isolation-myeloid-dendritic-cells-epithelial-cells-from-human

Isolation of Myeloid Dendritic Cells and Epithelial Cells from Human Thymus

In this protocol we provide a method to isolate dendritic cells (DC) and epithelial cells (TEC) from the human thymus. DC and TEC are the major antigen ...

University Clinic Tuebingen

Photodynamic therapy (PDT) using topical 5-aminolaevulinic acid (ALA) has been used to treat histologically confirmed cervical intraepithelial neoplasia (CIN-I and -I/II) in a randomised, double-blind, placebo-controlled protocol. Fluorescence microscopy revealed that topical application of 3% ALA in Intrasite Gel to the cervix for 3 hr resulted in the accumulation of protoporphyrin IX in the cervical epithelium. Treatment of CIN with ALA-PDT was well tolerated, with only 3/12 patients in the PDT arm (0/13 in the placebo arm) reporting any discomfort during illumination. Histologic examination of the treated tissue following loop excision 3 months post-PDT indicated that 33% of patients had no evidence of CIN, 42% had no change in the grade of their disease, whilst 25% exhibited an apparent progression of disease. In the control group, the respective figures were 31%, 38% and 31%. There was no significant difference in response between the groups receiving ALA-PDT and those receiving placebo treatment.

A randomised, double-blind, placebo-controlled trial of photodynamic therapy using 5-aminolaevulinic acid for the treatment of cervical intraepithelial neoplasia.

Two membrane compartments of Escherichia coli ghosts, representing empty bacterial cell envelopes, were investigated as carriers of foreign antigens. By subcutaneous immunisation of mice the immunogenicity of bacterial ghosts carrying the Hepatitis B virus core 149 protein (HBcAg-149) as model antigen anchored either in the inner or the outer membrane of E. coli was compared. Both systems induced significant immune responses against the foreign target antigen, the HBcAg-149, in mice. Results indicate that bacterial ghosts provide an excellent carrier system for antigen delivery.

Comparative immunogenicity of the hepatitis B virus core 149 antigen displayed on the inner and outer membrane of bacterial ghosts.

The bacterial ghost system is a novel vaccine delivery system unusual in that it combines excellent natural intrinsic adjuvant properties with versatile carrier functions for foreign antigens. The efficient tropism of bacterial ghosts (BG) for antigen presenting cells promotes the generation of both cellular and humoral responses to heterologous antigens and carrier envelope structures. The simplicity of both BG production and packaging of (multiple) target antigens makes them particularly suitable for use as combination vaccines. Further advantages of BG vaccines include a long shelf-life without the need of cold-chain storage due to their freeze-dried status, they are safe as they do not involve host DNA or live organisms, they exhibit improved potency with regard to target antigens compared to conventional approaches, they are versatile with regards to DNA or protein antigen choice and size, and as a delivery system they offer high bioavailability.

Bacterial ghosts as antigen delivery vehicles.

Enterohemorrhagic Escherichia coli (EHEC) is a bacterial pathogen that is associated with several life-threatening diseases for humans. The combination of protein E-mediated cell lysis to produce EHEC ghosts and staphylococcal nuclease A to degrade DNA was used for the development of an oral EHEC vaccine. The lack of genetic material in the oral EHEC bacterial-ghost vaccine abolished any hazard of horizontal gene transfer of resistance genes or pathogenic islands to resident gut flora. Intragastric immunization of mice with EHEC ghosts without the addition of any adjuvant induced cellular and humoral immunity. Immunized mice challenged at day 55 showed 86% protection against lethal challenge with a heterologous EHEC strain after single-dose oral immunization and 93.3% protection after one booster at day 28, whereas the controls showed 26.7% and 30% survival, respectively. These results indicate that it is possible to develop an efficacious single-dose oral EHEC bacterial-ghost vaccine.

Bacterial ghosts as an oral vaccine: a single dose of Escherichia coli O157:H7 bacterial ghosts protects mice against lethal challenge.

The introduced common brushtail possum (Trichosurus vulpecula) is a major pest in New Zealand and immunocontraceptive vaccines are being developed for biocontrol of possum populations, with bacterial ghosts (BGs) being evaluated as a means of oral delivery. Recombinant BGs expressing possum zona pellucida 3 protein (ZP3) as an L' membrane-anchored protein (ZP3-L') or as an S-layer SbsA-fusion protein (MBP-SbsA-ZP3) were produced by the expression of the cloned bacteriophage phiX174 lysis gene E in E. coli NM522. The humoral immune responses of possums immunised with BGs expressing possum ZP3 were investigated following oral, intranasal/conjunctival, parenteral, and intraduodenal administration to evaluate the BG-ZP3 system for possum fertility control. Antibodies to possum ZP3 were detected in the serum, oviduct secretions, and follicular fluid of immunised animals. Intranasal/conjunctival immunisation elicited reliable antibody immune response in serum and at a key effector site, the ovarian follicular fluid. Intraduodenal administration of possum ZP3 BG vaccine as a priming immunisation elicited significant systemic immune responses, but oral immunisation did not, indicating that protection of BG vaccines from degradation by gastric acidity would enhance the effectiveness of orally delivered vaccines. The detection of antibodies at elevated levels at target sites in the reproductive tract following mucosal delivery demonstrates, for the first time, the potential of BGs as an effective system for vaccine delivery to wild animals, and intranasal/conjunctival immunisation as a promising means for delivery of immunocontraceptive vaccines to wild animals.

Humoral immune responses in brushtail possums (Trichosurus vulpecula) induced by bacterial ghosts expressing possum zona pellucida 3 protein.

Concomitant mitral valve replacement and re-re-repair of severe pectus deformity correction in a patient with Marfan syndrome.

A driveline exit site infection is a serious and common complication in long-term left ventricular assist device (LVAD) support. To reduce the incidence and severity of late driveline infections, we modified our surgical technique (double tunnel), and compared it to the conventional short and straight driveline tunnel technique (conventional). We analyzed 43 consecutive patients (37 HeartMate II; 6 Ventrassist) regarding late onset driveline exit site infections after using the surgical driveline tunnel technique after successful LVAD implantation. Of these 43 patients, 11 were treated with the conventional short and straight driveline tunnel technique (conventional), while 32 patients were treated with the modified long subfascial, C-shaped technique (double tunnel). We observed slightly fewer superficial driveline exit site infections in the double tunnel group, even though the difference was not statistically significant (0.638 vs. 1.148 infections/1,000 patient-days; P = 0.22). There were also insignificantly fewer surgical interventions because of exit site infections in the double tunnel group (0.159 vs. 0.581 revisions/1,000 patient-days; P = 0.18). The double tunnel technique offers more surgical options in the case of driveline exit site infections. Due to the long subfascial tunnel, the infected site can be separated from the new driveline exit site, and vacuum-assisted closure therapy can be applied to the infected area. In conclusion, we recommend using the double tunnel driveline technique because of the low infection rate and better treatment options in the case of driveline exit site infection.

Article	Year
A randomised, double-blind, placebo-controlled trial of photodynamic therapy using 5-aminolaevulinic acid for the treatment of cervical intraepithelial neoplasia. International journal of cancer. Journal international du cancer\| PubMed ID: 12516106	2003
Comparative immunogenicity of the hepatitis B virus core 149 antigen displayed on the inner and outer membrane of bacterial ghosts. Vaccine\| PubMed ID: 15855021	2005
Bacterial ghosts as antigen delivery vehicles. Advanced drug delivery reviews\| PubMed ID: 15878634	2005
Bacterial ghosts as an oral vaccine: a single dose of Escherichia coli O157:H7 bacterial ghosts protects mice against lethal challenge. Infection and immunity\| PubMed ID: 16040994	2005
Humoral immune responses in brushtail possums (Trichosurus vulpecula) induced by bacterial ghosts expressing possum zona pellucida 3 protein. Vaccine\| PubMed ID: 20434548	2010
Concomitant mitral valve replacement and re-re-repair of severe pectus deformity correction in a patient with Marfan syndrome. The Journal of thoracic and cardiovascular surgery\| PubMed ID: 20633902	2010
Double tunnel technique for the LVAD driveline: improved management regarding driveline infections. Journal of artificial organs : the official journal of the Japanese Society for Artificial Organs\| PubMed ID: 21987183	2012

Christoph Haller

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