Streptomyces as a host for the secretion of heterologous proteins for the production of biopharmaceuticals

The commercial production of therapeutic or diagnostic proteins in recombinant microorganisms is of considerable interest. Several microbial protein production systems have been developed. So far, Escherichia coli have been the commonly employed host. However, proteins expressed in this host remain intracellular and often precipitate as inclusion bodies, which may seriously complicate downstreamprocessing. Faced with this problem, several genera of Gram-positive bacteria are being tested as host for the production of heterologous proteins due to their ability to efficiently secrete proteins in the culture medium. Among them is the genus Streptomyces since several of its species are known to secrete high amounts of proteins. Due to the absence of an extensive restriction-modification system, limited protease activity and the availability of suitable vector systems, Streptomyces lividans is the host of choice for the secretory production of heterologous proteins. The presented results show, that S. lividans can act as an interesting host to produce a number of proteins useful in several disease areas important in the worldwide pharmaceutical sales: i.e. oncology, immunology, cardiovascular diseases and infectious diseases

Optimum ground states for spin-32\frac{3}{2} chains

We present a set of {\em optimum ground states} for a large class of spin-$\frac{3}{2}$ chains. Such global ground states are simultaneously ground states of the local Hamiltonian, i.e. the nearest neighbour interaction in the present case. They are constructed in the form of a matrix product. We find three types of phases, namely a {\em weak antiferromagnet}, a {\em weak ferromagnet}, and a {\em dimerized antiferromagnet}. The main physical properties of these phases are calculated exactly by using a transfer matrix technique, in particular magnetization and two spin correlations. Depending on the model parameters, they show a surprisingly rich structure.Comment: LaTeX, 22 pages, 6 embedded Postscript figure

Colonisation of Clostridium in the body is restricted to hypoxic and necrotic areas of tumours

The use of gene therapy is one of the most recent molecular strategies for the treatment of cancer. It is essential, however, to have an efficient transfer system by which the desired gene can be delivered to the correct environment. The experiments described in this report investigate apathogenic Clostridium as a possible vector to transfer a specific gene product into the extracellular microenvironment of the tumour which is hypoxic/necrotic in parts, using WAG/Rij rats with transplantable rhabdomyosarcomas as a model. Our data show that Clostridium, after systemic administration of at least 10(7) spores, specifically colonises the hypoxic/necrotic areas of our tumour model, the most efficient species being C. acetobutylicum (NI-4082) and C. oncolyticum. Although spores were also detected in normal tissues for up to 4 weeks, they did not germinate in these tissues. We conclude that it seems likely that these bacteria can be used as a selective transfer system into the extracellular environment of tumours which have hypoxic regions. This strategy would be more tumour-specific than various other strategies that are currently being investigated in anti-cancer gene therapy. (C) 1998 Academic Press.</p

Explicit hypoxia targeting with tumor suppression by creating an “obligate” anaerobic Salmonella Typhimurium strain

Using bacteria as therapeutic agents against solid tumors is emerging as an area of great potential in the treatment of cancer. Obligate and facultative anaerobic bacteria have been shown to infiltrate the hypoxic regions of solid tumors, thereby reducing their growth rate or causing regression. However, a major challenge for bacterial therapy of cancer with facultative anaerobes is avoiding damage to normal tissues. Consequently the virulence of bacteria must be adequately attenuated for therapeutic use. By placing an essential gene under a hypoxia conditioned promoter, Salmonella Typhimurium strain SL7207 was engineered to survive only in anaerobic conditions (strain YB1) without otherwise affecting its functions. In breast tumor bearing nude mice, YB1 grew within the tumor, retarding its growth, while being rapidly eliminated from normal tissues. YB1 provides a safe bacterial vector for anti-tumor therapies without compromising the other functions or tumor fitness of the bacterium as attenuation methods normally do

Tumor Invasion of Salmonella enterica Serovar Typhimurium Is Accompanied by Strong Hemorrhage Promoted by TNF-α

BACKGROUND:Several facultative anaerobic bacteria with potential therapeutic abilities are known to preferentially colonize solid tumors after systemic administration. How they efficiently find and invade the tumors is still unclear. However, this is an important issue to be clarified when bacteria should be tailored for application in cancer therapy. METHODOLOGY/PRINCIPAL FINDINGS:We describe the initial events of colonization of an ectopic transplantable tumor by Salmonella enterica serovar Typhimurium. Initially, after intravenous administration, bacteria were found in blood, spleen, and liver. Low numbers were also detected in tumors associated with blood vessels as could be observed by immunohistochemistry. A rapid increase of TNF-alpha in blood was observed at that time, in addition to other pro-inflammatory cytokines. This induced a tremendous influx of blood into the tumors by vascular disruption that could be visualized in H&E stainings and quantified by hemoglobin measurements of tumor homogenate. Most likely, together with the blood, bacteria were flushed into the tumor. In addition, blood influx was followed by necrosis formation, bacterial growth, and infiltration of neutrophilic granulocytes. Depletion of TNF-alpha retarded blood influx and delayed bacterial tumor-colonization. CONCLUSION:Our findings emphasize similarities between Gram-negative tumor-colonizing bacteria and tumor vascular disrupting agents and show the involvement of TNF-alpha in the initial phase of tumor-colonization by bacteria