Alberto Bartorelli

The primary structure of UK 114 tumor antigen

UK114 is a tumor antigen expressed by various malignant neoplasms. The complete amino acid sequence of UK114 purified from goat liver has been determined by automated Edman degradation of CNBr and endoproteinase Lys‐C peptides. The protein contains 137 amino acid residues, which corresponds to a molecular mass of 14 229 Da. MALDITOF analysis resulted in a molecular weight of 14 290, suggesting that the N‐terminal Met residue is acetylated. Sequence comparison shows that UK114 from goat liver (1) has 77% identity with a previously described 23 kDa protein from rat liver (Levy‐Favatier et al. (1993) Eur. J. Biochem. 212, 665–673), (2) shares a very high degree of similarity with a family of prokaryotic and eukaryotic hypothetic proteins whose function have not yet been characterized, and (3) exhibits a significant similarity to a group of tumor‐associated antigens which belongs to a superfamily of heat shock proteins, acting as possible targets for the hosts antitumor immunity.

Biological components in a standardized derivative of bovine colostrum

Products of different origin, time of collection, and activities fall under the general term of colostrum and, therefore, great variability in composition as well as in the concentration of its components has been reported in the literature. In the present study, we describe the standardization of a bovine colostrum derivative and the characterization of its bioactive components. Evaluation of the most representative agents (lactoferrin, transferrin, IL-2, IFN-γ, tumor necrosis factor, IgG, and IgA) showed that a marked decrease in active components occurs after the first few hours. Bovine colostrum was, therefore, collected up to the fifth hour after delivery from Holstein cows, in the presence of preservatives, and immediately frozen. A protocol of centrifugation, filtration, and lyophilization was then applied to pools of colostrum from at least 30 cows to obtain a stable, sterile, standardized product. Preservatives were removed by dialysis. Evaluation of the active biological components of colostrum showed that the final product of colostrums contained significant and reproducible amounts of bioactive factors, including cytokines, immunomodulating factors, growth factors, and immunoglobulins. The final product appeared, therefore, as a sterile, pyrogen-free, standardized derivative of bovine colostrum with a high concentration of bioactive components.

Growth inhibition of DMBA-induced rat mammary carcinomas by UK 114

Abstract A perchloric acid-soluble protein extracted from goat liver and designated as UK 114 is known to be expressed over the cell membrane of (some) human cancer cell lines. This protein is antigenic, and specific antibodies elicit complement-dependent cytolysis of neoplastic target cells. In this study we demonstrate that administration of UK 114, either pure or as a crude extract (designated UK 101), inhibits the growth of mammary carcinomas induced in female Sprague-Dawley rats by dimethylbenzanthracene (DMBA). The mechanism of the tumour inhibitory activity of UK 114 is probably related to induction of immunosurveillance.

cDNA cloning and Escherichia coli expression of UK114 tumor antigen

Experimental evidence indicates that the antineoplastic effects of UK101, a goat liver perchloric acid extract, is likely due to one of its constituent proteins: the 14 kDa protein named UK114. The cDNA encoding UK114, obtained by PCR methodologies, contains an open reading frame coding for a protein of 137 amino acids with a theoretical molecular mass of 14298 Da. It shows high sequence homology with a 14 kDa protein identified in human, rat and Mus musculus tissues which is likely involved in the inhibition of cell-free protein synthesis. Northern blot analysis indicated that the transcript is present in variable amounts in a wide range of human tissues. Genomic Southern blots revealed that the UK114 mRNA in goat as well as in human is encoded by a single gene, as is the case in rat. The expression system for UK114 was constructed under the control of the PL promoter from bacteriophage lambda and the cDNA coding region has been highly expressed in Escherichia coli as a thioredoxin fusion protein. The recombinant UK114, purified to homogeneity, is immunoreactive to rabbit antisera prepared against UK101 or native UK114, as well as to sera of UK101-treated cancer patients. It inhibits cell-free protein synthesis at 8 microM concentration.

Perchloric acid-soluble proteins from goat liver inhibit chemical carcinogenesis of Syrian hamster cheek-pouch carcinoma

SummaryChemically induced Syrian hamster cheek-pouch squamous cell carcinoma is very similar to the corresponding human tumour. This paper describes a blind study in which inhibition of dimethylbenzanthracene-induced cheek-pouch tumours by a goat liver extract denominated UK101 was investigated. Less than 40% of animals treated with UK101 developed tumours compared with 100% of the controls. Intermediate results (80%) were noted in a positive control group treated with Calmette–Guérin bacillus. Immunocytochemical testing of cheek-pouch mucosa by Mib5 showed significantly less proliferating cells in UK101 animals than in the controls. The effect of UK101 was completely reversed when dexamethasone was added in a third control group. A significant difference in complement-mediated cytotoxicity was noted in the sera of UK101-tested and control animals. These findings suggest that an immune mechanism is responsible for the inhibition of hamster cheek-pouch carcinoma by UK101.

Prevention and Treatment of Lethal Murine Endotoxemia by the Novel Immunomodulatory Agent MFP-14

ABSTRACT Multifunctional protein 14 (MFP-14) is a ubiquitous protein that inhibits the production of tumor necrosis factor alpha (TNF-α) and gamma interferon (IFN-γ), which are involved in the pathogenesis of sepsis. Here, lipopolysaccharide (LPS)-induced lethality in mice was markedly reduced by MFP-14. The treatment also lowered LPS-induced levels of TNF-α and IFN-γ in the blood.

Immunological and radioimmunological studies of membrane antigen(s) from human breast carcinomas and non-tumoral breast tissues. II

The authors extracted and partially purified a pool of antigens from primary breast carcinomas. The antigens responded to anti-CEA antibody in a radioimmunoassay (R. I. A.) and were not detected in non-tumoral breast tissues used as controls. Antisera were obtained by immunizing rabbits.

Anti-angiogenic properties of calcium trifluoroacetate

Endothelial cell proliferation and the formation of new vessels are strictly regulated by angiogenic factors (e.g., VEGF) that induce the activation of signal transduction pathways controlled by calcium dynamics. Using in vitro and in vivo experiments, we investigated the effect of calcium trifluoroacetate (CaTFAc), a complex, poorly dissociated salt that is characterized by its low toxicity, on angiogenesis. In vitro, CaTFAc inhibited VEGF-induced effects on endothelial cell proliferation. In two in vivo models of angiogenesis, a Matrigel plug in mice and a chick embryo chorioallantoic membrane, CaTFAc inhibited the VEGF-induced formation of new vessels. The exact mechanism of action is still under investigation, but in vitro experiments demonstrate that CaTFAc induced a reversible increase in the levels of intracellular calcium under basal conditions and prevented calcium signaling induced by VEGF. These results are the first to suggest that CaTFAc may be useful for the treatment of diseases caused by enhanced angiogenesis.

Adenocarcinoma Polyclonal Antibody: as a Diagnostic Tool of High Specificity

Publisher Summary Studies have revealed the presence of carcinoembryonic antigen (CEA) not only in the adenocarcinomas but also in normal tissues and biological fluids. CEA extracted from adenocarcinomas is identical to that which is present in non-tumor tissue and in normal human plasma. Many immunological physicochemical parameters express the diversity of CEA from the various cross-reacting antigens that can be extracted from healthy tissues and normal biological fluids. By absorbing the antiCEA antisera with the cross-reacting antigens and in general with tissue and/or plasma extracts, it has been shown that it was not possible to make the antiCEA antisera tumor specific. Experiments show that the success of the absorption of the antiCEA antisera with plasma extracts does not depend on the material employed for the absorption but rather on the choice of the antisera. Only some of the antisera appear to be able to resist absorption, and to reveal a number of tumor specific immunological sites sufficient for diagnosis.

Radioimmunoassay of plasma carcinoembryonic antigen (CEA): Consideration of the results of two methods

SummaryThe two methods used to analyze 384 plasma samples were the carcinoembryonic antigen test (CRT) performed at the Roche laboratories and the direct radioimmunoassay plasma carcinoembryonic antigen (DRPC) technique. The first test, which is quantitative, gave many false positives below the 5 ng/ml level (also when used in a larger series of 4,628 subjects), thus invalidating its use for detection of gastrointestinal neoplasms. The DRPC technique was positive in 62.3 % of 101 cases of gastrointestinal adenocarcinoma and gave very few false positives.