Melanoma-associated antigens recognized by cytotoxic T lymphocytes

During the last 7 years significant progress has been made in the identification of melanoma-associated antigens recognized by cytotoxic T lymphocytes (CTL). These antigens belong to three main groups: cancer/testis-specific antigens (MAGE, BAGE, GAGE, PRAME and NY-ESO-1), melanocyte differentiation antigens (tyrosinase, Melan-A/MART-1, gp100, TRP-1 and TRP-2), and mutated or aberrantly expressed antigens (MUM-1, CDK4, beta-catenin, gp100-in4, p15 and N-acetylglucosaminyltransferase V). In this review we have summarized the available data concerning the characterization of melanoma-associated antigens, focusing on their immunogenic and protective properties. The development of a strong immune response to differentiation antigens is limited by the existence of tolerance to these "self"-antigens, permitting the involvement of only T cells with low affinity T-cell receptors. Among the melanoma differentiation antigens, only gp100 has been shown to be a tumor regression antigen. The cancer/testis-specific antigens such as MAGE and PRAME should potentially be highly immunogenic antigens. They contain several potential HLA class I binding epitopes and are present only in the testes, which are not accessible to the cells of the immune system owing to the lack of direct contact with the immune cells and the lack of HLA class I expression on the surface of germ cells. But only two patients have been found who responded to these antigens in vivo, indicating their genuinely low immunogenicity. A comparison of the predicted secondary structures of these two groups of antigens (cancer/testis-specific and differentiation antigens) revealed enrichment of long alpha-helical stretches in the cancer/testis-specific antigens. We hypothesize that such highly organized stable structures could, first, reduce denaturation of the protein and, thus, ubiquitinylation as a degradation signal, and, second, diminish the efficiency of the protein unfolding - a necessary step in the proteolytic cleavage by proteasomes. High structural stability could therefore be responsible for the low immunogenicity of these proteins. In this case, modifications decreasing the stability of these proteins might be a means of improving the immune response to these potentially therapeutically useful antigens.     

Generation and characterization of gp100 peptide-specific NK-T cell clones

MHC-restricted cytotoxic T lymphocytes (CTLs) specific for antigens expressed by malignant cells are important components of immune responses against human cancer. Peripheral blood monocytes of HLA-A2+ healthy donors were used to induce dendritic cells (DCs) by granulocyte-macrophage colony-stimulating factor and interleukin-4 and loaded with a gp100 peptide (YLEPGPVTA). By applying these peptide-loaded DCs, a CTL line that displayed high cytotoxic reactivity with peptide-loaded target cells was generated. A total of 11 gp100 peptide-specific CTL clones were generated from this cell line. Several of these CTL clones were studied in detail. Of particular interest was clone CTL-45, which, contrary to the parental cell line, displayed strong NK activity and, by flow-cytometric analysis, revealed a CD3+, TCR BV17, CD8+ and CD56+ phenotype. This clone was strictly peptide-specific and effectively killed a panel of melanoma cells expressing HLA-A2 and gp100. Tumor-specific T cells with this kind of dual function are potentially of great clinical importance as they have a backup mechanism that may go into action when tumor cells escape specific killing by losing their HLA-class I molecules.     

Preservation of precut white cheese by modified atmosphere packaging

Effects of modified atmosphere packaging (MAP) on storage stability and quality of precut fresh and aged white cheese were investigated. Fresh or aged white cheese was cut into small cubes and packaged in five different atmospheres [0% O₂ + 0% CO₂ + 100% N₂ (MAP1), 10% O₂ + 0% CO₂ + 90% N₂ (MAP2), 0% O₂ + 75% CO₂ + 25% N₂ (MAP3), 10% O₂ + 75% CO₂ + 15% N₂ (MAP4) and aerobic (air)]. Control samples were packaged in brine and vacuum for fresh and aged white cheese, respectively. Changes in gas composition, total plate count, lactococci, lactobacilli, yeast and mould counts, proteolysis, lipolysis, pH, colour, texture and sensory properties were investigated during refrigerated storage. The best packaging treatment for the fresh cheese was MAP3, as it inhibited mould growth and protected the hardness. MAP2 can be recommended for the packaging of the aged cheese, as it decreased lipolysis.     

Disruption of the MMAC1/PTEN gene by deletion or mutation is a frequent event in malignant melanoma

The MMAC1/PTEN gene, located at 10q23.3, is a candidate tumor suppressor commonly mutated in glioma. We have studied the pattern of deletion, mutation, and expression of MMAC1/PTEN in 35 unrelated melanoma cell lines. Nine (26%) of the cell lines showed partial or complete homozygous deletion of the MMAC1/PTEN gene, and another six (17%) harbored a mutation in combination with loss of the second allele. Mutations could also be demonstrated in uncultured tumor specimens from which the cell lines had been established, and cell lines derived from two different metastases from one individual carried the same missense mutation. Collectively, these findings suggest that disruption of MMAC1/PTEN by allelic loss or mutation may contribute to the pathogenesis or neoplastic evolution in a large proportion of malignant melanomas.     

Bran in the treatment of irritable bowel syndrome

The paper deals with the problem of using wheat bran in the treatment of patients with irritable colon syndrome expressed as spastic constipation. The effectiveness of the treatment with the wheat bran only (in a dose of 30-35 g/day), and in combination with drugs was comparatively studied in 105 patients. The bran fractions differing in the particle size, in the content of cellulose, starch and vitamins were used in the treatment. The combined therapy proved to be advantageous only in the rate of the clinical effect, while the acceleration of the movement along the large intestine did not depend on the treatment type. A long-term (during one year) follow-up of the patients showed that the bran intake led to the cessation of the disease relapse; when the bran was abolished the symptoms of the disease appeared in 11 out of 12 cases. The highest effect was recorded with the bran fraction containing 55.3% cellulose, 18.3% lignin, 157 micrograms tocopherol and the lowest amount of starch--18.0%. A conclusion has been made that the wheat bran are effective in the treatment and prevention of intestinal diseases, the effectiveness of the treatment depends on the summary content of food fibers in the nutrition.