The level of IgG antibodies reactive to TF, Tn and alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival.

BACKGROUND: The serum levels of IgG antibodies reactive to glycoconjugates (TF, Tn and αGal) were found to be associated with prognosis of gastrointestinal cancer patients. AIM: To study the relation between the levels of serum antibodies to TF-pAA, Tn-PAA and αGal-PAA polyacrylamide-based glycoconjugates and survival in breast cancer. MATERIALS AND METHODS: The preoperative level of IgG antibodies was analysed in the serum of patients (n = 59) using ELISA with polyacrylamide-glycoconjugates namely, TF-pAA (amide-type), and ethanolamide-conjugates Tn-PAA and αGal-PAA. Survival rate and hazard ratio (HR) were assessed by the Kaplan - Meier method and Cox univariate analysis in different pathomorphological groups. RESULTS: Significantly better survival was observed in patients with an increased level of anti-TF-pAA antibodies both for all patients in total and groups in stages II-III; N1-2 and G3 (p = 0.008-0.021, HR = 0.18-0.23, mean survival time in months 164-186 vs 69-121). A trend to worse survival was observed in increased level of anti-Tn IgG (stages II-III) and anti-αGal IgG (G3): p = 0.075, HR = 2.49 and p = 0.066, HR = 3.27, respectively. CONCLUSION: The method for the determination of circulating anti-TF-pAA IgG may be a useful supplement in long-term prognostic assessment of patients with breast cancer.

The characterization of IgG antibodies to GalNAc beta-terminated glycans of gastric cancer survivors.

UNLABELLED: The elevated anti-GalNAcß IgG level of serum was shown to be associated with the significantly better survival of patients with gastrointestinal cancer. AIM: To characterize the specificity of IgG antibodies to GalNAcß-terminated glycans of long-term gastric cancer survivors. METHODS: Serum antibodies and affinity-isolated antibodies were analysed by the indirect and competitive ELISA using glycan-polyacrylamide (PAA) conjugates as well as by isoelectric focusing and Western blotting. RESULTS: In the serum probes, a partial cross-reactivity of antibodies to GalNAcß, GalNAcß1-3Galß (X2di), GalNAcß1-3GalNAcß (PFdi) and GlcNAcß was observed. The isolated anti-GalNAcß IgGs demonstrated the cross-reactivity to the X2di glycan mainly. The affinity of the X2di-PAA to anti-GalNAcß IgGs was 11-21 times lower than that of the GalNAcß-PAA. Anti-X2di and anti-PFdi IgGs demonstrated monoreactivity to their key glycans-PAA used in isolation. The IC50 values of key glycoconjugates ranged from 1 to 5 · 10(-7) M. No polyreactivity of antibodies to the unrelated antigens (ferritin, casein and DNA) was found. The polyclonal or oligoclonal distribution of IgG bands was established and the monoreactivity of antibodies was not associated with the clonal distribution of bands. CONCLUSION: The cross-reactivity of anti-GalNAcß antibodies to X2di and related glycans deserves attention in the clarification of the role of antibodies in cancer progression and enhancement of the prognostic potential in the combined determination of antibody markers.

The level of anti-(GalNAc beta) and anti-para-Forssman disaccharide IgG antibodies in patients with gastrointestinal cancer: relation to survival.

UNLABELLED: Serum anti-(GalNAcß) and anti-para-Forssman disaccharide (PF(di), GalNAcß1--3GalNAcß) IgG levels were earlier found to be related to histological grading and progression of gastrointestinal cancer. AIM: To study the relation of serum antibodies level to survival in patients with gastrointestinal cancer. METHODS: The level of anti-GalNAcß, and PF(di) IgG was analysed in the serum of patients with gastric (n = 78) and colorectal (n = 48) cancers in the long-term follow-up using ELISA with polyacrylamide glycoconjugates. Survival rate and hazard ratio (HR) were assessed by the Kaplan -- Meier method and Cox univariate analysis in different patho-morphological groups. RESULTS: Better survival was observed in patients with an increased preoperative level of GalNAcß antibodies. These were the gastrointestinal group in stages II, III or tumors T2--4 (n = 90--104, P = 0.007, HR = 0.48--0.49, 95% CI 0.27--0.83, and the group with gastric cancer in stages I, II (n = 49, P = 0.051, HR = 0.39, 95% CI 0.14-1.04). The survival time was significantly longer in the gastrointestinal group in patients whose GalNAcß antibodies level rose in dynamics (stage III or N1--2: p = 0.031--0.039, HR = 0.29--0.31, 95% CI 0.09--1.00). No significant difference in survival of patients was observed in the evaluation of PF(di) antibodies. We suggest that the level of antibodies and its change reflect the enteric microbiota colonization, which may influence cancer progression via different interrelations between microbiota, the tumor and immune system. CONCLUSION: The preoperative level of GalNAcß antibodies and its dynamics may be of prognostic significance for clinical outcome assessment.

The relation of serum anti-(GalNAc beta) and -para-Forssman disaccharide IgG levels to the progression and histological grading of gastrointestinal cancer.

UNLABELLED: Earlier we found two unusual IgG-antibody specificities to GalNAc beta and GalNAc beta1-3GalNAc beta (para-Forssman disaccharide, PFdi) carbohydrate ligands in human serum. The aim of the study was to evaluate whether elevated antibody levels are related to the progression of gastrointestinal cancer and the histopathological grading. METHODS: Specific IgG levels were tested in 159 patients with gastric cancer, 88 patients with colorectal cancer and 96 blood donors by the ELISA using synthetic polyacrylaamide (PAA) conjugates, GalNAc beta-PAA and PFdi-PAA. Biochemical and haematological analyses were performed using automatic equipment. RESULTS: The anti-PFdi IgG levels were significantly higher in patients with gastric and colorectal cancer than in donors: in stages II-IV, P = 0.0002 - 0.04 (U-test). The elevated anti-PFdi IgG level was associated with the advanced gastric cancer: in stages II, III, IV vs stage I (P = 0.004 - 0.06) and in case of the tumor size T2 + T3 vs T1 (stages I, II; P = 0.03). Differences in anti-GalNAc beta IgG level were insignificant. No relation between antibody levels and the regional and distant metastases of gastric or colorectal cancer was found. The lower anti-GalNAc beta IgG level was associated with lower-differentiated carcinomas (P = 0.01 - 0.04). Prolonged postoperative changes in the levels of both antibodies during the follow-up were established. An elevation of both antibody levels in patients with gastrointestinal cancer was revealed after a surgical removal of G3-tumors (P = 0.003 - 0.01). The anti-PFdi IgG levels correlated with the levels of the C-reactive protein: r = 0.50, P = 0.003. The anti-GalNAc beta IgG levels correlated with the percentage of peripheral blood monocytes: r = 0.42, P = 0.002. CONCLUSION: The association of the anti-PFdi IgG level with cancer progression suggests the implication of antibodies in the pathogenesis of gastrointestinal cancer. Further studies are required to identify natural targets of antibodies, their relation to other diseases, prognostic significance in cancer.

The isolation and characterization of human natural alphaGal-specific IgG antibodies applicable to the detection of alphaGal-glycosphingolipids.

The Galalpha1-3Galbeta (alphaGal) hapten is xenogeneic for humans; natural anti-alphaGal antibodies are present in human serum. To study the possible abnormal expression of the alphaGal in humans and the pathophysiological role of antibodies, the method of affinity purification of human anti-alphaGal IgG was developed. The specificity of antibodies was evaluated using polyacrylamide (PAA)-based glycoconjugates in direct and competitive enzyme-linked immunosorbent assays (ELISA). The purified antibodies exhibited alphaGal-restricted specificity. The IC50 value for alphaGal-PAA was equal to 4 x 10(-8) M. In a competitive assay, the Galalpha1-3(Fucalpha1-2)Galbeta-PAA (trisaccharide of blood group B) was found to be one hundred times less active inhibitor than alphaGal-PAA. The multivalent alphaGal-PAA was 1100 times more potent an inhibitor than the monovalent spacered alphaGal-saccharide. The antibodies did not show any reactivity to the negatively charged antigens (DNA, human tumor-derived mucins). At a concentration of 2 microg/mL, the antibodies agglutinated rabbit erythrocytes but not hare erythrocytes. The high reactivity of antibodies to the alphaGal-glycosphingolipids of rabbit erythrocytes and the pig kidney was shown by a modified sensitive method of thin-layer chromatography with immunodetection.

Specificity of human anti-carbohydrate IgG antibodies as probed with polyacrylamide-based glycoconjugates.

The TF, Tn, and SiaTn glycotopes are frequently expressed in cancer-associated mucins. Antibodies to these glycotopes were found in human serum. A set of polyacrylamide (PAA)--based glycoconjugates was applied to the direct and competitive enzyme-linked immunosorbent assays (ELISA) to characterize the specificity of serum IgG antibodies. The anti-TF, -Tn and -SiaTn IgG were affinity purified from serum of cancer patients and characterized using PAA-conjugates and free saccharides. The anti-TF and -Tn antibodies were shown to be specific. The anti-TF IgG bound both Galbeta1-3GalNAcalpha- and Galbeta1-3GalNAcbeta-PAA, the latter was three-four times more effective inhibitor of antibody binding. The anti-Tn IgG reacted only with GalNAcalpha-PAA. The anti-SiaTn IgG cross-reacted with Tn-PAA but SiaTn-PAA was five-six times more effective inhibitor in a competitive assay. The IC50 values for PAA-conjugates with the corresponding antibodies typically ranged from 2 to 5 x 10(-8) M. The antibodies display a low specificity to mucin-type glycoconjugates in comparison with PAA-conjugates as was shown for mucins isolated from human malignant tumor tissues, ovine submaxillary mucin (OSM) and asialo-OSM. The unusual IgG-antibody specificity to GalNAcbeta and GalNAcbeta1-3GalNAcbeta ligands was found in human serum.

[ELISA of IgM antibodies to Thomsen-Friedenreich (TF) hapten in cancer diagnostics: comparison of data obtained with four TF-glycoconjugates]. / Immunofermentnyi analiz IgM antitel k gaptenu Tomsena-Fridenreikha (TF) v onkodiagnostike. Sravnenie dannykh, poluchennykh s pomoshch'iu chetyrekh TF-glikokon"iugatov.

The level of IgM antibodies to the Thomsen-Friedenreich hapten (TF) relative to the total IgM level in the blood sera of gastric and breast carcinoma patients and healthy persons was determined using enzyme-linked immunosorbent assay. The following TF glycoconjugates were tested: TF-polyacrylamide (PAA)(with 10 mol.% of TF hapten Gal beta 1-3GalNAc alpha 1-O(CH2)3NH per number of monomeric units in the polyacrylamide), TF-human serum albumin (HSA)(Gal beta 1-3GalNAc alpha 1-O-p-C6H4-HSA containing approximately 15 carbohydrate residues per HSA molecule), asialo-kappa-caseinoglycopeptide, and asialoglycophorin. The total IgM level was determined using antibodies to the mu-chain of human IgM. The statistically significant difference between cancer patients and healthy donors was revealed with two conjugates: TF-PAA and TF-HSA. In the case of TF-PPA, the sensitivity of the assay was 75-83%, and the specificity was 77%. Thus, TF-PAA is the most suitable conjugate for measuring the level of serum anti-TF-IgM antibodies.

The complex of alpha 2-macroglobulin with CD2 in the plasma of gastric carcinoma patients.

alpha 2-macroglobulin (alpha 2M) preparations purified from the plasma of healthy persons and gastric carcinoma patients were further analysed by immunochemical techniques. It was found that only alpha 2M from the plasma of gastric carcinoma patients gave a positive reaction with E-receptor sheep antiserum in rocket immunoelectrophoresis. The alpha 2M fraction giving the positive reaction with E-receptor sheep antiserum was isolated and analysed by the immunoblotting technique with monoclonal antibodies to T11 antigen. The presence of the alpha 2M-CD2 complex in the plasma of gastric carcinoma patients was demonstrated. This finding may explain the immunosuppressive effect of the plasma alpha 2M in cancer patients.

[The dynamics of the kallikrein-kinin system indices in patients with eczema vera during treatment by the laser photophoresis method with ortofen]. / Dinamika pokazatelei kallikrein-kininovoi sistemy u bol'nykh istinnoi ékzemoi v protsesse lecheniia metodom lazernogo fotoforeza s ortofenom.

A drastic increase (by 30.6 times) vs. the norm) of kallikrein activity was revealed in 70 patients with true eczema during exacerbation; blood plasma amidase activity was elevated 4.4-fold, serum alpha 2 macroglobulin antiprotease activity 1.4-fold. Laser photophoresis combined with application of a new Soviet nonsteroid anti-inflammatory agent orthofen reduced the parameters of the kallikrein-kinin system, and a tendency to their normalization could be observed.

[Experience in using laser photophoresis in dermatological practice]. / Opyt primeneniia lazernogo fotoforeza v dermatologicheskoi praktike.

A rationale is given for the use of a new physiotherapeutic method for the treatment of dermatologic patients: laser photophoresis. Experiments have demonstrated a better diffusion of the drug during photophoresis than through semipermeable membranes. The method has been employed in therapy of 27 patients with chronic eczemas with ortafen, a new Soviet anti-inflammatory nonsteroid drug.

[Characteristics of the carbohydrate structure of alpha 2-macroglobulin in stomach cancer]. / Osobennosti uglevodnoi struktury al'fa-2-makroglobulina pri rake zheludka.

A comparative study was made for alpha 2M preparations obtained from plasma of patients with gastric carcinoma and healthy individuals. No significant differences were found in trypsin-binding activity of alpha 2M, in isoelectric focusing microheterogeneity and in binding with lectins of Limulus polyphemus hemolymph. The study of alpha 2M-phytohemagglutinin (PHA) binding in sera of patients with gastrointestinal carcinoma and healthy people revealed highly significant differences. The alpha 2M preparations from the gastric carcinoma patients had less affinity for PHA. The data suggest the existence of an abnormal carbohydrate structure of alpha 2M in gastric carcinoma patients.