Mobilan: a recombinant adenovirus carrying Toll-like receptor 5 self-activating cassette for cancer immunotherapy.

Toll-like receptor 5 (TLR5) is considered an attractive target for anticancer immunotherapy. TLR5 agonists, bacterial flagellin and engineered flagellin derivatives, have been shown to have potent antitumor and metastasis-suppressive effects in multiple animal models and to be safe in both animals and humans. Anticancer efficacy of TLR5 agonists stems from TLR5-dependent activation of nuclear factor-κB (NF-κB) that mediates innate and adaptive antitumor immune responses. To extend application of TLR5-targeted anticancer immunotherapy to tumors that do not naturally express TLR5, we created an adenovirus-based vector for intratumor delivery, named Mobilan that drives expression of self-activating TLR5 signaling cassette comprising of human TLR5 and a secreted derivative of Salmonella flagellin structurally analogous to a clinical stage TLR5 agonist, entolimod. Co-expression of TLR5 receptor and agonist in Mobilan-infected cells established an autocrine/paracrine TLR5 signaling loop resulting in constitutive activation of NF-κB both in vitro and in vivo. Injection of Mobilan into primary tumors of the prostate cancer-prone transgenic adenocarcinoma of the mouse prostate (TRAMP) mice resulted in a strong induction of multiple genes involved in inflammatory responses and mobilization of innate immune cells into the tumors including neutrophils and NK cells and suppressed tumor progression. Intratumoral injection of Mobilan into subcutaneously growing syngeneic prostate tumors in immunocompetent hosts improved animal survival after surgical resection of the tumors, by suppression of tumor metastasis. In addition, vaccination of mice with irradiated Mobilan-transduced prostate tumor cells protected mice against subsequent tumor challenge. These results provide proof-of-concept for Mobilan as a tool for antitumor vaccination that directs TLR5-mediated immune response toward cancer cells and does not require identification of tumor antigens.

Follow-up of 11-16 years after modular fixed-bearing TKA.

To present representative data of long-term survivorship and clinical outcome for the PFC total knee arthroplasty (PFC-TKA). A consecutive series of 141 TKA was followed for a mean of 13 years (range, 11-16 years). Sixty-five knees were evaluated, 30 of these clinically and radiographically. Twenty-eight knees could only be assessed with the use of a questionnaire. Six patients were living in nursing homes. Fifty-four patients (65 knees) had died. Eleven had undergone a revision. One patient was considered lost to follow-up. With re-operation for any reason as the endpoint, the 10-year survival rate was 92% (n = 91 patients at risk), and the 14-year survival rate was 91% (n = 12). With aseptic loosening of the implant as the endpoint, the 10- and 14-year survival rates were 97%. The mean Knee Society and function scores were 76 and 48 points, respectively. In this multi-surgeon series modular fixed-bearing TKA had good clinical and radiographic results with excellent long-term survivorship.

Cancer-related fatigue in patients attending oncological rehabilitation programs: prevalence, patterns and predictors.

BACKGROUND: Fatigue is one of the most frequent symptoms in cancer patients. It is the aim of medical and psychosocial rehabilitation programs to reduce physical impairments and psychosocial distress. There are no reliable data on the prevalence or the pattern of fatigue in patients attending such rehabilitation programs. We therefore initiated a cross-sectional study to evaluate this phenomenon. PATIENTS AND METHODS: Using the EORTC QLQ-C30, HADS and MFI questionnaires in 144 patients with different malignant diseases the incidence, patterns and necessity of treatment of fatigue were evaluated. RESULTS: One third of the patients were diagnosed with breast cancer, 27% with gastrointestinal cancers and 20% suffered from haematologic/lymphatic diseases. There was a good correlation between fatigue levels in the 3 instruments used. 22% of all patients showed significant symptoms of fatigue. The univariate analysis did not reflect any significant differences in severity of fatigue with respect to diagnosis, haemoglobin levels, medical and therapeutic history or sociodemographic data. However, using cluster analysis we could identify 3 groups of patients that differ significantly in fatigue and quality-of-life measurements. CONCLUSION: We conclude from our data that fatigue is an important phenomenon in cancer patients who attend medical and psychosocial rehabilitation programs. This has an impact on the ability and degree to reintegrate these patients into everyday life.

T cell infiltration of the prostate induced by androgen withdrawal in patients with prostate cancer.

Manipulations capable of breaking host tolerance to induce tissue-specific T cell-mediated inflammation are of central importance to tumor immunotherapy and our understanding of autoimmunity. We demonstrate that androgen ablative therapy induces profuse T cell infiltration of benign glands and tumors in human prostates. T cell infiltration is readily apparent after 7-28 days of therapy and is comprised predominantly of a response by CD4+ T cells and comparatively fewer CD8+ T cells. Also, T cells within the treated prostate exhibit restricted TCR Vbeta gene usage, consistent with a local oligoclonal response. Recruitment/activation of antigen-presenting cells in treated prostate tissues may contribute to local T cell activation. The induction of T cell infiltration in prostate tissues treated with androgen ablation may have implications for the immunotherapeutic treatment of prostate cancer as well as other hormone-sensitive malignancies, including breast carcinoma.

Alterations in peripheral B cells and B cell progenitors following androgen ablation in mice.

The production of B lymphocytes is regulated in part by physiologic levels of androgens and estrogens. While these sex hormones down-regulate B lymphopoiesis, augmentation of B lymphopoiesis occurs under conditions where androgen or estrogen levels are decreased. In this study we examine the effect of androgen ablation of male mice on B lymphopoiesis and on the phenotypic composition of peripheral B lymphocyte populations. Spleen and thymic weights are significantly increased following castration, as is the total number of peripheral blood lymphocytes. However, the absolute numbers of B cells in the periphery are selectively increased following castration; the numbers of T cells, NK cells and granulocytes remain unchanged. The increase in circulating B cells is due largely to increases in the numbers of recent bone marrow emigrants expressing a B220(lo+)CD24(hi+) phenotype and these cells remain significantly elevated in castrated mice for up to 54 days post-castration. Similar increases in the percentages of newly emigrated B cells are observed in mice that lack a functional androgen receptor (TFM:). Finally, assessments of B cell progenitors in the bone marrow revealed significant increases in the relative numbers of IL-7-responsive B cell progenitors, including cells in Hardy fractions B (early pro-B cells), C (late pro-B cells), D (pre-B cells) and E (immature B cells). These findings demonstrate that androgen ablation following castration significantly and selectively alters the composition of peripheral B cells in mice. Further, these alterations result from the potentiating effects of androgen ablation on IL-7-responsive pro-B cell progenitors.

Inhibition of pseudomonal ulceration in rabbit corneas by a synthetic matrix metalloproteinase inhibitor.

PURPOSE: To evaluate the effect of the synthetic matrix metalloproteinase inhibitor, Galardin, on proteases produced by Pseudomonas aeruginosa (PA) and on a rabbit model of Pseudomonas keratitis. METHODS: Protease activities of culture broths from Pseudomonas strains PA-28 and W-186 were characterized in vitro by gelatin zymography and by digestion of Azocasein in the presence and absence of Galardin and the serine protease inhibitor, aprotinin. In a noninfectious in vivo experiment, sterile PA culture broth from W-186 was injected intrastromally into rabbit corneas that were treated topically with Galardin or vehicle, then evaluated clinically and histologically. In an infectious in vivo experiment, rabbit corneas were injected with washed PA-28, then treated topically with Galardin or vehicle and clinically scored. RESULTS: Gelatin zymography of culture broth from W-186 and PA-28 detected two proteases that were both inhibited by Galardin. Galardin reduced the digestion of Azocasein by both PA culture broths by 99%, whereas aprotinin did not significantly reduce the protease activity of PA-28 conditioned broth. Intrastromal injection of sterile W-186 culture broth caused rapid corneal destruction that was prevented by topical treatment with Galardin. Intrastromal injection of washed PA-28 bacteria resulted in progressive corneal melting that was significantly (P < 0.005) delayed, but ultimately not prevented, by topical treatment with Galardin. CONCLUSIONS: Pseudomonal protease activity in culture broth consisted predominantly of metalloproteinases and were effectively inhibited by Galardin in vitro. Topical treatment with Galardin prevented destruction of rabbit corneas by bacterial products present in culture broth, and it delayed corneal destruction after injection of PA bacteria. Galardin may be a useful adjuvant when corneal destruction proceeds despite prompt antibiotic treatment.

Effect of recombinant alpha interferons on fertility and interestrous interval in sheep.

In Experiment 1, 12 unmated cyclic ewes received twice-daily intrauterine injections on Days 12 to 14 of one of the following treatments: 1) ovine conceptus secretory proteins (oCSP) containing 25 microg of ovine trophoblast protein-1 (oTP-1) as determined by RIA; 2) 25 or 50 microg recombinant human interferon alpha1 (rhlFN); or 3) 1500 microg of serum proteins (oSP) from a Day-16 pregnant ewe (estrus = Day 0) per uterine horn. Ewes receiving oCSP had longer interestrous intervals (27 +/- 2 days; P<0.05) than ewes receiving oSP (17 +/- 2 days). Ewes receiving either dose of rhlFN had an interestrous interval of 16 +/- 2 days which did not differ (P>0.10) from that of oSP-treated ewes. In Experiment 2, 59 normally cycling ewes, mated on Day 0, received twice-daily intramuscular injections of either 2 mg recombinant bovine interferon alpha1 (rblFN) or placebo on Days 12 to 15 post estrus. On Day 16, pregnancy was confirmed by flushing a morphologically normal conceptus from the uterus. Pregnancy rates for rblFN-treated (80%) and placebo-treated (62%) ewes were not different (P>0.10). Uterine flushings and conceptus-conditioned medium were assayed for oTP-1. Total oTP-1 in conceptus-conditioned culture medium was higher (P<0.02) when conceptuses were from placebo-treated (104 +/- 14 microg/conceptus) than from rblFN-treated (56 +/- 12 microg/conceptus) ewes; while total oTP-1 in uterine flushings was similar (P>0.10) for placebo-treated (132 +/- 15 microg/conceptus) and rblFN-treated (147 +/- 17 microg/conceptus) ewes. The interval from mating to subsequent estrus following conceptus removal was 31 +/- 1 and 28 +/- 1 days for pregnant ewes treated with rblFN and placebo, respectively. Interestrous intervals for nonpregnant ewes were longer (P<0.02) for rblFN-treated (27 +/- 3 days) than for placebo-treated (18 +/- 2 days) ewes.

Luteal function after intrauterine infusion of recombinant bovine interferon-alpha I1 into postpartum beef cows expected to have short or normal luteal phases.

This study was conducted to determine whether intrauterine infusion of recombinant bovine interferon-alpha I1 (rboIFN-alpha I1), which has 70% sequence identity to bovine trophoblast protein-1, will prevent regression of corpora lutea anticipated to have a short lifespan. Twenty-six beef cows in good body condition were allotted to four treatment groups at parturition in a 2 x 2 factorial design. Treatments were: group 1, saline; group 2, rboIFN-alpha I1; group 3, norgestomet-saline; and group 4, norgestomet-rboIFN-alpha I1. Norgestomet implants were inserted on days 21-24 postpartum and removed 9 days later (before injection of human chorionic gonadotrophin (hCG)). Ovulation was induced 30 to 33 days postpartum with 5000 or 10,000 iu hCG. Groups 1 (n = 7) and 3 (n = 5) were given intrauterine infusions (rectocervical approach) twice daily with saline on days 1-12 or 13-24 after hCG injection, respectively. Cows allotted to groups 2 (n = 8) and 4 (n = 6) were given intrauterine infusions (rectocervical approach) of 2 mg rboIFN-alpha I1 twice daily on days 1-12 or 13-24 after hCG injection, respectively. Treatment with both norgestomet and rboIFN-alpha I1 delayed (P less than 0.01) luteolysis. Lengths of luteal phases (days; mean +/- SEM) were 8.4 +/- 0.7 (group 1, saline), 14.1 +/- 1.0 (group 2, rboIFN-alpha I1), 18.6 +/- 1.3 (group 3, norgestomet-saline) and 20.8 +/- 1.2 (group 4, norgestomet-rboIFN-alpha I1). Concentration of progesterone in serum was similar among all groups the first 6 days following hCG-induced ovulation, but differed (P less than 0.01) thereafter.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of bovine follicular fluid and follicle-stimulating hormone on follicular growth in unilaterally ovariectomized prepuberal heifers.

A study was conducted to determine if charcoal-extracted follicular fluid inhibits FSH-induced follicular development in prepuberal heifers. Thirty-six prepuberal heifers were allotted by breed and weight to a 2 x 2 factorial experiment involving charcoal-extracted follicular fluid and FSH treatments. Heifers were unilaterally ovariectomized and injected (intravenously; 10 ml) every 8 h for 88 h with either charcoal-extracted follicular fluid or saline. Follicle-stimulating hormone (2 mg) or saline was injected (intramuscularly) every 8-h starting 24 h after initiation of charcoal-extracted follicular fluid to 88 h following unilateral ovariectomy. Plasma samples were collected at 8-h intervals from 48 h prior to unilateral ovariectomy to 96 h following unilateral ovariectomy when the remaining ovary was removed. Follicular fluid and total ovarian weight increased following FSH treatment. The increases were not inhibited by charcoal-extracted follicular fluid. Total number of surface follicles was similar among treatments. However, FSH induced a shift in follicular diameter from small (less than or equal to 3 mm) to medium (7 to 9 mm) or large (10 to 13 mm) follicles, which was unaffected by charcoal-extracted follicular fluid. Plasma concentration of FSH, but not LH, declined following charcoal-extracted follicular fluid administration. In summary, charcoal-extracted follicular fluid did not inhibit FSH-induced follicular development in prepuberal heifers when charcoal-extracted follicular fluid was administered at a dosage that reduced circulating concentration of FSH by approximately 40%.