Immune responsiveness of patients with malignant melanoma and carcinoma of the breast.

Lymphocyte blastogenesis (LB) with PHA, CON-A, PWM, VII (homologous tumor extract) plus T/B ratio for pre-IT, post-IT (2 months), and 3, 6, 9, 12, 18, and 24 months prior to each VII booster are reported. Pre-IT and post-IT LB-PHA were similar for Stage I (ST I) or Stage II (ST II) breast cancer (BC) patients who expired or survived. The pre-booster results were similar except that prior to death, stimulation with PHA decreased. Melanoma (MM) patients LB-PHA pre-IT showed less stimulation for ST I patients than for ST II patients (survivors and expired). LB-CON-A and LB-PWM showed similar results with slight variations. When compared to survivors, LB-VII for BC patients who expired showed no stimulation. MM patients showed different results in LB-VII. T/B ratios for BC patients during the first 3 months showed 72% normal (N), 14% below N, and 14% above N for survivors and 40% normal (N), 60% below N, and 0 above N for expired. MM T/B ratio were not remarkable. LB anamnestic response to immune stimulation evaluated in 16 of the 62 BC and 10 of 81 MM patients showed increased peak levels over trough for survivors and a decrease for expired. The significance of data as in vitro correlates of clinical response is discussed.

Ultrastructural evidence of large granular lymphocyte (LGL) activity in lesions of chronic adult periodontitis.

The purpose of this investigation was to determine, on the basis of ultrastructural evidence, whether large granular lymphocyte (LGL)-mediated cytotoxic activity could be identified in lesions of chronic adult periodontitis. 18 gingival papilla biopsies were obtained from 8 adult patients, each satisfying the clinical criteria for chronic adult periodontitis. One-half of each biopsy was processed for examination by transmission electron microscopy (TEM). B-lymphocyte, T-lymphocyte, and NK-cell contributions to the inflammatory cell infiltrate were determined by subjecting the remaining one-half to the 3-layer avidin-biotin affinity immunoperoxidase technique. Immunohistochemistry using the anti-human leu-11b monoclonal antibody (representative of NK-cells which are known to comprise approximately 80% of the LGL population) showed that the Leu-11b+ population comprised 3-7% of the total monocytic infiltrate. The Leu-11b+ cells tended to occur as single cells or in small clusters of 3-12 cells. Generally, the Leu-11b+ cells exhibited perivascular locations situated subjacent to the epithelial basal lamina. TEM observations showed LGLs in intimate contact with fibroblasts that exhibited morphologic changes consistent with cellular damage or degeneration. In addition, LGLs were observed to exhibit apparent non-cytotoxic contacts with plasma cells, macrophages and other monocytic cells.

Comparison of NK-cell (Leu-7+ and Leu-11b+) populations in clinically healthy gingiva, chronic gingivitis and chronic adult periodontitis.

Various investigations have reported the presence of cytotoxic lymphocyte activity in inflammatory periodontal disease. The collective evidence indicates that the inflammatory infiltrates of gingivitis and periodontitis should feature a major component of large granular lymphocytes (NK-cells) possessing cytotoxic potential. Thus, the purpose of this study was to determine and compare, by use of immunohistochemical methods, the numbers of NK-cells in biopsies of clinically healthy gingiva, chronic gingivitis and chronic adult periodontitis and their relationship, if any, to the T- and B-lymphocyte populations. Gingival biopsies were obtained from 8 patients in each of three disease groups selected on the basis of predetermined clinical criteria. Using the avidin-biotin immunoperoxidase technique, four consecutive serial sections from each biopsy specimen were stained with a panel of antihuman monoclonal antibodies for T-lymphocytes (UCHL-1) B-lymphocytes (CD-45R), and NK-cells (Leu-7 and Leu-11b). Analyses of variance yielded a statistically significant main effect for each cell immunophenotype. The Newman-Keuls Sequential Range Test showed statistically significant differences for all but two mean comparisons (p less than 0.01). The comparisons for UCHL-1 and Leu-7 between chronic gingivitis and periodontitis specimens did not demonstrate significance. Although T- and B-lymphocyte populations increased approximately 20 x progressing from healthy to gingivitis to periodontitis specimens, the NK-cell population showed only a 3 x increase which represented 19%, 6.6% and 7% of the total of all positively stained lymphocytes across biopsy groups.

Adjuvant immunotherapy for melanoma.

Adjuvant immunotherapy was administered to 84 lymph-node-negative and 25 lymph-node-positive melanoma patients. This active specific homologous cell protein preparation was given after aggressive surgery and given over 2 years. Projected and observed survival rates are presented as well as other clinical and pathologic characteristics such as female-to-male ratio, site of primary, level and depth of invasion. The projected 5-year survival rates are 90% for all stage I with 89% for females and 94% for males. Lower extremity stage I projected 5-year survival rate was 88% for all, 87% for females, and 100% for males. The projected 5-year survival rate for stage II was 68% overall and 100% for lower extremity. Only two of five patients with an unknown primary have expired. All of these results are improved over expected survival. Hopefully a randomized prospective study will be stimulated to ascertain the basis for this improvement.

The role of the regional lymph node in breast cancer: a comparison between nodal and systemic reactivity.

Confusion remains concerning the role of the regional lymph node in the containment of cancer. Numerous investigators using a variety of assays have reported often conflicting results concerning the immunocompetency of lymphocytes residing in regional nodes. Forty-two axillary lymph nodes from ten stage I and stage II breast cancer patients were studied in lymphocyte blast assays using mitogens and breast cancer antigen (BCA). Three general response patterns to BCA were identified which were related primarily to tumor size. In the patients with the smallest primary tumor (0.5 cm), lymphocytes in the nodes reacted to a much greater extent than peripheral blood lymphocytes (PBL). In two of three patients with intermediate-size tumor (1.0 to 1.5 cm), a mixed pattern of responses was seen with both stimulation suppression occurring within the nodes of the same patient. In the four patients with the largest tumors (2.0 to 3.0 cm), 15 of 19 nodes had a lower stimulation index (SI) than the corresponding PBL. From the results of this study it appears that regional lymph nodes are dynamic immunologic structures which regress in responsiveness as tumor burden increases.

Immunologic responsiveness of patients with cancer: relationship to tumor type, stage and prognosis.

Data on cellular immunity of 39 patients with breast cancer and 29 patients with malignant melanoma, is presented. T/B lymphocyte ratios, lymphocyte blastogenesis (LB) with PHA and with cancer antigen preparations were carried out from two to four weeks after cancer surgery, and immediately after eight weeks of immunostimulation with a tumor-specific vaccine. Distinct differences were observed. Small breast cancers do not seem to evoke a host immune response, while small melanomas are associated with a state of immune stimulation. After immunotherapy LB in these breast cancer patients shows significant stimulation while LB in the melanoma patients changed from stimulation to normal response. Quite different results were obtained in patients with Stage II cancers; breast cancer patients showed immune stimulation which did not change after immunotherapy. The melanoma patients showed low normal immune response, which changed to high normal after immunotherapy. Patients with Stage III breast cancer and melanoma were similar in their poor immune responsiveness. Possible mechanisms are offered for these differences.

Immunologic responsiveness of the breast cancer patient.

Lymphocytic infiltration of the primary breast cancer and sinus histiocytosis of the axillary lymph nodes are indications of a favorable prognosis. Similarly, skin test responsiveness such as with DNCB or with tumor extracts correlates in general with stage of disease. This presentation will bring forth preliminary data on cellular immunity of breast cancer patients. Circulating lymphocytes (PBL) were stimulated with mitogens and a breast cancer antigen. PBL from patients with a primary tumor less than 2.4 cm in size reacted as though no immune stimulus existed. PBL from patients with a lump from 2.5 to 5.0 cm in size showed evidence of immune stimulation. An increase in size of the primary tumor over 5 cm and an increase in the number of axillary lymph nodes with metastasis were associaed with a diminution in cellular immunity. However, data from an adjuvant immunotherapy program show that cellular immunity can be improved in certain patients by immunization. Such patients continued to remain disease free, while patients whose cellular immunity was poor or not improved by adjuvant immunotherapy tended to develop recurrent disease.

Suppressor T cells and host resistance to tye 111 pneumococcus after treatment with antilymphocyte serum.

The antibody response to type III pneumococcal polysaccharide (SS-II) was significantly increased in mice treated with antilymphocyte serum (ALS). BALG/c mice given 0.25 ml of ALS on days -1, 0, and 1 relative to the days of immunization with 0.5 mug of SSS-II had a 20-fold increment (11,383 increased to 199,917) in the number of splenic plaque-forming cells enumerated on day 5 compared with untreated, immunized controls. This effect has been attributed to the elimination of subpopulation of thymus-derived lymphocytes (T cells) that has suppressor function. The present series of experiments relate the augmented antibody response to SSS-II in mice treated with ALS to increased host resistance after infection with Streptococcus pneumoniae, type III (Pn-II). The 50% lethal dose of Pn-III in niminnunized mice was 102 and the 100% lethal dose was 103 organisms. Mice immunized with 0.5 mug of SSS-III and challenged 5 days later with Pn-III were completely protected against a dose of up to 108 organisms. Mice treated with 0.25 ml of ALS on days -1, 0, and 1, immunized with SSS-III on day 0, and challenged with 2.5 X 10(9) Pn-III on day 5 had a mean survival time of greater than 100 h compared with 16 h for immunized non-serum-treated controls. Animals given a single injection of ALS before immunization showed no increase in resistance, whereas mice treated after immunization had significant prolongation of survival times. Untreated, immunized mice challenged with 5 X 10(9), 1 X 5 X 10(8) Pn-II survived 14 to 19 h, whereas ALS-treated animals had mean survival times of 48, 174, and 222 h, respectively. These findings suggest that immunoregulatory T cells may have a biologically significant effect in a narrow zone in which the normal host immune response is insufficient but still potentially capable of providing some additional degree of protection if suppressor cells are elimated.

Organ distribution of 99m-Tc-and 51-Cr-labelled thymocytes.

We have employed 99mTc as a radioisotopic label to study the organ distribution of murine thymocytes. The compartmentalization of 99mTc-labeled cells that had not been reduced by treatment with stannous chloride was similar to that of 51Cr-labeled cells and was characterized initially by 48-50% uptake of the injected radioactivity by the lungs. Increased hepatic and splenic and decreased pulmonary localization were noted at 1 hr and these shifts were more pronounced by 4 hr. Technetium-99m-labeled cells reduced by stannous chloride had significantly different patterns of hepatic, pulmonary, and splenic localization and at 4 hr the lungs still retained 24% of the injected radioactivity compared with only 3% in the spleen. Size distribution studies revealed that unlabeled as well as unreduced and reduced 99mTc-labeled thymocytes were almost identical to one another so that differences in compartmentalization could not be attributed to this factor. Since reduction with stannous chloride did not alter the distribution of 51Cr-labeled cells, this suggested some type of complex interaction between stannous ions and the labeling species of 99mTc. The in vivo localization of intravenously administered Na99mTcO4 and Na2CrO4 was markedly different from the corresponding radiolabeled cells thereby indicating that the distribution patterns that we observed truly represented cell-associated radioactivity. Although it may not necessarily be the proper reference point, the similarity in organ distribution of 99mTc- and 51Cr-labeled cells should allow direct comparison of previously reported data employing this radionuclide and that obtained in future studies with 99mTc.