Leukocyte Interferon Research Articles

P.267 Therapy with natural leukocyte interferon and ribavirin in “difficult to treat” patients infected with HCV

P.267 Therapy with natural leukocyte interferon and ribavirin in “difficult to treat” patients infected with HCV

Interferon alpha-2b as adjuvant treatment of recurrent respiratory papillomatosis in Cuba: National Programme (1994–1999 report)

Respiratory papillomatosis is a life-spoiling disease due to its high recurrence rate. Interferon (IFN)alpha-2b treatment, adjuvant to surgery, was assessed for its contribution to disease control and patient quality of life improvement. One hundred and sixty-nine patients (85 children and 84 adults) were included after surgical removal of the lesions followed by intramuscular IFN alpha-2b (Heberon alfa R, Heber Biotec), starting with 10(5) IU/Kg weight in children or 6 x 10(6) IU in adults, three times per week. The dose was reduced monthly, if no relapses occurred, until a monthly maintenance with 5 x 10(4) IU/Kgof weight in children or 3 x 10(6) IU in adults up to two years. In case of relapse, it was surgically removed and the patient returned to the higher dose level. The relapse frequency decreased significantly in 77 percent (69/90) of the recurrent patients both in children (34/46, 74 per cent) and adults (35/44, 79 per cent). Among patients included after their first papilloma, 67 per cent (44/66) had complete (no relapses) or partial (only one relapse) responses (children: 15/33, 45 per cent; adults 29/33, 88 per cent). One hundred and eighteen patients (73 per cent) concluded the treatment without lesions (children: 58 per cent; adults 82 per cent), while the rest showed a significant reduction in the number and size of lesions. IFN was well tolerated. Sixty-two patients (38 per cent) did not have adverse events. The main adverse reactions were fever (59 per cent), chills (24 per cent), arthralgias and myalgias (14 per cent) and headache (10 per cent). One patient developed anti-IFN alpha neutralizing antibodies and became resistant to treatment with recombinant IFN alpha-2b; he responded to natural leucocyte IFN alpha. Treatment with IFN alpha-2b, as an adjuvant to surgery represents a favourable and safe therapeutic alternative for patients with recurrent respiratory papillomatosis.

Inhibition of SARS coronavirus infection in vitro with clinically approved antiviral drugs.

Severe acute respiratory syndrome (SARS) is an infectious disease caused by a newly identified human coronavirus (SARS-CoV). Currently, no effective drug exists to treat SARS-CoV infection. In this study, we investigated whether a panel of commercially available antiviral drugs exhibit in vitro anti–SARS-CoV activity. A drug-screening assay that scores for virus-induced cytopathic effects on cultured cells was used. Tested were 19 clinically approved compounds from several major antiviral pharmacologic classes: nucleoside analogs, interferons, protease inhibitors, reverse transcriptase inhibitors, and neuraminidase inhibitors. Complete inhibition of cytopathic effects of SARS-CoV in culture was observed for interferon subtypes, β-1b, α-n1, α-n3, and human leukocyte interferon α. These findings support clinical testing of approved interferons for the treatment of SARS.

Natural leukocyte interferon alfa for the treatment of chronic viral hepatitis in heart transplant recipients.

A more rapid and aggressive course of hepatitis B virus (HBV)-related and hepatitis C virus (HCV)-related infection in organ transplant recipients has been described. Interferon alfa is the most accepted drug for treating HBV and HCV chronic infections. However, the use of interferon alfa-N3 has been contraindicated in heart transplant (HTx) recipients because of the hypothesized greater risk of triggering acute cellular rejection. The aim of this clinical pilot study was to evaluate tolerability, safety, and efficacy of natural leukocyte interferon alfa in the treatment of chronic HBV and HCV in HTx recipients. Seven HTx recipients were enrolled in the study: two with HBV, four with HCV, and one with combined HBV-HCV chronic infection. The patients had a mean follow-up after heart transplantation of 8.5+/-3 years, before starting interferon alfa-N3 treatment at a dose of 6 MU three times per week, intramuscularly for 12 months. All patients completed the treatment with no major side effects. No unexpected episodes of acute cellular rejection were observed during the treatment. Mean aminotransferase serum levels were significantly lower than before transplantation at 3 (P<0.03), 6 (P<0.02), and 12 (P<0.02) months of treatment and at the 12-month follow-up (P<0.02). A complete and sustained response was achieved in all subjects with HBV-related chronic hepatitis, whereas sustained virologic response was observed in one of four HCV patients. The preliminary data emerging from our study indicate that natural leukocyte interferon alfa-N3 can be safely administered in HTx recipients with chronic HBV or HCV viral hepatitis. Further studies with larger numbers of patients are needed to assess the efficacy of interferon alfa-N3 on HCV virologic response.

Bleomycin, vincristine, lomustine and dacarbazine (BOLD) in combination with recombinant interferon alpha-2b for metastatic uveal melanoma

This EORTC multicentre study analysed the efficacy and tolerability in patients with metastatic uveal melanoma of BOLD chemotherapy in combination with recombinant interferon alpha-2b. The dose of bleomycin was 15 mg on days 2 and 5, of vincristine 1 mg/m2 on days 1 and 4, of lomustine 80 mg on day 1, and of dacarbazine (DTIC) 200 mg/m2 on days 1–5, given every 4 weeks for a minimum of two cycles. Subcutaneous (s.c.) interferon alpha-2b at a dose of 3×106 IU was initiated on day 8 of the first cycle, and continued at a dose of 6×106 IU three times per week after 6 weeks. A median of two cycles were administered to 24 patients (median age 60.5 years). None achieved an objective response (0%; 95% Confidence Interval (CI): 0–14), 2 (8.3%) remained stable, 20 showed progression, and 2 (8.3%) were inevaluable. The median progression-free survival was 1.9 months (95% CI: 1.8–3.4) and overall survival 10.6 months (95% CI: 6.9–16.4). Overall survival improved with increasingly favourable pretreatment characteristics (median, 14.7 versus 6.9 versus 6.0 months for Helsinki University Central Hospital (HUCH) Working Formulation stages IVBa, IVBb and IVBc, respectively; P=0.018). Grade 3 alopecia and neurotoxicity occurred in 13% of the patients. This multicentre study did not confirm earlier reports that BOLD with human leucocyte or recombinant interferon would induce at least 15% objective responses in metastatic uveal melanoma.

The transfusion medicine program in Cuba

Cuba's Transfusion Medicine Program (TMP) is a subsystem of the country's National Health System. The TMP's objective is to ensure hemotherapy with blood that is safe and sufficient for all the individuals who need it. The TMP subsystem is made up of the National Commission on Transfusion Medicine, the Institute of Hematology and Immunology, 37 clinical services, 44 blood banks, 120 collection centers, 19 mobile units, and 37 blood certification laboratories. Additional facilities include a laboratory for plasma separation, a laboratory that produces leukocyte interferon and transfer factor, and two laboratories that produce reagents for blood classification and blood diagnosis symptoms. In Cuba, blood donation is voluntary. Since 1997 approximately 5% of the population per year has donated blood, thus meeting the goal recommended by the Pan American Health Organization of one voluntary blood donation annually for every 20 persons. During 2002, 563,204 blood donations were received, and there were 445,898 transfusions of blood or blood components. All donations are individually screened for HIV 1 and 2, hepatitis B, hepatitis C, and syphilis, thus meeting the country's current regulations. In 2002 these screening measures led to discarding, respectively, 0.12%, 0.60%, 0.71%, and 1.8% of the blood donations. Although the prevalence of human T-cell lymphotropic virus I and II in Cuba is very low, this test will soon be added to the screening process.

Comparison between leukocyte interferon alpha plus ribavirin and R-alpha2B interferon plus ribavirin for treatment of CHC in naive patients: a multicenter randomized Italian study

Comparison between leukocyte interferon alpha plus ribavirin and R-alpha2B interferon plus ribavirin for treatment of CHC in naive patients: a multicenter randomized Italian study

Metabolic flux analysis for human therapeutic protein productions and hypothesis for new therapeutical strategies in medicine

This work may be considered as a model study for therapeutic protein production, and a theoretical approach to hypothesise new medical strategies to further applied medical questions. A comprehensive generalised metabolic reaction network of Bacillus licheniformis that considers 149 reaction fluxes and 106 metabolites was used in the mass flux balance-based stoichiometric model for the analysis of human leukocyte interferon (IFN-α 1) and erythropoietin (EPO) production capacities of recombinant Bacillus species. The importance of cellular energetics on optimum performance was quantitatively assessed. The metabolic pathways leading to optimised IFN-α 1 and EPO overproduction were determined for the two carbon sources that have different reduction degrees ( γ), i.e. glucose ( γ=4.0) and citrate ( γ=3.0), and the variation of the fluxes were obtained. Metabolic capacity analyses showed that maximum IFN-α 1 and EPO synthesis rates were, respectively, 0.062 and 0.055 mmol g −1 DW h −1 at μ=0 h −1 when glucose uptake rate was 10 mmol g −1 DW h −1; and IFN-α 1 and EPO synthesis rates decreased, respectively, 1.70- and 1.75-fold when citrate was used as the carbon source. The flux distributions showed that the amino acid composition of the proteins influence the production. Leucine appears to be the most important amino acid for both IFN-α 1 and EPO production. Consequently, pyruvate seems to be the critical main branch point and B. pasteurii seems to be the favourable host for therapeutical protein production due to the high leucine uptake capacity. The results encourage the discussion on the potential strategies for improving production of IFN-α 1 and EPO, and further enable us to assert medical hypothesis in order to support the immune system of the human body against the deficiencies of the synthesis of IFN-α 1 and EPO in the human cells.

Presence of Cytokines in Biological Preparations

In vaccines produced in eukaryote cells as well as in commercial medical preparations of leukocyte interferon a number of cytokines such as IL-1β, IL-6 and TNF-α have been detected. Among the vaccines examined in this study the highest level of IL-1β was demonstrated in inactivated hepatitis A vaccine prepared in the green monkey kidney cell line 4647, that of IL-6 in inactivated rabies vaccine produced in Syrian hamster kidney (SHK) cell culture, and that of TNF-α in live poliomyelitis vaccine manufactured in VERO cells. A spontaneous and poliovirus-induced capacity of cell cultures to produce cytokines was detected. The level of cytokines produced depend on the kind of cell culture and the type of virus, a more pronounced effect being generated by types 1 and 2 poliovirus as compared with type 3. The presence of highly active cytokines in virus vaccines and interferon preparations points to the necessity of investigating the influence of the presence of cytokines on the biological activity of these preparations and to the advisability of standardizing and controlling the cytokine content.

Induction of Resistance to Phytophthora in Tubers of Transgenic Potato

Resistance of transgenic cultivars based on the expression of one or more resistance genes is sooner or later broken by pathogens whose race-producing rates are high. Thus, combining transgenesis with elicitor-induced resistance is a promising approach. The elicitor-induced resistance is based on the expression of multiple resistance genes, which can prevent the adaptation of pathogens to transgenic cultivars, maintain the stability of cultivars, and increase their lifespan. In this work, we used transgenic potato cultivars Temp and Superior transformed with Bacillus thuringiensis Δ-endotoxin gene and Luk'yanovskii transformed with leukocyte interferon gene. Arachidonic acid (10–8 M) and soluble chitosan (5 kDa, 100 μg/ml) were used as elicitors for tuber treatment. Our data showed that pretreatment with elicitors causes a 15–25% increase in both the systemic prolonged resistance of potato tubers to Phytophthora infestansand their ability to repair mechanical damage.

A Novel Class of Growth Factors Related to Herpesviruses

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Zinc potentiates the antiviral action of human IFN-alpha tenfold.

During the last decade, various zinc salts have been used against the common cold syndrome, which is known to be initiated by respiratory viruses, particularly rhinoviruses. Using rhinovirus as the challenge virus, we investigated whether zinc salts (Zn) could potentiate the antiviral action of native human leukocyte interferon (HuIFN-alpha) and rHuIFN-gamma. We found that HuIFN-alpha was potentiated tenfold at rather low levels of IFN activity (0.6-0.8 U/ml), resulting in 100% protection. Zn alone gave only marginal protection, if any. In contrast to HuIFN-alpha, rHuIFN-gamma directly increased the cytopathic effect of rhinovirus at low levels (<2 U/ml) but protected the cells at higher IFN levels (5-20 U/ml). No potentiation was seen with Zn. HuIFN-beta protected against rhinovirus at the same doses as used with HuIFN-alpha, but in contrast to HuIFN-alpha, no potentiation was noted.

Human leucocyte interferon-alpha in the treatment of chronic hepatitis C.

To assess the efficacy of different schedules of human leucocyte interferon alpha in chronic hepatitis C. A total of 213 naive patients with chronic hepatitis C were treated with 4 different schedules of human leucocyte interferon alpha. Sustained response was defined as persistently normal alanine amino transferase values with negative serum hepatitis C virus-RNA up to 12 months after therapy withdrawal. Rates of sustained response were 16% with 3 MU tiw for 6 months, 33% with 6 MU tiw for 5 months after a priming dose of 9 MU tiw for a month, 32% with 3 MU tiw for 12 months and 20% with 3 MU daily for 6 months. The major factors affecting the response rate were age and the hepatitis C virus genotype, as a sustained response was significantly higher in patients under 45 years and infected by hepatitis C virus types other than hepatitis C virus-1. Treatment was well tolerated and side-effects and drop-out events were similar to those described with other types of alpha-interferons. Human leucocyte interferon alpha appears to be equivalent to recombinant interferon-alpha in the treatment of chronic hepatitis C.

Detection of hepatitis C virus RNA in alpha interferon derived from in vitro culture of leukocytes of human origin

In order to assure the virological safety of blood products, in addition to serological testing of individual donations and virus inactivation steps undertaken during manufacture, routine PCR testing for HCV RNA of starting materials (plasma, cells), intermediates or final product is necessary. The aim of this study was to determine the rate of HCV RNA positive batches of human native leukocyte interferon during large-scale production. Our findings indicate the presence of HCV RNA in 6·1% batches despite acidification of intermediates in order to inactivate Sendai virus.

Interleukin-18 expression and modulation in human corneal epithelial cells

Purpose. The interferon-?-inducing factor Interleukin-18 (IL-18) is a recently described cytokine that appears to have multiple important pro-inflammatory effects including the induction of interferon-? (IFN-?) by activated T-cells. The expression of IL-18 by human cornea has not been previously reported. In the present study, we examine the possibility that human corneal epithelial cells are capable of producing this leukocyte-activating factor which may play an important role in IFN-?-dependent inflammation responses in the cornea. Methods. Northern blot analysis and ELISA were used to investigate the in vitro expression of IL-18 mRNA and protein respectively in primary (HCEC) and transformed human corneal epithelial cells (HCET). To determine if IL-18 expression was modulated by pro-inflammatory mediators, cells were treated with lipopolysaccharide (LPS), phorbol 12-myristate 13-acetate (PMA) or synthetic double stranded RNA (poly dI : dC). IL-18 bioactivity was determined in a leukocyte interferon-? induction assay and IL-18 was immunolocalized in whole human cornea by immunohistochemistry using a specific anti-IL-18 antibody. Results. IL-18 mRNA and bioactive protein was constitutively expressed by human corneal epithelial cells and upregulated by PMA, LPS and poly dI : dC. The constitutive expression of IL-18 protein immunoreactivity was also demonstrated in the epithelial cells of whole human cornea tissue. Conclusions. This is the first study demonstrating that corneal epithelial cells are capable of producing the IFN-? inducing factor IL-18. Increased bioactive corneal IL-18 production can be induced by a number of pro-inflammatory agents and may play an important role in initiating ?-interferon-mediated inflammatory responses in the cornea.

Failure to confirm major objective antitumor activity for streptozocin and doxorubicin in the treatment of patients with advanced islet cell carcinoma

We read with interest the article by Cheng et al. addressing the antitumor effect of streptozocin and doxorubicin in the treatment of patients with advanced islet cell carcinoma.1 They concluded that they were unable to confirm the findings from the Eastern Cooperative Oncology Group trial published in 1992.2 In this trial of the 38 patients who received streptozocin and doxorubicin, 36 were evaluable and 69% showed any regression with 14% complete regression. For this study, criteria for response evaluation were a > 50% decrease in hormone production; a > 30% reduction in hepatomegaly; and measurable tumor reduction, with the product of the perpendicular diameters reduced > 50%. Routine use of computed tomography (CT) scan was not indicated. From this study it is in retrospect difficult to calculate how many patients had a response based only on hormone reduction or reduced liver size. Cheng et al. used CT scan, magnetic resonance imaging, or X-rays of bidimensional measurable lesions as the only criteria for response as defined by Miller et al.3 With the same streptozocin and doxorubicin regimen2 given to all of their 16 patients with islet cell tumors, Cheng et al. observed 1 objective response, 9 patients with stable disease, and 6 patients with progressive disease. The median overall survival was not yet reached, with > 60% alive after follow-up ranging from 10 to 67+ months. We would like to add six islet cell tumor patients to this series. From 1995–1999, we treated these patients according to exactly the same chemotherapy scheme. The patients were evaluated as described by Cheng et al. None of the patients were hormone producers. One patient experienced a partial response, three had stable disease, and two had progressive disease. Overall survival after initiation of chemotherapy ranged from 4 to 17+ months. Apart from cardiotoxicity (n = 1), which was most likely due to doxorubicin treatment (with cardiac failure responsive to antidiuretic treatment), no severe toxicity of the regimen was observed. Combining the data from Cheng et al. and ours results in 2 objective tumor responses out of 22 (9%; 95% confidence interval, 1–29%). Although we find with these criteria a lower response rate than Moertel et al., it is also true that in these two series a good effect on survival cannot be denied. In islet cell tumor studies, to report precisely on the objective regression of tumor size may be of major relevance. If there is a regimen that results in a realistic option to create a reduction in tumor size, this would potentially allow surgical tumor resection and thus increase the potential for curative tumor resection. If the main goal of this regimen is quality of life rather than remission, the objective response rate of only 9% with the streptozocin/doxorubicin regimen justifies more interest in interferon treatment of patients with islet cell tumors. After an initial study with leukocyte interferon,4 interferon-α doses of 5–6 MU 3–5 times per week resulted in biochemical responses in 50% of the patients and tumor responses in 12%.5 Surprisingly, this option is not even mentioned in the National Cancer Institute's PDQ information for health care professionals on islet cell carcinoma.6 Treatment with somatostatin analogs is mentioned in the PDQ, reporting improvement of symptoms but little to no effect on tumor size. Attempts to combine the several options have been initiated.5 Because of the rarity of the disease, it is difficult to conduct large studies to confirm earlier interesting results or to design studies to improve these results. It will be of major importance to consider future multicenter studies in order to improve the outcomes of patients in this category. H. de Vries*, N. H. Mulder*, E. G. E. de Vries*, * Division of Medical Oncology, Groningen University Hospital, Groningen, The Netherlands

Leukocyte Interferon-alpha in the Treatment of Chronic Hepatitis C in Finland

To evaluate the efficacy of leukocyte interferon in previously untreated patients with chronic hepatitis C, 97 patients were enrolled in a prospective study in Finland with a stepped-care management protocol. The treatment was initiated with 3 million units of interferon-alpha subcutaneously three times a week. At 3 months, if the serum alanine aminotransferase was still abnormal, the dose was doubled. If serum hepatitis C virus (HCV) RNA had turned negative at 6 months, the treatment was stopped; if it was still positive, treatment was continued for up to 12 months. All patients were followed up after treatment for 6 months. Altogether, 74 patients completed the treatment and follow-up periods. Of all the originally enrolled patients 36% (35 of 97) achieved sustained virologic response, defined as HCV RNA negativity 6 months after the end of treatment. The commonest HCV genotype among these patients was 3a, and as many as 52% of such patients achieved sustained virologic response. Thirty-two per cent of the patients had HCV genotype 1a, 1b, or a mixture of these; a sustained response was achieved in only 6% of such patients but in 50% of patients with a non-1 genotype. Adverse effects caused treatment cessation for 10% of the patients and IFN dose reduction for 20%. Monotherapy with human leukocyte interferon resulted in sustained virologic response in 36% of patients with chronic hepatitis C. In those infected with a HCV genotype other than 1, the sustained virologic response rate was 50%.

Editorial: A tribute to Sidney Udenfriend

Editorial: A tribute to Sidney Udenfriend

The human interferon alpha species and receptors.

Interferon (IFN) was approved by the U.S. Food and Drug Administration on June 5, 1986. As the first biotherapeutic approved, IFN-alpha paved the way for development of many other cytokines and growth factors. Nevertheless, we have just touched the surface of understanding the multitude of human IFNs. This paper reviews the history of the purification of human leukocyte IFN and key aspects of our current state of knowledge of human interferon alpha genes, proteins, and receptors.

Efficacy and safety of two different doses of human leukocyte interferon alpha for treatment of hepatitis C virus infection: A pilot study

The purpose of this pilot study was to assess the efficacy and safety of a 6-month course of 6 million units (MU) of leukocyte interferon alfa (IFN-α) 3 times a week (TIW) in naive patients with chronic hepatitis C virus (HCV) infection. Nine patients were treated with leukocyte IFN-α at a dose of 6 MU TIW; a control group of 30 patients received the standard regimen of leukocyte IFN-α 3 MU TIW. All patients were treated for 6 months and followed up for an additional 6 months. Biochemical (alanine aminotransferase [ALT] values) and virologic (HCV-RNA) responses to treatment were assessed. At the end of the treatment period, 6 (67%) of 9 patients treated with IFN-α 6 MU showed normal ALT levels compared with 14 (47%) of 30 patients treated with 3 MU. The complete (biochemical and virologie) response was slightly greater in patients treated with 6 MU (3/9, 33%) than in those treated with 3 MU (7/30, 23%), both at the end of treatment or after 6 months of follow-up. Both of the IFN regimens were well tolerated. Results of this pilot study suggest that treatment with 6 MU of leukocyte IFN-α TIW for 6 months is associated with a slightly better biochemical and virologic response than is treatment with the standard regimen of 3 MU TIW for 6 months. These results, however, should be confirmed in a larger trial with a high power.