Role of levamisole immunotherapy as an adjuvant to radiotherapy in oral cancer--immune responses

Investigations were carried out to assess the effect of levamisole immunotherapy as an adjuvant to radiotherapy on the immune response of patients with squamous cell carcinoma of the oral cavity. Parameters assessed were leukocyte migration inhibition, response to PPD and oral cancer extract (OCA), lymphocyte transformation to PHA, circulating antibodies to OCA and circulating immune complexes (CIC). Comparisons were made between groups receiving levamisole (L), those receiving placebo (P) and normal controls. The results of a thirty-month follow-up are presented. Radiotherapy resulted in a depression of cell-mediated functions, reduction in antibody titer also showed a gradual increase with time of follow-up. Levamisole, however, appeared to reduce the levels of CIC.     

Lymphocyte subpopulations in an experimental model of axial and peripheral enthesiopathies.

We report a study of lymphocyte subsets in experimental arthritis induced in Wistar Furth rats by native human type II collagen and muramyl dipeptide. This experimental arthritis shares similarities with both the spondyloarthropathies and rheumatoid arthritis. Peripheral blood T lymphocytes, primarily the CD4+ cells (p = 0.01), were lower in arthritic rats than in the controls, although the difference in the CD4/CD8 ratio was not statistically significant. Splenic CD4 cells were significantly (p = 0.03) more numerous in arthritic rats, while the numbers of MHC class II positive cells (p = 0.002) and kappa-bearing B-cells (p = 0.0004) were significantly lower. Determination of peripheral blood and spleen lymphocytes subsets could therefore be used for the assessment of arthritis and for the evaluation of therapeutic agents. Thymic T-cell differentiation does not appear to be impaired in this model. These results differ from the peripheral blood disturbances described in the active stages of human rheumatoid arthritis and are more similar to those reported in ankylosing spondylitis patients. However, the absence of alterations in the Peyer's patches suggests that pathogeneic mechanisms involving mucosal areas and exogenous intestinal antigens are not reproduced in this model.     

Lymphocyte subpopulations in adult coeliac disease.

Rosetting techniques were used to estimate T and B cell subpopulations in the peripheral blood in patients with treated and untreated adult coeliac disease and in control subjects. In patients with untreated coeliac disease, T cell numbers were significantly lower than in controls or treated patients, although there was no difference in total lymphocyte counts. There was no significant difference in B cell numbers between treated and untreated patients, and the subpopulation which increased to replace the T cells in untreated patients comprised cells not identified by B or T cell markers. Total lymphocyte counts and lymphocyte subpopulations were affected by splenic atrophy. It is suggested that these effects might be caused by the loss of lymphocytes from the gastrointestinal tract in untreated coeliac disease.     

Lymphocyte subpopulations in rheumatoid synovial tissue.

Synovial tissue obtained at synovectomy of the knee joint in 21 patients with rheumatoid arthritis contained a significantly lower proportion of T lymphocytes as measured by spontaneous rosette formation with nonsensitized sheep red blood cells than did synovial fluid or blood from the same patients. There was on concomitant increase in synovial tissue lymphocytes with B-cell markers such as surface immunoglobulin or Fc fragment receptors. Removal of lymphocyte receptors with trypsin followed by culture to allow new receptors to form, led to an increase in rosette forming cells, suggesting that part of the synovial cells without B- or T-cell markers may be T lymphocytes with blocked receptors.     

Effect of levamisole as a surgical adjuvant therapy for malignant melanoma.

The preliminary results of a trial in progress are reported. Two hundred patients have been enrolled in a randomized, double-blind, placebo-controlled trial of levamisole in malignant melanoma. A median followup of 20 months has been reached. Endpoints of the study include disease-free interval, interval free of visceral disease, and survival. Overall, there is a trend in favor of the levamisole group with regard to all three endpoints, but the difference is not significant. Significant differences between the control and levamisole groups are found at some, but not all, time intervals when the analysis includes only patients with stage II disease (positive lymph nodes at the initiation of the study). Although these preliminary results are encouraging, at the present time the overall curve for patients with stage II disease is not statistically significantly different when analyzed by the method of Mantel.     

Cimetidine as an adjuvant treatment in colorectal cancer

PURPOSE: To evaluate the influence of a H ₂ receptor antagonist (cimetidine) on survival in patients with colorectal carcinoma, a randomized, controlled pilot study was performed in three university hospitals in Copenhagen, Denmark. METHODS: A total of 192 patients, who had undergone a resection or an exploratory operation for adenocarcinoma of the colon or rectum between May 1988 and May 1991, were enrolled in the study. After a median observation time of 40 months, outcome was noted for each patient concerning cancer-specific mortality rate. RESULTS: In patients operated with curative intent (n=148), no difference was found in cancer-specific mortality between the two treatments. However, a tendency toward reduction in mortality rate was found in patients with curatively operated Dukes Stage C carcinoma (P=0.11, log-rank test; difference, 29 percent; 90 percent confidence interval, 2 to 57 percent) in the cimetidine-treated group. In patients with disseminated disease no total difference was found between the two treatment groups. CONCLUSIONS: Cimetidine does not seem to reduce mortality in patients with colorectal cancer, but there seems to be a tendency toward a survival benefit in patients undergoing surgery for Dukes Stage C carcinoma. Results seem to justify trials in this patient catagory to reveal a benefit of H2 receptor antagonists in adjuvant therapy of colorectal carcinoma.Read at the XII International Congress of Pharmacology, Winnipeg, Montreal, Canada, July, 1994.Cimetidine and placebo tablets were supplied by GEA Ltd., Copenhagen, Denmark.     

Lymphocyte and macrophage subpopulations in pelvic ileal pouches.

This study aimed to characterise the mucosal cellular infiltrate in ileal reservoirs with and without pouchitis (reservoir ileitis). Intraepithelial lymphocyte counts were performed in biopsy specimens obtained from ileal pouches and compared with counts in normal ileum and normal colon. T lymphocyte and macrophage subpopulations were characterised immunohistochemically in pouch biopsy specimens using a panel of monoclonal antibodies. Normal ileum was used as a control. Intraepithelial lymphocyte densities (expressed as intraepithelial lymphocyte/100 epithelial cells) in pouches with and without pouchitis were significantly less than in normal ileum, and approached counts found in normal colon. There was no significant increase in counts even in pouchitis. There were no significant differences in the helper/inducer to suppressor/cytotoxic T cell (CD4:CD8) ratios between normal ileum and pouches with or without pouchitis, either in the epithelium or in the lamina propria. The proportions of RFD9+ (epithelioid cells and tingible body macrophages) and 3G8+ (CD16) macrophages were significantly higher in pouchitis compared with pouches without pouchitis or normal ileum. There were no significant differences between the three groups in the proportions of cells positive for the other macrophage markers (CD68, RFD1-dendritic cells, and RFD7-mature macrophages). The significance of low intraepithelial lymphocyte counts in ileal pouches is unknown, but this may be an adaptive response to the new luminal environment. Since an increase in RFD9+ macrophages occurs in inflammatory bowel disease, but does not occur as a non-specific response to an acute infective process, their presence in pouchitis suggests that effector mechanisms similar to those triggering the original ulcerative colitis may be operating in pouchitis.     

Lymphocyte subpopulations in malaria infected individuals living in an endemic area

Development of partial immunity in people living in malaria endemic area is complex. For better understanding, the lymphocyte subpopulations from infected patients were evaluated by flow cytometer before any antimalarial treatment. In P. vivax infection, the frequency of T-helper type 1 (Th1) was decreased significantly (p = 0.042). In contrast, the number of T- helper type 2 (Th2) was increased significantly (p = 0.001). These trends have also been observed in P. faciparum infection. The Th2 predominant response to the natural malaria infection is likely due to persistent stimulation by Plasmodium species. In P. falciparum infection, CD8+ cytotoxic lymphocytes were significantly reduced (p = 0.007). However, such changes were not found in P. vivax infection. This might suggest that CD8+ cell responses to different Plasmodium spp in a different way. Both Th2 activation and CD8+ cell suppression may reflect less protective effects and chronic malaria infection could be established.     

Lymphocyte subpopulations of sheep in protective immunity to Taenia hydatigena

Lymphocyte subpopulations entering the liver and surrounding the rejection sites during a 9-day period after infection of immune sheep with Taenia hydatigena were identified with the aid of monoclonal antibodies against lymphocyte cell surface markers. Viable lymphocytes were isolated from the liver tissue and stained by indirect immunofluorescence for subsequent flow cytometry analysis. Over the first 6 days after challenge infection a marked increase in the ratio of T-helper to T-suppressor/cytotoxic lymphocytes was observed. SBU-T19+ lymphocytes, a CD5+ T-cell subpopulation uniquely identified in the sheep, were present in small numbers in sheep liver both before and after infection. There was a large, continuous increase of sIg+ B-cells over the 9-day observation period after infection. Eosinophils were the predominant granulocytes in the liver of infected sheep. The exact location of the leucocyte subpopulations in respect to the rejection sites in infected liver was determined by in-situ immunoperoxidase staining of frozen liver sections. The evolution of the parasite-induced leucocyte response was characterized by the appearance of a central core of eosinophils surrounded by increasing numbers of CD4+ helper T-cells. CD8+ (suppressor/cytotoxic) and SBU-T19+ T-lymphocytes were present in much smaller numbers and by day 9 after infection were located predominantly around the periphery of the lesions. Distinct foci of tightly packed B-cells developed within the lesions and increased dramatically in size over the 9-day observation period. At this time, lesions appeared as compact aggregations of leucocytes encircled by a second band of eosinophils. Both T- and B-lymphocytes within the lesions stained positive for class II major histocompatibility antigens.     

Lymphocyte subpopulations of intestinal mucosa in inflammatory bowel disease.

Lymphocyte subpopulations in peripheral blood (PBL) and intestinal mucosa (IML) of 10 patients with inflammatory bowel disease (IBD) were compared with those of 11 non-IBD controls. PBL were separated on Ficoll/hypaque gradients, and IML were isolated by incubation in dithiothreitol, EDTA, and collagenase. These methods yielded cells of good viability and with intact HLA A and B-antigens. T-cells, identified by neuraminidase-treated sheep RBC rosettes and non-specific esterase staining, comprised approximately 91% of the IML from normal mucosa of all groups. B-cells, identified by erythrocyte-antibody-complement rosettes and surface immunoglobulins, were only 7% of these IML populations. Cell yields were two-fold or more greater from abnormal IBD mucosa, with T-cells ranging from 55 to 95% and B-cells from 2 to 36%. The percentage of Fc receptor bearing cells was low in all specimens. By these methods, T-lymphocytes predominated in intestinal mucosa of both IBD and non-IBD patients, but there is marked increase in the percentage of B-cells isolated from abnormal mucosa in IBD.     

Whole abdominal radiotherapy and concomitant 5-fluorouracil as adjuvant therapy in advanced colon cancer

PURPOSE: This analysis was undertaken to assess whole abdomen radiation therapy and concurrent 5-fluorouracil for toxicity and patterns of failure in high-risk colon cancer patients after curative surgical resection. METHODS: Eighteen patients were treated adjuvantly after curative resection. Four patients (22 percent) had Stage B and 14 (78 percent) had Stage C disease. Histology was poorly differentiated in 4 (22 percent) and moderately differentiated in 14 (78 percent) patients. Four patients received whole abdominal radiation only, 30 Gy at 1 Gy/day. Fourteen patients had an additional locoregional boost of 9.6 to 16 Gy at 1.6 Gy/day. The liver received 19.8 Gy at 0.67 Gy/day. 5-Fluorouracil was given as a continuous infusion during therapy. RESULTS: With a median follow-up of three years, 6 of 18 (33 percent) patients have relapsed. Failure occurred locally in 3 of 18 (17 percent) and distantly in 4 of 18 patients (22 percent). Four of six (67 percent) failures occurred in the liver. The five-year actuarial survival and disease-free survival were 78 percent and 66 percent, respectively. Median elapsed time on radiotherapy was 73 days, with 5 of 18 patients (28 percent) requiring two or more weeks of unplanned treatment breaks. Acute Grade 3 to 4 toxicity (diarrhea, leukopenia) occurred in 3 of 18 patients (17 percent), with late complications (bowel obstruction) occurring in 2 of 18 patients (11 percent). CONCLUSIONS: Whole abdominal radiotherapy with concomitant 5-fluorouracil appears to improve local control but not to prevent liver metastases. Significant toxicity resulted in frequent interruption of therapy and protracted its course. Whether this adjuvant regimen impacts on survival or offers an advantage over locoregional irradiation remains to be studied.     

Outcome of patients with clinical stage II or III rectal cancer treated without adjuvant radiotherapy

Background  To clarify the indications for preoperative adjuvant radiotherapy for rectal cancer, the outcome of patients who underwent curative surgery without adjuvant radiotherapy was investigated. Methods  A total of 817 consecutive patients who underwent curative surgery for clinical stage II or III rectal cancer without preoperative adjuvant radiotherapy between 1988 and 2002 were reviewed. Results  The actuarial 5-year local recurrence rate in the examined patients was 6.2%. Univariate analysis showed that sex, pathological T classification (pT), clinical N classification (cN), pathological N classification (pN), tumor site, distance from the anal verge, type of surgery, pathological stage, a positive radical margin, lymphatic invasion, and venous invasion were significantly correlated with local recurrence. Multivariate analysis of preoperative factors identified cN, distance from the anal verge, and sex as statistically significant risk factors for local recurrence. In patients with rectal cancer located less than 5 cm from the anal verge and with positive cN, the local recurrence rate was more than 10%. Conclusions  Patients with rectal cancer located less than 5 cm from the anal verge and with clinically positive lymph nodes should be given preoperative adjuvant radiotherapy.     

Dendritic cells as potential adjuvant for immunotherapy in adrenocortical carcinoma

Objective Adrenocortical carcinoma (ACC) is a rare malignancy associated with a dismal prognosis. Dendritic cells (DCs) are professional antigen‐presenting cells leading to an antitumour immune response. The aim of this study was to elaborate two methods of antigen delivery to DCs and to evaluate an immunotherapy protocol in ACC patients. Design/patients Autologous DCs were pulsed with autologous tumour lysate (TL). Fusion of DCs with tumour cells was based on a polyethylene glycol method. Two patients with metastasized hypersecretory ACC were vaccinated twice. Measurements In vitro data were quantified by measurement of PBMC (peripheral blood mononuclear cell) responses and cytokine secretion and by flow cytometry analyses. Clinical response was monitored by CT scan of tumour mass and measurement of angiogenic factors. Results The maximum loading of TL was obtained at 24 h as 48·2% (± 26·8%) of DCs were TL‐positive. The DC/tumour cell fusion efficacy was ～45% as shown by double positive staining for ACTH receptor and DC‐specific CD83. In vivo DC vaccination resulted in positive delayed‐type hypersensitivity skin reactions reflecting specific memory T‐lymphocyte reaction. In vitro analyses revealed specific T‐cell proliferation in patient 1 (stimulation index: 5·7 compared to pretreatment) and induction of cytotoxic granzyme B secreting T cells in patient 2 (0·41% CD8 + cells vs. 0·06% pretreatment) as indicators of specific cytotoxic T cells. Although angiogenic serum markers could be stabilized, no impact on tumour growth could be observed. Conclusion Our data demonstrate that autologous dendritic cells induce antigen‐specific Th1 immunity in adrenocortical carcinoma. The clinical outcome, however, was not improved in the patients studied here.     

Lymphocyte subpopulations of peripheral blood in tsutsugamushi disease

We examined lymphocyte subpopulations in the peripheral blood and serum concentrations of immunoglobulin (Ig) in 8 patients with tsutsugamushi disease. In 7 of the 8 cases, there was a 4-fold or greater increase in IgG and IgM antibody titers between the initial and convalescent serum specimens. In one case, there was no increase, but IgM antibodies were detected with diagnostically significant antibody titers. The percentages of CD8- and CD2-positive lymphocytes measured before treatment were found to be significantly higher than during the recovery stage of patients with tsutsugamushi disease. The CD4/CD8 ratio in the peripheral blood calculated before treatment was significantly lower than that during the recovery stage. Serum concentrations of IgG and IgM in patients during the recovery stage were significantly higher than pretreatment levels, respectively.     

Lymphocyte subpopulations in megaloblastic anaemia due to vitamin B-12 deficiency

Lymphocyte subpopulations were measured in the blood of 17 patients with megaloblastic anaemia due to vitamin B-12 deficiency. 14 patients had pernicious anaemia and 3 others were gastrectomized. By using monoclonal antibodies recognizing T cell surface markers and immunofluorescence microscopy, we found a significant decrease in the number of circulating suppressor T cells and an increase in the ratio of helper to suppressor T lymphocytes in pernicious anaemia patients. This finding may be related to other immune abnormalities found in pernicious anaemia, e.g. the presence of multiple autoantibodies.