Adenovirus-triggered innate signalling pathways.

Adenoviruses are important infectious agents and also emerging vectors in different biomedical applications. These viruses elicit a strong innate and adaptive immune response, which influences both the course of disease and the success of the applied vectors. Several Toll-like Receptor (TLR)-dependent and -independent mechanisms contribute to these responses. Understanding of the involved viral and cellular factors is crucial for the treatment of various adenovirus diseases and the optimal design of adenovirus vector applications. Here we summarize our current understanding of the complex nature of adenovirus-induced innate immune mechanisms.

Characterization of transgenic mice containing adenovirus early region 3 genomic DNA.

Human adenoviruses (Ad) contain a complex transcription region (E3) which codes for proteins that interact with several arms of the immune system. However, E3 genes are not essential for replication in tissue culture. An E3-encoded 19,000-molecular-weight (19K) glycoprotein (gp19K) binds to the class I major histocompatibility complex (MHC) in the endoplasmic reticulum and prevents MHC transport to the cell surface. Three other E3 proteins are involved in the inhibition of apoptosis by tumor necrosis factor alpha. The entire E3 genomic DNA was utilized to produce transgenic mice to study the effect of the E3 proteins on pathogenesis of various infectious agents and to investigate the in vivo synthesis and processing of the multiple E3 mRNAs and proteins. There was basal expression of the E3 promoter in the thymus, kidneys, uterus, and testes and at all levels of the gastrointestinal tract. In addition, the E3 promoter of the transgene could be activated in some other organs, including the liver, by infection of these animals with an E3-deficient Ad (Ad7001) which contains a functional E1A region. Transactivation in vivo could also be demonstrated by infusion of bacterial lipopolysaccharide. There appeared to be differential ratios of expression between several of the E3 mRNAs in transgenic lung fibroblasts and primary kidney cells cultured from the transgenic animals. This observation suggested that there was differential mRNA splicing that was organ specific. These transgenic animals should provide a useful model for studying the effects of the E3 proteins on the immune system and on diseases affected either by control of MHC or by selected functions of tumor necrosis factor that are inhibitable by Ad E3 proteins.

Laparoscopic gastric surgery. Early experiences.

In the Department of General Surgery the authors performed 12 elective laparoscopic gastric operations for gastric pathologies. The indications for the procedures were recurrent or therapy resistant and complicated peptic ulcer in 9 cases, benign gastric tumors in 2 cases and early gastric cancer in 1 case. Operative procedures were the next: posterior truncal vagotomy with anterior lesser curve seromyotomy (5 patients), total truncal vagotomy with gastrojejunostomy (2 patients), total truncal vagotomy with pyloroplasty (1 patient), total truncal vagotomy with antrectomy and Billroth-II reconstruction (1 patient), resection of benign gastric tumor by the transgastric approach (1 patient), Billroth-II resection for benign gastric tumor (1 patient), wedge resection of gastric wall for early gastric cancer (1 patient). Intraoperative gastroscopy was used for location of the lesion in 4 of 12 cases. Apart from delayed gastric emptying (2 cases), patients recovered without any problem. The mean hospital stay was 5.7 days. Early experiences with laparoscopic gastric surgery has shown that there are certain important advantages to the approaches. They hold the promise of less pain, less immobility, quicker alimentation, shorter hospitalization, less wound and respiratory complications and an earlier return to normal activities.

Positive regulatory domain I binding factor 1 silences class II transactivator expression in multiple myeloma cells.

The major histocompatibility complex (MHC) class II transactivator (CIITA) acts as a master switch to activate expression of the genes required for MHC-II antigen presentation. During B-cell to plasma cell differentiation, MHC-II expression is actively silenced, but the mechanism has been unknown. In plasma cell tumors such as multiple myeloma the repression of MHC-II is associated with the loss of CIITA. We have identified that positive regulatory domain I binding factor 1 (PRDI-BF1), a transcriptional repressor, inhibits CIITA expression in multiple myeloma cell lines. Repression of CIITA depends on the DNA binding activity of PRDI-BF1 and its specific binding site in the CIITA promoter. Deletion of a histone deacetylase recruitment domain in PRDI-BF1 does not inhibit repression of CIITA nor does blocking histone deacetylase activity. This is in contrast to PRDI-BF1 repression of the c-myc promoter. Repression of CIITA requires either the N-terminal acidic and conserved PR motif or the proline-rich domain. PRDI-BF1 has been shown to be a key regulator of B-cell and macrophage differentiation. These findings now indicate that PRDI-BF1 has at least two mechanisms of repression whose function is dependent on the nature of the target promoter. Importantly, PRDI-BF1 is defined as the key molecule in silencing CIITA and thus MHC-II in multiple myeloma cells.