Peripheral blood lymphocytes from a total of 373 tumor patients were tested by either a colony inhibition or a cytotoxicity test for cell-mediated immunity against human neoplasms of various histological types. Lymphocytes from 51 of 59 patients studied (88%) either reduced colony numbers formed by plated autochthonous tumor cells or were cytotoxic to them, and lymphocytes from 78 of 87 patients tested (89%) had a similar effect on allogeneic tumor cells of the same histological type as those of the lymphocyte donors. Evidence indicating antigenic cross-reactivity between tumors of the same histological type was obtained for the following seven groups of neoplasms: malignant melanomas, carcinomas of the colon, breast, testis, endometrium and ovary, and various sarcomas. Lymphocytes affecting tumor cells had no effect on normal cells from the same patient, or on cells from other types of neoplasms than the target cells under study.



The degree of cell-mediated immunity, as detectable with the techniques employed, was approximately the same in patients having active neoplastic disease as in patients who were clinically symptom-free. Eleven of 12 patients who were tested after having been symptom-free for more than 2 years had a lymphocyte-mediated anti-tumor immunity.



Mise en Evidence D'une Immunite A Mediation Cellulaire Envers les Neoplasmes Humains de Divers Types Histologiques



Nous avons etudie des lymphocytes du sang peripherique de 373 cancereux au moyen de tests d'inhibition des colonies et de cytotoxicite en vue de mettre en evidence une immunite a mediation cellulaire envers les neoplasmes humains de divers types histologiques. Les lymphocytes de 51 des 59 sujets etudies (88%) ont fait diminuer le nombre de colonies formees par les cellules tumorales autochtones ou se sont montres cytotoxiques a l'egard de ces cellules; les lymphocytes de 78 des 87 sujets testes (89%) ont eu un effet analogue sur les cellules tumorales allogeneiques du měme type histologique que celles des donneurs de lymphocytes. Nous avons obtenu la preuve de l'existence d'une reactivite antigenique croisee entre les tumeurs du měme type histologique pour les 7 groupes de neoplasmes suivants: melanomes malins, epitheliomas du colon, du sein, des testicules, de l'endometre et des ovaires, et divers sarcomes. Les lymphocytes agissant sur les cellules tumorales n'ont pas eu d'effet sur les cellules normales du měme malade, ni sur les cellules de neoplasmes de types histologiques differents.



Le degre d'immunite a mediation cellulaire decele au moyen des techniques utilisees etait a peu pres le meme chez les malades atteints d'une neoplasie active que chez les sujets ne presentant aucun symptǒme clinique. Nous avons observe une immunite antitumorale a mediation lymphocytaire chez 11 des 12 malades qui ont ete testes apres ětre restes asymptomatiques pendant plus de deux ans.

Demonstration of cell-mediated immunity to human neoplasms of various histological types.

Publisher Summary This chapter reviews the evidence for cell-mediated immune reactions against tumors in animals and man, particularly, the reactions that can lead to destruction of neoplastic cells cultivated in vitro. Various mechanisms by which tumor cells can escape from immunological destruction have been discussed, most notably one involving blocking factors, present in the serum. Findings analogous to those obtained when studying tumor immunity (coexistence of cell-mediated reactivity and blocking serum activity) have been summarized from three other areas (pregnancy, allografting, and chimeras). The studies reviewed in the chapter have given a relatively large amount of evidence for various cell-mediated and humoral immunological reactions against growing tumors, and at least some of the reactions observed in vitro appear to be able to influence tumor growth in vivo .

Lymphocyte-Mediated Cytotoxicity And Blocking Serum Activity To Tumor Antigens

Blood lymphocytes from tumor patients can specifically destroy cultivated neoplastic cells of the same histological origin as the tumors of the lymphocyte donors, irrespective of whether or not the donors have symptoms of growing tumor.



The purpose of the present study was to investigate whether sera from tumor patients could block the cytotoxic effect of lymphocytes immune to the specific antigens of the respective neoplasms. A wide variety of tumors were included in the tests, namely malignant melanomas, carcinomas of the colon, breast, ovary, endometrium, kidney, cervix uteri, lung, larynx, bladder, Fallopian tube, lip, seminomas and sarcomas.



Sera from 67 out of 81 patients with growing neoplasms were found to block the cytotoxic effect of specifically immune lymphocytes. A blocking effect was seen both when the tumor cells, lymphocytes and sera were derived from the same patients and when the lymphocytes and sera were taken from different donors who had the same types of tumor as the target cells. No blocking was seen when the same sera were tested on tumors of histological types other than those of the respective serum donors.



A blocking serum activity was seen in only three of 19 patients who were symptomfree after tumor therapy. The findings thus suggest that there is a correlation between tumor growth in vivo and the presence of a blocking serum activity in vitro.



Blocage de L'immunite Tumorale A Mediation Cellulaire au Moyen de Serums de Malades Porteurs de Neoplasmes Evolutifs



Les lymphocytes du sang de cancereux peuvent detruire specifiquement des cellules neoplasiques en culture dont l'origine histologique est la měme que celle des tumeurs des donneurs de lymphocytes, que ces donneurs presentent ou non des symptǒmes de tumeurs en pleine evolution.



Nous avons cherche a savoir si les serums de cancereux peuvent bloquer l'effet cytotoxique des lymphocytes immuns envers les antigenes specifiques de chaque neoplasme. Nous avons choisi pour ces etudes une large gamme de tumeurs: melanomes malins, carcinomes du colon, du sein, des ovaires, de l'endometre, du rein, du col de l'uterus, du poumon, du larynx, de la vessie, des trompes de Fallope, de la levre, seminomes, sarcomes.



Nous avons constate que le serum de 67 des 81 malades porteurs de neoplasmes evolutifs bloque l'effet cytotoxique des lymphocytes specifiquement immuns. l'effet de blocage se produit aussi bien lorsque les cellules tumorales, les lymphocytes et les serums proviennent des měmes malades que lorsque les lymphocytes et les serums proviennent de differents donneurs atteints du měine type de tumeur que les cellulescibles. Nous n'avons observe aucun blocage lorsque nous avons teste les měmes serums sur des tumeurs de type histologique different de celles de chaque donneur de serum. Nous avons remarque un blocage chez 3 malades seulement sur les 19 qui etaient devenus asymptomatiques apres traitement de la tumeur. Ces constatations donnent a penser qu'il existe une correlation entre la croissance tumorale in vivo et l'activite de blocage des serums in vitro.

Blocking of cell-mediated tumor immunity by sera from patients with growing neoplasms.

Over the past 7 years, 151 patients with malignant melanoma have been treated with BCG immunotherapy alone or as an adjunct to surgical therapy. Direct injection of metastatic melanoma lesions limited to skin resulted in 90% regression of injected lesions and 17% regression of uninjected lesions in immunocompetent patients. Approximately 25% of these patients remained free of disease for 1 to 6 years. Direct injections of BCG into nodules of patients with subcutaneous or visceral metastases resulted in a lower incidence of local control and no long term survivors. Attempts to improve the results of immunotherapy in these patients by palliative surgical resection of large metastatic lesions to lower tumor burden followed by BCG immunotherapy significantly improved the results although many patients still developed recurrent disease. Early results of a clinical trial combining BCG immunotherapy with regional lymphadenectomy in patients with melanoma metastatic to lymph nodes have been encouraging and promising. Further controlled clinical trials are necessary to elucidate the role of BCG in immunotherapy. However, since BCG is but one of a number of potential immunologic adjuvants, even more effective immunotherapy will be possible as further knowledge of the interactions of cellular and humoral immunity is acquired.

BCG Immunotherapy of Malignant Melanoma: Summary of a Seven-year Experience

Leukocytes from 17 of 19 patients with urinary bladder carcinomas were cytotoxic for autochthonous as well as allogeneic bladder carcinoma cells in vitro. The cells from all 11 individual bladder carcinomas studied were susceptible to this cytotoxic action of patients' leukocytes. In contrast, no cytotoxicity was observed when cells from unrelated tumours or various normal tissues were used as target cells. Purified blood lymphocytes from patients whose leukocytes were active, were also cytotoxic.



Sera of 8 out of 13 patients with bladder carcinomas contained complement-dependent antibodies which were cytotoxic for autochthonous and allogeneic bladder tumour cells. When serum was taken from the same patient on different occasions over a period of 8 months cytotoxicity was found in some samples but not in others. Sera from 4 out of 9 patients exhibited a blocking activity, which completely abolished the cytotoxic effects of patients' leukocytes. The serum of one out of 9 patients contained antibodies which were not cytotoxic without leukocytes but which conferred a cytotoxic activity onto leukocytes from control subjects. This opsonizing antibody was complement-dependent and probably of IgM nature.



The effect of hydrostatic pressure therapy on the immune state of the patients was followed by examination of the leukocytes from 16 patients with bladder carcinoma. Leukocytes from 10 patients were cytotoxic before as well as after the pressure treatment. Leukocytes from six patients did not react before the treatment while those from four became active after treatment. A more detailed study of three individual cases indicated that cytotoxicity of the leukocytes appeared after pressure treatment and was of relatively short duration (approximately 1 month). In one case, the development of cytotoxic antibodies was similar. The patient who had opsonizing antibodies had no cytotoxic antibodies. The opsonizing antibodies appeared when the cytotoxicity of the leukocytes decreased.

Cellular and humoral immune responses to human urinary bladder carcinomas.

Biopsy specimens and sera were obtained from 103 melanoma patients. Autoantibodies were demonstrated by (1) complement-dependent cytotoxicity of autologous melanoma cells in short-term culture; (2) complement-dependent inhibition of ribonucleic acid synthesis; (3) immunofluorescent staining of the cytoplasm of killed melanoma cells and of the surface membrane of viable melanoma cells. Over one-third of the sera studied had antibodies to autologous melanoma cells. Although for technical reasons all three tests could not be performed with the cells from every melanoma, whenever multiple testing was possible there was complete concordance. The autoantibodies were virtually confined to patients in whom the disease was not widely disseminated, and over 80% of such patients had positive sera. In a limited number of patients who have been followed autoantibodies disappeared as the disease progressed to become widely disseminated. Two patients with generalized disease developed autoantibodies following inoculation by their own irradiated tumour cells. Two types of autoantibodies were recognized: one, active against antigen(s) in the cell surface membrane, was specific for each tumour—that is, only the autologous serum reacted—and was concerned in the cytotoxic activity; the other reacted with cytoplasmic antigens which appeared to be present in most or all melanoma cells.

https://www.bmj.com/content/bmj/3/5670/547.full.pdf

Tumour-specific Antibodies in Human Malignant Melanoma and their Relationship to the Extent of the Disease

Thirteen patients have been immunized with their own irradiated melanoma cells on one or more occasions. Following these autografts tumour-specific cytotoxic antibodies were produced, the longest response lasting 14 days. This procedure had no apparent effect on the course of the disease in these patients.

/pdf/autoimmunization-with-irradiated-tumour-cells-in-human-17o0bor4io.pdf

Autoimmunization with Irradiated Tumour Cells in Human Malignant Melanoma

For many years evidence has been accumulating that there may be a host reaction to malignant melanoma and that this reaction could be immunological. First, melanoma is one of the malignant diseases that occasionally undergo spontaneous r e g r ~ ~ i ~ n . ~ . ~ . ~ ~ ~ ~ Second, the disease may remain localized for a long time before it becomes disseminated?' Third, even in patients with disseminated disease, some metastatic deposits can regress at the same time as new ones appear and others continue to grow.* Fourth, Lewis4 has shown in a series of Ugandan patients that those with localized tumors often had cytotoxic serum antibody active only against the patient's own melanoma cells. This suggested that there may be tumor-specific antigens in human melanoma of a type similar to those found in chemically induced tumors in animals. Morton and colleagues,' Oettgen and and Muna and co-~orkers ,~ using immunofluorescence, demonstrated cross-reactivity between melanoma cell preparations and the sera from a number of different melanoma patients. For these reasons we decided to make an intensive study of antibodies with the cooperation of many centers in London, Bristol, and elsewhere and have collected sera and biopsy material from over 100 patient^.^ The presence of autoantibodies was sought by three techniques: the complement-dependent cytotoxicity test, the complement-dependent inhibition of RNA synthesis using tritiated uridine, and immunofluorescent staining of the cytoplasm of killed melanoma cells and of the surface membrane of viable melanoma cells. I t was not possible to carry out all the tests on each patient, but whenever this could be done there was nearly always complete concordance. All those who had antibodies demonstrable in the cytotoxic test gave positive membrane immunofluorescence, but membrane immunofluorescence was occasionally positive even though cytotoxic antibodies were absent. By these techniques, autoantibodies were demonstrated in 35 out of the 119 sera tested, and two types could be recognized: one active against antigen(s) on the cell surface membrane that was specific for each tumor, and was also cytotoxic, the other active against cytoplasmic antigens present in most or all melanoma cells. In the cytotoxic test, only the autologous serum was cytotoxic to the melanoma cells. As with the patients from Uganda, the presence of cytotoxic autoantibodies was virtually confined to those who had localized disease. We divided the patients into three clinical grades: grade I-those with malignant melanoma confined to the primary anatomical site; grade II-those with invasion of regional lymph nodes with or without local satellite nodules; and grade IIIthose with tumor spread beyond the regional lymph nodes. The results are shown in TABLE 1. In three patients in whom the serum was originally positive, sera taken later, when the disease became widespread, failed to inhibit growth of the cultured

Ikonopisov Rl

Papers

Tumour-specific Antibodies in Human Malignant Melanoma and their Relationship to the Extent of the Disease

Autoimmunization with Irradiated Tumour Cells in Human Malignant Melanoma

Detection of tumor specific immune reactions in human melanoma.