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The use of viable frozen lymphocytes for studies in human tumor immunology.

Viable frozen lymphocytes displayed activity in blastogenesis assays that was indistinguishable from freshly prepared lymphoid cells. Similarly, cytotoxic activity of lymphocytes against melanoma target cells from melanoma patients was only slightly affected by the freezing procedure. Frozen lymphocytes provided a highly reproducible source of cells in these assays. The use of viable frozen peripheral blood lymphoid cells for the retrospective analysis of a cancer patient's immune response is described.

Adult↗

Inflammatory and immune cell function in psoriasis--a subtle disorder I. In vivo and in vitro survey.

Intradermal skin testing of normal and psoriatic subjects with common antigens, SKSD, Derm-O and PPD, reveals psoriatic subjects to have a decrease in both the amount (not incidence) of erythema (p less than 0.005) and in-duration (p less 0.005) to SKSD. Among all subjects having more than 10 mm erythema to Derm-O and SKSD, 49% of psoriatic and 77% of normal subjects have more than 10 mm induration (p less than 0.001). After sensitization, the response to 30 microgram challenge dose of dinitrochlorobenzene is positive in 50% of psoriatic and 88% of normal subjects (p less than 0.02). Uptake of (3)H thymidine by mitogen stimulated lymphocytes from psoriatic subjects is suppressed at each point of the linear component of a dose response curve. The mitogen dose to produce peak responses in psoriatics was 125% greater than that for normal subjects. In one-way mixed lymphocyte responses to pooled allogeneic stimulator lymphocytes, peripheral blood mononuclear cells from psoriatic subjects show suppression, the mean stimulation index was 55% of that of normal (p less than 0.01). Finally, in vitro polymorphonuclear leukocyte functions (chemotaxis, phagocytosis, and NBT reduction) appear to be within normal limits. When the foregoing parameters were compared with disease activity, there was no correlation.

Dinitrochlorobenzene↗

An animal model for chronic ulceration.

Few techniques are available for the study of persistent ulceration. In this report we describe a suitable animal model. Intracutaneous injection of sodium tetradecyl sulphate (STD) in guinea-pig flank skin caused a reproducibly sized and shaped superficial ulcer that healed approximately three times more slowly than punch biopsies of comparable size. Histological, immunofluorescent and autoradiographic studies showed that it possesses many of the morphological and kinetic features of human stasis ulceration. STD ulcers treated with a streptokinase/streptodornase solution healed with less slough and tended to re-epithelialize more rapidly than controls, but ulcers treated with dextran polymer beads or a stabilized hydrogen peroxide cream healed at the same rate as their control.

Adult↗

Bee venom immunotherapy induces a shift in cytokine responses from a TH-2 to a TH-1 dominant pattern: comparison of rush and conventional immunotherapy.

BACKGROUND: The mechanism of immunotherapy is unclear. Allergic disease is known to involve enhanced TH-2 cytokine responses to allergen. OBJECTIVE: In order to investigate the mechanisms of immunotherapy, we have examined changes in cytokine secretion before (13 patients) and during (nine patients) both rush and conventional venom immunotherapy (VIT) in bee venom allergic patients. METHODS: Peripheral blood mononuclear cells were stimulated in vitro with bee venom, non-specific antigen or mitogen and secretion of IL-4 (TH-2) and IFN gamma (TH-1) over the culture period measured. RESULTS: Untreated patients had TH-2 responses to venom and TH-1 responses to antigen and strong proliferative responses to venom. Controls showed no response (proliferation or cytokines) to venom and the normal TH-1 response to antigen. VIT resulted in marked changes in cytokine secretion to venom, with reduction of the abnormal TH-2 response and induction of a TH-1 response. The pattern differed in rush and conventional VIT. One day after rush VIT there was a significant fall in IL-4 secretion (P < 0.01), which rose by 3 weeks then declined. In conventional VIT there was a gradual reduction of IL-4 production significant after 2 months and undetectable by 6 months. IFN gamma secretion was induced by VIT. Proliferative responses mirrored the IL-4 changes. One day after rush VIT there was a loss of T cells, monocytes and NK cells from peripheral blood. CONCLUSION: This study shows that immunotherapy shifted cytokine responses to allergen from a TH-2 to a TH-1 dominant pattern, suggesting direct effects on T cells. How these cytokine changes relate to clinical desensitization is not clear. In the longer term they would result in an isotype switch from IgE to IgG. Early changes in cytokine or chemokine production might downregulate mast cell or basophil reactivity and explain the rapid desensitization in rush VIT.

Bee Venoms↗

Clonal priming of human lymphocytes with soluble microbial antigens: high-dose paralysis, restoration, and autologous leukocyte preference.

Lymphocytes stimulated in appropriate leukocyte cultures undergo blastogenesis and proliferation for a finite period of time. With specific antigens the proliferative response peaks usually between 4 and 8 days, after which the blastoid cells revert to small lymphocytes. Lymphocytes "primed" in this manner can be restimulated to proliferate only by the same antigen with which they were incubated and only with an adequate amount of a self-specific, autologous, somatic product(s). First or "primary" leukocyte cultures stimulated by optimal or supraoptimal concentrations of soluble protein antigens (purified protein derivative (PPD), tetanus toxoid, Candida albicans) will undergo proliferation in the first culture, but the increased number of small lymphocytes that can be visualized after 10--14 days often fail to respond to any stimulus in second (secondary) or "primed" cultures. However, when fresh X-irradiated autologous cells are re-added in appropriate amounts, vigorous accelerated proliferation takes place. Addition of allogeneic cells to antigen-primed populations has one of three effects: (1) no effect (complete restriction); (2) in some instances allogeneic cells restore a significant response to the specific antigen but almost never to the same degree as autologous cells; and (3) allogeneic cells can also induce high levels of accelerated responsiveness without added antigen. These findings are discussed in the context of a working hypothesis that self-specific factors are involved in all specific immune responses. The combination of antigen and self-specific factors may lead to a quantitatively unique immune response to all antigens in each individual. The preferential response to antigen in conjunction with autologous rather than allogeneic leukocytes suggests that self-specific products are required for recognition of soluble microbial antigens.

Antigens, Bacterial↗

Reversal reaction in patients with lepromatous leprosy after transplantation of human fetal thymic grafts.

An attempt has been made to reconstitute impaired cell-mediated immunity in 1 patient with indeterminate, 4 patients with borderline and 2 patients with polar lepromatous leprosy by grafting three thymus glands obtained from human fetuses of 14--19 weeks gestation. Most of these patients had severe ulcerative erythema nodosum leprosum (ENL) and were intolerant to dapsone. After thymus transplantation these patients were followed for 1 1/2 years. During this period, all conventional chemotherapy had been withdrawn. In most cases, there was dramatic improvement of the clinical condition, resolution of skin lesions, subsidence of ENL, clearance of bacteria from skin and reconstitution of several immunologic deficits; but late lepromin reactivity returned in none, which indicated permanent lose of resistance to Mycobacterium leprae.

Adult↗

The diagnostic and prognostic significance of delayed hypersensitivity skin testing in patients with urological cancer.

The responses to several skin test antigens of patients with bladder or prostatic cancer has been compared with responses of normal controls. All of the controls and all of the patients with prostatic cancer (irrespective of the stage of the tumour or the method of treatment) showed responses to dinitrochlorbenzene. 80% of all patients with bladder cancer responded to DNCB, but a highly significant number of patients with advanced disease showed no response. This test appears especially useful in predicting the prognosis in patients with bladder tumours and its routine use is recommended. Response to candida extract, streptokinase/streptodornase (SKSD) and purified protein derivative (PPD) were also studied. No conclusion could be drawn as to their value as a prognostic index.

Aged↗