concanavalin-a and Uterine-Cervical-Neoplasms

The concept of Ayurvedic expert guided drug discovery and development is defined and put to test systematically for the first time in literature. Western Science has explored only ~5% of the approximately 25,000 species of higher plants for drug leads. The ancient medical science of Ayurveda has however employed a much larger spectrum of plants for clinical treatment. Clerodendrum viscosum (CV), a commonly growing weed in the Indian subcontinent has been employed by S. Nirmalananda (Ayurvedic expert) for the treatment of cervical cancer. Here we isolate and characterize a water extract fraction (Cv-AP) from the root of CV and evaluate its anticervical cancer cell bioactivity. Our results indicate that Cv-AP possesses pro-apoptotic, anti-proliferative, and anti-migratory activity in a dose-dependent fashion against cervical cancer cell lines. In contrast, primary fibroblasts (control healthy cells), when exposed to similar concentrations of this extract, fail to undergo apoptosis and remain relatively unaffected. These findings suggest that Clerodendrum viscosum (CV) is a readily available source of components with potent anti-cancer activity and selective bioactivity against cervical cancer cells. The major component in CV-AP was identified as a glycoprotein via SDS Page and Concanavalin-A binding studies. This study serves to illustrate that systematic collaboration with Ayurveda is a practical and powerful strategy in drug discovery and development.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Cell Movement; Cell Proliferation; Clerodendrum; Concanavalin A; Dose-Response Relationship, Drug; Drug Discovery; Female; Fibroblasts; Glycoproteins; HeLa Cells; Humans; Medicine, Ayurvedic; Organ Specificity; Plant Extracts; Plant Proteins; Plant Roots; Plants, Medicinal; Primary Cell Culture; Uterine Cervical Neoplasms

Trichomonas vaginalis, a flagellated protozoan parasite causes a variety of adverse health consequences in both men and women. The parasite exists in the trophozoite and the pseudocystic stage. The study reports for the first time that pseudocyst forms of T. vaginalis isolated from cervical neoplasia (CN) patients demonstrated distinct, different and significant in vitro growth profiles when grown in vitro cultures from day 1 up to day 5 (p<0.05, Mann-Whitney test) when compared with the same life cycle stages isolated from non-cervical neoplasia but symptomatic patients (NCN). Pseudocysts from CN and NCN isolates remained viable in distilled water until 3 h 10 min and 2 h 10 min, respectively. The nucleus of pseudocysts in CN isolates using acridine orange and DAPI showed more intense staining revealing higher nuclear content. The FITC-labeled Concanavalin A stained stronger green fluorescence with surface of pseudocysts in CN isolates showing more rough and creased surface with higher numbers of deep micropores with larger numbers of chromatin masses, vacuoles, and hydrogenosomes. The study confirms that pseudocystic stage from CN, despite the uniformity in appearance of being rounded and showing no motility without a true cyst wall under light microscopy, demonstrated different biochemical, surface, and ultrastructural properties. The study provides evidence that phenotypic variant forms of pseudocysts does exist and possibly does play a role in exacerbating cervical cancer.

Topics: Cell Survival; Concanavalin A; Female; Humans; Staining and Labeling; Trichomonas Infections; Trichomonas vaginalis; Uterine Cervical Neoplasms

Endocervical and endometrial tissues were stained with four lectins to determine the difference in staining pattern between non-neoplastic and neoplastic conditions of these tissues.. The lectins used were Ulex europaeus agglutinin (UEA), Dolicho biflorus agglutinin (DBA). Concanavalin A (Con A), and Phaseolus vulgaris agglutinin (PHA). Endocervical tissues included normal endocervical glands, microglandular hyperplasia, minimal deviation adenocarcinoma and endocervical adenocarcinoma, well to poorly differentiated types. Endometrial tissues were collected from normal endometrium, simple glandular hyperplasia, complex hyperplasia, atypical hyperplasia and adenocarcinoma grades 1-3. Non-neoplastic and neoplastic endocervical and endometrial glandular epithelium showed positive reaction for UEA, Con A and PHA. Non-neoplastic glands showed mild to moderate intensity and apical and/polar type of staining pattern for all lectins. Endocervical adenocarcinoma including minimal deviation adenocarcinoma (MDC) and adenocarcinoma well to moderately differentiated type showed diffuse cytoplasmic type of staining pattern for all lectins, but poorly differentiated adenocarcinoma of endocervix showed only a stromal pattern for all lectins. Endometrial hyperplasia and adenocarcinoma grades 1-3 showed positive reaction for all lectins except for DBA. The staining pattern of endometrial hyperplasia was variable, but adenocarcinoma grades 1-3 showed diffuse type.. Intensity and staining patterns of lectins are helpful in distinguishing between endocervical and endometrial non-neoplastic and neoplastic lesions. Intense positive reaction of MDC, especially for Con A and PHA, can differentiate this lesion from normal endocervical glands. The stromal type of staining pattern of poorly differentiated endocervical adenocarcinoma can also have diagnostic significance. Negative reactions of DBA lectin for endometrial adenocarcinoma can be used for differentiating it from endocervical adenocarcinoma.

Topics: Adenocarcinoma; Adult; Aged; Cervix Uteri; Concanavalin A; Diagnosis, Differential; Endometrium; Epithelium; Female; Humans; Hyperplasia; Lectins; Middle Aged; Phytohemagglutinins; Plant Lectins; Staining and Labeling; Uterine Cervical Neoplasms; Uterine Neoplasms

Glycosyl-phosphatidylinositol-anchored hydrophobic placental folate receptors (PFRs), which have an important functional role in maternal-to-fetal transplacental folate transport, can be converted to soluble hydrophilic forms by a placental metalloprotease. Using a Triton X-114 temperature-induced phase separation assay to monitor enzyme-mediated conversion of radiolabeled hydrophobic PFR into hydrophilic PFR, a metalloenzyme was isolated to apparent homogeneity from Triton X-114-solubilized human placenta using concanavalin A-Sepharose and reverse-phase high performance liquid chromatography (HPLC) as major purification steps. The purified hydrophobic enzyme eluted as a single protein peak on reverse-phase HPLC and SDS-polyacrylamide gel electrophoresis revealed a single 63,000 M(r) species, which was reduced to 58,000 M(r) following deglycosylation, findings comparable with amino acid analysis (M(r) approximately 59,000). The metalloenzyme was activated by Mg2+, Zn2+, Mn2+, and Ca2+, optimally at physiologic pH; it also exhibited EDTA-sensitive endoproteolytic cleavage of [3H]leucine-labeled full-length nascent PFR polypeptide generated in vitro in the absence of microsomes. Rabbit polyclonal anti-metalloprotease antiserum specifically immunoprecipitated 125I-metalloprotease and recognized cross-reacting moieties on plasma membranes of normal human hematopoietic progenitor cells and human cervical carcinoma cells, both of which also express FR.

Topics: Amino Acids; Animals; Antibodies; Carrier Proteins; Cations, Divalent; Chromatography, Affinity; Chromatography, Gel; Chromatography, High Pressure Liquid; Concanavalin A; Cross Reactions; Electrophoresis, Polyacrylamide Gel; Enzyme Activation; Female; Folate Receptors, GPI-Anchored; Humans; Kinetics; Metalloendopeptidases; Molecular Weight; Placenta; Pregnancy; Rabbits; Receptors, Cell Surface; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Human chorionic gonadotropin (hCG) is a glycoprotein of which sugar chains are considered to show structural changes with malignancy. To study the sugar chain heterogeneity of urinary hCG in patients with gynecological disease, we employed serial lectin affinity chromatography (LAC) using concanavalin A (Con A) and phytohemagglutinin-E (PHA-E) which can separate N-glycoside-linked sugar chains, and Jacalin lectin which is specific for O-glycoside-linked sugar chains. The proportion of hCG which did not bind to Con A was clearly higher in patients with cervical cancer than in healthy pregnant women. The complex-type sugar chains bearing bisecting (beta 1-4) N-acetylglucosamine which bound to PHA-E increased in the early stage of cervical cancer, and tri- and tetra-antennary complex type sugar chains also increased in the advanced stages. In addition, the Jacalin-bound hCG increased significantly along with the stage of the cancer, especially in advanced cervical cancer with distant metastases. Taken together, these results show that alteration in sugar chain structures of hCG reflect the advanced stage of cervical cancer.

Topics: Carbohydrates; Chorionic Gonadotropin; Chromatography, Affinity; Concanavalin A; Diagnosis, Differential; Endometriosis; Female; Humans; Phytohemagglutinins; Pregnancy; Uterine Cervical Neoplasms; Uterine Neoplasms

The invasive potential of a set of HPV-33- and HPV-33 + ras-transfected cervical keratinocytes was investigated. These cell lines were previously separated into 2 groups according to their behavior on collagen rafts. Cell lines from the first group reconstituted CINIII-like lesions, whereas cell lines from the second group reconstituted epithelia comparable to micro-invasive carcinomas. They were thus postulated to represent distinct stages of cervical carcinogenesis. The present results have shown that lines from group I, which have conserved an epithelial morphology in monolayer, (i) could not invade matrigel when tested in a modified Boyden chamber assay, (ii) produced solely gelatinase B and (iii) were unable to activate exogenous gelatinase A. On the other hand, lines from group II associated epithelial-to-mesenchymal transition (acquisition of elongated morphology, vimentin positivity) with high in vitro invasive potential and with the ability both to produce and to activate gelatinase A. These results strongly support the hypothesis that the epithelial-to-mesenchymal transition and the associated events might be implicated in the progression to the metastatic phenotype.

Topics: Blotting, Northern; Cell Line; Cervix Uteri; Collagen; Concanavalin A; DNA, Viral; Drug Combinations; Enzyme Activation; Epithelium; Female; Gelatin; Gelatinases; Humans; Keratinocytes; Laminin; Matrix Metalloproteinase 2; Mesoderm; Metalloendopeptidases; Neoplasm Invasiveness; Papillomaviridae; Proteoglycans; Transfection; Uterine Cervical Neoplasms

Various immune parameters, i.e., peripheral T and B cell distribution, T cell subpopulation, interleukin 2 (IL-2) activity and in vitro lymphocyte responses to phytohemagglutinin (PHA) and concanavalin A (Con A), were investigated in 24 pretreated patients with cervical intraepithelial neoplasia (CIN) and in 35 normal controls. Of the measured parameters, lymphocyte response to PHA was significantly lower in the CIN than control group (P less than 0.05). The values of all other measured parameters were similar in both groups.

Topics: B-Lymphocytes; Carcinoma in Situ; Concanavalin A; Female; Humans; Immunity, Cellular; Interleukin-2; Leukocyte Count; Phytohemagglutinins; T-Lymphocytes; Uterine Cervical Neoplasms

Glycoproteins from normal and malignant human cervix were studied using an organ culture system and compared by gel electrophoresis and autoradiography. Five glycoproteins of 178 kDa, 95 kDa, 93 kDa, 82 kDa and 38 kDa and 1 glycolipid (46 kDa) were detected more frequently in squamous carcinomas. Certain glycoproteins were shown to be oncofoetal and some had affinity for Concanavalin A (Con A). The 82 kDa glycoprotein was present in 16/17 squamous carcinomas but in only 1/13 normal cervices. This band represented a glycoprotein containing glucosamine, mannose, small quantities of methionine and no fucose. These preliminary results suggest that these glycoproteins and in particular the 82-kDa glycoprotein are worthy of further investigation and characterisation.

Topics: Adenocarcinoma; Adult; Aged; Biopsy; Blotting, Western; Carcinoma, Squamous Cell; Chromatography; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Female; Glycoproteins; Humans; Lectins; Middle Aged; Tumor Cells, Cultured; Uterine Cervical Neoplasms

The lectin binding capacity of the exfoliated epithelial cells of the uterine cervix was investigated with a modified concanavalin A - horse radish peroxidase labelling procedure. The normal healthy females exhibited a distinct rise in the percentage of labelled cells in comparison to those bearing squamous cell carcinoma. In the treated group, the responsive patients again exhibited a higher labelling percentage than the non-responsive patients - showing that the method is of both diagnostic and prognostic significance.

Topics: Adult; Cells, Cultured; Cervix Uteri; Concanavalin A; Female; Follow-Up Studies; Horseradish Peroxidase; Humans; Middle Aged; Peroxidases; Tumor Cells, Cultured; Uterine Cervical Neoplasms

We have investigated the effects of various interferons on the receptors for recombinant tumor necrosis factor-alpha (rTNF-alpha) and also their effects on rTNF-alpha-mediated cytotoxicity on human cervical carcinoma cell line ME-180. Preincubation of cells with interferon (IFN)-gamma causes a concentration- and time-dependent increase in rTNF-alpha receptor number without any change in the affinity constant of the receptors. The increase in receptor number is caused only by IFN-gamma and not by IFN-alpha or IFN-beta. Approximately 4-6 h of preincubation with IFN-gamma are required for maximum increase in rTNF-alpha binding to the cells, and this increase can be abolished by inhibitors of protein synthesis, suggesting de novo synthesis of rTNF-alpha receptors. The half-life of both uninduced and induced receptors of rTNF-alpha is approximately 2 h, indicating a rapid turnover. The binding of rTNF-alpha to the cells can also be eliminated by pretreatment of cells with trypsin. Following the removal of trypsin, binding of rTNF-alpha gradually increases, and this requires the synthesis of new proteins. The cytotoxic effect of rTNF-alpha on ME-180 cells is potentiated severalfold by the addition of either IFN-alpha, -beta, or -gamma. However, at similar concentrations, relatively higher potentiation of rTNF-alpha cytotoxicity is observed with IFN-gamma as compared to IFN-alpha and IFN-beta. The pre-exposure of cells to IFNs is as effective as co-exposure in enhancing cytotoxic effects of TNF-alpha. The induction of TNF-alpha receptors by IFNs is observed in different cell types regardless of their sensitivity to TNF-alpha, suggesting that increase in receptor number alone is not sufficient for the enhanced cytotoxic response. Because the enhancement of cytotoxic effects of TNF-alpha is observed by all IFNs but receptor induction in ME-180 cells occurs only with INF-gamma and because metabolic inhibitors which down-regulate TNF-alpha receptors also enhance cytotoxic response, we suggest that the induction of TNF-alpha receptor by IFNs is not a major mechanism of synergism between these cytokines.

Topics: Cell Line; Cell Survival; Chromatography, High Pressure Liquid; Concanavalin A; Cycloheximide; Dactinomycin; Dose-Response Relationship, Drug; Drug Synergism; Female; Glycoproteins; Humans; Interferon-gamma; Interferons; Receptors, Cell Surface; Receptors, Tumor Necrosis Factor; Recombinant Proteins; Tumor Necrosis Factor-alpha; Uterine Cervical Neoplasms

Topics: Antibodies, Monoclonal; Concanavalin A; Female; Humans; Lymphocyte Activation; Phytohemagglutinins; T-Lymphocytes, Regulatory; Uterine Cervical Neoplasms

The effects of various lectins on the interaction of the human cervical carcinoma cell line ME-180 with recombinant human tumor necrosis factor-alpha (rTNF-alpha) was investigated. rTNF-alpha is known to have cytotoxic effects on this tumor cell line and has been reported to interact with these cells through a single class of specific high affinity receptors (Kd = 0.45 nM; approximately 1790 binding sites/cell). Exposure of cells to concanavalin A (ConA) causes an approximately 2-fold increase in rTNF-alpha receptors without any significant change in their affinity constant (Kd = 0.36 nM; approximately 3662 binding sites/cell). This increase in receptor number is dependent on temperature, the time of exposure and dose of ConA, and does not require the synthesis of new proteins. In spite of an increased binding of rTNF-alpha to cells, the cell killing induced by rTNF-alpha is totally blocked by ConA. Cells are also protected by this lectin from the synergistic cytotoxic effects of rTNF-alpha and recombinant human interferon-gamma. Furthermore, it was also found that ConA decreases the rate of internalization and dramatically inhibits the release and degradation of rTNF-alpha by the cells. These results, overall, demonstrate that ConA increases total number of binding sites for rTNF-alpha but blocks the transduction of the signal for the cytotoxic response.

Topics: Cell Line; Cell Membrane; Cell Survival; Concanavalin A; Dactinomycin; Female; Glycoproteins; Humans; Kinetics; Lectins; Receptors, Cell Surface; Receptors, Tumor Necrosis Factor; Recombinant Proteins; Structure-Activity Relationship; Tumor Necrosis Factor-alpha; Uterine Cervical Neoplasms

For the detection of cancerous and precancerous lesions in cervical cytopathology, the feasibility of a concanavalin A-peroxidase labeling procedure was tested and compared with the Papanicolaou method. To this end, the percentage of labeled flattened epithelial cells with a morphologically normal appearance present in cervical cell suspensions was determined. It was found that the mean labeling percentage of the control group was 71% (SD, 11%). The means for mild, moderate, and severe dysplasia groups were, respectively, 54% (SD, 19%), 48% (SD, 13%), and 44% (SD, 16%). The mean for the carcinoma in situ group was 32% (SD,11%), and for the squamous cell carcinoma group 16% (SD, 5%). It appeared that the labeling percentage gradually decreases with increasing atypia of the epithelium as confirmed by histological observation. A complete dissimilarity was found between healthy individuals and cancer patients. In a follow-up study it was found that the mean labeling percentage did not alter in cases of an unchanged stage of disease. A reestablishment of the normal concanavalin A-peroxidase labeling percentage often appeared once the cancerous or precancerous lesion was treated. In conclusion, the concanavalin A-peroxidase labeling method can be considered as a supplementary technique to the Papanicolaou method for the early detection of cervical cancer. It reduces the effect of sampling and screening errors of the Papanicolaou method, and it allows a more objective cytological diagnosis. In addition, it may possess prognostic significance.

Topics: Carcinoma in Situ; Carcinoma, Squamous Cell; Cell Adhesion; Concanavalin A; Female; Horseradish Peroxidase; Humans; Precancerous Conditions; Uterine Cervical Neoplasms

The lectin binding capacity of the cell surface of normal flattened exfoliated epithelial cells of the uterine cervix was investigated looking for differences between specimens from normal and cancer patients. The method used was a modified concanavalin A-horseradish peroxidase (Con A-HRP) labeling procedure. Both normal and cancer specimens contain labeled as well as unlabeled usual flattened cells. There is a distinct difference between the labeling intensity of labeled and that of unlabeled cells. Quantification of the labeling results has been achieved using a light microscope equipped with a computerized video system. Apparently healthy persons, having a percentage of labeled flattened cells between 54 and 94% (mean = 73%, SD = 10%, N = 40), were totally discriminated by this method from the cancer patients. These patients with a histologically confirmed squamous cell carcinoma, showed a labeling percentage between 10 and 22% (mean = 15%, SD = 4%, N = 10). Hormonal factors, such as phase of cycle and pill use, appeared to have no significant influence. Statistical analysis revealed that at least 99% of all healthy persons will have a labeling percentage above 45%, while at most 1% of the cancer patients will show a labeling percentage above 30%. When choosing the labeling percentage of 45% as critical value, the Con A-HRP labeling might serve as an additional detection method for cancer of the uterine cervix. Moreover, as it is based on the abundantly present normal cells, and not on the often scarce abnormal cells, the method is not liable to sampling and screening errors.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Cervix Uteri; Computers; Concanavalin A; Contraceptives, Oral; Epithelial Cells; Epithelium; Female; Horseradish Peroxidase; Humans; Menstrual Cycle; Microscopy; Microscopy, Electron; Middle Aged; Peroxidases; Statistics as Topic; Uterine Cervical Neoplasms

Concanavalin A-peroxidase (Con A-HRP) labeling of exfoliated cells of the uterine cervix have been shown to possess clinical significance in the detection of cancer. In the present study, a more simple method is used instead of the earlier applied complicated method. These two procedures for the assessment of the labeling results are compared. The first method is an objective machine-aided assessment procedure, consisting of a light microscope connected with a video system used in the sliced mode. The second is a more subjective method using human visual assessment with a light microscope only. The latter method would be suitable for routine use, if it shows similar Con A-HRP labeling results as obtained with the machine-aided procedure. In comparison with the machine-aided assessment procedure, the visual assessment procedure is less accurate. Moreover, the visual assessment is accompanied by intraobserver (between day) and interobserver variations. Although the discriminatory capacity in the detection of cancer patients is significantly lower for the human visual assessment procedure, this difference is small. It is of clinical relevance that in general a complete discrimination of healthy individuals and cancer patients is still possible. Therefore, visual assessment with a light microscope only, is preferred because of its simple equipment, which allows this procedure to be used as a routine method.

Topics: Carcinoma, Squamous Cell; Concanavalin A; Female; Horseradish Peroxidase; Humans; Microscopy; Peroxidases; Uterine Cervical Neoplasms; Vaginal Smears

Using the methods described, it is not possible to determine the number of N- and O-linked oligosaccharides on ectopic hCG beta. On standard hCG beta there are two NeuAc residues on each N- and O-linked oligosaccharide, so that the number of NeuAc residues is proportional to the number of oligosaccharides. Ectopic hCG beta and desialylated ectopic hCG beta are of similar molecular size to the standard preparations (gel filtration and RIA with anti-CTP antisera, data not presented). This suggests that ectopic hCG beta is sialylated to a similar extent as standard hCG beta, so the number of oligosaccharides on ectopic hCG beta could be similar to the number on standard hCG beta. There is a Fuc attached to the N-linked oligosaccharides of standard hCG beta (Fig. 3). Using the methods described, it was not possible to determine if this residue is also found on the N-linked oligosaccharides of ectopic hCG beta. Recently, a second form of ectopic hCG beta was identified (22). This form lacks the characteristic hCG beta carboxyterminal peptide, and as such is unrecognized by the RIA used in this study. Like the ectopic hCG beta described herein, and that produced by other cancers, this molecule only partially binds to Con A, and binds to Ricinus communis-120 following neuraminidase digestion. Intact hCG and free hCG subunits, which only partially bind to Con A, are found in cancer tissues, cancer sera, and the medium of cultured trophoblastic and nontrophoblastic cancer cells (Table 1). Our studies with DoT cancer of the cervix cells clearly indicate that the partial binding could be the consequence of the linkage of extra beta G1cNAc residues.

Topics: Carcinoma, Squamous Cell; Cell Line; Chorionic Gonadotropin; Chorionic Gonadotropin, beta Subunit, Human; Chromatography, Affinity; Concanavalin A; Female; Glycoside Hydrolases; Hormones, Ectopic; Humans; Lectins; Oligosaccharides; Peptide Fragments; Uterine Cervical Neoplasms

Topics: Adolescent; Adult; Carcinoma; Concanavalin A; Female; Humans; Immune Tolerance; Immunoglobulins; Kenya; Lymphocyte Activation; Middle Aged; Monocytes; Phytohemagglutinins; Skin Tests; T-Lymphocytes; Uterine Cervical Neoplasms

T gamma cells in peripheral blood and regional lymph nodes of 9 patients with uterine cervical cancer were studied. The results obtained were as follows: 1) The ratio of T gamma to total T lymphocytes in the peripheral blood was 12.2 +/- 3.2% in patients with uterine cervical cancer, being 11.2 +/- 2.3% at stage I and 14.5 +/- 3.8% at stage II, respectively. This value is not statistically different when compared with that of healthy donors. 2) The ratio of T gamma to total T lymphocytes of regional lymph nodes was 3.2 +/- 1.5%, much less than that of peripheral blood and showing that there were few T gamma cells in regional lymph nodes of patients with uterine cervical cancer. 3) T gamma cells were very few even in the regional lymph nodes with metastasis as well as without metastasis. These results are very interesting in connection with the immunological role of regional lymph nodes, but the significance requires further investigation.

Topics: Concanavalin A; Female; Humans; Lymph Nodes; Lymphocyte Activation; Phytohemagglutinins; Receptors, Fc; Receptors, IgG; T-Lymphocytes; Uterine Cervical Neoplasms

Human peripheral blood lymphocytes were sensitized in vitro against cervical cancer cell lines and the effect of immunostimulants, such as BCG, streptococcal preparation (OK-432), yeast cell wall, and Concanavalin A, on the generation of cytotoxic lymphocytes was examined. The cytotoxic activity of sensitized lymphocytes was augmented by the addition of OK-432 (0.01 KE/ml) during in vitro sensitization, but the induction of sensitized lymphocytes was inhibited by the addition of yeast cell wall (10 micrograms/ml). When the lymphocytes from cervical cancer patients free from tumor after surgical operation were stimulated on the autologous tumor cell monolayer, the cytotoxic lymphocytes could be generated. In vitro primed lymphocytes could be reactivated following the stimulation with OK-432 (0.01 KE/ml) for 48 hr and the requirement for a proliferative trigger seems to be important for the in vitro generation of cytotoxic lymphocytes. The role of nonspecific stimulation by LD-like products in the in vitro generation of cytotoxic lymphocytes was discussed.

Topics: Adjuvants, Immunologic; Antigens, Fungal; Cell Line; Concanavalin A; Cytotoxicity, Immunologic; Female; Humans; Lymphocyte Activation; Lymphocytes; Mycobacterium bovis; Phytohemagglutinins; Picibanil; Uterine Cervical Neoplasms

We have investigated the spleen cell response to Con A in terms of the blastoformation response of lymphocytes, which is known to be an index of the nonspecific immune response of the host organism, in various stages of experimentally induced lesions of the murine uterine cervix. It was found that, under the controlled conditions of the present experiment, suppressor T cells are induced in histological samples exhibiting AT III or severer lesions, which correspond to carcinoma of the uterine cervix. This fact suggests the possibility that antibody synthesis is suppressed. In animals administered the immunopotentiator, ATSO, the development of malignant lesions of the uterine cervix was suppressed, the induction of suppressor T cells was suppressed and the response of spleen cells to Con A was found to decrease.

Topics: Animals; Concanavalin A; Female; Lymphocyte Activation; Mice; Neoplasms, Experimental; Spleen; T-Lymphocytes, Regulatory; Uterine Cervical Neoplasms

Recent studies have demonstrated that ectopic glycoprotein hormones only partially bind Con A. To investigate the basis for these findings, the Con A binding of ectopic hCG beta from DoT cervical carcinoma cells was examined after digestion with various glycosidases. Ectopic hCG beta only partially bound Con A, as was the case after digestion with neuraminidase and beta-galactosidase. However, subsequent digestion with N-acetylhexosaminidase increased Con A binding to 96%. It is apparent that Con A binding of ectopic hCG can be inhibited by a residue removed by N-acetylhexosaminidase, probably extra beta-N-acetylglucosamine linked to beta-mannose on N-linked oligosaccharides. The method used involved the glycosidase digestion of glycoprotein alkylated under denaturing conditions and was first validated with milligram amounts of standard hCG beta.

Topics: Acetylglucosaminidase; beta-Galactosidase; Cell Line; Chorionic Gonadotropin; Chromatography, Affinity; Concanavalin A; Female; Glycoside Hydrolases; Humans; Macromolecular Substances; Methylation; Neuraminidase; Protein Binding; Uterine Cervical Neoplasms

Topics: Adult; Concanavalin A; Female; Fetus; Humans; Leukocyte Count; Lymphocyte Activation; Middle Aged; Neoplasm Staging; T-Lymphocytes; Thymus Extracts; Uterine Cervical Neoplasms

Topics: Adolescent; Adult; Concanavalin A; Female; Humans; Immunity, Cellular; Lymphocytes; Middle Aged; Phytohemagglutinins; Pokeweed Mitogens; Uterine Cervical Neoplasms

Fibroblasts underlying human uterine cervical dysplasia, carcinoma in situ, and invasive carcinoma are agglutinable by concanavalin A (Con A) but not by wheat germ agglutinin, except at very high concentration. Studies with low levels of Con A show that maximal agglutination is obtained with fibroblasts from invasive carcinoma, while the fibroblasts underlying dysplasia give minimal agglutination reactions. Fibroblasts underlying carcinoma in situ give agglutination reactions halfway between those obtained with fibroblasts underlying dysplasia and invasive carcinoma. An epithelial-like cell line obtained from a case of dysplasia shows agglutinability by Con A very similar to that obtained with fibroblasts underlying dysplasia. These epithelial-like cells are also not agglutinable by wheat germ agglutinin. Treatment of the cervical cells, both epithelial and fibroblasts, with neuraminidase leads to slight increase in agglutination by both Con A and wheat germ agglutinin. Marked increase in agglutination is not obtained even after treatment with high concentration of neuraminidase (10 units/10(6) cells). Marked agglutinability, however, is observed after trypsin treatment. The results suggest that, while the fibroblasts obtained from normal cervix are not agglutinable by Con A, surface alterations necessary for Con A-specific agglutination exist in fibroblasts during the early stage of development of uterine cervical epithelial neoplasia (dysplasia) and increase with the progression through carcinoma in situ to invasive carcinoma. Loss of cell surface sialic acids may result in a slight increase in agglutinability, but some other mechanism(s) is likely to be involved in alteration of surface properties that lead to marked agglutinability of the human uterine cervical cells obtained from cancer precursor lesions.

Topics: Agglutination; Carcinoma; Carcinoma in Situ; Cell Line; Cells, Cultured; Concanavalin A; Epithelial Cells; Epithelium; Female; Fibroblasts; Humans; Lectins; Neuraminidase; Precancerous Conditions; Uterine Cervical Neoplasms