concanavalin-a and Head-and-Neck-Neoplasms

We evaluated the efficiency of recombinant vaccinia virus expressing interleukin-2 (rvv-IL-2) as a tumor vaccine in an immunocompetent mouse model of head and neck squamous cell carcinoma (SCC VII/SF). Mice with five-day-old tumors in the floor of the mouth were treated with rvv-IL-2 by intratumoral injections. These treated mice survived longer (P <.03) than mice treated with control vaccines. Splenocytes, bone marrow, and lymph node cells from tumor-bearing mice responded poorly to concanavalin A stimulation, suggesting induction of immunosuppression. The rvv-IL-2 virus grew for 7 days in the tumor following intratumoral injection. We did not detect any virus particles in several normal organs following rvv-IL-2 injection. Comparison of expression levels of several potential immune inhibitory mediators between the tumors growing in mice and cultured tumor cells demonstrated higher expression of IL-10, GM-CSF, TGF-beta, and NO synthetase in tumors. These results suggested possible roles for these molecules in immunosuppression. We conclude that rvv-IL-2 has potential as a therapeutic vaccine for head and neck cancer and that it can be more effective provided the immunosuppression is reversed.

Topics: Animals; Bone Marrow; Carcinoma, Squamous Cell; Concanavalin A; DNA Primers; Genetic Therapy; Genetic Vectors; Granulocyte-Macrophage Colony-Stimulating Factor; Head and Neck Neoplasms; Humans; Immunosuppression Therapy; Interleukin-10; Interleukin-2; Lymph Nodes; Mice; Neoplasm Transplantation; Nitric Oxide Synthase; Reverse Transcriptase Polymerase Chain Reaction; Spleen; Survival Rate; T-Lymphocytes; Transforming Growth Factor beta; Vaccinia virus

Seventy newly diagnosed Caucasian male patients with head and neck squamous cell carcinomas (HNSCC) were included in the study. All patients were less than 80 years of age, with no cachexia or auto-immune disease, and they were not taking immuno-active medications. Monocytes from these patients were cultured in vitro and supplemented with autologous serum under ex vivo conditions or cultured with serum-free medium. Comparison was made to monocytes from 59 patients with benign HN diseases. Similar physical activity levels prior to testing as well as a minimum stress load were ensured in both groups. Increased monocyte supernatant levels were determined under ex vivo conditions of interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and tumor necrosis factor (TNF)-alpha, but not of interleukin-12 (IL-12) with endotoxin stimulated monocytes of HNSCC patients compared to control conditions. Increased monokine levels were not present with mononuclear cell cultures stimulated with a T-cell general stimulatory agent or with purified monocytes not specifically stimulated. With endotoxin-stimulated monocytes under in vitro conditions, an increased supernatant was shown for TNF-alpha, but not IL-6. With serum from the different patients cultured with monocytes employed from a healthy control, no difference between the groups was shown in the IL-6 and TNF-alpha response to endotoxin stimulation. The differences in IL-1 beta and TNF-alpha, but not IL-6 levels were differentiated statistically from the smoking and alcohol histories of the patients. These findings suggest that the function of monocytes in general, and thus possibly all mononuclear phagocyte system cells in HNSCC patients, are altered.

Topics: Alcoholism; Carcinoma, Squamous Cell; Cell Movement; Concanavalin A; Head and Neck Neoplasms; Humans; Interleukin-1; Interleukin-12; Interleukin-6; Leukocytes, Mononuclear; Male; Personality Inventory; Retrospective Studies; Smoking; Tobacco Use Disorder; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

The in vitro responsiveness of peripheral blood mononuclear cells (PBMC) T lymphocytes was studied in 81 patients with limited or extended head and neck squamous cell carcinoma (HNSCC), as judged by T, N and T + N stages. Patients included in the study were males below 80 years of age, without auto-immune disease or cachexia, who were not taking any immuno-active medication at the time of diagnosis. The patients were divided into groups according to TNM stage T0-2 vs T3-4, N0-1 vs N2-3 or T + N0-3 vs T + N4-7. When cells from patients with early and late stage, according to T, N or T + N stage, were compared, we found a decreased level of mitogen stimulated T-cells and decreased spontaneous proliferation with increasing disease stage. The same was true if the in vitro mitogenesis of T-cells was analysed separately, depending on the laryngeal or oral cavity/pharyngeal origin of the patients' tumours. If the patients were divided into two groups based on N stage, decreased gamma-interferon, and to some extent interleukin (IL-2), but not IL-4 levels, were found to be related to the disease stage.

Topics: Aged; Analysis of Variance; Carcinoma, Squamous Cell; Cell Division; Concanavalin A; Head and Neck Neoplasms; Humans; Interferon-gamma; Interleukin-2; Interleukin-4; Laryngeal Neoplasms; Lymphocyte Activation; Male; Mitogens; Mouth Neoplasms; Multivariate Analysis; Neoplasm Staging; Pharyngeal Neoplasms; T-Lymphocytes

Immunosuppression in patients with head and neck cancer is well recognized. Previous investigations have demonstrated graded immunosuppression that becomes more pronounced as lymphocyte activity is measured closer to the primary neoplasm. In fresh tumors, a soluble factor has been identified that may partly account for the observed graded immunosuppression.. To examine the effect of soluble factors produced by head and neck sqamous cell carcinoma cell lines on the generation of lymphokine activated killer cell cytotoxicity and peripheral blood lymphocyte proliferation induced by interleukin 2, concanavalin A, and phytohemagglutinin.. Conditioned supernatant fluids were generated in a 4-day incubation period, using 5 head and neck squamous cell carcinoma cell lines, and were assayed for the ability to inhibit the generation of lymphokine activated killer cell cytotoxicity and naive peripheral blood lymphocyte proliferation induced by interleukin 2, concanavalin A, and phytohemagglutinin.. All conditioned supernatant fluids significantly inhibited the generation of lymphokine activated killer cell cytotoxicity relative to controls, and this inhibition was dose dependent. In contrast, supernatant fluids from a myelogenous leukemic tumor cell line, K562, and an ovarian epithelial cell line, SKOV-3, failed to inhibit cytotoxicity. Supernatant fluids conditioned with head and neck squamous cell carcinoma cell lines also profoundly inhibited the naive peripheral blood lymphocyte proliferation induced by interleukin 2, concanavalin A, and phytohemagglutinin.. These studies demonstrate that the cell lines of head and neck squamous cell carcinoma produce soluble factors that inhibit lymphocyte function. Furthermore, these experiments suggest that the inhibition previously observed with enzymatically disaggregated fresh tumors is due to factors produced by the tumor cells rather than by other cells within the tumor matrix.

Topics: Biological Factors; Carcinoma, Squamous Cell; Concanavalin A; Culture Media, Conditioned; Cytotoxicity Tests, Immunologic; Cytotoxicity, Immunologic; Head and Neck Neoplasms; Humans; Immune Tolerance; Interleukin-2; Killer Cells, Lymphokine-Activated; Phytohemagglutinins; Tumor Cells, Cultured

Patients with head and neck cancer often have decreased local or regional immunocompetence. Lymphocytes obtained from tumor-involved or -uninvolved lymph nodes (LNL) of these patients showed low or undetectable levels of antitumor cytotoxicity and low proliferative responses in vitro to interleukin 2 (IL2) or mitogens in comparison to peripheral blood lymphocytes (PBL). Lymphokine-activated killer (LAK) cell activity of LNL was lower (P less than 0.05) than that of autologous PBL. Fresh LNL were neither enriched in cells with the CD8+ CD11b+ "suppressor" phenotype nor did they suppress proliferative or cytotoxic responses of autologous PBL in mixing experiments. LNL did not inhibit LAK cell generation from autologous PBL in the presence of IL2. Also, no evidence for the inhibition of autotumor-restricted responses by IL2-activated LNL was obtained. Spontaneous or in vitro-induced production of IL1 beta. TNF alpha, and IFN-tau was low or undetectable in LNL from tumor-involved and -uninvolved lymph nodes in comparison to that in normal or autologous PBL. Mitogen-induced IL2 production was normal in LNL. The depressed ability to produce certain cytokines may be in part responsible for a state of unresponsiveness present in lymph nodes obtained from patients with head and neck cancer. No evidence for the presence of lymphoid suppressor cell in LNL of these patients was obtained.

Topics: Aged; Carcinoma, Squamous Cell; Concanavalin A; Cytokines; Cytotoxicity, Immunologic; Female; Head and Neck Neoplasms; Humans; Immunophenotyping; Interleukin-2; Killer Cells, Lymphokine-Activated; Killer Cells, Natural; Lymph Nodes; Lymphatic Metastasis; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphocytes; Male; Middle Aged; Phytohemagglutinins; T-Lymphocytes, Regulatory

Approximately two thirds of patients with head and neck cancer have been shown to have peripheral mononuclear cells that exhibit a lowered blastogenic response to the T-cell mitogens, concanavalin A and phytohemagglutinin. To investigate the possible mechanisms of this phenomenon, we measured the amount of activated T-cell lymphokine interleukin-2 present in the supernatant of concanavalin A- or phytohemagglutinin-stimulated peripheral mononuclear cells taken from patients with squamous cell carcinoma of the upper aerodigestive tract. Concentrations were found that were similar to those of healthy subjects. The rate of interleukin-2 consumption and the degree of interleukin-2 receptor expression also were similar for patients and controls. In the course of these experiments, it was noted that differences in blastogenic response between patients and controls were abolished when, 24 hours after the beginning of either concanavalin A or phytohemagglutinin stimulation, the culture supernatant was removed and replaced by fresh medium, containing recombinant interleukin-2 to further sustain cell growth. This suggests that the lower blastogenic response found in patients with head and neck cancer is not due to global immune unresponsiveness, but instead, is caused by selective cell dysfunction(s), which may include the production of a suppressor factor following concanavalin A or phytohemagglutinin stimulation.

Topics: Adult; Aged; Concanavalin A; Female; Flow Cytometry; Head and Neck Neoplasms; Humans; Interleukin-2; Leukocytes, Mononuclear; Lymphocyte Activation; Male; Middle Aged; Phytohemagglutinins; Receptors, Interleukin-2; Suppressor Factors, Immunologic

Altered cellular immunity in patients with advanced head and neck cancer includes impairments in lymphokine production, blastogenesis, in vitro cytotoxicity, and T-cell levels. Recent evidence for the potential importance of in lymphokine interleukin 2 (IL-2) in patients with cancer prompted a study of the kinetics of IL-2 receptor expression on lymphocytes from patients with untreated advanced head and neck cancer and normal subjects and an evaluation of the in vitro effects of the T-cell immune-reconstituting peptide, thymosin alpha 1. Concanavalin A-stimulated IL-2 receptor expression was maximal after 72 hours and was higher in normal subjects than in patients. This was due to lower levels of helper/inducer (CD4) cells expressing IL-2 receptors in the patients compared with the normal subjects. Thymosin alpha 1 further decreased levels of IL-2 receptor-positive (both CD4 and CD8) cells at 48 and at 72 hours. At 96 hours, levels of IL-2 receptor-positive cells and proportions of cells in G2 and M phases of the cell cycle were similar among both groups of subjects. Simultaneous cell kinetic studies indicated that thymosin alpha 1 down regulation of IL-2 receptors was not due to an effect on proliferation and that differences in IL-2 receptor expression at 72 hours among normal subjects and the patients with cancer were more likely related to differences in cell proliferation kinetics.

Topics: Carcinoma, Squamous Cell; CD4 Antigens; Concanavalin A; Down-Regulation; Flow Cytometry; Head and Neck Neoplasms; Humans; Kinetics; Receptors, Interleukin-2; Thymosin

The present study suggests a correlation between concanavalin A-driven blastogenesis and the clinical course of head and neck cancer. Blastogenesis assays were conducted on peripheral blood lymphocytes from controls and from patients with squamous cell carcinoma (SCC) of the head and neck. Our results indicated that 3H-thymidine incorporation in response to concanavalin A and phytohemagglutinin stimulation were significantly lower for patients' than for controls' lymphocytes, whereas PWM stimulation was not statistically different in these two groups. Differences between patients and controls were most notable with concanavalin A stimulation. Five of seventeen patients had a response to concanavalin A stimulation that was in the normal range when expressed as relative to control values. The clinical course of these five patients seems to point to a better prognosis than that of the remaining patients who had below-normal mitogenic responses.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Concanavalin A; Female; Head and Neck Neoplasms; Humans; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Phytohemagglutinins; Prognosis

In earlier experiments chemotactic responsiveness of peripheral blood monocytes obtained from patients with head and neck cancers was found to be markedly depressed. In an attempt to attribute this defect in migration to an influence excited by low molecular weight factors of less than 25,000 daltons, derived from the tumor, Amicon filtrates of head and neck cancer cells were administered subcutaneously to C3H mice 24 hrs. before the intraperitoneal injection of concanavalin A. Subsequent macrophage accumulation into the peritoneal cavity was quantified. A clear inhibition of macrophage infiltration was found, particularly when filtrates of poorly differentiated tumors were used. Injection of filtrates from healthy oral mucosa were negative, whereas mouse mammary carcinoma filtrates strongly inhibited accumulation.

Topics: Adult; Aged; Animals; Carcinoma, Squamous Cell; Cell Extracts; Cell Movement; Concanavalin A; Female; Head and Neck Neoplasms; Humans; Macrophages; Male; Mice; Mice, Inbred C3H; Middle Aged; Molecular Weight; Phytohemagglutinins; Tissue Extracts

This study is, to our knowledge, the first attempt to evaluate cellular immune mechanisms in regional lymph nodes of patients with head and neck cancer. Twenty lymph nodes from eight patients with stage III-IV squamous cell carcinoma were evaluated using an in vitro culture system. The T-cell mitogenic (concanavalin A) response of patients' peripheral blood mononuclear cells was modulated by the addition of cells from regional lymph nodes removed at neck dissection. Modulatory activity showing augmentation was significantly correlated with the size of the primary tumor and histopathologic grade of the tumor. Modulatory activity did not correlate with the histologic pattern of lymph node reactivity. Although these relationships suggest that regional immunity may be important in tumor-host interactions, further study is necessary to establish their biologic and prognostic importance.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Concanavalin A; Female; Head and Neck Neoplasms; Humans; Immunity, Cellular; In Vitro Techniques; Lymph Nodes; Male; Middle Aged; T-Lymphocytes

The effects of the addition of indomethacin to PHA or Con A stimulated lymphocytes from patients with untreated squamous cell carcinoma of the head and neck or from patients with the disease who have just finished irradiation therapy from the disease was quantitated and compared to those of the control group. Lymphocytes from eight of 26 patients with untreated carcinoma were significantly augmented by the addition of indomethacin. The remaining eighteen patients were equal to the controls. For all 17 patients who had just finished extensive field irradiation therapy, significant enhancement of PHA and Con A reactivity by indomethacin was found, which did not appear to be solely a function of low baseline mitogen reactivity. In additional studies, stimulated lymphocytes of irradiated patients were tested for their sensitivity to the inhibitory effect of PGE2. The mitogen treated lymphocytes from all patients that had just finished irradiation therapy were found to be significantly more sensitive to the inhibition by PGE2 as compared to the normal lymphocyte response. This effect was also found not to be related merely to a low PHA or Con A reactivity of the lymphocytes. In both patient groups there was a striking correlation between the percent augmentation of indomethacin and the percent inhibition of PGE2 in that when the percent augmentation values were low so were percent inhibition values and when the degree of augmentation by indomethacin was elevated so was the inhibition by PGE2. This data suggests that increase sensitivity of stimulated lymphocytes to PGE2 may be responsible, at least in part, for the depressed mitogen response and the significant augmentation of this immune response by indomethacin in about 1/3 of the untreated patients with advanced head and neck carcinoma and in those patients who have just finished irradiation therapy. The results of this study support the hypothesis that perhaps patients receiving irradiation therapy may benefit by the oral administration of indomethacin, an approach that needs further consideration.

Topics: Carcinoma, Squamous Cell; Concanavalin A; Head and Neck Neoplasms; Humans; Indomethacin; Lymphocyte Activation; Lymphocytes; Reference Values

The mean number of lymphocytes, response to phytohemagglutinin (PHA), and response to concanavalin A (Con A) in whole-blood cultures for 106 patients with head and neck cancer were 83%, 73%, and 64%, respectively, of values for healthy control individuals. During radiotherapy, lymphocyte counts declined to 44% and PHA and Con A responses declined to about one third of control values. Lymphocyte counts slowly increased after treatment to 77% of control values after two years, but responses to mitogens remained at about 40%. Responses to PHA and Con A for 38 patients who lived beyond 18 months were significantly greater before and after treatment than responses for 39 patients who died within 18 months. In general, a poor pretreatment response to PHA and Con A correlated with a poor clinical course, whereas responses near the control level indicated a good clinical course.

Topics: Carcinoma, Squamous Cell; Concanavalin A; Head and Neck Neoplasms; Humans; Leukocyte Count; Lymphocytes; Neoplasm Staging; Phytohemagglutinins; Pokeweed Mitogens; Prognosis

Regional lymph node lymphocytes from patients with squamous cancer of the head and neck were tested in vitro for their ability to proliferate in response to phytohemagglutinin, concanavalin A, and allogeneic stimuli in one-way mixed lymphocyte culture. Their ability to act as cytotoxic effectors in phytohemagglutinin-dependent cellular cytotoxicity was also evaluated, and all results were compared to normal lymph node or blood lymphocytes. The regional lymph node lymphocytes retained proliferative capabilities equal to those in control lymph nodes or blood, whereas they were unable to mediate phytohemagglutinin-dependent cellular cytotoxicity. However, this was not a tumor-related effect because normal lymph node lymphocytes were also ineffective in this assay. The failure of the regional immune response to control early tumor growth could not be accounted for by generalized nonspecific immunosuppression in regional lymph node lymphocytes, inasmuch as these cells demonstrated normal in vitro activity.

Topics: Adult; Aged; Antigens; Carcinoma, Squamous Cell; Concanavalin A; Cytotoxicity Tests, Immunologic; Head and Neck Neoplasms; Humans; Immunity; Immunity, Cellular; Kinetics; Lectins; Lymph Nodes; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphocytes; Middle Aged; T-Lymphocytes

Patients with mucosal squamous cell carcinoma of the head and neck generally have suppressed T cell function, and this suppression tends to be more pronounced with progression of disease. The prognostic relationships of multiple tests of immune function were analyzed in 183 clinically staged patients. Correlation of immune parameters with prognosis was evident only with the DNCB response, which correlated with recurrence in stages I and II disease (but not in stages III and IV). There was no correlation of any of the in vitro tests of immunofunction with recurrence in any stage. There were no correlations between any of the immune parameters and age or sex nor between DNCB reactivity and any of the in vitro responses.

Topics: Carcinoma, Squamous Cell; Concanavalin A; Dinitrochlorobenzene; Female; Head and Neck Neoplasms; Humans; Immunity, Cellular; Lectins; Leukocyte Count; Lymphocyte Activation; Male; Middle Aged; Mitogens; Neoplasm Staging; Prognosis; Recurrence; Risk; T-Lymphocytes

Ffty-three patients with head and neck cancer tested before radiation treatment to determine numbers of blood lymphocytes and immunologic responses to mitogens of lymphocytes in whole-blood cultures had mean values that were 19% to 26% less than values for healthy individuals. Thirty patients whose disease was in stages III or IV had values similar to those in 23 patients whose disease was in stages I or II. Values for 45 patients tested at end of radiotherapy decreased to about 50% of pretreatment values; however, patients with advanced lesions experienced greater decreases (to 24% to 50%) than patients with localized lesions (to 71% to 84%). Patients with advanced lesions usually received radiation to larger areas than patients with localized lesions; therefore, the extent of decline in laboratory values was most likely dependent on volume of tissue treated.

Topics: Concanavalin A; Head and Neck Neoplasms; Humans; Hydroxyurea; Lectins; Leukocyte Count; Lymphocyte Activation; Lymphocytes; Radiotherapy Dosage

Lymphocytes isolated from the peripheral blood and from tumor tissues of patients with African Burkitt's lymphoma have been studied for cap formation and agglutinability by Concanavalin A (Con A). Peripheral blood from healthy adult persons served as a normal control and blood from patients with carcinoma served as a non-lymphoma control. These studies included 29 patients with Burkitt's lymphoma, 93 with carcinoma, and 105 healthy adult persons, as well as tumor tissues from 13 patients with Burkitt's lymphoma. The great majority of the carcinomas were from the face and neck regions. Lymphocytes from the blood of the majority of patients with Burkitt's lymphoma, as well as those from tumor tissues, exhibited a reduced cap-forming ability (2-6%) and increased Con-A-induced agglutinability compared to lymphocytes from healthy normal donors and from patients with carcinoma, although some of the lymphocytes from patients with carcinoma had a somewhat lower range of cap formation than the lymphocytes from healthy donors. No difference was observed in the interaction with Con A of lymphocytes from the different types of carcinoma studied. Eight lymphoid cell lines were established in our laboratory from the tumor tissues of patients with Burkitt's lymphoma. The cap-forming ability and agglutinability by Con A of these lines was examined and compared to those of the "classical" lymphoma lines: Raji, Daudi and P3HR1. All cell lines exhibited an increased Con-A-induced agglutinability and a reduced cap-forming ability compared to normal lymphocytes, except for P3HR1 cells which exhibited a cap-forming ability of 15-20%. These findings are discussed in relation to the association of the lymphocytes with malignancy and as a possible aid in the differential diagnosis between malignant lymphomas and other diseases.

Topics: Agglutination; Burkitt Lymphoma; Cell Line; Cell Membrane; Concanavalin A; Head and Neck Neoplasms; Humans; Lymphocytes; Lymphoma; Receptors, Drug

Cellular immunity was assessed in patients with operable squamous cell cancer of the head and neck using in vivo skin tests and in vitro lymphocyte stimulation tests. An expansion of a previous study continued to show that 30 per cent of patients with T1N0M0 lesions were DNCB-negative and that with more advanced lesions there was further impairment. A similar finding was observed in the blastogenic response to phytohemagglutinin and concanavalin A but not pokeweed mitogen. Overall, 40 per cent of patients with resectable cancer had a significant depression of the blastogenic responses to conconavalin A and phytohemagglutinin. This depression ranged from 15 per cent in patients with T1N0M0 lesions to 71 per cent in those with T3N0M0 lesions. Although this depression was more severe in patients with palpable cervical node metastases, it was related more to the size of the primary tumor than to the nodes per se. An exception occurred in patients with large fixed nodes in whom the depression of lymphocyte stimulation was most severe. The absolute T-cell count was also depressed in patients with head and neck cancer. This depression parallelled the lymphocyte stimulation results with phytohemagglutinin and conconavalin A and was progressive with increasing stage of disease. A correlation exists between DNCB negativity and early recurrence and shortened survival. Clinical follow-up study is too short to assess the correlation of in vitro immune function with these clinical prognostic factors.

Topics: Alcoholism; B-Lymphocytes; Carcinoma, Squamous Cell; Concanavalin A; Dinitrochlorobenzene; Fluorescent Antibody Technique; Head and Neck Neoplasms; Humans; Immune Adherence Reaction; Immunity, Cellular; Immunologic Deficiency Syndromes; Leukocyte Count; Lymphatic Metastasis; Mitogens; Skin Tests; T-Lymphocytes