Cytokine production in whole blood cell cultures of patients undergoing therapy with biological response modifiers or 5-fluorouracil

Abstract

We have measured the levels of interferon gamma (IFN gamma), tumor necrosis factor alpha (TNF alpha), interleukin-1 alpha (IL-1 alpha), IL-1 beta, and IL-2 in the whole blood cell culture supernatants of 43 tumor patients undergoing a treatment with biological response modifiers or a conventional therapy with 5-fluorouracil and leucovorin. In the blood cell cultures of the 16 patients who received 5-fluorouracil and leucovorin IFN gamma levels decreased (P < or = 0.01) and TNF alpha levels rose (P < or = 0.05) during each therapy cycle. However, in the blood samples a declining number of total leukocytes and lymphocytes was measured (P < or = 0.05). Progressive disease could be correlated to a tendency towards lower IFN gamma levels in the pretherapeutic cultures of these patients. The second group analyzed consisted of 8 patients receiving a low-dose IL-1 beta therapy. In this group we found either an unchanged or an augmented IFN gamma production of the blood cells during treatment. In the group of 13 patients receiving low-dose recombinant IL-2 (< or = 4.5 x 10(6) IU m-2 day-1) significantly increasing IFN gamma levels were seen in the blood cell cultures during the therapy (P < or = 0.05), although total leukocyte counts decreased. In this group, 4 had stable disease for at least 2 months and 9 patients had tumor progression under therapy. In the cultures of the latter a tendency towards lower IFN gamma values was found. Finally, the cytokine production in the blood cell cultures of 6 patients receiving a combination therapy of IFN alpha and high-dose IL-2 was studied. During this therapy a dramatically reduced production not only of IFN gamma but also of all other measured cytokines was found. In this group all patients had tumor progression under therapy. It is concluded that the measurements of cytokine production in a reproducible whole blood culture system may be useful for monitoring immunological therapies and may help us to find out which doses of biological response modifiers have enhancing or suppressive effects on the functions of the immune cells.

Immunotoxicology: challenges in the 21st century and in vitro opportunities

“…A very interesting opportunity, as mentioned above, is the fact that the very same test can be performed ex vivo after immunomodulatory treatment or intoxication (Hartung et al, 1995a;von Aulock et al, 2004;elsässer-Beile et al, 1993). the example of the whole blood immunotoxicity assay shows that with the relatively easy access to human primary cells, the field is predestined to use cells from the target species of interest.…”

Tumour necrosis factor, a key role in obesity?

“…When TNF is bound to these soluble receptors, it can no longer interact with the membrane forms and, therefore, it has been speculated that the presence of the soluble forms may constitute a way of regulating TNF actions [8]. Highly elevated concentrations of the soluble receptors are found in the course of AIDS [9], endotoxinemia [10] or cancer [11], which suggests a regulatory mechanism to counteract TNF production. Interestingly, Hotamisligil et al [12] have shown that TNFR2 is overexpressed in adipose tissue of obese humans (with a strong correlation with BMI, hyperinsulinaemia and TNF mRNA levels in adipose tissue) and that the levels of the soluble receptor (sTNFR2) are elevated six‐fold in the obese subjects as compared with the healthy controls.…”