Role of interleukin-2 (IL-2) in cancer-related immune deficiency: in vitro response to IL-2, production of IL-2, and IL-2 receptor expression in patients with advanced cancer.

The aims of the investigation were 1) to determine if there are defects in interleukin-2 (IL-2) regulation on either phytohemagglutinin (PHA)-activated or non-PHA-activated peripheral blood mononuclear cells (PBMC) in cancer patients to ascertain the role of IL-2 in this disease; 2) to carry out preliminary experiments for a direct quantitative evaluation of endogenous IL-2 production by PBMC cultures; and 3) to evaluate the IL-2 receptor expression by PBMC of cancer patients. An assessment of lymphocyte subsets was also performed with monoclonal antibodies in a selected group of patients. A total of 170 patients entered the study. Cancer sites were larynx (48), breast (44), lung (30), colorectal(23), and gynecologic (25). Staging showed in the former three cancer sites predominantly localized or only locally advanced disease and in the latter two sites disseminated disease. PBMC cultures were performed with microtiter plate technique and 3H-thymidine uptake evaluation using polyclonal mitogens, IL-2, and a monoclonal antibody against IL-2 receptor. Our results provided evidence that the cancer patients exhibit a T-cell functional immunodepression, which, to some extent, progresses during tumor growth so that the localized disease shows a low-grade defect and advanced disease a high-grade defect. Our data also clearly suggest that IL-2 deficiency is the primary factor involved in this functional immune impairment. We found no significant defect in the IL-2 receptor expression by PBMC of cancer patients. Our data also seem to support the in vivo therapeutic administration of IL-2 and lymphokine-activated killer cells to cancer patients.

Interleukin 2 (IL 2) relationships with the cancer-related immunodeficiency: in vitro response to exogenous IL 2 by PHA-activated and non PHA-activated peripheral blood mononuclear cells from cancer patients.

The aims of the investigation were: 1) to determine if there are defects in interleukin 2 (IL 2) regulation either on phytohemagluttinin (PHA)-activated or non PHA-activated peripheral blood mononuclear cells (PBMC) in cancer patients, in order to ascertain the role of IL 2 in this disease, and 2) to carry out preliminary experiments for a direct quantitative evaluation of endogenous IL 2 production by PBMC cultures of cancer patients. An assessment of lymphocytes subsets was also performed with monoclonal antibodies in a selected group of patients. A total of 159 patients entered the study. Cancer sites were: larynx, 49; breast, 42; lung (NSC), 25; colorectal, 18; and gynecologic, 25. In the former 3 cancer sites, staging showed localized or only locally advanced disease and in the later 2 sites it showed disseminated disease. Our results provided evidence that the cancer patients exhibit a T cell functional immunodepression, which progresses during tumor growth, so that the localized disease shows a low-grade defect and advanced disease shows a high-grade defect. Our data also clearly suggested that the factor involved with a primary role in this functional immune impairment is the IL 2 deficiency. In our study we have not found a substantial difference of activity between recombinant and nonrecombinant IL 2, although the comparison of the relative activities of the two types of IL 2 is not easy to make, since they are expressed in different ways; however the recombinant one appeared to be slightly more active, probably for the higher purity. Our data also seem to support the perspective of the in vivo therapeutic administration of IL 2 in cancer patients.

Familial chronic B-cell malignancy. Hairy cell leukaemia in mother and daughter.

Two familial cases of hairy cell leukaemia are reported: a daughter, 44-years-old, with a very unusual ultrastructural pattern found in hairy cells, the "tubuloreticular inclusions", and her mother, 71-years-old, who was affected six years later. Routine laboratory investigations, cytochemical and cytogenetic studies including HLA typing, as well as in vitro proliferative response of peripheral blood mononuclear cells (PBMC) to polyclonal mitogens and to exogenous interleukin 2, were performed. The immunological characterization by assessing the cell surface phenotypic markers with monoclonal antibodies and transmission electron microscopy (TEM) investigations were also carried out. In case 2 all tests were performed both on PBMC and on the bone marrow cells. To the best of our knowledge this is one of the first reports of such familial association. The possibility that genetic factors might play a role in the etiology of leukaemia in man is discussed: in our two cases, however, cytogenetic studies did not support this, while HLA typing revealed a non-significant association of HCL with DQw3 allele. Alternatively, an environmental factor has been considered, and a viral infection-perhaps by a retrovirus of the HTLV family has been suggested as tubuloreticular inclusions have been found in both hairy cell leukaemia, as reported, by us, and AIDS-LAS. However, a long time elapsed between the manifestation of HCL in the daughter and in the mother, and as the two patients had not been living together at that time, the possibility of a viral transmission seems minimal. The results of TEM and of immunological investigations are presented and discussed. Both, but particularly the latter, support the B cell nature of the hairy cell.

Peripheral blood lymphocyte response to recombinant and non-recombinant interleukin 2 in previously treated patients with Hodgkin's disease, long-time off-therapy.

13 patients with Hodgkin's disease (HD) previously treated, 9 of whom were long-time (more than 2 yr) off-therapy, were studied for peripheral blood lymphocyte response to interleukin 2 and for lymphocyte subpopulations by means of in vitro cultures and monoclonal antibodies. The aim of the study was to ascertain the role played by interleukin 2 in the impaired cell-mediated immunity of HD patients. The results show a response of peripheral blood mononuclear cells of HD patients to either the T cell-specific polyclonal mitogens PHA and Con A or to the T cell-dependent, although B cell-specific, PWM, most significantly decreased compared to the normal response. As far as the interleukin 2 involvement in HD is concerned, our study suggests: an impaired endogenous interleukin 2 production by T lymphocytes, a most probable deficiency of the interleukin 2 receptor (Tac) expression and 3) a decrease of the number and/or of the function of NK cells no longer responsive in vitro to interleukin 2. The phenotypic analysis of peripheral blood mononuclear cells showed a slight decrease of total T cells (T3+), of the helper/inducer subset (T4+) and of the T4+/T8+ cells ratio. Our data seem to support the rationale for a therapeutical approach with interleukin 2 in controlled clinical trials also in HD patients, according to the experiments in progress in solid tumor patients.

Physiological and genetic characterization of the Gly40Ser mutation in the glucagon receptor gene in the Sardinian population. The Sardinian Diabetes Genetic Study Group.

A Gly40Ser amino acid substitution in the glucagon receptor gene has been associated with non-insulin-dependent diabetes mellitus (NIDDM), but the questions raised about its physiological implications have not been resolved. The aim of this study was to determine the frequency of the Gly40Ser mutation in different regions of Sardinia and to investigate the physiological implications of the mutation in glucose and insulin homeostasis. We studied a population of 691 subjects selected on the basis of their Sardinian origin. Only heterozygous subjects were found, 21 of 574 (3.6%) in NIDDM patients and 5 of 117 in non-diabetic subjects (4.2%). In northern Sardinia 3.4% of the NIDDM patients were carriers of the Gly40Ser substitution, 1.4% in central Sardinia, while 7.6% carried the substitution in the Southern part. No significant differences were found between hypertensive and normotensive subjects with respect to the presence of Gly40Ser. Ten subjects with Gly40Ser were carefully matched for diabetic state, BMI, age, sex, and geographical origin with 10 patients with Gly40, and a glucagon infusion test was performed using 1, 3, 9 and 27 ng glucagon kg-1.min-1 for 30 min. Blood for determination of glucose, glucagon, and insulin concentrations was drawn at 15-min intervals from the Controlateral arm. Plasma glucagon increased dose-dependently during the infusion with no significant difference between the two groups. Carriers of Gly40Ser had a significantly lower (p < 0.02) increase in plasma glucose concentration in response to glucagon infusion compared to Gly40 homozygous subjects at all times, while the plasma insulin increase was not significantly different at any time. In conclusion, our results indicate that the Gly40Ser variation is not associated with NIDDM in the Sardinian population and that its frequency varies in different parts of Sardinia. Moreover in vivo Gly40Ser plays a physiological role in the glucose homeostasis under glucagon control both in NIDDM and non-diabetic subjects. This latter result suggests that this amino acid substitution in the glucagon receptor may lead to a decreased blood glucose concentration because of the reduced stimulation of liver glucose output via the glucagon receptor.