Radioembolization of hepatocarcinoma with 90Y glass microspheres: treatment optimization using the dose-toxicity relationship.

AIM: Transarterial radioembolization (TARE) is, by all standards, a radiation therapy. As such, according to Euratom Directive 2013/59, it should be optimized by a thorough treatment plan based on the distinct evaluation of absorbed dose to the lesions and to the non-tumoural liver (two-compartment dosimetry). Since the dosimetric prediction with 99mTc albumin macro-aggregates (MAA) of non-tumoural liver is much more accurate than the same prediction on lesions, treatment planning should focus on non-tumoural liver rather than on lesion dosimetry. The aim of this study was to determine a safety limit through the analysis of pre-treatment dosimetry with 99mTc-MAA single photon emission computed tomography (SPECT/CT), in order to deliver the maximum tolerable absorbed dose to non-tumoural liver. METHODS: Data from intermediate/advanced hepato-cellular carcinoma (HCC) patients treated with 90Y glass microspheres were collected in this single-arm retrospective study. Injection was always lobar, even in case of bilobar disease, to avoid treating the whole liver in a single session. A three-level definition of liver decompensation (LD) was introduced, considering toxicity only in cases of liver decompensation requiring medical action (LD type C, LDC). We report LDC rates, receiver operating characteristic (ROC) analysis between LDC and NO LDC absorbed dose distributions, normal tissue complication probability (NTCP) curves and uni- and multivariate analysis of risk factors associated with toxicity. RESULTS: A 6-month timeline was defined as necessary to capture all treatment-related toxicity events. Previous transarterial chemoembolization (TACE), presence or extension of portal vein tumoural thrombosis (PVTT) and tumour pattern (nodular versus infiltrative) were not associated with tolerance to TARE. On the contrary, at the multivariate analysis, the absorbed dose averaged over the whole non-tumoural liver (including the non-injected lobe) was a prognostic indicator correlated with liver decompensation (odds ratio = 4.24). Basal bilirubin > 1.1 mg/dL was a second even more significant risk factor (odds ratio = 6.35). NTCP analysis stratified with this bilirubin cut-off determined a 15% liver decompensation risk at 50 Gy/90 Gy for bilirubin >/< 1.1 mg/dL. These results are valid for a 90Y glass microsphere administration 4 days after the reference time. CONCLUSION: Given the low predictive accuracy of 99mTc-MAA on lesion absorbed dose reported by several authors, an optimized TARE with 90Y glass microspheres with lobar injection 4 days after reference time should aim at an absorbed dose averaged over the whole non-tumoural liver of 50 Gy/90 Gy for basal bilirubin higher/lower than 1.1 mg/dL, respectively.

Altered cytokine production after human herpes virus type 6 infection.

Several strategies allow viruses to elude the surveillance of the immune system and to establish persistent infection in the host. One of such mechanisms is the immunosuppression caused by the direct infection and functional impairment of immune cells. Human Herpes virus type 6 (HHV-6) is a typical immunosuppressive agent, as suggested by its tropism for both CD4+ and CD8+ T cells, B cells, monocytes/macrophages, megakaryocytes and NK cells. In this study the production of IL-10 and IL-12 by peripheral blood mononuclear cells (PBMC) was evaluated during HHV-6 infection "in vitro". Our results demonstrate that HHV-6 up-regulates IL-10 production by PBMC. Furthermore, our data suggest that rhIFN gamma addition counteracts the effect of HHV-6 in promoting IL-10 release. To gain more insight into the role of IFN gamma, anti-IFN gamma monoclonal antibodies were added to PBMC stimulated with LPS. Neutralization of endogenous IFN gamma upregulated IL-10 release. Furthermore, HHV-6 infection inhibited IFN gamma release induced by LPS in PBMC. No basal production of IL-12 was found in PBMC. Moreover, HHV-6 infection did not induce IL-12 release by PBMC. On the contrary, IL-12 was detected in the supernatants of PBMC treated with LPS with or without rhIFN gamma. In these experimental conditions the further addition of HHV-6 markedly impaired IL-12 production. Moreover, the neutralization of IL-10 resulted in a significant up-regulation of IL-12. Finally our data suggest that the immunodysregulation induced by HHV-6 could be accounted for by a shift from a Th-1 to a Th-2 type cytokine profile.

Interleukin-12 and interleukin-10 production by mononuclear phagocytic cells from breast cancer patients.

Interleukin-10 (IL-10) and Interleukin 12 (IL-12) generation may be regulated by a complex monocyte and macrophage-derived cytokine network and an impairment of the immune system can be observed in neoplastic disease. In this study, we examined the production of these cytokines by phagocytic cells, obtained from breast cancer (BCa)-bearing patients. Our results suggest that an increased IL-10 formation may represent an important regulatory pathway of IL-12 production by BCa mononuclear cells. In this report, we show that mononuclear cells of patients affected by breast cancer have a defective IL-12 production capability while generating higher amounts of IL-10.

Granulocyte-macrophage colony stimulating factor modulates the production of TNF alpha by differentiated U937 cells infected with Leishmania major.

In this work, the production of tumor necrosis factor alpha (TNF alpha) during interaction of human phagocytes with the intracellular parasite Leishmania major was further investigated. The human monocytic cell line U937, differentiated with a combination of 1 alpha, 25 dihydroxyvitamin D3 (VD) and retinoic acid (RA), or with granulocyte macrophage colony stimulating factor (GM-CSF) was used. Differentiated U937 cells were infected with Leishmania major promastigotes, and TNF alpha was assayed in cell culture supernatants. It was found that the cytokine was produced only by U937 cells differentiated with VD/RA and further incubated with GM-CSF and LPS or interferon gamma (IFN gamma). L. major induced TNF alpha production only in the presence of GM-CSF. No direct relationship was found, however, between production of TNF alpha and resistance of differentiated U937 cells to infection with L. major.

Production of TNF alpha and interleukin 6 by differentiated U937 cells infected with Leishmania major.

Several cytokines play a crucial role in the defense of the host against protozoa belonging to the genus Leishmania. However, the role of tumor necrosis factor alpha (TNF alpha) and interleukin 6 (IL-6) in human leishmaniasis is still controversial. The aim of this work was to study, in an "in vitro" model, the interactions of human phagocytes with L. major. The U937 human monocytic cell line, differentiated with phorbol myristate acetate (PMA) or a combination of 1 alpha,25 dihydroxyvitamin D3 (VD) and retinoic acid (RA), was used in all the experiments. The rate of infection, as well as the production of TNF alpha and IL-6 by cells upon infection with promastigotes, were studied. It was found that, depending on the agent used for differentiation, U937 cells produced different patterns of cytokines. PMA differentiated cells produced significantly more TNF alpha, but less IL-6 than cells differentiated with VD-RA. No direct relationship was found between the ability of differentiated U937 cells to release TNF alpha or IL-6 and their leishmanicidal activity.

In vitro effects of lithium chloride on TNF alpha and IL-6 production by monocytes from breast cancer patients.

It is well known that lithium chloride (LiCl) is able to trigger human monocytes to release tumor necrosis factor alpha (TNF alpha). In this study we have evaluated the in vitro effect of LiCl on TNF alpha and interleukin-6 (IL-6) release by monocytes from patients affected by non-metastatic (BCa/M0) and metastatic breast cancer (BCa/M1), preincubated with autologous serum (sPt). Our data demonstrate that monocytes from cancer patients (BCa) treated with LiCl released lower amounts of TNF alpha compared to those from healthy donors (HD). Preincubation in autologous serum (sPt) impaired TNF alpha production by monocytes from BCa with LiCl. On the contrary, our data indicate that IL-6 production by monocytes treated was not impaired. Moreover, the results obtained from the same cells, preincubated in sPt and treated with LiCl, indicate that serum factors may synergize with LiCl treatment in releasing IL-6.

Productive HHV-6 infection in differentiated U937 cells: role of TNF alpha in regulation of HHV-6.

This study characterizes the effect of differentiation on the resistance of the human monocytic cell line U937 to human herpes virus type 6 (HHV-6). The use of monocytic cell line has the advantage of avoiding genetic variations among different donors. The HHV-6 infection was compared in undifferentiated U937 cells and U937 cells differentiated with a combination of vitamin D3 and retinoic acid. Undifferentiated U937 cells were highly resistant to HHV-6 infection. Differentiation of U937 cells was accompanied by an increase in permissiveness for HHV-6 demonstrated in terms of extracellular virus production and viral antigen positive immunofluorescent cells. Tumor necrosis factor alpha (TNF alpha) appears to be an essential mediator during the first line defences of the host against viruses, even though its role during viral infection remains controversial. For this reason we examined the behaviour of TNF alpha in differentiated U937 upon HHV-6 infection. No basal production of TNF alpha was found in culture supernatants, while HHV-6 infection up-regulated TNF alpha release. The addition of human recombinant-TNF alpha to HHV-6 infected cells induced a marked cytotoxic effect accompanied by an increased release of extracellular virus, whereas it did not affect viral replication, as shown by the unmodified percentage of antigen positive cells. In conclusion, TNF alpha acts as a soluble mediator of cytotoxicity against HHV-6 infected U937 cells, but it fails to induce an antiviral state.

Halogenated isoniazid derivatives VI. Evaluation of anti-HSV-1 and anti-HIV-1 in vitro activities of fluorinated analogues.

The synthesis and evaluation of antiviral in vitro activity are reported of some 2'-(1-arylethyl)isonicotinohydrazides (5a-d) and N-(1-arylethyl)isonicotinohydrazonic acids (6a-d), obtained by reducing fluorinated acetophenone isonicotinoylhydrazones (2a-d) with sodium cyanoborohydride. These INH analogues, along with other ones previously prepared, i.e. benzaldehyde isonicotinoylhydrazones 1, 4-aryl-1-methoxyl-1-(4-pyridyl)-2,3-diaza-1,3-butadienes 3 and 5-aryl-4-methyl-2-(4-pyridyl)-delta 2-1,3,4-oxadiazolines 4, were assayed for anti-HSV-1 activity on the monoblastoid cell line U937. Only some compounds (1b, 1d, 4d and 4e) displayed a moderate antiherpetic activity. In addition, the reduced compounds 5 and 6, submitted to the anti-HIV-1 screening, did not display significant effects in reducing virus-induced cytopathogenicity. The cytotoxicity of all compounds has been assayed on Vero cells and some considerations in correlation with structure are discussed.

Role of IFN gamma on TNF alpha, IL-1 beta and IL-6 release during HHV-6 infection.

The release of monokines such as Tumor Necrosis Factor a (TNF alpha), Interleukin-1 beta (IL-1 beta) and Interleukin-6 (IL-6) by activated monocytes/macrophages is an important step in the immune as well as in the inflammatory response. In this study the production of TNF alpha, IL-1 beta and IL-6 by human monocytes (HM) and peripheral blood mononuclear cells (PBMC) was evaluated after HHV-6 infection. Our results demonstrate that HHV-6 can selectively regulate monokine synthesis, in a time-dependent manner. Moreover, we observed a different response closely related to the cellular population (HM or PBMC) examined. The hypothesis we evaluated was that IFN gamma is an important factor triggering the activation of HHV-6 infected human monocytes, to release monokines.