William H. Adler

Transfer factor in the treatment of a case of chronic mucocutaneous candidiasis

Abstract A girl with a severe case of chronic mucocutaneous candidiasis in whom a partial defect in cell-mediated immunity to C. albicans was demonstrated is described. Chemotherapy with anti-Candida agents afforded only temporary clinical improvement. Although her delayed hypersensitivity skin test response to C. albicans was consistently negative, her lymphocytes underwent blast transformation when cultured in the presence of C. albicans and other mitogens. The addition of exogenous transfer factor to the culture medium increased her lymphocyte blast transformation response to C. albicans. Because of these findings she was treated with transfer factor with the aim of restoring expression of cutaneous hypersensitivity to the antigen, and providing her with clinically significant immunity to the antigen. Following a course of repeated injections of transfer factor given in conjunction with a short course of intravenous amphotericin B, her infection cleared and she has had no recurrence of infection for at least 6 months.

Selective effects of mitogens on subpopulations of mouse lymphoid cells

Abstract The mitogenic effects of phytohemagglutinin (PHA), pokeweed mitogen (PWM), staphylococcal enterotoxin B (SEB) and endotoxins upon mouse spleen cells and various subpopulations were examined. The differential responses to these mitogens by cells of varying density indicate that LPS stimulates a different subpopulation from that of PHA and SEB. The subpopulation containing cells responsive to PWM seemed to differ from both of these patterns. In contrast to the effect of PHA, PWM, and SEB, thymus cells were not stimulated by LPS. LPS- and PWM-stimulated cells taken from both neonatally thymectomized and adult-thymectomized, irradiated, and bone-marrow-reconstituted animals: SEB and PHA failed to do this. It is concluded that the subpopulation responding to SEB, like that to PHA, is thymus dependent but that the two are probably not completely overlapping. That containing PWM responsive cells involves both thymus-independent and thymus-dependent cell components. The subpopulation stimulated by LPS is contained in a low density sub-population, which appears to be completely T independent.

The effect of PHA, PPD, allogeneic cells, and sheep erythrocytes on albumin gradient-fractionated mouse spleen cell populations☆

Abstract Populations of spleen cells from immune and nonimmune mice were separated by albumin density-gradient centrifugation. Primary responsiveness to PHA, to alloantigens in mixed cell culture, and to SRBC was assayed in various fractions by tritiated thymidine incorporation. Changes in the antigen-sensitive cell population in various gradient fractions were also examined in BCG-, alloantigen-, and SRBC-immunized animals. Primary reactivities to PHA, SRBC, and alloantigens are properties of a small, dense lymphocyte population. The antigen-sensitive cell population from immune mice is augmented chiefly in the less dense, large lymphocyte populations of the spleen in the cases of alloantigen and SRBC immunizations. In contrast only the denser, small lymphocytes showed increased antigen sensitivity after BCG immunization. These data suggest that immunity to alloantigens involves augmentation of a second population of antigen-sensitive cells which may produce antibody whereas immunity to BCG under these conditions is limited to an antigenreactive small lymphocyte population.

The interactions of mitogens with lymphoid cells and the effect of neuraminidase on the cells' responsiveness to stimulation☆

Abstract Early events in the interaction between mouse spleen cell populations and two mitogenic agents, phytohemagglutinin and bacterial lipopolysaccharide, have been examined. Qualitative assessment by a fluorescent antibody technique indicated that these mitogens are first bound to cell surface membranes, then internalized. This process was maximal 30 min after PHA contact; a similar effect required up to 1 hr in the case of LPS. Internalization, but not binding, was essentially blocked by low temperature or by pretreatment of the cells with neuraminidase. More quantitative data were obtained through an absorption technique which assayed the amount of mitogen removed by cells through binding and internalization. Neuraminidase treatment almost completely blocked the PHA-absorptive capacity of such cells; however, recovery from this block occurred 4–6 hr after incubation without the enzyme. Similar treatment with neuraminidase also blocked the mitogenic effects of PHA. Prolonged incubation of cells alone in vitro resulted in progressive partial loss of their absorptive capacity; when they had been stimulatd by PHA and incubated for 24 hr or more, absorptive capacity for both mitogens was entirely lost. These studies show that binding and internalization of mitogens precede transformation and mitosis, and that both absorptive capacity and mitogenic effects are correlated with the capacity to internalize these mitogens.

Identification of mouse thymus antigen recognition function in a minor, low-density, low-Θ cell subpopulation

Abstract Using a one- or two-step syngeneic transfer system in bone marrow-restored, lethally irradiated mice, the anti-SRBC cooperative cell function of the thymus was identified as being in low-density cell subpopulations constituting less than 10% of total thymus cells. This “B layer” population consisted of about 30% small lymphocytes and 70% large lymphoid cells and blasts, which have an average Θ-antigen content more characteristic of peripheral lymphocytes than of thymocytes. The origin of these cells and some possible implications of these findings are discussed.

Strain specificity of monodisperse gamma-globulin appearance after immunization of inbred mice.

Summary Immunization of various inbred lines of young mice led to the appearance of monodisperse bands of gammaglobulin having antibody activity in the serum. Whereas the antibody titers achieved did not differ significantly, the proportion which developed these bands was as high as 60% in DBA/2 or C57BL/6 and F1 hybrids having these as one parent. This activity did not occur in A/J or CBA strains. These studies suggest an additional way in which genetic control might be exerted over the quantity of the immune response.

25 – PREPARATION AND ASSAY OF MOUSE MITOGENIC FACTORS

Publisher Summary The use of mouse lymphoid cell cultures to prepare mitogenic factors allows the investigation of the possible participation of histocompatibility antigens in the mitogenic stimulus. In some mixed spleen cell cultures, there appear to be histocompatibility antigens released into the medium. Other mixed-cell cultures and tuberculin-stimulated cultures appear to release nonspecific mitogens into the medium. This chapter describes an experiment for the preparation and assay of mouse mitogenic factors. In the experiment, mice were sacrificed by cervical dislocation. The abdomen was wiped with 70% ethyl alcohol and the skin incised with a small straight dissecting scissors. The skin was cut away from the left upper quadrant of the abdomen. The peritoneum was lifted with forceps and with a clean sterile scissors; the peritoneum was nicked and then incised over the area of the spleen. The supernate was poured off and 5 ml RPMI were again added. The cell button was dispersed by mixing and then recentrifuged. The supernatant was again poured off and the tubes drained for a minute. Untreated cells were then added to the mitomycin-treated cells.

Transfer factor in the treatment of chronic mucocuataneous candidiasis

The use of “transfer factor” to correct a partial defect in cellular immunity to Candida albicans was studied in an 8-year-old girl with chronic granulomatous mucocutaneous candidiasis.The patient first presented at age 5 years with extensive deforming encrusted lesions on her face, head, trunk and extremities. No evidence of endocrinopathy or antibodies to endocrine tissue was found. Her general humoral immunity was intact. She had a normal complement of granulocytes and lymphocytes. Candida aggregation activity was present in her serum. Her skin test response to C. albicans extract was consistently negative. Her lymphocytes underwent blastogenesis to PHA, diphtheria toxoid and C. albicans extract. However, in the presence of exogenous transfer factor, blastogenesis to candida increased. Transfer factor extracted from her cells did not transfer immunity to non-sensitized cells.Amphotericin B therapy cleared her skin lesions temporarily, but neither fresh frozen plasma injections nor 5-fluorocytosine was effective. Following 2 injections of transfer factor, she developed a positive skin test response to C. albicans extract, and her lymphocytes produced leukocyte inhibition factor (LIF) to candida. After a third injection there was appreciable clearing of the skin lesions.These findings indicate that exogenous transfer factor can restore cellular immunity to candida in a patient with chronic mucocutaneous candidiasis and may be an effective treatment for this disease.

PREPARATION AND ASSAY OF AN INHIBITOR OF DNA SYNTHESIS AND A NONSPECIFIC MITOGEN ELABORATED BY HUMAN LYMPHOBLASTS

Publisher Summary The long term cultures of human lymphoblast cell lines provide an excellent source for the study of certain biologically active, soluble factors produced by lymphoid cells, such as migration inhibitory factor, lymphotoxin, and others. Additional factors that have been recently found are the inhibitor of nucleic acid synthesis and a non-specific mitogen. These factors were discovered during attempts to determine whether the original donors of lymphoblast cell lines, derived from normal individuals and from individuals having infectious mononucleosis, were stimulated by their own long term cell lines. As the experiments eventually evolved, supernatants from these cell lines were incubated with the original donors peripheral leucocytes. Depending upon the culture conditions, either marked inhibition or stimulation was observed as detected by the incorporation of 3 H-thymidine or uridine.