Suppressor substance produced by the K562 cell line in vitro.

Supernatant fluids from three human cultured cell lines, K562, NALM-1, and Daudi, all isolated from patients with lymphoreticular malignancies, were tested for suppressive activity toward normal lymphocytes in vitro. It was found that the K562 line elaborated material which effectively suppressed the response of normal human blood lymphocytes to the T-cell mitogens phytohemagglutinin and concanaval in A as well as the mixed lymphocyte reaction. Little or no suppressive activity was evident when supernatants from Daudi or NALM-1 were used. In contrast, a recent report [29] indicates that K562 does not inhibit the induction of immunoglobulin synthesis of B lymphocytes by pokeweed mitogen. The findings are discussed in the context of the immunosuppression associated with cancer and the apparent selective inhibition of different lymphocyte functions by material elaborated by different neoplasms.

Mycosis fungoides: an overview.

Mycosis fungoides (MF) is a lymphoma that appears to have a unique relationship with the skin. In recent years our knowledge about this disease has increased considerably, in particular regarding the nature of the malignant cells and their functional characteristics. Furthermore, there is mounting evidence suggesting a viral etiology. Since its first description by Alibert in the early part of the past century, MF has been the subject of much controversy in regard to fundamental issues such as clinical and histological criteria for the establishment of the diagnosis [1] as well as its relationship to other lymphomas and skin diseases. Some of this controversy remains today. Mycosis fungoides is a relatively uncommon disease, the incidence being in the order of 1-2 million per year [1,2]. As is the case with most neoplastic diseases, the incidence increases with age; however, relatively speaking MF is not exceptional among individuals in their twenties or thirties. As for sex distribution there is a slight preponderance of men [3-5].

Factors affecting immune responsiveness in vitro of germfree allogeneic radiation chimeras. I. Differences between donor and host strains.

An important aspect of bone marrow transplantation has been focused upon. It has been demonstrated that significant differences between in vitro responses of germfree DBA/2 mice (donor strain) and of C3H/He (recipient strain) exist. The PHA response of spleen cells from germfree C3H/He mice is greater than that of DBA/2 mice. However, the reverse is true in regard to Con A responsiveness. In fact, calculation of a Con A/PHA ratio reveals a striking difference between strains. B cell reactivity as assessed by LPS mitogenic responsiveness is similar in both strains. The cell mediated responsiveness of DBA/2 mice is also diminished as measured by mixed lymphocyte reaction and cell mediated cytotoxicity. An inverse correlation between plaque forming cell (PFC) responses in vitro and the Con A/PHA ratio in DBA/2 mice is suggestive of a predominance of inherent suppressor cell activity in this strain. These characteristics of the DBA/2 immune responsiveness may be a factor in the apparent T cell unresponsiveness seen in DBA/2 leads to C3H/He mouse allogeneic bone marrow chimeras.

Treatment of Kaposi's sarcoma with vinblastine.

Fourteen patients with Kaposi's sarcoma (KSY were treated systemically with vinblastine sulfate in a low-dose regimen and compared with 23 patients reported in the medical literature. The therapeutic results in our series were excellent in terms of regression of cutaneous lesions. Vinblastine appears to be a drug that is well suited for the management of KS in an outpatient setting. Intravenous therapy may be supplemented with intralesional or intraarterial vinblastine.

Lymphotoxins: selective cytotoxic effects.

Materials with lymphotoxin activity produced by lectin stimulated primary cultures of human lymphocytes enriched for T cells or B cells, as well as material obtained from established tissue culture lines of human lymphoid cells were tested for cytotoxicity towards a large number of primary cultures and established lines of human and animal cells. Highly selective effects were found. The patterns of responses to the various lymphotoxin preparations indicate heterogeneity not only among the different cell cultures but also among the lymphotoxin preparations. In addition to the heterogeneity of the various cell cultures in their responses to the lymphotoxin preparations, a spectrum of susceptibilities was also noted among clonal derivatives of the same parental lines. In quantitative terms the responses of the most resistant and the most susceptible cell cultures spanned approximately a 200-fold range. These findings could have bearing upon the mechanism of tumor evasion of host antitumor immunity.

Stimulation of phagocytic activity of neutrophilic granulocytes by sera of patients with solid tumors.

The effect of serum of patients with solid tumors on the phagocytic activity of normal neutrophilic granulocytes was investigated. The largest groups of tumors studied were carcinoma of colon/rectum, sarcomas and melanomas. Sera from all three groups of patients were found to have highly significant (p < 0.001) stimulatory effects on granulocyte phagocytic activity. These findings contrast with previously reported depression of granulocyte phagocytic activity associated with various forms of leukemia and lymphomas.

Mycosis fungoides associated with florid sarcoid reactions.

We report a 59-year-old woman with disease manifestations diagnosed as mycosis fungoides (MF), as well as findings of sarcoidosis. The concurrence of these two disorders, both of which may be associated with basic immunopathogenic factors, has been reported. Histologically, the cutaneous findings of both non-caseating granulomas and an infiltrate compatible with MF in the same site leads to a discussion of granulomatous MF. Whether granulomatous MF simply represents sarcoidosis developing concomitantly with MF lymphoma or is, in fact, a distinct clinical and histologic type of MF is discussed. Reasons why granulomatous MF, like granulomators Hodgkin disease, might have a more favorable prognosis than the usual type of MF are considered.

cis-Dichlorodiammineplatinum(II) and DTIC in malignant melanoma.

Twenty-nine patients with advanced malignant melanoma were randomized to receive DTIC at a dose of 200 mg/m2 iv on Days 1-5 and cis-dichlorodiammine-platinum(II) at a dose of 40 mg/m2 iv on Days 1 and 4, repeated every 4 weeks (group A), or the same drugs plus procarbazine at a dose of 75 mg/m2 orally daily on Days 1-8 and vincristine at a dose of 1.4 mg/m2 iv on Day 1 (group B). These drugs were generally well-tolerated, but five of 16 patients in group A and six of 13 patients in group B required dose modification for either hematologic or renal toxicity. There were six objective responses among 16 patients in group A including one complete regression, while there were two objective responses among 13 patients in group B.

Cis-dichlorodiammineplatinum(II) in metastatic soft tissue sarcomas.

cis-Dichlorodiammineplatinum(II) was given to 13 patients with metastatic sarcomas following failure in two previous chemotherapy protocols. Two of these patients had complete regression which lasted 6 months in one patient and is still present after 8 months in the other. One patient had a greater than 50% regression which lasted 5 months. Three patients had disease stabilization for 2 months each. cis-Dichlorodiammineplatinum(II), as a single agent, was well-tolerated with no significant renal or auditory toxicity.

Tourniquet infusion chemotherapy in extremities with malignant lesions.

Tourniquet infusion chemotherapy involves the direct injection of a chemotherapeutic drug into the main artery of an extremity with prior application of an external tourniquet proximally on this extremity set at above the level of systolic pressure for ten minutes. Thus, the drug is not diluted and pushed by the blood into the venous circulation before diffusion into the tissues has occurred. This technique, applied in seven patients for a total of 40 instances, proved to be safe. Skin erythema and blisters, which are reversible, occur in the treated area. Complete clinical regression was achieved in all six patients with evaluable tumor, with high percentages of tumor necrosis. It was possible to avoid amputation in five of the seven patients treated. This technique appears superior to perfusion, but the long term permanence of regression has not yet been ascertained.

Explorations of antimitotic agents in the treatment of a congenital disease, ichthyosis linearis circumflexa.

Successful therapy of a genetic disorder, Ichthyosis Linearis Circumflexa, was achieved using a low dose systemic cyclophosphamide. Prior to such therapy, 80 to 90% of the body surface was affected; the use of antimitotic agents reduced the extent of the lesions to less than 15% of the body surface. As a result, the clinical status was changed from severely disabling to being compatible with a normal way of life. It is believed that this phenomenon is related to selective effects of cyclophosphamide, such as those on lymphocyte subpopulations. To our knowledge, successful chemotherapy for diseases of genetic or congenital origin has not been previously reported.

Activation of neutrophilic granulocytes by products of human lymphocytes.

Materials produced by stimulated primary cultures of human lymphocytes and by a continuous human lymphoid cell line, RPMI 1788, were tested for granulocyte phagocytosis promoting activity. Blood lymphocytes of healthy donors were stimulated with tuberculin (PPD) or concanavalin A (con A) and the culture fluids collected after 3 days of in vitro incubation. Fluids of the lymphoid cell line were collected from 2-day-old cultures. All three kinds of preparations were found to induce a significant increase in neutrophilic granulocyte phagocytic activity. Studies on the kinetics of granulocyte activation revealed that it is a relatively rapid process, being maximal within 60-120 min. This is in contrast to the considerably slower activation of monocytes in a comparable system. Intracutaneous inoculation of the active materials in human volunteers resulted in reactions that grossly and histologically resembled accelerated delayed type hypersensitivity reactions.

Enhancement of granulocyte phagocytosis by products of PPD stimulated human lymphocytes.

The effect of supernatant fluids from cultures of stimulated human lymphocytes on the phagocytic activity of human neutrophilic granulocytes was investigated. It was found that lymphocytes of tuberculin (PPD) positive donors after in vitro stimulation with PPD produced factors that significantly increased granulocyte phagocytosis of latex particles.

Nonspecific antigen reactions.

Cell-mediated immune challenge reactions to a number of antigens at tumor sites resulted in regressions of various types of benign, premalignant, and malignant lesions in man. Regressions varied from partial to complete and lasted from several months to more than 10 years. Increased levels of cell-mediated immunocompetence were attained by several means, including reduction of tumor burden, immunopotentiation, or transfer of immunity. Antitumor activities of immune challenge reactions are selective for tumor cells and appear to be independent of the nature of the antigens used or the type of neoplasm. Combinations of immunotherapy with other treatment modalities resulted in augmentation of antitumor effects. Preliminary studies indicate that cellular and noncellular mediators of delayed hypersensitivity induce antitumor activities that may be significant in the regressions of neoplasms induced by cell-mediated immune challenge reactions. Since cell-mediated immune reactions are frequent occurrences, they may be a factor in apparently spontaneous regressions of neoplasms.

Immunotherapy for accessible tumors utilizing delayed hypersensitivity reactions and separated components of the immune system.

Courses of repeated delayed hypersensitivity challenge reactions at the sites of tumors have been shown to eradicate malignant and premalignant epidermal neoplasms. Local immunotherapy produces therapeutic responses in malignant and premalignant lesions before they are clinically detectable. This leads to a reduced incidence and prevention of tumors. Immunotherapeutic approaches are effective in controlling the early stages of mycosis fungoides and may aid in the management of the cutaneous manifestations in the late stages of the disease. Immunotherapeutic methods induce regressions of soft tissue lesions of a number of multifocal or metastatic malignant diseases with or without concurrent chemotherapy. Immunotherapeutic effects on tumors are similar with primary and recall antigens. Separated components of the cell-mediated immune system induce regressions of tumors following intralesional or perilesional administration, indicating common factors in host defenses against malignant diseases.

Discussion paper: effect of supernatants from long-term lymphoid cell lines on metastatic cutaneous tumors following local injection.

A fraction with lymphokine properties was isolated from supernatant medium of the continuous cultured human lymphoblast cell line, 1788. Culture medium containing 2% human serum was used for cell growth in order to minimize antigenicity of supernatant fractions isolated from the medium. The culture medium was passed through an Amicon XM-100 membrane, concentrated over a PM-10 membrane, lyophilized, and reconstituted to a final concentration of approximately 40:1. Studies in vivo and in vitro showed that the active fraction contained skin reactive factor (when injected intradermally into guinea pigs and humans), lymphotoxin, migration inhibition factor, chemotactic factor, and macrophage activation factor. This same preparation, when injected intralesionally into cutaneous tumors, induced an inflammatory reaction followed by tumor regression. The fraction confined between membranes of pore size 10,000-100,000 daltons was active in promoting tumor regression, while the fraction less than 10,000 daltons was inactive. Patients with skin lesions from metastatic carcinoma of the breast and other malignancies were studied, and 16 out of 30 treated lesions were judged to have undergone either complete or greater than 50% regression. Of these, 8 were biopsied before and after lymphokine injection, and 6 out of 9 were negative for tumor cells. Additional studies in vitro with material fractionated on Sephadex G-200 indicated that the macrophage-activating component binds to alpha-2 macroglobulin in the culture medium.