opportunities for such a diversified agricultural land use, the scarcity of water at critical times in the cropping cycles has remained a major constraint to the districts economic and social advancement. Yet Batticaloas water resources in terms of total annua! precipitation and other natura! sources are not meagre by any standard. The water problem is caused primarily by the uneven distribution of rainfall and the inadequacies in conservation and management. Only 34 percent of the rainwater in an average year can be conserved in the existing irrigation tanks when they are fully functioning. However, until the mid 1980s these large and small irrigation schemes, which supplied water for 37.5 percent of the 57, 120 ha of paddy cultivated in the district, played a major role in making Batticaloa a surplus producer of paddy. With the advent of the civil war around 1985, the districts economy, largely dependent on agriculture and fisheries, came under growing stress leading to negative growth rates in several years. The irrigation infrastructure has been subject to damage and degeneration due to the effects of the war. Today, a majority of the minor irrigation schemes and many irrigation and drainage structures of the major schemes are in a bad state of disrepair. Moreover, the socio-economic dislocations caused by the war have seriously undermined local level institutions concerned with land water resources management and rural development