No abstract available.
Interferon-alpha* ; Interleukin-2* ; Killer Cells, Lymphokine-Activated*

PEGylation is considered one of the most successful techniques to improve the characteristics of protein drugs including to increase the circulating half-life of proteins in blood and to decrease their immunogenicity and antigenicity. One known PEG modification method is to attach PEG to the free amino group, typically at lysine residues or at the N-terminal amino acid with no selectivity, resulting in a heterogeneous product mixture. This lack of selectivity can present problems when a therapeutic PEGylated protein is being developed, because predictability of activity and manufacturing reproducibility are needed for regulatory approval. Enzymatic PEGylation of proteins is one route to overcome this limitation. Transglutaminases (TGase) are enzyme candidates for site-specific PEGylation. We use human interferon alpha 2a (IFN α2a) as a test case, and predict that the potential modification residues are Gln101 by computational approach as it contains 12 potential PEGylation sites. IFN α2a was PEGylated by Y shaped PEG40k-NH2 mediated by microbial transglutaminase. Our results show that the microbial transglutaminase mediated PEGylation of IFN α2a was site-specific only at the site of Gln101 in IFN α2a, yielding the single mono-conjugate PEG-Gln101-IFN α2a with a mass of 59 374.66 Da. Circular dichroism studies showed that PEG-Gln101-IFN α2a preserved the same secondary structures as native IFN α2a. As expected, the bioactivity and pharmacokinetic profile in rats of PEG-Gln101-IFN α2a revealed a significant improvement to unmodified IFN α2a, and better than PEGASYS.
Animals ; Antiviral Agents ; Humans ; Interferon alpha-2 ; metabolism ; Interferon-alpha ; biosynthesis ; pharmacokinetics ; Polyethylene Glycols ; pharmacokinetics ; Protein Structure, Secondary ; Rats ; Recombinant Proteins ; biosynthesis ; pharmacokinetics ; pharmacology ; Reproducibility of Results ; Transglutaminases ; metabolism

About 50% of renal cell carcinoma patients initially present with a regional or distant metastatic disease. Attempts to treat metastatic renal cell carcinomas have been directed at cytokine-based immunotherapy. Response rates of interleukin-2-based immunotherapy of 5 to 29% have been reported in this disease. Immunization as a mechanism to recruit host antitumor responses is increasingly being described as a potentially effective and less toxic approach for the treatment of metastatic and high-risk primary cancers. An autologous renal cell cancer vaccine has been applied at our institution for the prevention of recurrence or metastasis in locally advanced cases for more than one year. Herein, two metastatic renal cell carcinoma cases, which failed to show a response to initial immunotherapy or chemoradiotherapy, which were successfully treated with IL-2 (Aldesleukin) and interferon-alpha as autologous cancer vaccine adjuvants is reported.
Carcinoma, Renal Cell* ; Chemoradiotherapy ; Humans ; Immunization ; Immunotherapy* ; Interferon-alpha ; Interleukin-2 ; Neoplasm Metastasis ; Recurrence ; Vaccines

OBJECTIVE: To explore the synergistic immunomodulatory mechanism of interferon alpha-1b, interleukin-2 and thalidomide (ITI) regimen on patients with acute myeloid leukemia (AML). METHODS: Sixty eight untreated de novo or relapsed or refractory or maintenance therapy patients with AML admitted in the Affiliated Cancer Hospital of Zhengzhou University and the other 11 medical units from March 2016 to May 2019 were treated with ITI regimen. Peripheral blood specimen per patient was collected into EDTA-K3 anticoagulation vacuum tube before the administration of ITI and 3 months after the treatment; peripheral blood lymphocyte subsets and perforin and Granzyme B expression were analyzed by using flow cytometry; the levels of VEGF, IFN-γ, TNF-α and IL-6 in the plasma were detected by using a cytometric bead array. Thirty-five healthy subjects from the hospital physical examination centre were selected as normal controls. RESULTS: The ratio of CD4 CONCLUSION The ITI regimen can raise the ratio of CD4
CD8-Positive T-Lymphocytes ; Humans ; Interferon-alpha ; Interleukin-2 ; Leukemia, Myeloid, Acute/drug therapy* ; Perforin ; Thalidomide

PURPOSE: Despite numerous trials no various immunochemotherapeutic agents, advanced renal cell carcinomas (RCC) remain highly resistant to treatment. We report the results of immunochemotherapy to evaluate its efficacy, and to investigate factors modulating responses to treatment. MATERIALS AND METHODS: In this retrospective analysis, 54 patients with advanced RCC, having received either interferon-alpha (IFN-alpha) monotherapy, or a combination of interleukin-2 (IL-2), 5-fluorouracil (5-FU) and vinblastine, with IFN-alpha between January 1999 and March 2001, were reviewed. The patients were divided into 2 groups, either single or combined, therapy groups, which were further subdivided into locally advanced and metastatic disease groups. The nephrectomy and performance stati, cancer stage at the beginning of the immunotherapy, metastasized organs and interval to metastasis, were considered, and the responses to treatment and survival were evaluated accordingly in each subgroup. RESULTS: Of the 54 patients, 39 received monotherapy, 34 of which had a metastatic disease. Partial remission was observed in 1 patient (2.9%) with a duration of response of 52 weeks, a median survival of 47 weeks (4-323 wks) and a 1 year survival rate of 41.2%. Of the 15 patients in the combined therapy group, 12 with a metastatic disease, complete remission was noted in 1 patient (8.3%), with a continuous response at 57 weeks, a median survival of 34 weeks (4-204 wks) and a 1 year survival rate 41.7%. Of the prognostic factors, the nephrectomy status proved to be the only significant factor improving survival. CONCLUSIONS: Advanced RCC exhibited a very limited response, and nephrectomy status was revealed to be the only significant factor affecting survival. The efficacy of immunotherapy on locally advanced RCC needs further controlled evaluative studies.
Carcinoma, Renal Cell* ; Fluorouracil ; Humans ; Immunotherapy ; Interferon-alpha ; Interleukin-2 ; Neoplasm Metastasis ; Nephrectomy ; Retrospective Studies ; Survival Rate ; Vinblastine

OBJECTIVETo investigate the effects of interferon-α2b combined with thalidomid(THD) on the proliferation and apoptosis of HEL cells, and expression of JAK2V617F-mutated gene.

METHODSThe cell survival rates were assayed by CCK-8 after HEL cells were treated by interferon-α2b, thalidomid and thalidomid combined with interferon-α2b for 24, 48 and 72 hours, while the apoptosis and expression of JAK2V617F mutation gene were detected by flow cytometry and fluorescence quantitative PCR respectively after treatmemt for 48 hours.

RESULTSIFN-&alpha;2b combined with THD could more obviously inhibit the proliferation of HEL cells than IFN-&alpha;2b or THD alone for 24 and 48 and 72 hours, and the differences between groups were statistically significant(P<0.05). In addition, the apoptosis-inducing effect of IFN-&alpha;2b combined with THD on HEL cells was more obvious than that of IFN-&alpha;2b used alone, the differences was statistically significant(P<0.05). Although IFN-&alpha;2b combined with THD can decrease the JAK2V617F mutation gene expression more obviously than IFN-&alpha;2b alone,but the difference was no statistically significant (P>0.05). And the remaining groups showed statistically significance(P<0.05).

CONCLUSIONIFN-&alpha;2b combined with THD can obviously inhibit the proliferation and induce apoptosis of HEL cells more than that of IFN-&alpha;2b alone, but the effect of reducing JAK2V617F mutation gene expression is not different.

BACKGROUND: To evaluate the cellular immune response to interferon(IFN)-alpha and ribavirin combination therapy in patients with chronic hepatitis C, we monitored serum levels of soluble IL-2 receptor(sIL2R) before and after the therapy. METHODS: Serum sIL2R levels before and after the combination therapy were measured in 19 patients with chronic hepatitis C. IFN(3 MU/day, 3 times/week) and ribavirin 1000 mg/day were administered for 24 weeks to all patients. RESULTS: After the therapy, sIL2R levels were increased (before, 3.13 0.67 ng/m L, and after 4.08 2.13 ng/mL, p=0.059) but statistically insignificant(p>0.05). The patients were divided into two groups : the responder group who were negative for serum hepatitis C virus(HCV)-RNA after the therapy, and the non-responder group who were still positive for HCV-RNA after the therapy. Between these two groups, sIL2R levels before and after the therapy were not significantly different. The ratio of sIL2R levels before and after the therapy was calculated, although the ratio was higher in responder group, but there was no significant difference between the two groups(sIL2R after the therapy)/(sIL2R before the therapy) : 1.43 0.70 in the responder group and 1.04 0.28 in the nonresponder group, p=0.096). CONCLUSION: Although these results failed to demonstrate that sIL2R level was increased during the combination therapy in patients with hepatitis C, this study suggested that cytokines which mediate immune response may be involved in the pathogenesis of chronic heaptitis C virus infection.
Cytokines ; Hepacivirus ; Hepatitis C ; Hepatitis C, Chronic* ; Hepatitis, Chronic* ; Humans ; Immunity, Cellular ; Interferon-alpha ; Interferons* ; Interleukin-2* ; Receptors, Interleukin-2* ; Ribavirin*

PURPOSE: The role of immunotherapy in the treatment of patients with advanced renal cell carcinoma(RCC) is being evaluated by a number of investigators. In this study, we evaluated the efficacy and toxicity of a low-dose subcutaneous regimen of interferon-alpha(IFN-alpha) in combination with intravenous administration of 5-fluorouracil(5-FU) with or without interleukin-2 (IL-2) in patients with advanced RCC. MATERIALS AND METHODS: 16 patients with advanced RCC were treated with an 8-week cycle of 6 to 9 MU/M2 IFN-alpha given 1 to 3 times a week during the 8 week period, and sequentially combined with 5 to 20 MU/M2 IL-2 given 3 times a week for 4 weeks and 750mg/M2 5-FU once a week for 4 weeks. 5 patients were treated with only IFN-alpha and 5-FU. Treatment schedule was identical except for the exclusion of IL-2 in this group. Among those 16 patients treated with 3-drug regimen(including IL-2), 10 patients had measurable metastatic lesions, and 4 patients had measurable metastatic lesions among 5 patients treated with 2-drug regimen(without IL-2). RESULTS: Among 19 consecutive men and 2 women treated, 14 had measurable metastatic lesion. 10 out of 14 patients with measurable metastatic lesions were treated with 3- drug regimen, and objective tumor regressions were achieved in 4 patients(40%) consisting of complete response in 1(10%) and partial response in the other 3(30%). Metastatic sites were lung in 3 patients, bone in 1 patient. 3 of the 4(75%) patients with pulmonary metastasis showed complete or partial response. Median response duration(complete plus partial response) was 14.3 months(range 11 to 22+). Stable disease lasting 3 months was noted in 1 patients(10%). Other 4 patients out of 14 patients with measurable metastatic lesions were treated with 2-drug regimen, and partial remission lasting 9 months was noted in 1 patient(25%). There were only mild to moderate side effects; maximum toxicity grade of 0 in 10 patients, grade I in 3, II in 7, III in 1 according to the World Health Organization classification. None of the patients experienced major IL-2 related toxicity and no toxic deaths occurred. CONCLUSIONS: This combination immunochemotherapy may be a promising regimen with modest toxicity in advanced RCC. The most favorable response can be expected in pulmonary metastasis. Most of side effects were tolerable. Three-drug regimen including IL-2 showed better response rate than two-drug regimen, suggesting a major role of IL-2. A large prospective randomized trials are required to confirm whether the combination immunochemotherapy has an effect on advanced RCC or not.
Administration, Intravenous ; Appointments and Schedules ; Carcinoma, Renal Cell* ; Classification ; Female ; Fluorouracil* ; Humans ; Immunotherapy ; Interferon-alpha* ; Interleukin-2* ; Lung ; Male ; Neoplasm Metastasis ; Research Personnel ; World Health Organization

Renal cell carcinoma (RCC) is a malignancy arising from the tubular epithelial cell. RCC is a rare disease in children with an incidence of 0.1~0.3% of all childhood cancer. It is known that RCC in children is different from its adult counterpart in some aspects. Although conventional treatment of RCC is radical nephrectomy with lymph node dissection, we tried systemic treatment for advanced disease. Because metastasis is common at diagnosis and RCC is usually resistant to radiotherapy or chemotherapy, immunotherapy such as interferon-alpha (IFN-alpha) or interleukin-2 (IL-2) is investigated and clinically used. We have experienced 3 children with RCC and two with stage IV disease were treated with systemic immunotherapy. One who had stage IV, recurred RCC is followed-up for 16 months without the evidence of disease progression with IFN-alpha and IL-2. We report these 3 cases with a brief review of literatures.
Adult ; Carcinoma, Renal Cell* ; Child ; Diagnosis ; Disease Progression ; Drug Therapy ; Epithelial Cells ; Humans ; Immunotherapy* ; Incidence ; Interferon-alpha ; Interleukin-2 ; Lymph Node Excision ; Neoplasm Metastasis ; Nephrectomy ; Radiotherapy ; Rare Diseases

Spontaneous remission (SR) of leukemia is a rare event in clinic, which possibly correlated with severe infection and sepsis, but its exact mechanism has not been confirmed. Plasmacytoid dendritic cells (pDC) and myeloid dendritic cells (mDC) play a key role in innate and adaptive immunity respectively. A patient with severe infection of staphylococcus aureus acquired completely spontaneous remission (SR), moreover a increased number of pDC were observed, suggesting that bacteria-activated pDC may play an important role in SR. This study was purposed to explore if the bacteria can stimulate pDC successfully and get a functional pDC. Both pDC and mDC were isolated from freshly collected, leukocyte-rich buffy coats from healthy blood donor and leukemic patient with SR by using MACS and FACS. The pDC were cultured in RPMI 1640 medium and were stimulated with different kinds of bacteria and the expression of CD40, CD86 and HLA-DR on the cell surface was analyzed by flow cytometry. The cytokine (IFN-&alpha;, IL-12, IFN-γ, IL-2, IL-4, IL-10) production was measured by using ELISA kits. The results showed that the stimulation with staphylococcus aureus and pseudomonas aeruginosa resulted in the maturation of pDC, which secrete a large number of IFN-&alpha; and promote the differentiation of naive CD4⁺ T cells to Th1 cells. The activated pDC expressed high level of CD40 and CD86 and showed higher T cell stimulatory capacities. It is concluded that staphylococcus aureus and pseudomonas aeruginosa can activate pDC, the activated pDC secrete high quantity of IFN-&alpha;. This result suggests that bacteria stimulated pDC may play a key role in SR of leukemia following severe infections.
CD4-Positive T-Lymphocytes ; Dendritic Cells ; immunology ; microbiology ; Humans ; Interferon-alpha ; Interleukin-10 ; Interleukin-12 ; Interleukin-2 ; Interleukin-4 ; Leukemia ; diagnosis ; immunology ; microbiology ; Remission, Spontaneous ; Staphylococcus aureus