Inhibitory proteaz na powierzchni ciała pszczół (Apis mellifera L.) w klatce i w ulu

The aim of the work was to determine the activity of protease inhibitors sampled from the body surface of bee workers kept in a natural hive environment and in a cage. The samples were collected for five weeks. 40 cage samples and 50 hive samples were gathered, each containing 10 bees. Hydrophilic (water-treated) and hydrophobic (Triton-rinsed) proteins were isolated from the insects. The samples containing washed-out proteins were tested as follows: the activity of aspartic and serine protease inhibitors by the Lee and Lin method; electrophoretic analysis of proteins in a polyacrylamide gel for protease inhibitor detection by means of the modified Felicioli method; and in vivo tests of antifungal and antibacterial activity using the double application method. The cage environment had a destabilizing effect on the natural protease inhibitor system causing radical variation in its activity, which was not the case with the hive environment. The samples were not found to be active in relation to M. luteus and E. coli. The cage bees were less resistant to microorganisms. The results of the in vivo microorganismal test confirmed the fact of weaker protease inhibitor activity in the washed-out body-surface samples of the cage bees that was also observed in in vitro biochemical analyses. The results of cage-based analyses of non-specific apian resistance should be treated with caution when used in reference to hive bees.Określono aktywność inhibitorów proteaz wyizolowanych z powierzchni ciała robotnic utrzymywanych w naturalnym środowisku ula oraz w klatce. Próby pobierano przez pięć tygodni, pozyskując 40 prób z klatek i 50 prób z ula, w każdej po 10 pszczół. Z owadów wyizolowano białka hydrofilne (przy użyciu wody) oraz hydrofobowe (przy użyciu tritonu). Próbki z wypłukanymi białkami poddano następujący moznaczeniom: aktywność inhibitorów proteaz asparaginowych i serynowych wg metody Lee i Lina; analiza elektroforetyczna białek w żelu poliakrylamidowym do wykrywania inhibitorów proteaz wg zmodyfikowanej metody Felicioliego; aktywność przeciwgrzybowa i antybakteryjna w testach in vivo metodą płytek dwuwarstwowych. Środowisko klatki działało destabilizująco na system naturalnych inhibitorów proteaz wywołując duże wahania ich aktywności, co nie zdarzyło się w ulu. W próbkach nie zaobserwowano aktywności wobec M. luteus i E. coli. Pszczoły w klatce miały słabszą oporność przeciwko mikroorganizmom. Wyniki testu z mikroorganizmami in vivo potwierdziły słabszą aktywność inhibitorów proteaz w próbkach wypłukanych z powierzchni ciał pszczół w klatkach, wykazaną również w analizach biochemicznych in vitro. Uzyskane w klatkach wyniki badań oporności nieswoistej pszczół należy ostrożnie odnosić do pszczół w ulu

Oncolytic reovirus enhances rituximab-mediated antibody-dependent cellular cytotoxicity against chronic lymphocytic leukaemia

The naturally occurring oncolytic virus (OV), reovirus, replicates in cancer cells causing direct cytotoxicity, and can activate innate and adaptive immune responses to facilitate tumour clearance. Reovirus is safe, well tolerated and currently in clinical testing for the treatment of multiple myeloma, in combination with dexamethasone/carfilzomib. Activation of natural killer (NK) cells has been observed after systemic delivery of reovirus to cancer patients; however, the ability of OV to potentiate NK cell-mediated antibody-dependent cellular cytotoxicity (ADCC) is unexplored. This study elucidates the potential of oncolytic reovirus for the treatment of chronic lymphocytic leukaemia (CLL), both as a direct cytotoxic agent and as an immunomodulator. We demonstrate that reovirus: (i) is directly cytotoxic against CLL, which requires replication-competent virus; (ii) phenotypically and functionally activates patient NK cells via a monocyte-derived interferon-α (IFNα)-dependent mechanism; and (iii) enhances ADCC-mediated killing of CLL in combination with anti-CD20 antibodies. Our data provide strong preclinical evidence to support the use of reovirus in combination with anti-CD20 immunotherapy for the treatment of CLL