Gerald W. King

Evidence for hydroxyl radical generation by human Monocytes.

A number of highly reactive oxygen species have been implicated in the oxygen-dependent mechanisms involved in bactericidal activity of phagocytic leukocytes. Hydrogen peroxide and superoxide, two agents known to occur during phagocytosis, are thought to interact to generate hydroxyl radical, singlet oxygen, and other potentially reactive molecules. Using an assay system of ethylene generation from methional, cell preparations of human monocytes were demonstrated to generate hydroxyl radical or a similar agent during phagocytosis of zymosan particles. The generation of ethylene was impaired by agents which reduce superoxide or hydrogen peroxide concentrations as well as by agents reported to be hydroxyl radical scavengers. The ethylene generation did not appear to be dependent on myeloperoxidase in that azide enhanced ethylene generation. Monocytes from a patient with chronic granulomatous disease failed to generate ethylene during phagocytosis. This assay technique may be useful in exploring the metabolic events integral to the bactericidal and inflammatory activity of phagocytic leukocytes.

A comparison of the metabolic response to phagocytosis in human granulocytes and monocytes.

Recent studies indicate that oxygen radicals such as superoxide or singlet oxygen may be important in the functional activity of human granulocytes. We have examined the possible importance of these radicals in the functional capacity of human blood monocytes. Monocytes, like granulocytes, generate chemiluminescence during phagocytosis. Chemiluminescence is impaired 50-90% by superoxide dismutase, an enzyme which enhances the dismutation of superoxide to hydrogen peroxide. These results indicate that superoxide is related to the chemiluminescence generated by monocytes. Superoxide dismutase in a concentration which impaired chemiluminescence also impaired the staphylococcal killing by monocytes. Hexose monophosphate shunt activity and hydrogen peroxide production by granulocytes and monocytes were also evaluated. The oxidation of [1-14C]glucose was used as a measure of hexose monophosphate shunt activity and the oxidation of [14C]formate as an estimation of hydrogen peroxide production. The oxidation of both substrates by monocytes was increased during phagocytosis but, in contrast to results in granulocytes, was not further increased by the addition of superoxide dismutase. These data indicate that superoxide may be important in bactericidal activity of human monocytes. Our results also suggest that the metabolism of oxygen radicals in monocytes and granulocytes may be different.

Leukemic reticuloendotheliosis: A study of the origin of the malignant cell

A highly pure preparation of neoplastic cells from the spleen of a patient with leukemic reticuloendotheliosis was studied for function, membrane characteristics and glucose metabolism. Glass adherence and phagocytosis of small particles (latex and carbon black) were demonstrated with phase contrast microscopy. Staphylocidal activity was similar to that of normal monocytes. Immunofluorescent assays revealed nonspecific uptake of antiserums to immunoglobulins G (IgG), M (IgM), A (IgA) and kappa and kappa and lambda light chains. Rosette assays indicated the presence of receptors for IgG on the surface of all cells but no receptors for complement (C3) or sheep red blood cells. Glucose metabolic studies revealed a pattern that differed from that of normal monocytes or lymphocytes with intermediate values for glycolysis, low hexose monophosphate shunt activity and high Krebs cycle activity. Increments in tritiated (3H)-thymidine uptake and glucose metabolism in response to phytohemagglutinin stimulation were minimal (5 per cent of normal lymphocyte values) and no response was noted with pokeweed mitogen stimulation. These findings suggest that the leukemic reticuloendotheliosis cell most closely resembles cells of the monocyte-histiocyte series.

A retrospective and prospective study of serial CEA determinations in the early detection of recurrent colon cancer

The results of the present study emphasize the importance of minimizing the time delay between a significant elevation in CEA and a second-look operation. Equally important are the frequency of serial determinations, a thorough understanding of the limitations of the assay, and careful evaluation of the clinical condition of the patient.

5 FU infusion with mitomycin‐C vs. 5 FU infusion with methyl‐CCNU in the treatment of advanced upper gastrointestinal cancer. A Southwest oncology group study

A randomized trial was conducted by the Southwest Oncology Group (SWOG) in advanced carcinoma of the stomach and pancreas. Patients were assigned to receive monthly 5‐fluorouracil 96‐hour continuous infusions with either bolus mitomycin‐C or oral methyl‐CCNU. Mitomycin‐C and methyl‐CCNU were administered every eight weeks. The 5 FU‐mitomycin combination produced a 14% and 22% response rate in disseminated stomach and pancreatic carcinoma, respectively. The combination of infusion 5 FU and methyl‐CCNU achieved responses in 9% and 5% of stomach and pancreatic tumors, respectively. There was ho significant difference in survival between limbs for either tumor. Median survival in gastric carcinoma on the 5 FU‐mitomycin regimen was 25 weeks vs. 18 weeks on the 5 FU‐methyl‐CCNU arm. In pancreatic carcinoma median survival on the mitomycin limb was 19 weeks as compared to 17 weeks on the methyl‐CCNU program. Leukopenia was greater for the first course on the mitomycin limb. Regression analysis demonstrated that performance status was the most important pretreatment characteristic for predicting survival in both tumors. Neither 5 FU infusion combination appears to significantly alter the dismal prognosis of advanced upper gastrointestinal neoplasms.

Immune function of successfully treated lymphoma patients.

Immunologic function was evaluated in 12 patients with Hodgkins disease and 5 patients with lymphocytic lymphoma who had been successfully treated with either chemotherapy, radiation therapy, or both of these modalities 3-42 mo previously. Only two of the patients were found to have total anergy to a battery of six recall skin test antigens and all were responsive to skin testing with phytohemagglutinin. However, 10 of 16 patients were unable to develop delayed cutaneous hypersensitivity to either of the neoantigens dinitrochlorobenzene or keyhole limpet hemocyanin. Four other patients developed reactivity to only one of these neoantigens for a total of 14 of 16 (88%) of the patients demonstrating some impairment in neoantigen response. Total lymphocyte, T-lymphocyte, B-lymphocyte, and null cell numbers, as well as serum immunoglobulins were quantitatively normal. Monocyte numbers, chemotaxis, and Fc receptor activity were normal. Monocyte staphylocidal activity at 60 min was modestly depressed and candidacidal activity was depressed in those receiving both chemotherapy and radiation therapy. Spontaneous (unstimulated) lymphocyte [3H]thymidine incorporation was low in the patients as a group and lymphoblastic transformation to specific antigens was impaired in 11 of 17 patients who had positive skin test reactions to the same antigen. Highly significant suppression of lymphoblastic transformation was noted after stimulation by the mitogens phytohemagglutinin, pokeweed, and concanavalin-A. The greatest impairment of mitogen response was seen in those patients receiving both chemotherapy and radiation therapy. These data demonstrate specific impairments of neoantigen processing, lymphocyte function, and to a lesser extent monocyte function in successfully treated patients with lymphoma. These impairments may contribute to the increased incidence of infections and second primary malignancies in these patients.

The effect of tuberculosis and neoplasia on human monocyte staphylocidal activity

Abstract Monocytes from normal subjects and patients with tuberculosis, lymphoma, or solid tumors were evaluated for their ability to kill Staphylococcus aureus organisms in an in vitro system. Monocytes from patients with tuberculosis demonstrated enhanced staphylocidal activity as compared to normal monocytes after 30- and 60-min periods of incubation. Monocytes from lymphoma patients had normal killing rates after 30 min incubation but demonstrated significantly enhanced killing after 60 min incubation. Monocytes from patients with solid tumors demonstrated essentially normal killing rates. These data suggest functional activation of human monocytes by mycobacterial infection and lymphoma similar to that described in animal models.

Hodgkin's disease occurring in a patient with the Wiskott-Aldrich syndrome.

Hodgkins disease, nodular sclerosing, developed in a 16‐year‐old man with the Wiskott‐Aldrich syndrome. Two brothers and two nephews had documented Wiskott‐Aldrich syndrome and had died of infectious complications in childhood. While the patient reported here had lifelong thrombocytopenia and recurrent upper respiratory infections, he had no severe infection prior to the development of Hodgkins disease. Skin test sensitization with dinitrochlorobenzene was unsuccessful. No antibodies were found after immunization with pneumococcal polysaccharides. Platelet aggregation studies were abnormal in the patient, his mother, and one of his nephews. A complete response of short duration occurred after treatment with nitrogen mustard, vincristine, procarbazine, and prednisone. On recurrence, he proved unresponsive to further chemotherapy or radiation therapy. Infection with four different fungi was found at autopsy. This patient is the third recorded case of Hodgkins disease associated with the Wiskott‐Aldrich syndrome.

Neoantigen Response in Patients Successfully Treated for Lymphoma: A Southwest Oncology Group Study

To ascertain the cellular immune function of patients successfully treated for lymphoma, we measured skin-test reactivity to a battery of recall antigens, phytohemmagglutinin (PHA), and the neoantigens keyhole limpet hemocyanin (KLH) and dinitrochlorobenzene (DNCB). Seventy-four patients with Hodgkins disease and 31 patients with non-Hodgkins lymphoma were studied from 3 to 186 months after cessation of therapy for lymphoma. Although reactivity to recall antigens and PHA was normal, the number of patients responding to the neoantigens was significantly (P less than 0.01) lower than normal (KLH, 35%; and DNCB, 34%). This impairment in reactivity to neoantigens could not be correlated with specific diagnosis, stage of disease, or type of treatment. Reactivity to DNCB was significantly (P less than 0.01) improved in those patients studied more than 3 years after treatment, but the number who reacted was still markedly abnormal (17 of 33). Thus, successfully treated patients with lymphoma seem to have difficulty in responding to new foreign antigens.

Normal human monocytes inhibit tumor cell growth in vitro

Normal human monocytes were evaluated in an in vitro assay of growth inhibition of tumor cells. Monocytes were isolated from the blood of 6 normal subjects by Ficoll-Hypaque separation and adherence to plastic microtest wells. Cervical carcinoma cells (HeLa) were added to the microwells to result in a ratio of 50 monocytes to one HeLa cell. Cultures were then incubated for 6 to 46 hr. Growth inhibition was evaluated by measuring the uptake of 3H-thymidine over a 4-hr pulse period after 2, 18, or 42 hr of monocyte-HeLa interaction. Inhibition of HeLa growth by monocytes was 23.8% +/- 8.6% over 6 hr, 22.0% +/- 8.9% over 22 hr. and 68.3% +/- 7.5% over 46 hr. Growth inhibition of HeLa cells was confirmed by direct enumeration of HeLa cells at the end of coincubation. Attachment of monocytes to the HeLa cells was confirmed by light and scanning electron micrographs. Granulocytes, lymphocytes, and other cell lines did not comparably inhibit HeLa growth and media replenishment did not ablate the effect. These data demonstrate that normal human monocytes can inhibit the growth of a malignant cell line in vitro in the absence of overt activation procedures.