Archive of Medical Research

Therapeutic vaccination against malignant gliomas based on allorecognition and syngeneic tumor antigens: Proof of principle in two strains of rat.

In the present study we investigated whether allogeneic glioma cells can be utilized to evoke prophylactic or therapeutic immune-mediated elimination of syngeneic glioma in two rat strains. Fisher 344 and Sprague-Dawley (SD) rats were injected with two syngeneic glioma cell lines, 9L and C6, respectively, resulting in progressive tumor growth. 9L is syngeneic to the Fisher 344 and allogeneic to the SD rats, while C6 cells are syngeneic to SD rats and allogeneic to Fisher 344 rats. Both rat strains were subcutaneously injected with their respective allogeneic tumor cells, which proved unable to grow progressively. The allogeneic cells were either rejected immediately in SD rats or within 25 days in Fisher rats, after limited tumor outgrowth. Both rat strains were subsequently challenged with their respective syngeneic glioma tumor cells and once more 10 days later with a fivefold higher dose. SD rats, even after reinjection with five times the original dosage of C6 cells, remained tumor free for at least 360 days. Similarly, Fisher rats, after initially rejecting allogeneic tumors, failed to develop syngeneic tumors. To determine anti-tumor immunity against established glioma tumors under more demanding therapeutic conditions, rats were first injected subcutaneously with their respective syngeneic tumor and vaccinated once or repeatedly (at 5-day intervals) with a mixture of the allogeneic or xenogeneic cells, with or without a lysate from the same syngeneic tumor, which served as a therapeutic vaccine preparations. The control group received either no treatment or syngeneic instead of allogeneic cells. In both strains of rats, we demonstrated that the therapeutically vaccinated groups were able to significantly reduce tumor growth, while complete rejection of tumors was noted in the SD rats. Immunization with syngeneic tumor cells alone failed to evoke anti-tumor immunity. We conclude that therapeutic immunization with a combination of allogeneic cells and syngeneic lysates induces rejection of malignant gliomas and offers a protective effect against challenge with syngeneic tumor cells. This immunization approach may prove useful as a post-surgery adjuvant therapy in future cancer treatment protocols, or even as a stand-alone therapeutic tumor vaccination.

Stathopoulos A, Samuelson C, Milbouw G, Hermanne JP, Schijns VE, Chen TC.

Department of Neurosurgery, Les Cliniques du Sud Luxembourg (CSL), Arlon, Belgium; Department of Pathology, University of California, Los Angeles, CA, USA.

Long-term antibody persistence induced by a combined hepatitis A and B vaccine in children and adolescents.

Two cohorts, comprising of subjects aged 1-6 years and 6-15 years were vaccinated with Twinrixtrade mark according to a 0-, 1- and 6-month schedule. The 1-6 years cohort was followed up for 7.5 years and the 6-15 years cohort for 10 years. At the latest follow-up time point, all subjects were seropositive for anti-HAV antibodies, while 86.5% (32/37) and 95.5% (21/22) had anti-HBs>/=10mIU/ml in the 1-6 years and in the 6-15 years cohort. The geometric mean concentrations (GMCs; mIU/ml) were 233 and 680 for anti-HAV antibodies, and 147 and 165 for anti-HBs antibodies, in the 1-6 years and 6-15 years cohorts, respectively. The high persistence of circulating anti-HAV and anti-HBs antibodies in children and adolescents demonstrates the long-term protection offered by Twinrixtrade mark in these age groups.

Diaz-Mitoma F, Law B, Subramanya A, Hoet B.

Children\’s Hospital of Eastern Ontario, Ottawa, Ontario, Canada.

Development of Eurasian H7N7/PR8 high growth reassortant virus for clinical evaluation as an inactivated pandemic influenza vaccine.

Avian-to-human transmission of the high pathogenicity (HP) H7N7 subtype avian influenza viruses in the Netherlands during 2003 caused zoonotic infections in 89 people, including a case of acute fatal respiratory distress syndrome. Public health emergency preparedness against H7N7 avian influenza viruses with pandemic potential includes the development of vaccine candidate viruses. In order to develop a high growth reassortant vaccine candidate virus, low pathogenicity (LP) A/mallard/Netherlands/12/2000 (H7N3) and A/mallard/Netherlands/2/2000 (H10N7) strains were selected as donors of the H7 haemagglutinin and N7 neuraminidase genes, respectively. The donor viruses exhibited high amino acid sequence homology with the surface glycoproteins of A/Netherlands/219/03 H7N7 virus (NL219), an isolate recovered from the fatal human case. Adhering to the seasonal influenza vaccine licensure regulations, we generated a H7N7/PR8 reassortant containing desired surface glycoprotein genes from the mallard viruses and internal genes of A/Puerto Rico/8/34 human vaccine strain (H1N1). Antigenic analysis revealed that the vaccine candidate virus confers broad antigenic cross-reactivity against contemporary Eurasian and the North American H7 subtype human isolates. Mice immunized with formalin inactivated (FI) H7N7/PR8 whole virus vaccine with or without aluminum hydroxide adjuvant conferred clinical protection from mortality and reduced pulmonary replication of the NL219 challenge virus. The FI H7N7/PR8 whole virus vaccine also afforded cross-protection in mice at the pulmonary level against antigenically distinct North American LP A/Canada/444/04 (H7N3) human isolate. The vaccine candidate virus satisfied the agricultural safety requirements for chickens, proved safe in mice, and has entered in phase-I human clinical trial in the United States.

Jadhao SJ, Achenbach J, Swayne DE, Donis R, Cox N, Matsuoka Y.

Influenza Division, Centers for Disease Control and Prevention, 1600 Clifton Road, NE, Atlanta, GA 30333, USA.

Kinetics of asthma- and allergy-associated immune response gene expression in peripheral blood mononuclear cells from vaccinated infants after in vitro re-stimulation with vaccine antigen.

The global expression of immune response genes in infants after vaccination and their role in asthma and allergy is not clearly understood. Pharmacogenomics is ideally suited to study the involved cellular responses, since the expression of thousands of genes can be assessed simultaneously. Here, array technology was used to assess the expression kinetics of immune response genes with association to asthma and allergy in peripheral blood mononuclear cells (PBMC) of five healthy infants after vaccination with Infanrix-Polio+Hib. At 12h after in vitro re-stimulation of the PBMC with pertussis toxin (PT) antigen, 14 immune response pathways, 33 allergy-related and 66 asthma-related genes were found activated.

Lahdenperä AI, Nilsson LJ, Regnström K.

Division of Paediatrics, Department of Clinical and Experimental Medicine, Faculty of Health Sciences, Linköping University, 58185 Linköping, Sweden.

WHO Working Group on revision of the Manual of Laboratory Methods for Testing DTP Vaccines-Report of two meetings held on 20-21 July 2006 and 28-30 March 2007, Geneva, Switzerland.

This report reflects the discussion and conclusions of a WHO group of experts from National Regulatory Authorities (NRAs), National Control Laboratories (NCLs), vaccine industries and other relevant institutions involved in standardization and control of diphtheria, tetanus and pertussis vaccines (DTP), held on 20-21 July 2006 and 28-30 March 2007, in Geneva Switzerland for the revision of WHO Manual for quality control of DTP vaccines. Taking into account recent developments and standardization in quality control methods and the revision of WHO recommendations for D, T, P vaccines, and a need for updating the manual has been recognized. In these two meetings the current situation of quality control methods in terms of potency, safety and identity tests for DTP vaccines and statistical analysis of data were reviewed. Based on the WHO recommendations and recent validation of testing methods, the content of current manual were reviewed and discussed. The group agreed that the principles to be observed in selecting methods included identifying those critical for assuring safety, efficacy and quality and which were consistent with WHO recommendations/requirements. Methods that were well recognized but not yet included in current Recommendations should be taken into account. These would include in vivo and/or in vitro methods for determining potency, safety testing and identity. The statistical analysis of the data should be revised and updated. It was noted that the mouse based assays for toxoid potency were still quite widely used and it was desirable to establish appropriate standards for these to enable the results to be related to the standard guinea pig assays. The working group was met again to review the first drafts and to input further suggestions or amendments to the contributions of the drafting groups. The revised manual was to be finalized and published by WHO.

Corbel MJ, Das RG, Lei D, Xing DK, Horiuchi Y, Dobbelaer R.

National Institute for Biological Standards & Control, UK.

Augmentation of SIV DNA vaccine-induced cellular immunity by targeting the 4-1BB costimulatory molecule.

DNA vaccines are effective at inducing antigen-specific cellular immune responses. Approaches to improve these responses, however, are needed. We examined the effect of stimulating 4-1BB, an activation-inducible T-cell costimulatory receptor, by intravenously co-administering anti-human 4-1BB monoclonal antibody (mAb) in DNA-immunized cynomolgus macaques. Three groups of six cynomolgus macaques were immunized intramuscularly with a DNA vaccine encoding SIV Gag antigen (pSIVgag) at weeks 0, 4 and 8. At days 12, 15, and 19, six macaques received anti-4-1BB 4E9 mAb and six macaques received anti-4-1BB 10C7 mAb. Treatment with 10C7 mAb led to a significant augmentation of SIV Gag-specific IFN-gamma, granzyme B and perforin responses. Treatment with humanized 4E9 mAb also resulted in an enhancement of SIV Gag-specific cellular responses but the magnitude was lower compared to animals receiving 10C7 mAb. These responses persisted up to week 40 and were mostly mediated by CD8(+) T cells. Treatment with anti-4-1BB mAb was more effective in driving the CD8(+) T cells toward a more differentiated CCR7(-)/CD45RA(+) effector state. This study demonstrates that targeting the 4-1BB molecule in vivo results in an enhanced and long-lasting cellular immune response. 4-1BB stimulation may be a promising approach to enhance the effectiveness of DNA vaccines.

Calarota SA, Hokey DA, Dai A, Jure-Kunkel MN, Balimane P, Weiner DB.

Research Institute for Genetic and Human Therapy, Ploiclinico S. Matteo, 27100 Pavia, Italy; Research Institute for Genetic and Human Therapy, Cambridge, MA 02142, USA.

Clinical experience in mental health nursing: Determining satisfaction and the influential factors.

Clinical exposure to the mental health environment has been identified as a major factor in promoting a more favourable attitude towards mental health nursing. However, little attention has been devoted to measuring specific aspects of the clinical placement experience. This paper presents the findings of a survey of 703 undergraduate nursing students designed to measure satisfaction and identify factors influencing satisfaction (length and type of placement, time spent with a preceptor). A questionnaire was administered to undergraduate nursing students in Victoria, Australia, at the conclusion of their clinical experience in mental health. The findings demonstrate a high level of satisfaction, particularly in feeling welcomed, well oriented and supported, and opportunities for patient care. Students\’ views were influenced by the duration and type of placement and time spent with a preceptor. These findings provide valuable information about characteristics of a positive clinical experience that should be considered in structuring clinical placements in the mental health field.

Happell B.

Department of Health Innovation, Centre for Social Science Research, Central Queensland University, Bruce Hwy, Rockhampton, Queensland 4701, Australia.

Synthesis, crystal structure and anti-inflammatory properties of curcumin analogues.

Curcuminoids have been reported to possess multifunctional bioactivities, especially the ability to inhibit proinflammatory induction. Since it has been suggested that the seven-carbon beta-diketone linker in curcumin is responsible for its instability, nine mono-carbonyl five-carbon linker containing analogues were designed and synthesized. Their bioactivity against lipopolysaccharide-induced TNF-alpha amd IL-6 secretion was evaluated by using mouse J774.1 macrophages. The results showed that the 3\’-methoxyl plays an important role in bioactivity and cyclohexanone containing analogues exhibited stronger inflammatory inhibition than acetone and cyclopentanone analogues. Subsequently the most active analogue 3c was determined using single-crystal X-ray diffraction. X-ray analysis and comparison with curcumin reveals that the presence of cyclohexanone in 3c, which remotely resembles the 6-membered ring in the enol tautomer in curcumin, may play an important role in the bioactivity. It is suggested that five-carbon linker analogues containing a cyclohexane ring which are synthetically assessable may be pharmacologically important.

Liang G, Yang S, Zhou H, Shao L, Huang K, Xiao J, Huang Z, Li X.

School of Pharmacy, Wenzhou Medical College, Wenzhou 325035, China; College of Chemistry, Nanjing University of Science and Technology, Nanjing 210094, China.

New diketone based vanadium complexes as insulin mimetics.

Since 1985, when Heyliger et al. first reported the in vivo insulin mimetic activity of oral vanadate, extensive studies exploring vanadium chemistry, including the synthesis of novel complexes and their biological effects both in vitro and in vivo have been pursued. Such complexes have emerged as possible potential agents for diabetes therapy. Among the several existing compounds, diketone based vanadium complexes have been chosen for the current study. Two new complexes namely bisdimethylmalonatooxovanadium(IV) and bisdiethylmalonatooxovanadium(IV) have been synthesized and characterized by UV-visible, FTIR and mass spectral studies. The antidiabetic activity of the complexes was proved by animal study. The results show that the above complexes have comparable antidiabetic potential with respect to the standard drug as well as with bisacetylacetonatooxovanadium(IV) which has been studied earlier by Reul et al.

Sheela A, Roopan SM, Vijayaraghavan R.

Chemistry Division, School of Science and Humanities, VIT University, Vellore 632 014, Tamil Nadu, India.

No association of DAPK1 and ABCA2 SNPs on chromosome 9 with Alzheimer\’s disease.

Recently genetic variation in the DAPK1 and ABCA2 genes has been reported to be associated with late- and early-onset Alzheimer\’s disease (AD), respectively. We examined the most significant two single-nucleotide polymorphisms (SNPs) in DAPK1 in a large case-control cohort of late-onset subjects and matched controls and one of the most significant SNPs in ABCA2 in a small set of early-onset subjects as well. We did not detect associations with AD for any variation.

Minster RL, Dekosky ST, Kamboh MI.

Department of Human Genetics, University of Pittsburgh, Pittsburgh, PA 15261, United States.