Effect of B7.1-transfected human colon cancer cells on the induction of autologous tumour-specific cytotoxic T cells

Background : The induction of tumour-specific immunity is important for advanced cancer therapy. There are many molecules, including costimulatory molecules, that have been identified as the activator for tumour-specific T cells. Methods : To induce autologous tumour-specific cytotoxic T lymphocytes (CTL) more effectively, we studied whether the expression of the B7 gene may render human colon cancer cells able to stimulate autologous peripheral blood mononuclear cells (PBMC) to become tumour-specific cytotoxic T cells. After the establishment of a B7.1 gene transfected tumour cell line, Cw2/B7.1, we first examined its stimulatory effect on autologous PBMC and subsequently, its effect on the induction of parental cell (Cw2)-specific CTL. Results : The results showed that Cw2/B7.1 had a more potent stimulatory effect on PBMC for the induction of both proliferation and cytotoxicity than Cw2. By adding a low dose of interleukin-2, Cw2/B7.1-activated killer cell activity was significantly increased. The specificity of Cw2/B7.1-activated killer cells was demonstrated by the absence of their cytotoxicity to either human lymphocyte antigen (HLA)-A33 identical (ORF) or HLA-non-identical (MT) allogenic colon cancer cell lines. Furthermore, such Cw2-specific cytotoxic activity was significantly reduced by the deletion of CD8⁺ cells but not CD4⁺ cells, indicating that these killer cells were mainly CD8⁺ T cells. Conclusions : Thus, our results demonstrate that, by using B7.1 gene-transfected tumour cell lines, we effectively induced autologous tumour-specific CTL. These results will provide us with new tools for adoptive immunotherapy for colon cancer patients. © 1999 Blackwell Science Asia Pty Ltd     

Resistance of established porcine islet xenografts to humoral rejection by hyperimmune sera.

BACKGROUND: Although preformed natural antibodies cause hyperacute rejection of primarily vascularized xenografts, tissue grafts such as skin or islets are revascularized by in-growth of host capillaries and therefore might be resistant to circulating antibodies. We examined the effect of hyperimmune serum and primed T cells on the survival of long-term porcine islet xenografts in diabetic nude mice. METHODS: Porcine islets were transplanted beneath the kidney capsule of streptozotocin-induced diabetic BALB/c athymic mice. Hyperimmune serum and sensitized splenocytes were prepared by repeated immunization of BALB/c mice with porcine lymph node cells. Splenic T cells were enriched by nylon wool column separation. Tissues were examined by immunohistology using murine- and porcine-specific monoclonal antibodies. RESULTS: Porcine islets survived in nude mice for > 100 days with high levels of circulating porcine C-peptide and maintenance of normoglycemia. Injection of the hyperimmune sera (IgG) into normoglycemic nude mice bearing porcine islets for > 70 days failed to induce rejection despite the continued presence of circulating anti-porcine cytotoxic antibody. Injection of sensitized T cells caused acute rejection of long-term (>140 days) porcine islets, whereas injection of naive T cells had no effect. Histologically, porcine islets removed from mice treated with hyperimmune serum showed no staining for IgG. Long-surviving porcine islet grafts showed strong staining for interleukin (IL)-10 and a lesser amount of IL-4 but no staining for IL-2 or interferon-gamma. Although fresh porcine islets were positive for swine leukocyte antigen class 1 antigen and intercellular adhesion molecule (ICAM)-1 but negative for mouse platelet endothelial cell adhesion molecule and ICAM-2, long-surviving porcine islets showed positive endothelial staining for mouse platelet endothelial cell adhesion molecule and ICAM-2. CONCLUSIONS: Established islet xenografts are resistant to hyperimmune serum as a result of a lack of target endothelial antigens, whereas they remain susceptible to rejection caused by primed T cells. Local production of Th2 cytokines may explain the inability of long-surviving islet xenografts to activate injected naive T cells.     

Advanced emphysema: preoperative chest radiographic findings as predictors of outcome following lung volume reduction surgery.

PURPOSE: To determine whether preoperative chest radiographic findings alone can reliably predict which patients will achieve the best functional outcome of lung volume reduction surgery. MATERIALS AND METHODS: The preoperative chest radiographs obtained in 57 patients who had undergone lung volume reduction surgery were retrospectively scored by five blinded readers for severity and distribution of emphysema, evidence of lung compression, disease heterogeneity, and other features. Comparisons were made with the 3-6-month postoperative functional outcome for each patient. RESULTS: High disease heterogeneity (score > 2) and unequivocal lung compression (score 1) both were 100% predictive of a favorable outcome (FEV1 increase, > or = 30%). Low heterogeneity (score < 1) was 94% predictive of an unfavorable outcome (FEV1 increase < 30%), as was a lack of lung compression, which was 92% predictive of an unfavorable outcome. These two features also correlated with an improved 6-minute walk test result, although this correlation was weaker. CONCLUSION: Chest radiography alone may be sufficient for initial screening. High disease heterogeneity and lung compression on chest radiographs are highly predictive of a favorable functional outcome.     

Real World Treatment Practices for Chronic Lymphocytic Leukemia in Japan: An Observational Database Research Study (CLIMBER-DBR)

There are limited real-world data on the treatment practices and healthcare resource utilization associated with chronic lymphocytic leukemia (CLL) in Japan. In this study (CLIMBER-DBR), we performed retrospective analyses of the Japanese Medical Data Vision database, and extracted data for 2562 patients with newly diagnosed CLL (CLL-1 cohort) and 930 patients receiving CLL treatment (CLL-2 cohort) registered between March 1, 2013 and February 28, 2018. The median follow-up in the CLL-1 cohort was 721 (quartile 1–3: 363–1267) days and the median time to initial (first-line) treatment was 1331 (quartile 1–3: 189–not reached) days. In the CLL-2 cohort, the most frequently used regimens were fludarabine alone (17.7%), cyclophosphamide alone (13.7%), and bendamustine/rituximab (8.2%). The median (quartile 1–3) times to second-line and third-line treatments were 1066 (273–not reached) and 1795 (631–not reached) days, respectively. The CLIMBER-DBR was the first database research study to assess current treatment practices for CLL in Japan, where the treatment patterns were driven by the approval/reimbursement status of drugs in the study period. Our study provides an important benchmark for future studies of CLL in Japan.     

MK‐6, a novel not‐α IL‐2, elicits a potent antitumor activity by improving the effector to regulatory T cell balance

IL‐2 is a pleiotropic cytokine that regulates immune cell homeostasis. Its immunomodulatory function has been used clinically as an active immunotherapy agent for metastatic cancers. However, severe adverse effects, including the vascular leak syndrome and the preferential stimulation of anti‐immunogenic Treg rather than effector T cells, have been obstacles. We newly designed a mutein IL‐2, Mutakine‐6 (MK‐6), with reduced IL‐2Rα–binding capability. MK‐6 induced comparable cell growth potential toward IL‐2Rβγ–positive T cells but was far less efficient in in vitro Treg proliferation and STAT5 activation. Unlike IL‐2, in vivo administration of MK‐6 produced minimal adverse effects. Using CT26 and B16F10‐syngeneic tumor models, we found MK‐6 was highly efficacious on tumor regression. Serum albumin conjugation to MK‐6 prolonged in vivo half‐life and accumulated in CT26 tumors, showing enhanced antitumor effect. Tumor‐infiltrating leukocytes analysis revealed that albumin‐fused MK‐6 increased the ratio of effector CD8⁺ T cells to CD4⁺ Treg cells. These results demonstrated that MK‐6 is an efficient immunomodulator potentially used for improved immunotherapy with decreased adverse effects and attenuated Treg stimulation.     

Treatment of murine collagen‐induced arthritis by ex vivo extracellular superoxide dismutase gene transfer

Objective

Superoxide dismutase (SOD) is a potent antiinflammatory enzyme that has received growing attention for its therapeutic potential. This study was undertaken to examine the efficacy of extracellular SOD (EC‐SOD) gene therapy in murine collagen‐induced arthritis.

Methods

Embryonic DBA/1 mouse fibroblasts were infected with a recombinant retrovirus expressing human EC‐SOD. DBA/1 mice that had been treated with type II collagen were administered subcutaneous injections of 2 × 10⁷ EC‐SOD–expressing fibroblasts on day 29, when symptoms of arthritis were already present. The severity of arthritis in individual mice was evaluated in a double‐blind manner; each paw was assigned a separate clinical score, and hind paw thickness was measured with a caliper. Mice were killed on day 50 for histologic examination of the joints.

Results

High serum concentrations of EC‐SOD were maintained for at least 7 days. Mice treated with the transgene exhibited significant suppression of clinical symptoms such as disabling joint swelling, deformity, and hind paw thickness, compared with the untreated group (mean ± SD maximum clinical score in the untreated and the transgene‐treated groups 2.71 ± 1.08 and 1.35 ± 1.22, respectively; P < 0.01, and hind paw thickness 3.04 ± 0.18 mm and 2.56 ± 0.12 mm, respectively; P < 0.05). Histologic abnormalities, including destruction of cartilage and bone, infiltration of mononuclear cells, and proliferation of synovial cells, were also markedly improved in the EC‐SOD–treated mice compared with the control group (histopathologic score 7.50 ± 1.13 and 4.13 ± 1.88 in the untreated and transgene‐treated groups, respectively; P < 0.05).

Conclusion

These results indicate that EC‐SOD gene transfer may be an effective form of therapy for rheumatoid arthritis.

     

New chitin-based polymer hybrids, 3. Miscibility of chitin-graft-poly(2-ethyl-2-oxazoline) with poly(vinyl alcohol)

The miscibility of blends of poly(vinyl alcohol) (PVA) with chitin-graft-poly(2-ethyl-2-oxazoline) ( 1 ) and poly(2-ethyl-2-oxazoline) homopolymer (PEtOZO) was investigated. Calorimetric results showed a single glass transition temperature (T_g) in the entire range of compositions for both blend systems, which indicated that PVA is miscible with both the graft copolymer 1 and PEtOZO. The T_g of PVA is also shifted to lower temperature upon blending with the graft copolymer 1 . IR analysis revealed the existence of specific interactions via hydrogen bonding between the hydroxyl groups in PVA and the carbonyl groups in the poly(2-ethyl-2-oxazoline) side chain of graft copolymer 1 . The results show that the interaction of graft copolymer 1 with PVA is increased by introduction of longer poly(2-ethyl-2-oxazoline) side chains. Thermal decomposition (TG) measurements supported the compatibility of PVA with graft copolymer 1 and with PEtOZO, and showed that the thermal stability of PVA is improved upon blending with 1 or PEtOZO.