Th1 immune response induction by biogenic selenium nanoparticles in mice with breast cancer: preliminary vaccine model

Background: tumor associated antigens can be viably used to enhance host immune response

Objectives: the immunomodulatory effect of biogenic selenium nanoparticles [SeNPs] was compared between treated and untreated mice with crude antigens of 4T1 cells

Materials and Methods: female inbred BALB/c mice [60] were injected by cancinogenic 4T1 cells causing breast cancer. After 10 days, all tumor bearing mice were divided into 4 groups. Group 1 was daily provided oral PBS and injected by the same buffer after tumor induction and was considered as control. Group 2 received only 100 [micro]g/day SeNPs as an oral supplement for 30 days. Group 3 was only injected with 4T1 cells crude antigens with nil supplementation of SeNPs. Group 4 animals were supplemented 100 [micro]g/day SeNPs for 30 days and simultaneously injected with crude antigens of 4T1 cells. All antigens or PBS injections were introduced at 7, 14 and 28 days following tumor induction. Oral PBS and SeNPs supplementation initiated from the first day of tumor induction and continued up to 30 days. During tumor growth, animal weights and survival rates were monitored and at the end of the study the concentrations of different cytokines and DTH responses were measured

Results: data clearly showed that the levels of cellular immunomodulatory components [granzyme B, IL-12, IFN-[lambada], and IL-2] significantly increased [P < 0.05] in mice treated with both SeNPs and crude antigens of 4T1 cells in comparison to the other groups. In contrast, the levels of TGF-[beta] in these mice decreased

Conclusions: although SeNPs showed a noticeable boosting effect for the immune response in mice bearing tumor exposed to crude antigens of 4T1 cells, further complementary studies seem to be inevitable

Radioprotective effect of melatonin on the cervical spinal cord in irradiated rats

It has been suggested that the vascular endothelial growth factor [VEGF] gene expression plays an important role in radiation-induced injury to the spinal cord. This study assesses the radioprotective effects of N-acetyl-5-methoxytryptamine [melatonin] through its modulation of VEGF expression after localized irradiation of the cervical spinal cord. In this experimental study, we divided 192 male rats into four groups: 1. control [n=48]; 2. rats that received an intraperitoneal [IP] injection of melatonin [n=48]; 3. rats that received an IP injection of melatonin 30 minutes prior to cervical spinal cord gamma irradiation [dose: 22 Gy; [n=48]]; and 4. rats that received an IP injection of vehicle prior to spinal cord irradiation [n=48]. The changes in VEGF expression were assessed using real-time RT-PCR and enzyme-linked immunosorbent assays. Samples for light microscopy were stained with hematoxylin and eosin [H and E]. The differences among the groups were analyzed using the analysis of variance [ANOVA] test followed by Tukey's multiple comparisons test. Up-regulation of VEGF expression was observed from 8 to 22 weeks after irradiation [p<0.05]. Paralysis and other radiation-induced myelopathy manifestations developed within 22 weeks after irradiation. VEGF expression in the melatonin pre-treatment group significantly down-regulated in the 20th and 22[nd] weeks after irradiation compared to the radiation-only group. The results support the hypothesis that modulation of VEGF expression by melatonin administration may increase the survival rate of irradiated animals

Ligation independent cloning of polycistronic, genetically modified, HuMAb4D5-8 F [ab'] 2, in bacterial plasmid

In recent years, recombinant monoclonal antibodies and their derivatives have emerged as important targeted therapy agents. Monoclonal antibodies are ex-tremely difficult to produce. So, the cost of production is very high and many people cannot afford these drugs. In this regard, choosing inexpensive and easy ways to manipulate production systems such as bacterial hosts to reduce the cost of manufacturing these critical components are considered as vital step for developmental issues in recombinant expression systems. We, therefore, at-tempted to generate a polycistronic construct of anti HER-2 F[ab']2 fragment antibody for insertion in an expression bacterial plasmid. Also some modifica-tions were made in the hinge region to express antibody F[ab']2 fragment in its authentic form preventing from multiple varieties of disulfide bond formation. Finally, synthesized construct was cloned in pET-32 Ek/LIC vector without using restriction enzyme digestion or ligation reactions. The results of this study showed that modified F[ab']2 fragment was simply and successfully inserted in Escherichia coli [E.coli] using the Ligation Independent Cloning technology

A Recombinant Plasmodium vivax Apical Membrane Antigen-1 to Detect Human Infection in Iran

In Iran, Plasmodium vivax is responsible for more than 80% of the infected cases of malaria per year. Control interventions for vivax malaria in humans rely mainly on developed diagnostic methods. Recombinant P. vivax apical membrane antigen-1 (rPvAMA-1) has been reported to achieve designing rapid, sensitive, and specific molecular diagnosis. This study aimed to perform isolation and expression of a rPvAMA-1, derived from Iranian patients residing in an endemic area. Then, the diagnostic efficiency of the characterized Iranian PvAMA-1 was assessed using an indirect ELISA method. For this purpose, a partial region of AMA-1 gene was amplified, cloned, and expressed in pET32a plasmid. The recombinant His-tagged protein was purified and used to coat the ELISA plate. Antibody detection was assessed by indirect ELISA using rPvAMA-1. The validity of the ELISA method for detection of anti-P. vivax antibodies in the field was compared to light microscopy on 84 confirmed P. vivax patients and compared to 84 non-P. vivax infected individuals. The ELISA cut-off value was calculated as the mean+2SD of OD values of the people living in malaria endemic areas from a south part of Iran. We found a cut-off point of OD=0.311 that showed the best correlation between the sera confirmed with P. vivax infection and healthy control sera. A sensitivity of 81.0% and specificity of 84.5% were found at this cut off titer. A good degree of statistical agreement was found between ELISA using rPvAMA-1 and light microscopy (0.827) by Kappa analysis.

Rt-PCR cloning and expression of complementary DNA for human tumor necrosis factor alpha

U-937, a monocytic cell line was induced with Phorbol Myristate Acetate [PMA] for human tumor necrosis factor alpha [hTNF- alpha] production. An optimized RT-PCR was employed for construction of hTNF- alpha complementary DNA [cDNA]. The resulted fragment was verified by restriction digestion mapping with PvuII.The verified fragment was cloned in pUC18 plasmid and transformants were pelletted onto LB agar medium containing ampicillin, X-gal and IPTG. The resulting white colonies were verified by PCR and cultured in LB medium containing ampicillin and IPTG. The biological activity and the quantity of hTNF- alpha expression was assessed by an ELISA method using a monoclonal anti hTNF- alpha antibody together with a bioassay utilizing L-929 line as sensitive cells