In vitro and in vivo evaluation of two DNA vaccine candidates

DNA vaccinology, although still in its infancy, has raised a great wave of interest among researchers, due to its wide applicability and the potential advantages it appears to offer over traditional methods of vaccination. In this study two independent bacterially derived antigens were evaluated as DNA vaccine candidates. The main thrust being to determine the potential of these vaccines to express antigen in vitro and stimulate immune responses in vivo. In addition, the route of immunisation on the magnitude and orientation of the immune response to the encoded antigens was also considered, with mice immunized either intramuscularly or intradermally using a Gene Gun. One of the candidates, a DNA vaccine encoding a Mycobacterium bovis antigen (MPB-70) expressed very effectively in vitro when transfected into COS-7 cells. However, it failed to stimulate an immune responses in mice, regardless the route of immunisation used. In contrast, a second bacterial antigen. Fragment C of tetanus toxin produced by Clostridium tetani, expressed poorly in eukaryotic cells but elicited a specific immune response in vaccinated mice. Although, the magnitude of these responses did not appear to differ when mice were immunized using intramuscular or intradermal immunisation using the Gene-Gun, the quality of the immune response generated varied considerably. These results demonstrated that vaccination of pcDNA3/tetC by intramuscular route was associated with a specific anti-Fragment C humoral and cellular response associated with Th1 CD4 cells, whilst immunisation using the Gene-Gun appeared to bias the response toward a CD4 associated Th2 profile. Moreover, both methods were able to prime a specific immune response with a magnitude that could be improved when the mice were boosted using the purified Fragment C antigen. Boosting these animals in such a manner resulted in an improvement in the magnitude of the immune response generated, however, the orientation of this response remained faithful to that originally initiated by the original route of DNA vaccination. Interestingly, the Gene- Gun mediated immunisation appeared to be far more efficient at stimulating immune responses as the magnitude of the immune response to the antigen was unaffected despite only a fiftieth of the amount of DNA used for each immunisation. This work highlights the potential impact of immunisation route on the quality and magnitude of an immune response to Fragment C when delivered as a DNA vaccine

In Vitro Hypoxic Preconditioning of Bone Marrow Stromal Cells Triggers ERK-Mediated Signaling and Growth of L363 Myeloma Cells Celulele stromale ale măduvei hematopoietice precondiţionate la hipoxie in vitro induc proliferarea celulelor mielomatoase L363

Abstract Multiple myeloma (MM

Autoantibody subclass predominance is not driven by aberrant class switching or impaired B cell development

A subset of autoimmune diseases is characterized by predominant pathogenic IgG4 autoantibodies (IgG4-AID). Why IgG4 predominates in these disorders is unknown. We hypothesized that dysregulated B cell maturation or aberrant class switching causes overrepresentation of IgG4+ B cells and plasma cells. Therefore, we compared the B cell compartment of patients from four different IgG4-AID with two IgG1-3-AID and healthy donors, using flow cytometry. Relative subset abundance at all maturation stages was normal, except for a, possibly treatment-related, reduction in immature and naïve CD5+ cells. IgG4+ B cell and plasma cell numbers were normal in IgG4-AID patients, however they had a (sub)class-independent 8-fold increase in circulating CD20-CD138+ cells. No autoreactivity was found in this subset. These results argue against aberrant B cell development and rather suggest the autoantibody subclass predominance to be antigen-driven. The similarities between IgG4-AID suggest that, despite displaying variable clinical phenotypes, they share a similar underlying immune profile.</p

It's Never over until It's over: How Can Age and Ovarian Reserve Be Mathematically Bound through the Measurement of Serum AMH-A Study of 5069 Romanian Women.

Wide regional differences in the age-related Anti Mullerian hormone (AMH) regression patterns or age at onset of natural menopause have been reported, possibly reflecting genetic, socioeconomic, environmental, racial or ethnic peculiarities. Moreover, adaptation of AMH levels from different assays using regression functions may lack accuracy and externally defined references for AMH levels may not fully comply with a specific geographical area. The current study aimed to establish an accurate mathematical relationship between AMH serum values and age in a large group of women from Romania, as any consistent difference from previously reported regression models may aid in building specific profiles for the AMH decline with age in this geographical region. Our study pointed out to the quadratic regression as the most fitted pattern of correlation for all the age groups between 24 and 45. To our knowledge the current manuscript is based on the singular study carried out in this geographical region, generating a particular age-related pattern of association between age and serum AMH levels in women, regardless of their subjacent pathologies

The decline pattern of serum AMH levels in relationship to aging in the studied group.

<p>Patients (n = 5069) were grouped based on their age (green lines); the maximum AMH levels were given in numbers; the red line merges the mean value of every group.</p

Representation of age-specific AMH median, mean, and SD values at one year age intervals.

<p>Serum AMH values decline in direct relationship to aging. Mean (blue), median (green) and standard deviation (SD, orange) of AMH values are represented versus age (years).</p

The quadratic regression of the AMH mean values is more useful to illustrate the AMH decline with age (in years).

<p>The quadratic regression of the AMH mean values is more useful to illustrate the AMH decline with age (in years).</p

Correlation between the 3rd, 10th, 25th, 40th, 50th, 75th, 90th, and 95th percentiles of serum AMH level and age (n = 4888, age between 24 and 45).

<p>Correlation between the 3rd, 10th, 25th, 40th, 50th, 75th, 90th, and 95th percentiles of serum AMH level and age (n = 4888, age between 24 and 45).</p

The quadratic regression versus linear regression of the AMH median values.

<p>The quadratic regression versus linear regression of the AMH median values.</p