Functional analysis of mononuclear cells infiltrating into tumors: lysis of autologous human tumor cells by cultured infiltrating lymphocytes.

Lymphocytes separated from surgically resected tumor tissue, uninvolved lung tissue, and peripheral blood of lung cancer patients were investigated for cytotoxic potential and analyzed for their phenotypes at the time of surgery and after having been propagated for 4 to 5 wk in the presence of interleukin-2. Most of the tumor lymphocyte infiltrates examined were shown to have a shift in favor of T8 subsets from those found in peripheral blood. No natural killer activity and low cytotoxicity against the autologous tumor were found to characterize the tumor-derived lymphocyte population. Propagation of lymphocytes from the different tissues of the cancer patient in the presence of interleukin-2 preparation induced widespread lytic activity against K562 cells, autologous and allogeneic tumors, but not autologous normal lung or lymphoblasts. However, cytotoxic activity against autologous tumor cells exerted by cultured tumor-infiltrating lymphocytes was found to be significantly higher than the activity of cultured lymphocytes isolated from peripheral blood or uninvolved lung tissue of the same patient. The elevated lytic activity of cells derived from the tumor tissue indicates the accumulation at the tumor site of precursors of natural killer-like cells and specifically stimulated antitumor effectors. Our results suggest the coexistence of two types of anti-autotumor cytotoxic lymphocytes at the tumor site: natural killer-like and specific cytotoxic T-cells.

Interleukin-2 production and activity in aged humans.

Peripheral blood mononuclear cells (PBMC) from aged humans were found impaired in their ability to mount T mitogen induced proliferative responses and NK cell activity. The production of IL-2 in response to PHA has also been found decreased in the aged subjects. When the data from IL-2 production and cellular responsiveness of aged and young groups were pooled for regression analysis, a significant correlation was found both for mitogenic responses and NK activity. The reduced proliferative responses and NK cell activity can be enhanced by exposing them to exogenous IL-2. However, they were not fully reconstituted to the levels achieved in PBMC of the young subjects supplemented in vitro with IL-2. This would indicate that the reduced cellular responsiveness in aging may be related both to a defect of IL-2 receptor expression on T cells of aged subjects and to an impairment in the endogenous IL-2 synthesis in the aged individuals.

Augmentation of mitogen responsiveness in the aged by a special lipid diet AL 721.

Diminished immune responses in aging may be partially due to alteration of the lipid composition of membrane and a decrease in membrane fluidity. The effect of a daily diet of 10 g active lipid (AL 721) on the mitogen responsiveness of peripheral blood mononuclear cells (PBMC) from aged volunteers was studied. AL 721 is a mixture of lipids from hen egg yolk which was formulated for in vivo rectification of rigidified cell membranes in an attempt to restore proper physiological function. After 3 weeks on the diet, six aged participants displayed a significant increase in lymphocyte responsiveness to mitogens, while in four the responsiveness was unaffected, or increased insignificantly. In all six participants, a decline in lymphocyte responsiveness was observed a week after termination of the diet. Our preliminary results indicate that improvement of immune functions in the aged may be achieved by dietary manipulation of lipids. This innocuous approach seems to be of potential value and merits further studies.

Xenogenic response of human peripheral blood lymphocytes against murine cells in vitro.

Human peripheral blood lymphocytes (HPBL) were sensitized to mitomycin-treated murine spleen cells and mitomycin-treated mastocytoma P-815. A DNA proliferative response occurred to both cell types. The sensitized HPBL lysed chromium-51-labelled mastocytoma P-815 cells in a cell-mediated cytotoxicity assay (CMC). Evidence is presented that this human-murine xenogeneic system involves T cell response with specificity in CMC mainly toward H-2 antigens. DBA/2 splenocytes were superior to mastocytoma cells as sensitizers in the xenogeneic DNA proliferative assay.

Cellular and humoral suppressor activity induced by concanavalin A-stimulated human fetal liver cells.

Incubation of human fetal liver cells (HFLC), obtained from 8-12-week-old fetuses, with mitogenic doses of Concanavalin A (Con A) resulted in generation of a population that could suppress alloantigen-induced lymphoproliferation. The supernatant media in which these cells were activated suppressed DNA synthesis, both in mitogen and alloantigen induced peripheral blood lymphocyte cultures. Suppression was achieved only when the Con A-activated cells or the supernatants were added during the early stages of sensitization. Treatment of the cells with mitomycin C after Con A activation did not alter their ability to mediate suppression. This study indicates the presence of suppressor or presuppressor cells in the liver in the early period of fetal life. If the suppressor activity of HFLC in vitro is also induceable in vivo, it may contribute to the fetal defense mechanism against maternal rejection.

Autologous mixed lymphocyte reaction in man: further characterization of responding cells.

Experiments were performed to determine the type of cells undergoing thymidine incorporation in 7-day autologous mixed lymphocyte reactions (AMLR). Peripheral blood mononuclear cells (PBM) were separated into B cells, T cells, B + Null cells and T + Null cell-enriched populations. Cells were cultured in various combinations. Monocytes were removed to determine their influence on AMLR. The main thymidine-incorporating cells in cultures were shown to be Null cells and to a lesser extent T cells. Monocytes were found to have a more pronounced suppressor effect on stimulation of T cells by non-T cell populations in younger individuals than in the elderly. Whether Null cells undergo a spontaneous blastogenesis in culture or are stimulated by T or B cells, could not be answered by the present investigation.

Resistant hypertension associated with a renin-producing ovarian Sertoli cell tumor.

A 17-year-old girl with refractory, but nonmalignant, hypertension and hypokalemia is presented. A state of primary hyperreninism and secondary hyperaldosteronism was found to exist, together with a renin-producing Sertoli cell ovarian tumor. Upon removal of the tumor, this patient attained both a normotensive and a normokalemic state. A brief review is given of renal and nonrenal renin-producing tumors.

Secretion of suppressor factors by EBV infected B cell lines.

The infection of human peripheral blood B cells with Epstein-Barr Virus (EBV), induced the production of suppressor factor(s) which were released into the supernatant of the B-cell cultures. The induction of suppressive activity was independent of T-cell presence. The suppression was exhibited both against T-cell activity (MLR and mitogenic stimulation) as well as against B-cell mitogenic stimulation of human or murine B lymphocytes. The suppressive factor(s) was of a low molecular weight (equal or less than 5,000), resistant to trypsin and heating at 80 degrees C and its activity was partially inhibited by neuraminidase treatment. These findings indicate that the suppressive factor(s) is not correlated to immunoglobulin production, is not apparently of a protein nature, and might be of ganglioside or siaylated glycoprotein structure. Our present findings suggest that, in addition to T cells, B cells might also play an immunoregulatory role in the expression of immune response potential.

Immunosuppression in vitro and in vivo by supernatants from murine lymphoma cell lines.

Supernatants from four murine B and one T-cell lymphoma cultures were tested for their suppressive effect on allogeneic response in vitro (MLR), on their effect on mitogenic response to ConA and LPS and on their effect in vivo on allogeneic stimulation of BALB/c mice by C57BL/6J spleen cells. Two out of four B-cell supernatants (WEHI-231 and 24-666) were also tested for their effect on induction of IL-2 like activity by ConA in BALB/c spleen cells and on the effect of their injection on subsequent allogeneic response of spleen cells from BALB/c mice immunized against C57BL/6J spleen cells. The supernatants from all lymphoma lines inhibited MLR but only supernatants from the B-cell lymphoma lines also inhibited mitogenic stimulation in vitro and allogeneic immunization. The two B-cell supernatants tested also inhibited induction of IL-2 like activity and decreased the ability of spleen cells from the injected mice to respond to allogeneic stimulation. Partial characterization of supernatants from WEHI-231 and 24-666 lymphoma cultures revealed the non-protein nature of the suppressor factor(s), indicated a molecular weight of less than 5 Kd and suggested a ganglioside or a sialyated glycoproteinic nature.

Listeria monocytogenes pericarditis in a chronically hemodialyzed patient.

We describe the third documented case of Listeria monocytogenes pericarditis. This occurred in a 54-year-old woman with end-stage renal failure on chronic hemodialysis. Her initial presentation was one of Listeria monocytogenes bacteremia, which apparently responded to two weeks of cefazolin sodium therapy. After cessation of therapy the patient returned with Listeria monocytogenes pericarditis, this responding completely to four weeks of erythromycin therapy. We could not rule out coexisting endocarditis, especially since we found high levels of circulating immune complexes which subsided as the patient's condition improved. Further immunological studies displayed a decrease in cellular functions. This case illustrates the importance of Listeria monocytogenes as a human pathogen in immunocompromised patients. Listeria should be included among the potential causative agents of pericarditis in such patients.

Acute anuric bilateral ureteral obstruction in malignant lymphoma.

Two cases of bilateral ureteral obstruction in malignant lymphoma are reported. In one case, primary infiltration of the ureters by Hodgkin's disease was demonstrated, while in the other the obstruction was due to ureteral compression by enlarged lymph nodes. The diagnostic procedures and the management of such cases are discussed.

Cholesterol granuloma.

The "idiopathic haemotympanum" is in reality a persistent secretory otitis media. Histologically it demonstrates a chronic granular mastoiditis associated with a cholesterol granuloma. This, however, is not a very specific finding and may be frequently found in other types of the 'underaerated middle ear syndrome', wuch as ears with central perforations or indeed with cholesteatoma. In these granulomas, iron deposits are found, as well as the iron-chelating agent--lactoferin, which is known to be part of the defence mechanism of all mucosas. Lactoferrin is specifically secreted by the exocrine glands and it is not surprising to find much of it in the middle ear in a situation where mucus is being abundantly synthesized--as the case may be in any otitis media. There is no real clinical or histological evidence, so far, that neither the iron deposits nor the cholesterol crystals of these granulomas emanate from a previous hemorrhage.

Stevens-Johnson syndrome associated with Mycoplasma pneumoniae infection.

The Stevens-Johnson syndrome is a relatively uncommon, severe form of erythema multiforme. A case is presented in which Mycoplasma pneumoniae infection was established as the causative agent. Clinicians need to be aware of this uncommon association. We emphasize the need to consider M. pneumoniae infection as one of the many agents responsible for the Stevens-Johnson syndrome.

[Diabetes insipidus induced by post-resuscitation hypoxia].

A 13-year-old girl was admitted with meningeal signs. A lumbar puncture was followed shortly by cardiorespiratory arrest. In spite of intensive resuscitation she remained comatose and severe polyuria diagnosed as diabetes insipidus appeared. The differential diagnosis of dehydration and polyuria in patients after cardiorespiratory arrest and resuscitation should include hypothalamic injury causing diabetes insipidus.