concanavalin-a and Lung-Neoplasms

Lung cancer has high morbidity and mortality and small cell lung cancer (SCLC) is a highly invasive malignant tumor with a very unfavorable survival rate. Early diagnosis and treatment can result in better prognosis for the SCLC patients but current diagnostic methods are either invasive or incapable for large-scale screen. Therefore, discovering biomarkers for early diagnosis of SCLC is of importance. In this work, we covalently coupled Concanavalin A (ConA) to functionalized magnetic nanoparticles to obtain magnetic ConA-nanoparticles (ConA-NPs) for the enrichment of glycosylated proteins. We then purified glycosylated proteins in 36 urine samples from 9 healthy controls, 9 SCLC patients, 9 lung adenocarcinoma (LUAD) patients, and 9 lung squamous cell carcinoma (LUSC) patients. The purified glycosylated proteins were digested and analyzed by LC-MS/MS for identification and quantification. Among the 398 identified proteins, 20, 15, and 1 glycosylated protein(s), respectively, were upregulated in the urine of SCLC, LUAD, and LUSC patients. Immunoblotting experiments further demonstrated that cathepsin C and transferrin were significantly upregulated in the ConA-NP purified urine of SCLC patients. This work suggests that glycosylated cathepsin C and transferrin might be able to serve as potential biomarkers for the noninvasive diagnosis of SCLC patients.

Topics: Biomarkers; Biomarkers, Tumor; Chromatography, Liquid; Concanavalin A; Humans; Lung Neoplasms; Magnetic Phenomena; Nanoparticles; Proteomics; Small Cell Lung Carcinoma; Tandem Mass Spectrometry

Malignant melanoma is one of the most dangerous skin cancer originating from melanocytes. Thus, an early and proper melanoma diagnosis influences significantly the therapy efficiency. The melanoma recognition is still difficult, and generally, relies on subjective assessments. In particular, there is a lack of quantitative methods used in melanoma diagnosis and in the monitoring of tumour progression. One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides.

Topics: Biomarkers, Tumor; Biosensing Techniques; Concanavalin A; Glucose; Glycosylation; Gold; Humans; Lung Neoplasms; Mannose; Melanocytes; Melanoma; Microscopy, Atomic Force; Polysaccharides; Quartz Crystal Microbalance Techniques; Surface Properties

Polymyxin B (PmB) belongs to the group of cyclic peptide antibiotics, which neutralize the activity of LPS by binding to lipid A. The aim of this study was to analyze the effect of PmB on the biological activity of lipooligosaccharide (LOS E. coli B,rough form of LPS) in vitro and in experimental metastasis models.. Cultures of murine macrophage J774A.1 cells and murine bone marrow-derived dendritic cells (BM-DC) stimulated in vitro with LOS and supplemented with PmB demonstrated a decrease in inflammatory cytokine production (IL-6, IL-10, TNF-α) and down-regulation of CD40, CD80, CD86 and MHC class II molecule expression. Additionally, PmB suspended in drinking water was given to the C57BL/6 mice seven or five days prior to the intravenous injection of B16 or LLC cells and intraperitoneal application of LOS. This strategy of PmB administration was continued throughout the duration of the experiments (29 or 21 days). In B16 model, statistically significant decrease in the number of metastases in mice treated with PmB and LOS (p<0.01) was found on the 14th day of the experiments, whereas the most intensive changes in surface-antigen expression and ex vivo production of IL-6, IL-1β and TNF-α by peritoneal cells were observed 7 days earlier. By contrast, antigen expression and ex vivo production of IL-6, IL-10, IFN-γ by splenocytes remained relatively high and stable. Statistically significant decrease in LLC metastases number was observed after the application of LOS (p<0.01) and in the group of mice preconditioned by PmB and subsequently treated with LOS (LOS + PmB, p<0.01).. In conclusion, prolonged in vivo application of PmB was not able to neutralize the LOS-induced immune cell activity but its presence in the organism of treated mice was important in modulation of the LOS-mediated response against the development of metastases.

Topics: Administration, Oral; Animals; Carcinoma, Lewis Lung; Concanavalin A; Cytokines; Dendritic Cells; Disease Models, Animal; Escherichia coli; Female; Injections, Intravenous; Lipopolysaccharides; Lung; Lung Neoplasms; Lymphocytes; Macrophages, Peritoneal; Melanoma, Experimental; Mice, Inbred C57BL; Phenotype; Polymyxin B; Spleen

Lung cancer is one of the most important avoidable causes of death around the world, the most widespread carcinoma, with a very poor prognosis, and is the leading cause of cancer death in both developed and developing countries. We report morphological and biological behavior characteristics of a tumor that arose in only one BALB/c mouse of an experimental group treated with urethane, a chemical lung-tumorigenic agent. Morphological and immunochemical analysis indicated phenotypic compatibility with a lung adenocarcinoma. The tumor was named LAC1 (lung adenocarcinoma 1). Implant success in eight LAC1-bearing mice generations was 100%, with a fast evolution (58 survival days) and good metastatic capacity (41% of animals with metastases). The tumor induced a paraneoplastic syndrome characterized by anemia, neutrophilia, cachexia, splenomegaly and thymic atrophy. The lymphoproliferation to Con A was altered in tumor-bearing mice. This lung adenocarcinoma may be a useful experimental model for studying tumor progression, paraneoplastic syndromes and immunology in carcinogenic studies.

Topics: Adenocarcinoma; Animals; Cell Proliferation; Cells, Cultured; Concanavalin A; Erythrocyte Count; Hematocrit; Immunohistochemistry; Leukocytes, Mononuclear; Lung Neoplasms; Mice; Mice, Inbred BALB C; Models, Animal; Neoplasm Transplantation; Neutropenia; Organ Size; Splenomegaly; Thymus Gland; Urethane

Combinations of cellular immune-based therapies with chemotherapy and other antitumour agents may be of significant clinical benefit in the treatment of many forms of cancer. Gamma delta (gammadelta) T cells are of particular interest for use in such combined therapies due to their potent antitumour cytotoxicity and relative ease of generation in vitro. Here, we demonstrate high levels of cytotoxicity against solid tumour-derived cell lines with combination treatment utilizing Vgamma9Vdelta2 T cells, chemotherapeutic agents and the bisphosphonate, zoledronate. Pre-treatment with low concentrations of chemotherapeutic agents or zoledronate sensitized tumour cells to rapid killing by Vgamma9Vdelta2 T cells with levels of cytotoxicity approaching 90%. In addition, zoledronate enhanced the chemotherapy-induced sensitization of tumour cells to Vgamma9Vdelta2 T cell cytotoxicity resulting in almost 100% lysis of tumour targets in some cases. Vgamma9Vdelta2 T cell cytotoxicity was mediated by perforin following TCR-dependent and isoprenoid-mediated recognition of tumour cells. Production of IFN-gamma by Vgamma9Vdelta2 T cells was also induced after exposure to sensitized targets. We conclude that administration of Vgamma9Vdelta2 T cells at suitable intervals after chemotherapy and zoledronate may substantially increase antitumour activities in a range of malignancies.

Topics: Adenocarcinoma; Antineoplastic Agents; Apoptosis; Breast Neoplasms; Burkitt Lymphoma; Carcinoma; Cell Line, Tumor; Cisplatin; Colorectal Neoplasms; Concanavalin A; Cytotoxicity, Immunologic; Diphosphonates; Doxorubicin; Drug Screening Assays, Antitumor; Drug Synergism; Etoposide; Female; Genes, T-Cell Receptor delta; Genes, T-Cell Receptor gamma; Humans; Imidazoles; Interferon-gamma; Lovastatin; Lung Neoplasms; Male; Membrane Glycoproteins; Neoplasms; Perforin; Pore Forming Cytotoxic Proteins; Prostatic Neoplasms; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocyte Subsets; Urinary Bladder Neoplasms; Vincristine; Zoledronic Acid

We have shown that the sera of lung cancer patients affect the response of ConA-stimulated normal peripheral blood mononuclear cells by decreasing the expression of IL-2Ralpha and inhibiting the release of IL-1beta and IL-2. A tendency to enhance the release of IL-6 was also observed. We conclude that an imbalance in the Th1/Th2 cytokine response, typical for cancer patients, may at least partly be related to soluble factors circulating in the patients' blood. We discuss a putative role of serum IL-10, IL-1ra, and soluble IL-2Ralpha in the effects observed.

Topics: Adenocarcinoma; Adult; Aged; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Concanavalin A; Cytokines; Female; Humans; Interleukin-2 Receptor alpha Subunit; Lung Neoplasms; Male; Middle Aged; Monocytes; Receptors, Interleukin; Receptors, Interleukin-2; Serum

We have characterized the regulation of expressed human ecto-ATPase (E-NTPDase 2), a cell surface integral membrane glycoprotein. Ecto-ATPase activity is inhibited by parameters that decrease membrane protein interaction, i.e., detergents and high temperatures. These inhibitory effects are overcome when membranes are pretreated with concanavalin A or chemical cross-linking agents that increase the amounts of ecto-ATPase oligomers. Cross-linking agents also abrogate substrate inactivation of the ecto-ATPase, a unique characteristic of the enzyme. These effects indicate that the magnitude of negative substrate regulation is dependent on quaternary structures of the protein, which likely involves interaction of transmembrane domains. The importance of transmembrane domains of ecto-ATPase in activity modulation is demonstrated further by the stimulatory effect of digitonin, a steroid glycoside that preferentially interacts with cholesterol in the membranes but does not promote oligomer formation. These results indicate that ecto-ATPase activity is regulated by a multitude of mechanisms, some of which may have physiological significance. Ecto-ATPase is also susceptible to transcriptional regulation. Ecto-ATPase gene expression is increased in a human hepatoma whereas it is undetectable in the normal liver.

Topics: Adenosine Triphosphatases; Carcinoma, Hepatocellular; Carcinoma, Small Cell; Cell Membrane; Concanavalin A; Enzyme Activation; Gene Expression Regulation, Enzymologic; Humans; Liver Neoplasms; Lung Neoplasms; Protein Conformation; Protein Structure, Quaternary; Recombinant Proteins; Transcription, Genetic; Tumor Cells, Cultured

The activity principle of the mistletoe (Viscum album L.) phytotherapeutics could be considered as combined cytotoxic and "biological response modifying" activities (increasing host defense against cancer) that result from the activities of the plant lectins and the other biologically relevant substances. We found before that the aqueous extract Isorel, produced by Novipharm GmbH (Pörtschach, Austria) from the entire plant (planta tota) of fresh mistletoe under standardized conditions with bioassay validated batch consistency, can be valuable in experimental adjuvant cancer therapy increasing efficiency of cyclophosphamide chemotherapy. In current study we found that Isorel increases the reactivity of the tumor-bearing mice lymphocytes to the mitogens (ConA and LPS) in vitro, thus indicating its immune stimulating effects for the cancer-immunosuppressed lymphocytes. Moreover, Isorel inhibited the incorporation of 3H-labelled amino acids (protein synthesis) in various malignant cell lines. For the growth inhibition mostly higher MW components were responsible, although even less than 500 Da components were also active. We further analyzed the effects of drug application in vicinity of tumor (murine mammary carcinoma) and compared it with systemic effects. The animals carried mammary carcinoma in both hind limbs and were also injected with tumor cells i.v. to develop artificial lung metastases. Isorel was applied only at the right side (in the limb distal from the tumor) and caused persistent and almost complete inhibition of the tumor growth for 2/7 animals. Anticancer effects were less pronounced on the contralateral side tumors, although tumor growth rate was transiently reduced for some mice. Histology revealed that Isorel treatment, both at the side of tumor and systemically, increased the incidence of apoptosis and necrosis in the tumors, while reduction of mitosis was noticed only for the tumors in vicinity of the tumor exposed to Isorel. Finally, animals treated with Isorel had, on the average, three times less lung metastases than the controls. Thus, we conclude that both local and systemic effects of the application of Isorel could be of benefit for the tumor-bearing organism resulting in immunomodulation combined with tumor growth inhibition and reduction of metastases. According to the in vitro results, antitumorous effects could be the result not only of the mistletoe lectins and the other high MW factors, but also of the very low MW (< 500 Da)

Topics: Adjuvants, Immunologic; Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Concanavalin A; Drug Synergism; Female; Lipopolysaccharides; Lung Neoplasms; Lymphocyte Activation; Lymphocytes; Mammary Neoplasms, Experimental; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Necrosis; Phytotherapy; Plant Extracts; Plant Lectins; Plants, Medicinal; Tumor Cells, Cultured

Nitric oxide (NO) is known to facilitate tumour metastasis through the promotion of angiogenesis, vascular dilation, platelet aggregation, etc. In the present study we explored its novel role in producing dysfunction of the host immune system in the metastasis of murine metastatic melanoma B16-BL6 cells. A significant reduction in the mixed lymphocyte reaction (MLR) was observed in the spleen cells from B16-BL6-bearing mice, but not in those from mice bearing the parent cell B16. When B16-BL6 cells were added in vitro to the MLR, a significant decrease was also found, even when they were co-cultured with the lymphocytes in two compartments of a Transwell chamber separated by an 8.0 microm filter. The supernatant from cultured B16-BL6 but not B16 cells, which had a greatly increased NO activity, significantly inhibited concanavalin A- and lipopolysaccharide-induced lymphocyte proliferation. A remarkably higher expression of inducible NO synthase (iNOS) was detected in B16-BL6 cells than in B16 cells. Nomega-Nitro-l-arginine (l-NNA), a NO synthase inhibitor and superoxide dismutase, significantly antagonized the above inhibition by B16-BL6 cells, while l-arginine, a NO precursor, and S-nitroso-N-acetyl-d,l-penicillamine, a NO donor, strengthened the inhibition. Furthermore, l-NNA significantly inhibited lung metastasis of B16-BL6 cells, while l-arginine tended to enhance the metastasis. The cytotoxicity of B16-BL6-specific T-cells was significantly decreased by pre-culture with B16-BL6 cells in a Transwell chamber or the culture supernatants of B16-BL6 cells, whereas l-iminoethyl-lysine, a selective inhibitor of iNOS, showed a significant recovery from the disease. These results suggest that NO released by metastatic tumour cells may impair the immune system, which facilitates the escape from immunosurveillance and metastasis of tumour cells.

Topics: Animals; Arginine; Blotting, Western; Coculture Techniques; Concanavalin A; Cytotoxicity Tests, Immunologic; Female; Lipopolysaccharides; Lung Neoplasms; Lymphocyte Activation; Male; Melanoma; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; T-Lymphocytes; Tumor Cells, Cultured; Tumor Escape

Mice lacking beta2-microglobulin (beta2m- mice) express greatly reduced levels of MHC class I molecules, and cells from beta2m- mice are therefore highly sensitive to NK cells. However, NK cells from beta2m- mice fail to kill beta2m- normal cells, showing that they are self tolerant. In a first attempt to understand better the basis of this tolerance, we have analyzed more extensively the target cell specificity of beta2m- NK cells. In a comparison between several MHC class I-deficient and positive target cell pairs for sensitivity to beta2m- NK cells, we made the following observations: First, beta2m- NK cells displayed a close to normal ability to kill a panel of MHC class I-deficient tumor cells, despite their nonresponsiveness to beta2m- concanavalin A (Con A)-activated T cell blasts. Secondly, beta2m- NK cells were highly sensitive to MHC class I-mediated inhibition, in fact more so than beta2m+ NK cells. Thirdly beta2m- NK cells were not only tolerant to beta2m- Con A blasts but also to Con A blasts from H-2Kb-/Db- double deficient mice in vitro. We conclude that NK cell tolerance against MHC class I-deficient targets is restricted to nontransformed cells and independent of target cell expression of MHC class I free heavy chains. The enhanced ability of beta2m- NK cells to distinguish between MHC class I-negative and -positive target cells may be explained by increased expression of Ly49 receptors, as described previously. However, the mechanisms for enhanced inhibition by MHC class I molecules appear to be unrelated to self tolerance in beta2m- mice, which may instead operate through mechanisms involving triggering pathways.

Topics: Animals; Antigen Presentation; beta 2-Microglobulin; Concanavalin A; Crosses, Genetic; Cytotoxicity, Immunologic; Genes, MHC Class I; H-2 Antigens; Histocompatibility Antigen H-2D; Interferon-gamma; Killer Cells, Natural; Lung Neoplasms; Lymphocyte Activation; Lymphoma; Mice; Mice, Inbred BALB C; Mice, Knockout; Neoplasm Transplantation; Recombinant Proteins; Self Tolerance; T-Lymphocytes; Tumor Cells, Cultured

The lymphocyte proliferative response to PHA, Con A and PWM was studied in patients with lung tumors (stage I-IV) and other histological types. Disorders in the mitogen-induced lymphocyte activation were identified in more advanced tumors. Certain peculiarities of lymphocyte proliferative activity were found to be determined by histological pattern and localization. A method of evaluation of blasttransformation consequences with due account of lymphocyte functional activity in cancer patients is suggested.

Topics: Adult; Aged; Cell Division; Concanavalin A; Female; Humans; Lectins; Lung Neoplasms; Lymphocyte Activation; Male; Middle Aged; Mitogens

We investigated the expression of differential lambda light chains in human B cell lines secreting immunoglobulin (Ig). When these cell lines were cultured with concanavalin A for a long period of time, a subpopulation of some but not all of these cell lines was induced to express new lambda light chains replacing the original lambda chain (light chain shifting). Production of the new lambda chain, which replaces the original lambda chain, results from a VJ rearrangement at a previously excluded allele and a dramatic reduction of the original lambda chain transcript, although no difference was found in the level of heavy chain transcript. Recombination activating genes RAG-1 and RAG-2, which are normally expressed during specific early stages of lymphocyte development, were expressed in not only the light chain shifting-inducible lines but also in the non-inducible cells. Treatment of these Ig secreting cell lines with dibutyryl cAMP, which is known to enhance expression of the RAG genes, could not induce the creation of new lambda light chain-producing cells from the inducible lines, suggesting that the expression of the two RAG genes is not sufficient for inducing new lambda light chain production. Concanavalin A induced a gradual but significant production lost of the original lambda chain in a subpopulation of the light chain shifting-inducible cells but not in the non-inducible cells. Association of new lambda light chain production with loss of original lambda chain raises the possibility that, when the RAG genes are expressed, concanavalin A may act on a novel intracellular mechanism controlling lambda light chain allelic exclusion in these plasma cell lines.

Topics: Adenocarcinoma; Alleles; B-Lymphocytes; Base Sequence; Burkitt Lymphoma; Cell Line; Concanavalin A; DNA Primers; DNA-Binding Proteins; Flow Cytometry; Gene Rearrangement; Genes, Immunoglobulin; Homeodomain Proteins; Humans; Hybridomas; Immunoglobulin lambda-Chains; Lung Neoplasms; Molecular Sequence Data; Nuclear Proteins; Polymerase Chain Reaction; Protein Biosynthesis; Transcription, Genetic

Tumor cells interact with the hemostatic system in various ways and may thus influence malignant growth and spread. MC28 fibrosarcoma cells possess a potent procoagulant activity (PCA) and form lung tumors following intravenous injection. The aim of this work was to study the relationship between PCA, intravascular coagulation and lung seeding in the MC28 model. MC28 cells were injected into control, warfarinized and heparinized hooded Lister rats. Coagulation changes were monitored by thromboelastography (TEG) and Sonoclot analysis (SA), lung fibrin formation by light and electron microscopy, tumor seeding by macroscopic counting and tumor cell and platelet deposition in the lungs by radiolabelling. PCA was measured by chromogenic assay. MC28 PCA was characterized as a tissue factor-factor VIIa complex that probably arose during cell culture or disaggregation of solid tumors. Injection of tumor cells caused marked coagulopathy and was rapidly (within 30 min) followed by fibrin deposition in the lungs and accumulation of radiolabelled platelets. Heparin and warfarin significantly reduced lung seeding (p < 0.001) and reduced retention of radiolabelled tumor cells in the pulmonary circulation (p < 0.01). Inhibition of cellular PCA by prior treatment with concanavalin A markedly reduced intravascular coagulation and lung seeding. We conclude that MC28 cells cause intravascular coagulation as a direct result of their procoagulant activity. The data suggest that tumor cells form complexes with platelets and fibrin which are retained in the lungs long enough for extravasation and seeding to occur.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Blood Coagulation; Blood Coagulation Tests; Blood Platelets; Cell Adhesion; Concanavalin A; Culture Media, Serum-Free; Cysteine Endopeptidases; Disseminated Intravascular Coagulation; Fibrin; Fibrosarcoma; Heparin; Injections, Intravenous; Lung; Lung Neoplasms; Macrophage Activation; Male; Neoplasm Proteins; Neoplasm Transplantation; Neoplastic Cells, Circulating; Pulmonary Circulation; Rats; T-Lymphocytes; Tumor Cells, Cultured; Warfarin

Analysis of cell surface glycosylation not only provides information about cell properties such as their state of differentiation or histogenetic lineage. The carbohydrate chains also provide potentially functional binding sites to endogenous carbohydrate-binding proteins. This interaction can elicit consequent signalling processes. Because of the importance of neutrophils in the host defence system, we monitored the effect of the binding of such sugar receptors to their cell surface on the release of the enzymatic activities of lysozyme, elastase, and myeloperoxidase. Besides the mannose-binding lectin concanavalin A and the immunomodulatory alpha/beta-galactoside-binding lectin from Viscum album L., three preparations of human sugar receptors - beta-galactoside-binding lectin (M(r) 14 kDa) and two affinity-purified polyclonal IgG fractions from serum with the capacity to recognize alpha- or beta-galactosides, respectively - were used. Two animal lectins from chicken liver and intestine that bind beta-galactosides, as well as the lectin-like human serum amyloid P component, were included in order to assess the importance of slight differences in ligand recognition. Cytochalasin B-enhanced enzyme release was invariably seen with the two plant lectins and the chicken liver beta-galactoside-binding lectin, but the related intestinal lectin did not increase enzyme release. The mammalian homologue of these avian lectins triggered lysozyme secretion, and the lactoside-binding IgG fraction enhanced the amount of extracellular elastase activity slightly but significantly. Thus, the actual lectin, not the nominal specificity of sugar receptors, is crucial for elucidation of responses. Due to the highly stimulatory activity of the two plant lectins, neutrophils from patients with non-cancerous diseases and from patients with lung cancer were monitored for the extent of lectin-mediated enzyme release. Only the concanavalin A-mediated reactivity of the neutrophils was associated with the type of disease.

Topics: Adult; Aged; Aged, 80 and over; Agglutinins; Autoantibodies; Carbohydrate Metabolism; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Concanavalin A; Female; Galactosides; Glycosides; Humans; Immunoglobulin G; Lectins; Leukocyte Elastase; Lung Neoplasms; Male; Middle Aged; Mistletoe; Muramidase; Neutrophils; Pancreatic Elastase; Peroxidase; Plant Lectins; Plant Proteins; Plants, Medicinal; Protein Binding; Serum Amyloid P-Component

Maintaining B16F10 tumor cells in stirring culture for 48 h leads to an increase in lung and liver colonizing capacity in comparison with cells in adherent culture. Parallel to the increased metastatic capacity, we have observed a decrease in the proliferative rate of tumor cells (as the percentage of proliferating cell nuclear antigen-positive cells) and an increase in the population of tumor cells expressing Ia antigen. These results are not exclusive to B16F10 cells, since the same results were obtained when we analyzed 3LL cells maintained in identical culture conditions. In all the tumor lines tested, we found an association between the nonproliferating and the Ia-positive cell populations. We induced Ia expression by treating B16F10 cells in adherent culture with the lectin concanavalin A and again, coincident with an increase in metastatic capacity, we found the same association between the two parameters analyzed--nonproliferating state and Ia antigen expression. In addition, it was found that B16F10 cells induce lymphocytic proliferation, and a direct relationship was established between the number of Ia+ cells and lymphocytic proliferation.

Topics: Animals; Concanavalin A; Culture Media; Histocompatibility Antigens Class II; Liver Neoplasms; Lung Neoplasms; Lymphocyte Activation; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Mitomycins; Proliferating Cell Nuclear Antigen; T-Lymphocytes; Tumor Cells, Cultured

Treatment of squamous cell carcinoma (SCCVII) bearing mice with the immunostimulant schizophyllan (SPG) raised the relative content of Mac-1 positive host cells infiltrating the tumor and increased photofrin retention in these tumors. In vitro colony formation assay following photofrin-based photodynamic therapy (PDT) in vivo revealed a greater killing of tumor cells in the SPG pre-treated group, particularly pronounced when the tumor excision was delayed for 8 h after PDT. The tumor cure rate increased approximately three times when PDT was preceded by the SPG therapy. In contrast, the administration of SPG after PDT was of no benefit for tumor control.

Topics: Animals; Carcinoma, Squamous Cell; Cell Death; Cell Migration Inhibition; Clone Cells; Combined Modality Therapy; Concanavalin A; Cytokines; Disease Models, Animal; Female; Hematoporphyrin Derivative; Immunotherapy; Leukocytes; Lung Neoplasms; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Neoplasm Recurrence, Local; Neoplasm Transplantation; Photochemotherapy; Sizofiran; Tumor Cells, Cultured

Immunomodulating properties of ceruloplasmin were determined in vivo on healthy mice line C57BL/6 and mice with Lewis carcinoma. It was established, that ceruloplasmin at the concentration of 1-40 mg/kg stimulated both T- and B--systems of immunity and prevented immunodepression activity of tumor growth.

Topics: Adjuvants, Immunologic; Animals; B-Lymphocytes; Ceruloplasmin; Concanavalin A; Lung Neoplasms; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Neoplasms, Experimental; T-Lymphocytes; Tumor Cells, Cultured

Lectin binding to tumor cells in tissue sections of nonmetastatic and metastatic murine Lewis lung carcinoma (LLC) was assessed by light and electron microscopy using a lectin-gold technique. Ulex europaeus agglutinin-I (UEA-I) and peanut agglutinin (PNA) showed no binding, whereas concanavalin A (Con A), soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), Maclura pomifera agglutinin (MPA), and Ricinus communis agglutinin-I (RCA-I) bound equally to the transplanted sites and metastases. However, wheat germ agglutinin (WGA) bound to metastases more highly than to the transplanted sites and there was a statistically significant difference (P less than 0.01) between the transplanted sites and metastases with regard to pre-embedding method. The tumor cells binding to WGA clearly decreased in number after sialic acid pretreatment and were rich in more well-differentiated organelle. In the bromodeoxyuridine (BrdUrd) labeling in vivo, cell proliferation was greater in the metastatic sites than in the transplanted sites. The above findings suggest that glycoconjugates on the tumor cell surface are altered in the process of metastasis and correlate with metastatic potential and cell proliferation.

Topics: Animals; Bromodeoxyuridine; Cell Count; Cell Division; Cell Transformation, Neoplastic; Concanavalin A; Histocytochemistry; Immunohistochemistry; Lectins; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Microscopy, Electron; Neoplasm Metastasis; Neoplasms, Experimental; Peanut Agglutinin; Plant Lectins; Tumor Cells, Cultured

Progressive growth of metastatic Lewis lung carcinoma (LLC) tumors results in a concurrent stimulation of myelopoiesis and the appearance of immune-suppressive bone marrow cells. The present study has shown that normal bone marrow cells could be induced to become immune-suppressive by 3 days of culture with supernatants of cloned metastatic LLC-LN7 variant cells. The capacity of the LLC-LN7 supernatants to stimulate the appearance of suppressor cells was directly proportional to the concentration of supernatant used in the bone marrow culture. When adoptively transferred with a LLC-LN7 tumor inoculum, the supernatant-induced suppressor bone marrow cells increased the rate of appearance of palpable tumors and the frequency of tumor establishment. The LLC-LN7 supernatants containing suppressor-cell-inducing activity also had colony-stimulating factor (CSF) activity. The CSF activity produced by the LLC-LN7 cells could be diminished with neutralizing antibodies to either granulocyte/monocyte(GM-) CSF or to interleukin-3 (IL-3). Likewise, the suppressor-inducing activity in the LLC-LN7 supernatants was diminished by pretreatment with anti-GM-CSF or anti-IL-3. The combination of anti-GM-CSF and anti-IL-3 completely neutralized all suppressor-inducing activity produced by the LLC-LN7 cells. These results suggest that the secretion of IL-3 and GM-CSF by LLC-LN7 tumor cells is a mechanism by which the tumors stimulate myelopoiesis and induce normal bone marrow cells to become immune-suppressive. Bone marrow cells that are induced to become immune-suppressive by culture with LLC-LN7 supernatants can, in turn, facilitate the establishment of tumor in vivo.

Topics: Animals; Bone Marrow; Colony-Stimulating Factors; Concanavalin A; Dose-Response Relationship, Drug; Immune Tolerance; In Vitro Techniques; Lewis X Antigen; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; T-Lymphocytes, Regulatory; Tumor Cells, Cultured

The carcinoma SC42 was transplanted into the liver of its syngeneic mice DS, and the immunological integrity of the spleen and the effects of splenectomy on the growth and pulmonary metastasis of the liver tumor were assessed. On day 7 after liver tumor transplantation, the natural killer (NK) activity of the splenocytes was significantly elevated; it subsequently decreased at a later stage of the tumor. The response of the splenocytes to PHA and Con-A decreased significantly from the early stage of the tumor. However, the mixed lymphocyte-tumor cell reaction increased significantly from day 14 to day 28. The survival rate of the mice, which had undergone simultaneous splenectomy and liver tumor transplantation, was significantly lower than that of sham-operated control mice. The number of pulmonary metastases in splenectomized mice was significantly greater than in the control mice. There was, however, no difference between the two groups in the weight of the liver tumor. By contrast, splenectomies performed 14 days before or 14 days after tumor transplantation had no significant influence on the survival of the mice. Splenectomies performed on day 0 and on day 3 after tumor transplantation significantly increased the number of pulmonary metastases. Furthermore, the intravenous injection of anti-asialo GM1 antisera on day 0 and day 3 significantly increased the number of pulmonary metastases, but injection of anti-Thy 1.2 antisera had no effect. These results suggest that splenic NK cells may play an important role in the suppression of pulmonary metastasis at early stages of the liver tumor.

Topics: Animals; Concanavalin A; Killer Cells, Natural; Lectins; Liver Neoplasms, Experimental; Lung Neoplasms; Lymphocytes; Male; Mice; Neoplasm Transplantation; Organ Size; Spleen; Splenectomy

Anti-tumour effector cells were generated through 4 days culture of normal C57BL/6 splenocytes in a medium containing concanavalin A supernatant and then fractionated with Dolichos biflorus lectin (DBA) into DBA+ (agglutinable with DBA) and DBA- (non-agglutinable with DBA) cells. The DBA- cells, infused intravenously into mice together with B16 melanoma cells, or adoptively transferred into mice 3 days after the injection of B16 cells, caused a marked decrease in the number of lung nodules, while the DBA+ cells exerted no effect. On the other hand, the DBA+ cells exhibited higher cytolytic activity in vitro than the DBA- cells in short-term 51Cr-release assays. Then, we analysed the mechanism of the strong anti-tumour activity of DBA- cells in vivo. We found that DBA- cells showed higher response to recombinant interleukin-2 (rIL-2) than DBA+ cells and proliferated very well with a small amount of IL-2. In addition, DBA- cells adhered more strongly to lung endothelial cells than DBA+ cells in response to rIL-1 or rTNF. Furthermore, DBA- cells produced larger amounts of macrophage activating factor (MAF) including IFN-gamma when cultured with B16 melanoma. Taken together, our results show that DBA- cells are effective in reducing experimental pulmonary metastases not only by the direct lytic activity but also by the indirect killing activity through the activated macrophage.

Topics: Animals; Cell Adhesion; Cell Separation; Concanavalin A; Endothelium, Vascular; Female; Immunization, Passive; Interleukin-2; Killer Cells, Lymphokine-Activated; Lectins; Lung Neoplasms; Lymphokines; Macrophage-Activating Factors; Melanoma; Mice; Mice, Inbred Strains; Plant Lectins; Spleen

118 surgical specimens were obtained from patients with lung cancer treated in our hospital during the period of 1977-1983 and tested with carcinoembryonic antigen (CEA), ABO (H) isoantigen and Concanavalin A (ConA). The results showed that the CEA assessment, which give no prognostic significance for long-term survival (P greater than 0.05), would be useful in predicting short-term (6 month) out come (P less than 0.05). All patients with positive ABO (H) test for squamous cell lung carcinoma survived at the end of five years. As for ConA, no definite link has been found with the prognosis.

Topics: ABO Blood-Group System; Adenocarcinoma; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Concanavalin A; Humans; Isoantigens; Lung Neoplasms; Prognosis

The freshly separated indicator cells (suspension of leukocytes) used in humoral leukocyte adherence inhibition test were labeled either with 14C-amino acid mixture or 3H-concanavalin A (3H-ConA). Instead of counting the adherent cells, the amount of 'adherent' radioactivity was measured by a liquid scintillation spectrometer. By the modified method, sera from 25 lung-carcinoma-bearing patients as well as serum samples from 21 healthy persons were examined in the presence of crude antigens prepared from 'normal' lung tissue or lung tumors of various histologic types. Although the results demonstrated high specificity and reproducibility of both methods, the binding of 3H-ConA to the surface of adherent cells is more expressed and assures higher levels of radioactivity.

Topics: Adenocarcinoma; Adult; Aged; Amino Acids; Carcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Concanavalin A; Female; Humans; Immunologic Techniques; Indicators and Reagents; Leukocyte Adherence Inhibition Test; Lung Neoplasms; Male; Middle Aged

Peripheral blood samples from six cancer patients (five colon cancer, one lung cancer) and six healthy volunteers were tested in vitro for oxygen radical production by phagocytic cells and in assays of mitogen-induced lymphoblastogenesis at physiologic and pharmacologic concentrations of pyridoxine (PN, 1.8-96 nmol/ml) or pyridoxal (PL, 0.08-90 nmol/ml). Plasma levels of pyridoxal-5'-phosphate (PLP), 4-pyridoxic acid (4PA), pyridoxamine phosphate (PMP), and PL were also determined. Phagocytic cells from three patients showed significantly increased capacity for oxygen radical production when incubated in PL-, but not PN-supplemented media. Oxygen burst capacity of cells from healthy subjects was significantly enhanced by PN-, but not PL-enriched media. Lymphocyte responsiveness to phytohemagglutinin or pokeweed mitogen (PWM) stimulation showed a modest increase in cell activation in three patients as the concentration of PN was increased; with concanavalin A, two showed enhanced responsiveness. On the other hand, PL-supplementation resulted in greater cell proliferation only with PWM. The cancer patients had significantly lower plasma PLP, 4PA, and PMP levels when compared with the healthy volunteers. These data indicate that in the cancer patients and in a majority of the healthy volunteers, vitamin B-6 status was marginal or deficient and suggest that increasing PN or PL in vivo levels may augment functions related to cell-mediated immunity.

Topics: Adult; Aged; Colonic Neoplasms; Concanavalin A; Female; Free Radicals; Humans; Leukocytes; Lung Neoplasms; Lymphocyte Activation; Male; Middle Aged; Oxygen; Phytohemagglutinins; Pokeweed Mitogens; Pyridoxal; Pyridoxine

A 67-year-old man presented with a pulmonary atypical carcinoid tumor with marked elevation of the serum alpha-fetoprotein (AFP) level to 181,000 ng/ml and no hepatic metastases. Immunohistochemistry revealed AFP-positive fine granules, sparsely distributed in some cells. The proportion of the concanavalin A nonbinding subfraction was 33.7%. Light microscopy revealed hyaline globules within or outside the clear and reticular cytoplasm of a few cells. These were ultrastructurally electron-dense materials similar to the hyaline bodies observed in yolk sac tumors. The Grimelius silver method stained only a few cells and very few cells showed a positive Masson-Fontana reaction. Electron microscopy revealed secretory granules measuring 220 nm on the average in scattered cells. Immunohistochemical studies showed 5-hydroxytryptophan in many cells and 5-hydroxytriptamine or serotonin in only a few cells. As for polypeptide hormones, gastrin was detected and in autopsy specimens carcinoembryonic antigen (CEA) immunoreactive cells were observed. Past case reports on the coexistence of carcinoid tumors and adenocarcinomas in the digestive tract suggest that the tumor cells in our case are also derived from primitive or stem cells of endodermal origin and expressed unusual differentiation in the course of treatment.

Topics: Adenocarcinoma; Aged; alpha-Fetoproteins; Carcinoembryonic Antigen; Carcinoid Tumor; Concanavalin A; Cytoplasmic Granules; Humans; Immunoelectrophoresis, Two-Dimensional; Immunohistochemistry; Lung Neoplasms; Male; Microscopy, Electron

1. alpha 1-acid glycoprotein (AAG) concentration and molecular heterogeneity, and oxprenolol protein binding were studied in serum of 15 healthy volunteers, 14 patients with lung carcinoma and 17 patients with liver cirrhosis. 2. The AAG serum concentration was increased to 180.7% in patients with lung cancer and decreased to 73.4% in cirrhotic patients as compared with controls (P less than 0.05). 3. The concanavalin A (conA) dependent heterogeneity of serum AAG was very similar in controls and patients with lung cancer: a ratio of 9/9/2 was obtained for the conA nonreactive, the conA weakly reactive and the conA strongly reactive subfraction respectively; in cirrhotic patients, the ratio shifted to 11/7/1. 4. The heterogeneity in electric charge, demonstrated by isoelectric focusing, was similar in the three groups of subjects: 70-80% of the focussed bands were found in the main three bands. 5. The binding of oxprenolol to serum proteins was increased in lung tumour patients and decreased in liver cirrhotic patients as compared with controls (P less than 0.05). There was no change in binding affinity and oxprenolol binding was significantly correlated to total AAG serum concentration and to the concentration of each of the conA dependent subtypes, in controls as well as in both patients groups.

Topics: Adult; Blood Proteins; Concanavalin A; Electrophoresis; Female; Humans; Immunoelectrophoresis; Isoelectric Focusing; Liver Cirrhosis; Lung Neoplasms; Male; Middle Aged; Orosomucoid; Oxprenolol; Protein Binding

Human lung lavage proteins were fractionated by centrifugation and molecular sieving. An antiserum to the post-albumin fraction of the soluble proteins reacted with a 10 KD protein and this protein was isolated by conventional chromatography. The protein, which has a pI of 4.8, consists of two 5 KD polypeptides and is rich in glutamic acid, leucine, serine, and aspartic acid amino acids. The protein does not bind to concanavalin A, pancreatic elastase, leukocyte elastase, or trypsin, and lacks anti-protease activity. It constitutes about 0.15% of the soluble proteins in lung lavage. Antibodies to the 10 KD protein specifically and exclusively stain Clara cells in human, dog, and rat. Staining of granules of Clara cells was prominent in the distal bronchioles; however, the non-ciliated cells of respiratory bronchioles did not stain for the 10 KD protein. This 10 KD protein appears in fetal lungs at 21 weeks of gestation, and was present in about 10% of the primary pulmonary adenocarcinomas. As a specific marker for Clara cells, this protein could be useful in the study of development, regulation of secretion, and pathobiology of these cells.

Topics: Aged; Amino Acids; Bronchoalveolar Lavage Fluid; Cadaver; Chromatography, Gel; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Histocytochemistry; Humans; Immunoenzyme Techniques; Lung; Lung Neoplasms; Male; Molecular Weight; Pancreatic Elastase; Protease Inhibitors; Proteins; Tissue Distribution

This article documents a patient with lung carcinoma that produced three oncofetal antigens including alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and human chorionic gonadotropin (hCG). Serum AFP, CEA, and hCG-beta-subunit were extremely high--118,000 ng/ml, 133 ng/ml and 0.9 ng/ml, respectively. Immunohistochemical staining of these tumor markers revealed that these proteins were present in different cells. The pattern of lectin affinity electrophoresis of AFP resembled that of hepatocellular carcinoma. Also investigated was the reactivity of serum CEA to monoclonal antibodies against peptide or sugar moieties. Serum CEA values measured by antipeptide monoclonal antibodies were higher than those measured by antisugar monoclonal antibodies. The demonstration of AFP, CEA, and hCG in different tumor cells suggests that three genomes were not reactivated together in a cell, and the lung carcinoma probably consisted of at least three clones of cancer cells with different phenotypes.

Topics: alpha-Fetoproteins; Antibodies, Monoclonal; Carcinoembryonic Antigen; Carcinoma; Chorionic Gonadotropin; Concanavalin A; Electrophoresis; Female; Histocytochemistry; Humans; Immunoenzyme Techniques; Lung Neoplasms; Middle Aged

Peritoneal adherent cells (PAC) obtained from Propionibacterium granulosum KP-45-treated or Lewis lung carcinoma-bearing BDF1 mice suppressed in vitro the NK-like cytotoxic activity of murine splenocytes against YAC-1 tumor target cells. Maximum inhibition occurred when suppressor and effector cells were preincubated together for 18 h, but the effect was demonstrable also when the two groups of cells were mixed only at the onset of the 4-h cytotoxic assay (i.e. without previous contact). Inhibitory cells appeared to be mostly macrophages, as judged by adherence to plastic and morphologic features, and as little as 5 to 20% of PAC, relative to the total number of co-incubated cells, were required for the clear demonstration of the effect. In addition to activated also normal, resident PAC obtained from untreated animals inhibited the NK cell-mediated cytotoxicity, but the effect was significantly pronounced only when 20% of suppressor cells were incubated overnight with effector splenocytes. The results favor the hypothesis that both functionally activated as well as resting macrophages operate as important regulators of the activity of NK cells in vivo.

Topics: Animals; Concanavalin A; Cytotoxicity, Immunologic; Female; Immunosuppression Therapy; In Vitro Techniques; Killer Cells, Natural; Lung Neoplasms; Macrophages; Mice; Propionibacterium; T-Lymphocytes, Regulatory

This study has examined cells from naturally-occurring murine mammary tumours to ascertain whether cell surface glycoproteins play a significant role in colonisation of the lungs after intravenous inoculation. It was found that gel electrophoretic analysis of membrane extracts and lectin adsorption studies did not reveal any consistent differences in glycoprotein composition of cells from tumours which can heavily colonise the lungs relative to ones from tumours which cannot do so or to cells from pulmonary metastases. Also, alteration of structural and functional properties of surface glycoproteins by treatment with succinylated lectins or with drugs such as tunicamycin and swainsonine, which inhibit glycosylation of membrane proteins, had no specific effects on metastatic colonisation of the lungs. Tunicamycin apparently decreased capability to form experimental metastases but also diminished tumourigenicity on subcutaneous inoculation, although it did not affect tumour cell viability in vitro. This information supports earlier studies from this laboratory involving enzymic digestion of the surface of living tumour cells before inoculation and demonstrates that the pulmonary colonisation capability of these mammary tumour cells can withstand global disorganisation of membrane glycoprotein structure and composition. This implies that either the surface glycoproteins are not important in the colonisation process, or that these tumour cells have great capability for rapid repair of their surfaces. It is concluded that a clear answer to whether surface glycoprotein composition has a decisive role in pulmonary colonisation by these mammary tumour cells requires introduction of stable heritable traits into tumour cell populations by genetic manipulation.

Topics: Alkaloids; Animals; Cell Membrane; Concanavalin A; Glycoproteins; Lung Neoplasms; Mammary Neoplasms, Experimental; Membrane Proteins; Mice; Mice, Inbred C3H; Neoplasm Metastasis; Neoplasm Proteins; Swainsonine; Tunicamycin; Wheat Germ Agglutinins

Serum concentrations of serum amyloid A protein (SAA), peripheral blood lymphocytes (PBL) mitogenic response to phytohemagglutinin (PHA) and Concanavalin A (Con A), numbers of circulating T- and B-lymphocytes and length of survival after diagnosis were measured in 50 patients with cancer of the lung. SAA levels were significantly elevated when compared to 50 control subjects (P less than 0.001), but did not correlate with state of tumor spread at the time of diagnosis. Mitogenic responses of PBL from the cancer patients to PHA and Con A were significantly depressed (P less than 0.001), but also did not predict state of metastatic spread. The percentage of circulating T-lymphocytes was also significantly depressed in cancer patients when compared to controls. In six patients who survived tumor-free for greater than 2.5 years, SAA serum concentrations returned to normal. Statistical analysis showed a significant correlation between SAA serum concentrations and PBL mitogenic response to Con A. In addition, both high SAA concentrations and depressed PBL responses to Con A correlated with shortened survival. Therefore, these parameters may be of value in evaluating prognosis in patients with lung cancer. In addition, serial monitoring of SAA concentrations may be of value in evaluating recurrence or cure of lung cancer.

Topics: Adenocarcinoma; Adult; Aged; Amyloid; Carcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cells, Cultured; Concanavalin A; Humans; Leukocyte Count; Lung Neoplasms; Lymphocytes; Male; Middle Aged; Phytohemagglutinins; Prognosis; Prospective Studies; Serum Amyloid A Protein

Quantitative studies on the binding of concanavalin A (Con A) and wheat-germ agglutinin (WGA) to a series of rat hepatocarcinoma metastatic variants revealed a positive correlation between the amount of cell-surface-bound lectin and lung colonization potential. Scatchard analysis of Con A and WGA binding to 10 individual clones isolated from a subcutaneous (s.c.) tumor transplant and to tumor-cell isolates from 10 individual spontaneous lung metastases from the same animal showed diverse binding characteristics for these cell populations. Nevertheless, the expression of Con A receptor sites accurately predicted the lung colonization potential of 3 isolates from the lung metastases. Higher lectin binding curves were observed for the clones from the subcutaneous tumor than for the isolates from lung metastases. These data suggest that a high Con-A binding potential is indicative of a high lung colonization potential for these hepatocarcinoma cells, but that this phenotype may be rapidly lost during tumor outgrowth in the lungs. The binding of tumor cells to vascular endothelial cell monolayers was inhibited in the presence of Con A; however, no inhibition was observed with 2 other lectins. Attachment of tumor cells to endothelial cell monolayers was also inhibited by the monosaccharides methyl alpha-D-mannopyranoside and N-acetyl-D-galactosamine. Other monosaccharides tested did not alter the attachment of tumor cells to endothelial cell monolayers.

Topics: Animals; Blood Vessels; Carbohydrates; Cell Adhesion; Cell Line; Concanavalin A; Endothelium; Glycopeptides; Iodine Radioisotopes; Lectins; Lung Neoplasms; Mannose; Monosaccharides; Neoplasm Metastasis; Neoplasms, Experimental; Rats; Skin Neoplasms; Wheat Germ Agglutinins

To evaluate the efficacies of various administration routes of the streptococcal preparation, OK-432, we studied the lymphocyte proliferation responses to lectins in patients with malignancies. OK-432 was administered intravenously (I.V.), intramuscularly (I. M.) or intradermally (I.D.) for 2 weeks. Lymphocyte proliferation responses to concanavalin A, PHA and SuM (crude extract of OK-432) were studied before and after OK-432 administration. Enhanced responses were observed in 7 out of 9 patients (77.8%) in the I.V. group compared with 3 out of 7 (44.2%) in the I.M. group and 4 out of 9 (44.4%) in the I.D. group. Ratios of stimulation index (S.I.) after administration over S.I. before administration were highest in the I.V. group. These results suggest that I.V. administration of OK-432 is most effective for stimulating host immune systems.

Topics: Adenocarcinoma; Adult; Aged; Biological Products; Carcinoma, Squamous Cell; Concanavalin A; Female; Humans; Infusions, Parenteral; Lung Neoplasms; Lymphocyte Activation; Male; Middle Aged; Phytohemagglutinins; Picibanil

Topics: Adenocarcinoma; alpha-Fetoproteins; Animals; Carcinoma, Hepatocellular; Concanavalin A; Diagnosis, Differential; Humans; Isoenzymes; L-Lactate Dehydrogenase; Liver Neoplasms; Lung Neoplasms; Mediastinal Neoplasms; Mesonephroma; Mice; Protein Binding

The lectin (PHA and Con A) induced agglutination of a human tumour cell line (lung adenocarcinoma and osteosarcoma) was estimated by the spectrophotometric method. A decrease in the optic density for 1 min (delta D) of cell suspension and the time (t0) necessary for a complete sedimentation of cells (at delta D = 0) were used as quantitative indicators of agglutination. An increase in the concentration of tumour cells and lectins resulted in an increase of delta D and a decrease of t0. The results of spectrophotometry were correlated with the microscopic study data for tumour cells agglutination.

Topics: Adenocarcinoma; Agglutination; Cell Aggregation; Cell Count; Cells, Cultured; Concanavalin A; Humans; Lectins; Lung Neoplasms; Neoplasms; Osteosarcoma; Phytohemagglutinins; Spectrophotometry

The concentration of alpha 1-acid glycoprotein (AGP, orosomucoid) was measured in sera from 19 patients with primary squamous cell carcinoma of the lung, 16 patients with an inflammatory lung disease and 17 persons with normal health. All sera were further subjected to crossed immuno-affinoelectrophoresis with addition of Con A in the first dimension and sugar in the second dimension. The distribution of AGP into four microheterogeneity forms, which were the result of this analysis, was estimated by measuring the area under the precipitation curve. The microheterogeneity patterns of AGP in the three groups were significantly different from each other (p less than 0.001). The total concentration of AGP in the two groups of patients was significantly different from the concentration in the healthy group (p less than 0.001).

Topics: Carcinoma, Squamous Cell; Concanavalin A; Humans; Immunoelectrophoresis, Two-Dimensional; Lung Diseases; Lung Neoplasms; Orosomucoid

The suppressor activity induced by Concanavalin A (Con A) was evaluated in peripheral lymphocytes from 20 patients with solid malignant tumors of different origin, that is: 9 lung epidermoid carcinomas; 6 breast adenocarcinomas and 5 melanomas. Simultaneously 10 normal control subjects were studied. No significant differences in the percentage of suppression were found in patients bearing breast adenocarcinoma and lung epidermoid carcinoma as compared to normal subjects. Melanoma patients, on the contrary, showed significant differences on the same test. On the other hand, the Con A lymphoproliferative response was found to be only significantly increased in the melanoma patients compared to normal controls. No differences were found in the percentage of peripheral blood lymphocytes between patients and controls.

Topics: Adenocarcinoma; Adult; Aged; Breast Neoplasms; Carcinoma, Squamous Cell; Concanavalin A; Humans; In Vitro Techniques; Lung Neoplasms; Lymphocyte Activation; Lymphocytes; Melanoma; Middle Aged; T-Lymphocytes, Regulatory

When the tumor-bearing leg of C57BL/6J mice was amputated 16 days after SC inoculation of 10(6)B16 melanoma cells, all the amputated mice died of pulmonary metastases. Transfer of lungs from the amputated to normal syngeneic mice revealed tumor cells in the lungs just after amputation. Repeated weekly injections of BCG and irradiated tumor cells, beginning 24 h after amputation of the tumor-bearing limb, prolonged the survival only of mice presensitized to BCG. Injections of BCG or irradiated melanoma cells alone, or neuraminidase- and mitomycin C-treated tumor cells or of Levamisole had no effect, but injections of ConA-coated tumor cells slightly prolonged the survival of the amputated mice. Both BCG and B16 cells induced humoral and cell-mediated immunity but there was no cross-reactivity between BCG and B16 cells.

Topics: Animals; Antibody Formation; BCG Vaccine; Concanavalin A; Disease Models, Animal; Immunity, Cellular; Immunization; Immunotherapy; Levamisole; Lung Neoplasms; Melanoma; Mice; Neoplasm Metastasis; Neoplasms, Experimental; Neuraminidase

Proliferation of B lymphocytes is depressed in Lewis lung tumor bearing mice. Treatment of these mice with Wy-18,251 (5 mg/kg) significantly increased the responsiveness of their splenic T cells to concanavalin A in cell culture. Levamisole (5 mg/kg) acted more weakly than Wy-18,251. Neither Wy-18,251 nor levamisole elevated the depressed B cell mitogenesis. Wy-18,251 significantly increased macrophage phagocytosis against 51chromium labeled opsonized chicken red blood cells. Levamisole behaved differently: it either had no effect on phagocytosis, or depressed it.

Topics: Adjuvants, Immunologic; Animals; B-Lymphocytes; Benzimidazoles; Concanavalin A; Levamisole; Lipopolysaccharides; Lung Neoplasms; Lymphocyte Activation; Macrophages; Male; Mice; Phagocytosis; T-Lymphocytes

C57Bl/6 mice bearing a palpable Lewis lung carcinoma (LLC) were immunized against prostaglandin E2 (PGE2) in order to prevent the immune suppression typical of tumor bearers. Proliferation in response to concanavalin A (Con A) by normal spleen cells cultured in the presence of PGE2 or by spleen cells of LLC-bearing mice cultured with Con A alone was suppressed. However, the mitogenic response of spleen cells from PGE2 immunized LLC-bearing mice was only partially suppressed. The random migration of normal macrophages cultured in the presence of PGE2 or of macrophages from LLC-bearing mice cultured in medium alone was enhanced when compared to the random migration of normal macrophages cultured in the absence of PGE2. In contrast, the random migration of macrophages obtained from PGE2 immunized LLC-bearing mice was the same as that of normal macrophages cultured in the absence of PGE2.

Topics: Animals; Cell Movement; Concanavalin A; Dinoprostone; Immune Tolerance; Immunity; Lung Neoplasms; Lymphocyte Activation; Macrophages; Mice; Mice, Inbred C57BL; Neoplasms, Experimental; Prostaglandins E; Spleen

Topics: Adult; Aged; Carcinoembryonic Antigen; Concanavalin A; Female; Humans; Immunosuppression Therapy; Leukocyte Count; Lung Neoplasms; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Phytohemagglutinins

A method is described for using a fluorescein isothiocyanate concanavalin A conjugate to stain human cell membranes in formalin fixed paraffin embedded tissue. 57 neoplastic and normal tissue sites were examined. In 54 malignant tumours, bright green fluorescence was confined to the cell membranes while in 23 benign tumours and normal tissue sites, the membranes were unstained or showed a diminished level of fluorescence. The distinction between malignant and hyperplastic or normal cells was clear cut and definite.

Topics: Breast Neoplasms; Concanavalin A; Female; Fluorescein-5-isothiocyanate; Fluoresceins; Fluorescent Antibody Technique; Humans; Liver Neoplasms; Lung Neoplasms; Neoplasms; Pancreatic Neoplasms; Skin; Skin Neoplasms; Thiocyanates

To clarify immune response of regional lymph nodes in 30 patients with lung cancer, the T cell population, phytohemagglutinin (PHA) and Concanavalin A (ConA) stimulation and the T cell subpopulation bearing Ig-G Fc receptors (Tg cells) were tested concerning the clinical stage, histology and the lymph nodes locations. The T cell population, and PHA and ConA response rates in lymph nodes of stage I patients were grossly the same as those of benign lung diseases, but were statistically higher than those of stage III patients (p less than 0.025, p less than 0.005). Among the three histological types of lung cancer, regional lymph nodes of adenocarcinoma had the greatest T cell populations, and PHA and ConA responses. Concerning the location of lymph nodes, the closer the lymph nodes was to the tumor, the greater was its T cell population, and vice versa. However there was no difference in PHA and ConA responses. All lung cancer patients revealed a significant increase of Tg cells in regional lymph nodes and in their peripheral blood lymphocytes compared with peripheral blood lymphocytes in normal subjects (p less than 0.005).

Topics: Adenocarcinoma; Concanavalin A; Humans; Leukocyte Count; Lung Neoplasms; Lymph Nodes; Lymphocyte Activation; Phytohemagglutinins; T-Lymphocytes

Human leucocyte migration inhibiting factor (LIF) form Con A stimulated lymphoid cells inhibited the migration of human and chicken peripheral blood leucocytes. The evaluation of human LIF activity on chicken cells is easy to perform as a routine test and has the advantage that large quantities of indicator cells are readily available. The chicken cell LIF test can be performed in the absence of serum, so avoiding effects due to plasminogen activator, and does not require the use of concentrated supernatants. LIF activity against both human and chicken targets was absent in supernatants from lung cancer subjects.

Topics: Animals; Chickens; Concanavalin A; Fucose; Humans; Isoflurophate; Leukocyte Migration-Inhibitory Factors; Leukocytes; Lung Neoplasms; Lymphokines

Suppressor cells in spleens of mice bearing the Lewis lung carcinoma (3LL) suppressed the anti-tumor immune response in vivo but not in vitro. The hydrocortisone (HC)-resistant fraction of spleen cells from tumor-bearing mice (TBM) was, on the other hand, suppressive in vitro, due to enrichment for HC-resistant suppressor cells. This cellular fraction suppressed the tumor-induced differentiation of memory cells as well as the tumor-independent in vitro activation of TBM spleen cells. The in vitro lymphoproliferative response of TBM spleen cells to mitogens was also subjected to suppression: these cells showed a lower response to a mitogen such as Concanavalin A (Con A) and were capable of suppressing the response of normal spleen cells to this mitogen. HC treatment of TBM 2 days prior to spleen harvest enriched the splenic population for suppressor cells that suppressed Con A-induced activation.

Topics: Animals; Cell Transformation, Neoplastic; Concanavalin A; Cytotoxicity, Immunologic; Drug Resistance; Hydrocortisone; Lung Neoplasms; Male; Mice; Neoplasms, Experimental; Spleen; T-Lymphocytes; T-Lymphocytes, Regulatory

Indomethacin (prostaglandin synthetase inhibitor) was found to be capable of enhancing the mitogen-induced lymphocyte proliferative responses of healthy subjects and patients with lung cancer. A whole-blood culture technique was used. Indomethacin had no mitogenic activity. We observed a greater enhancement of lymphocyte response by indomethacin in weak responders as compared with strong responders in healthy subjects and lung cancer patients. A greater enhancement was also noted in lung cancer patients with active disease as compared with lung cancer patients in remission. In a separated cell culture system, the indomethacin exerted no effect on purified T cells in the absence of monocytes, while this agent exerted its enhancement effect on T lymphocyte response in the presence of autologous monocytes of lung cancer patients. This suggests that monocytes (suppressor cells) may secrete prostaglandins, which are responsible for the impairment of T lymphocyte response in lung cancer patients.

Topics: Adult; Aged; Carcinoma; Concanavalin A; Dose-Response Relationship, Drug; Female; Humans; Indomethacin; Lung Neoplasms; Lymphocyte Activation; Male; Middle Aged; Phytohemagglutinins; Pokeweed Mitogens; T-Lymphocytes

Topics: Breast Neoplasms; Cell Adhesion; Concanavalin A; Female; Humans; Immunosuppression Therapy; In Vitro Techniques; Lung Neoplasms; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Macrophages; Male; Monocytes; Phytohemagglutinins; T-Lymphocytes

Concanavalin A and wheat germ agglutinin, lectins that interact with serum glycoprotein in a manner similar to the antigen--antibody reaction, were used as "antibodies" in a single radial immunodiffusion technique to test a coded serum panel (from the National Cancer Institute, Bethesda, Md., and the Mayo Clinic, Rochester, Minn.) containing a) 99 serum samples from patients with different types of malignant neoplasms of the gastrointestinal tract, prostate gland, and lung, b) 50 samples from patients with benign diseases of the same organs as those affected in the cancer patients, and c) 50 samples from apparently healthy smokers. The resulting precipitation rings were not correlated to serum protein concentration, and the differences (demonstrated by Student's t-test and with a generalization of the one-sided two-sample Kolmogorov-Smirnov statistic for evaluating diagnostic tests) established that serum glycoproteins are glycosylated differently in cancer patients than in people without cancer.

Topics: Blood Proteins; Concanavalin A; Female; Gastrointestinal Neoplasms; Glycoproteins; Humans; Immunodiffusion; Lectins; Lung Neoplasms; Male; Neoplasm Proteins; Neoplasms; Prostatic Neoplasms

The comparative aspects of cell growth, i.e., [3H]thymidine and [14C]leucine uptake of low-cancer (P4Bis) and high-cancer (P4BisT) cell lines and of their normal counterparts, have been studied in the presence and absence of concanavalin and Robinia lectins. These lectins have similar effects on cell growth, on thymidine and leucine uptake, and on incorporation of these precursors. The growth of normal cells is stimulated by both lectins, whereas the growth of transformed cells is inhibited. In all cases the uptake of both leucine and thymidine by cells is increased by the lectins, but the percentage of incorporation of the precursors is affected in a different manner. The percentage of thymidine incorporated by normal and transformed cells increases or decreases in direct proportion to cell growth; leucine incorporation is not affected significantly. The reversibility of these lectin effects by specific inhibitors shows that cell membranes are implicated in these phenomena. Our study with normal and transformed cells suggests that cell surface may play a role in the process of malignant transformation and that P4Bis cells are "transitory" between PB1 normal cells and P4BisT high-cancer cells.

Topics: Animals; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Lectins; Leucine; Lung; Lung Neoplasms; Mice; Neoplasm Transplantation; Neoplasms, Experimental; Thymidine; Time Factors; Transplantation, Isogeneic

Lymphocyte proliferation (LP) assays were performed in microculture using the T-cell mitogens phytohemagglutinin (PHA) and concanavalin A (Con A); the T + B cell mitogens, pokeweed mitogen (PWM) and staphylococcal phage lysate (SPL); and a pool of allogeneic stimulating leukocytes in one-way mixed leukocyte cultures (MLC) in lung and breast cancer patients and normal individuals. The resultant data were expressed in three different ways: (1) as mean counts per minute (CPM) of tritiated thymidine incorporation; (2) as a stimulation index (SI) and (3) as a relative proliferation index (RPI). The RPI is defined as the ratio of net CPM (nCPM) in experimental cultures with stimulant (E) minus medium control cultures (C) of a test individual to the mean nCPM of three or more normal individuals examined in the same assay on the same day. These expressions were then compared for their ability to discriminate between LP responses in cancer patients and normal individuals. The RPI value and selected cut-off values gave the most sensitive measure for the determination of depressed proliferative responses. These analyses demonstrated that lung carcinoma patients were depressed to PHA (50%), MLC (47%), PWM (43%) and Con A (40%). To a lesser degree, breast carcinoma patients were also depressed to MLC (36%), PHA (31%), PWM (27%) and Con A (19%). Our data indicate that the use of the RPI in the analysis of LP response represents an improved method for detecting impaired response of lymphocytes to general mitogens and alloantigens which can consistently reveal immunosuppression in many cancer patients and may be useful for serial monitoring of individual patients.

Topics: Antigens, Bacterial; Breast Neoplasms; Concanavalin A; Female; Humans; Isoantigens; Lung Neoplasms; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Male; Mitogens

Lymphocyte mitogen transformation has been studied before and after treatment with sugery and radiation in lung cancer patients, in surgically treated patients with non-malignant lesions, and in healthy persons. We have shown a fall in lymphocyte response following operation, even in patients with benign lesions. Patients with cancer have a lymphocyte response which is depressed following operation more than the patients with benign lesions. Following cobalt irradiation there is a further fall in response. We were not able to identify a factor in the serum of patient with depressed lymphocyte transfromation which affected lymphocyte response to Phytohemagglutinin. A recovery in response towards pre-treatment levels occurs earliest in patients with benign lesions. In the patients who had not received irradiation, lymphocyte transformation did not have any prognostic value. In patients with advancing cancer, who ultimately die following irradiation, there is no recovery of response. This lack of recovery signals a very poor prognosis.

Topics: Concanavalin A; Humans; Lectins; Lung Neoplasms; Lymphocyte Activation; Lymphocytes

The responses of lymphocytes to stimulation by three common plant mitogens (PHA, Con A, and PWM) have been studied prior to irradiation treatment in 65 patients with bronchogenic carcinoma. The lymphocyte mitogen stimulation (LMS) responses of these patients were determined to be normal or abnormal based on data obtained from similar studies in healthy volunteers. The data for the patients with lung cancer were analyzed for correlations between the lymphocyte responses and (1) the stage of disease, (2) prognostic significance, and (3) period of survival. Statistically significant correlations were observed between the responses of lymphocytes and the stage of disease and the period of survival. However, this study indicates that these correlated responses will be of limited prognostic clinical value for individual patients.

Topics: Aged; Carcinoma, Bronchogenic; Concanavalin A; Humans; Lectins; Lung Neoplasms; Lymphocyte Activation; Mitogens; Prognosis

The properties of the amylase produced by carcinoma of the lung were studied. The abnormal amylase recognized in the serum of a patient with carcinoma of the lung had a mobility with beta-position and showed reduced migration to the cathodic side after neuraminidase digestion. This abnormal amylase had a close affinity for Concanavalin A and this affinity was not retarded by the neuraminidase digestion. However, the purified, tumor-extracted, amylase from the same patient had the same electrophoretic migration as normal human salivary amylase and was not affected by neuraminidase treatment. The abnormal affinity for Concanavalin A was not observed in this purified tumor-extracted amylase. It is suggested that some transglycosidation steps are needed for the appearance of the abnormal amylase in the patient's serum, and that the terminal sialic acid is independent of the affinity for Concanavalin A. The dissociation constants of the tumor amylase for several substrates were smaller than those of normal pancreatic or salivary amylases. Moreover, maltotriose had no affinity for the tumor-extracted amylase and it was not digested to maltose and glucose by the purified tumor extracted amylase. These differences in the kinetic properties and in the mode of digestion were of interest in the study of tumor-produced amylases.

Topics: Adenocarcinoma; Adult; Amylases; Chromatography, Affinity; Concanavalin A; Humans; Kinetics; Lung Neoplasms; Male; Organ Specificity; Protein Binding; Saliva; Starch

Topics: Animals; B-Lymphocytes; Binding Sites, Antibody; Bone Marrow Cells; Cell Differentiation; Concanavalin A; Immunity, Cellular; Lung Neoplasms; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred C57BL; Mononuclear Phagocyte System; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Radiation Chimera; Rats; Rats, Inbred Lew; Spleen; T-Lymphocytes; Thymectomy; Thymidine; Transplantation, Heterologous; Tritium

Concanavalin A (Con A) injected intraperitoneally at a dose of 50 mg per kg was not lethal for male BALB/c mice. Six hours after administration of 5 mg Con A/kg, the proportioy 24 hr, the proportion of granulocytes had decreased to 56%. Adiministration of 5 mg Con A/kg 24 hr before 200 mg of 5[3,3-bis(2-chloroethyl)-triazeno]-imidazole-4-carboxamide per kg, or 100 mg of 5-fluorouracil per kg resulted in a significant enhancement of lethality. Simulatenous administration of 5 mg Con A/gm and 10 mg of daunomycin per kg also resulted in enhanced lethality. Administration of 5 mg Con A/kg 24 hr before 40 mg of 1,3-bis(2-chloroethyl)-1-nitrosourea per kg, 200 mg of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea per kg, 1000 mg of cytosine arabinoside per kg, 0.1 mg of mithramycin per kg, 2 mg of pactamycin per kg or 1 mg of vincristine per kg did not result in enhanced lethality. Lipid A prepared from Escherichia coli 0127:B8 Boivin lipopolysaccharide has been complexed to Con A. The lipid A-Con A complex (5mg/kg) was no more, or less effective in enhancing the lethality of 5-fluorouracil than 2.5 mg Con A/kg. The lipid A-Con A complex (40 mg/kg), given simultaneously with drug, enhanced lethality per kg. In this regard, the lipid A-Con A complex had vincristine per kg. In this regard, the lipid A-Con A complex had activity comparable to the complex formed between lipid A and bovine serum albumin. Conceivably, Con A can be used to enhance the susceptibility of neoplastic cells to phase-specific antitumor drugs, especially those acting on deoxyribonucleic acid synthesis.

Topics: Animals; Antineoplastic Agents; Concanavalin A; Cyclic AMP; Drug Synergism; Granulocytes; Leukocyte Count; Liver; Lung Neoplasms; Malaria; Male; Mice; Mice, Inbred BALB C; Neoplasms, Experimental; Plasmodium berghei

Syngeneic chimeric (lethally irradiated and reconstituted with syngeneic bone marrow cells) mice manifested an increased resistance to the development of Lewis lung carcinoma. In addition, these mice had a higher response to polyvinylpyrrolidone and a reduced reactivity to T mitogens. The present findings suggest that syngeneic chimeric mice lack suppressor T cells shown to regulate the development of Lewis lung tumor and the response to polyvinylpyrrolidone. Other components of the T cell population, such as helper cells responding to sheep red blood cells or cells involved in allograft rejection, assayed in these syngeneic chimeras were found unaffected. The fact that chimeric mice are deficient in a certain suppressor T cell population whereas other T activities are normal suggests the existence of different cell lines within the T cell population.

Topics: Animals; Antibody Formation; Bone Marrow; Bone Marrow Cells; Concanavalin A; Escherichia coli; Female; Graft Rejection; Hemolytic Plaque Technique; Lectins; Lipopolysaccharides; Lung Neoplasms; Lymphocyte Activation; Lymphocyte Depletion; Mice; Neoplasm Transplantation; Polysaccharides, Bacterial; Radiation Chimera; Skin Transplantation; T-Lymphocytes; Transplantation, Homologous

A strain-specific transplantable melanoma (S-91) growing progressively in DBA/1 mice and metastasizing selectively to the lungs was maintained for 16 days in organ culture before being grafted to syngeneic (DBA/1) and allogeneic (BALB/c and C57BL/6) recipients. The cultured S-91 grew progressively in the syngeneic mice and to a moderate degree in the allogeneic strains; it showed an increased tendency to metastasize in both the DBA/1 and C57BL/6 recipients. Heterophilic cytoagglutination assays of cultured S-91 were less apt to aggregate in the presence of concanavalin A than were their noncultured counterparts, which suggested alteration of the plasma membrane. Organ culture explantation appeared to alter phenotypically the cell-surface membrane and thus increase the cell's ability to metastasize while possibly reducing the immunogenicity of the cultured tumor cells.

Topics: Agglutination Tests; Animals; Cells, Cultured; Concanavalin A; Lung Neoplasms; Melanoma; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred DBA; Neoplasm Metastasis; Neoplasms, Experimental

The immunological response of mice submitted to hydantoin treatment was determined. Hydantoin reduced the absolute number of spleen cells in treated animals and did not modify spleen cells reactivity to concanavalin A, although the response to SRBC challenge, as measured by the Jerne plaque-forming cell technique, was significantly decreased. Following these findings, the influence of hydantoin on carcinogenesis was evaluated by using the model of urethane-induced lung adenomas in SWR mice. Treatment with hydantoin significantly reduced the incidence of the induced adenomas. We confirmed that hydantoin modifies the immune response of the host mainly by depressing its humoral function and we have shown that this effect was associated with an inhibitory effect on tumour induction.

Topics: Adenoma; Animals; Concanavalin A; Female; Immune Adherence Reaction; Immunity; Immunosuppression Therapy; Lung Neoplasms; Lymphocyte Activation; Mice; Phenytoin; Spleen; Urethane

The present work investigates the influence of autosensitized lymphocytes on the carcinogenic response of the host. Urethane treated SWR mice received 6 fortnightly injections of lymphocytes sensitized in vitro against syngeneic fibroblasts. An increased incidence of lung adenomata was found in these mice compared with controls injected with unsensitized lymphoid cells or with lymphoid cells sensitized against unrelated transplantation antigens. Autosensitized lymphocytes also modified the response of host lymphoid cells to concanavalin A or to stimulation in a mixed lymphocyte culture assay. These results indicate that autoimmune lymphocytes may increase susceptibility of a host to the induction of tumours.

Topics: Adenoma; Animals; Autoantibodies; Autoantigens; Concanavalin A; Female; Fibroblasts; Graft vs Host Reaction; Immunity; Lung Neoplasms; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphocytes; Mice; Mice, Inbred BALB C; Mice, Inbred Strains; Neoplasms, Experimental; Spleen; T-Lymphocytes; Transplantation, Autologous; Tritium; Urethane