Effects of Atorvastatin on Atherosclerosis and Atherogenesis in Systemic Lupus Erythematosus: A Pilot Study

Objective. The effect of statins on atherogenesis in systemic lupus erythematosus (SLE) is poorly known. To inform a wider trial we performed a pilot study evaluating the intima-media thickness of the common carotid artery (CIMT) and some oxidative [beta-2-glycoprotein-1 complexed with oxidised low density lipoprotein (β2GPIoxLDL)], metabolic [paraoxonase (PON), nitrate (NO3 −), nitrite (NO2 −) and nitrotyrosine (NT)], inflammatory [C-reactive protein (CRP) and serum amyloid A (SAA)], and lipid markers before and after 1 year of treatment with 40 mg of oral atorvastatin (AT). Methods. Randomised, double blind, placebo controlled pilot study on consecutive SLE patients: 17 SLE patients were randomised into the AT arm and 20 into the placebo arm. CIMT was measured by high-resolution sonography, PONa by a spectrophotometric method, NO3 − and NO2 − by a colorimetric assay and oxLDL-β2GPI, NT, CRP, and SAA by Elisa. Results. After correction for age and disease duration oxLDL-β2GPI decreased by 27% (P=0.002) and PON/HDL ratio increased by 12% (P=0.01) but CIMT did not change. Conclusion. This pilot study revealed a decrease of oxLDL-β2GPI (oxidant marker) and an increase of PON/HDL ratio (antioxidant activity) after AT indicating a favourable effect of the drug on atherogenic pathways that should be explored on larger trials.

Comparative Analysis of the Informative Value of Radioimmunoassay and Laser Correlation Spectroscopy in Myasthenia Gravis

The objective of this study was to compare informative value of traditional approach (anti-AChR antibody radioimmunoassay) and evaluation of metabolic shifts by laser correlation spectroscopy in myasthenia gravis. The method based on changes in spectral characteristics of laser radiation caused by scattering in a disperse system yields a histogram reflecting particle contribution into light scatter as a function of particle radius in nanometers. The spectrum of anti-AChR-positive serum is characterized by appreciably increased contribution of particles with a radius 4.6–6.2 nm. Binding of serum components with solubilized AChR confirms that this peak is determined by elevated concentration of antibodies to this receptor. The search for the relationship between the disease severity and the distribution pattern of subfraction serum components revealed three informative zones: 6–15, 27–67, and 127–223 nm. In patients without disturbances of vital functions, the contribution of the first zone particles into light scatter increases and that of the third zone particles decreases. Considerable differences attaining the level of statistical significance in zones 4–6 and 20 nm were revealed in the spectra of serum from patients with myasthenia gravis of the same severity with and without thymoma. This opens prospects for dynamic monitoring of the efficiency of therapy.

Effect of Analgesics on Monoclonal Antibody Ascites Production in Mice Administered Upon Recognition of Pain

Monoclonal antibody (mAb) ascites fluid production in mice is a well described method of antibody production, although ethical questions regarding the pain and distress of the animals utilized in this process have been raised. In this study, mice were injected with pristane to initiate granuloma formation, followed by an injection of murine hybridoma PA 2II 2F9-1-1 (2F9) to produce IgG1 subclass mAb directed against protective antigen (PA) protein of Bacillus anthracis. Upon the recognition of pain or distress, characterized by well accepted clinical signs, analgesics were administered by treatment group. The control group (A) received saline, group (B) received meloxicam, group (C) received buprenorphine, and group (D) received both meloxicam and buprenorphine. Analgesics were administered by group for a total of 36–48 hours prior to the second ascites fluid collection. There was no statistical difference in the antibody titer or functionality (P>0.05) between treatment groups at the first or the second collection time points. As reported here, analgesics may be administered upon recognition of pain in mice used for mAb ascites fluid production without affecting antibody concentration or quality and may warrant further evaluation as a refinement in other hybridoma cell lines.

Molecular and Cellular Pathways of Immunoglobulin G Activity In Vivo

In retrospect, the therapeutic potential of immunoglobulins was first demonstrated by von Behring and Kitasato in the late nineteenth century by protecting mice from the lethal effects caused by tetanus and diphtheria toxin via injection of a hyperimmune serum generated in rabbits. Even today, hyperimmune sera generated from human donors with high serum titers against a certain pathogen are still in use as a means of providing passive protection. More importantly, therapeutic antibodies specific for malignant or autoreactive cells have become included in the standard of care in diseases such as breast cancer and malignant lymphoma. Despite this clinical success, we are only at the beginning of understanding the precise molecular and cellular pathways responsible for immunoglobulin G (IgG) activity in vivo. Since then, an enormous amount of information about the mechanism of IgG activity has been obtained in various model systems. The aim of this review is to provide a comprehensive overview of our current understanding of how IgG antibodies mediate their activity in vivo and how we can use this knowledge to enhance the activity of therapeutic antibodies or block the proinflammatory and tissue pathology inducing activity of autoantibodies.

Coevolution of Mucosal Immunoglobulins and the Polymeric Immunoglobulin Receptor: Evidence That the Commensal Microbiota Provided the Driving Force

Immunoglobulins (Igs) in mucosal secretions contribute to immune homeostasis by limiting access of microbial and environmental antigens to the body proper, maintaining the integrity of the epithelial barrier and shaping the composition of the commensal microbiota. The emergence of IgM in cartilaginous fish represented the primordial mucosal Ig, which is expressed in all higher vertebrates. Expansion and diversification of the mucosal Ig repertoire led to the emergence of IgT in bony fishes, IgX in amphibians, and IgA in reptiles, birds, and mammals. Parallel evolution of cellular receptors for the constant (Fc) regions of Igs provided mechanisms for their transport and immune effector functions. The most ancient of these Fc receptors is the polymeric Ig receptor (pIgR), which first appeared in an ancestor of bony fishes. The pIgR transports polymeric IgM, IgT, IgX, and IgA across epithelial cells into external secretions. Diversification and refinement of the structure of mucosal Igs during tetrapod evolution were paralleled by structural changes in pIgR, culminating in the multifunctional secretory IgA complex in mammals. In this paper, evidence is presented that the mutualistic relationship between the commensal microbiota and the vertebrate host provided the driving force for coevolution of mucosal Igs and pIgR.

Cytomorphological and Cytochemical Identification of Microglia

Microglia is one of the major resident immune cells in the central nervous system and is considered to be the key cellular mediator of neuroinflammatory processes. Identification of different Microglial states of activation by morphologic means has been one of the major challenges in the field of neurobiology of diseases. Therefore, microglial biology demands techniques to identify differing stages of microglia in different neuroanatomic locations as well as understanding the role of Microglia in different Neurological diseases. This present study is aimed towards summarizing the literature and for understanding the progress made in different Cytomorphological and Cytochemical techniques of identifying Microglia. This study also review recently used Immunohistochemistry techniques, along with Ultrastructural studies determining different morphological features of resting to activated phagocytic Microglia in a viral induced experimental animal model of neuroinflammation. Results revealed that chronic Microglial activation is considered to be an important component of neuronal dysfunction, injury, and loss (and hence to disease progression). Thus, Microglial research with special emphasis on identification of different activation states of Microglia has gradually become significant.

Retrospective Analysis of Efficacy and Safety of Tocilizumab Treatment in Patients with Severe Systemic-Onset Juvenile Idiopathic Arthritis Followed for 12 Months

The results of the retrospective study evaluating efficacy and safety of tocilizumab treatment in 75 patients with severe systemic-onset juvenile idiopathic arthritis refractory to standard immunosuppressive therapy are presented in the paper. Inactive disease was documented in 64% of patients after 6 months of treatment and in 73% of patients after 12 months. Adverse events manifested as mild and moderate infections as well as laboratory abnormalities: leukopenia, neutropenia, and elevated aminotransferase levels.

Increased Frequency of Antigen-Specific Polyfunctional T Cells in Tuberculosis Patients

This study assessed the polyfunctional T cells in healthy household contacts (HHCs) and TB patients. This study also assessed the memory subsets responsible for the secretion of IFN-γ during the short-term culture with Mycobacterium tuberculosis antigens. Frequencies of CD4+IFN-γ+TNF-α+ T cells and CD8+IFN-γ+TNF-α+ T cells specific to M. tuberculosis antigens were significantly higher in TB patients compared to HHC. IFN-γ-secreting T cells, during overnight stimulation with M. tuberculosis antigens, belonged to effector memory subset with a CD45RA−CD27− phenotype. However, the number of IFN-γ-secreting effector memory cells did not differ between HHC and TB patients.

Partial Characterization of Immunoglobulin Cμ Gene of Water Buffalo (Bubalus bubalis) Predicts Distinct Structural Features of C1q-Binding Site in Cμ3 Domain

Partial characterization of immunoglobulin Cµ gene of water buffalo (Bubalus bubalis) revealed high amino acid sequence identity with Cµ of cattle (94.28%) and sheep (91.71%). Four amino acid replacements (Met-301, Val-310, Asn-331, and Thr-432) in Cµ2, Cµ3, and Cµ4 of buffalo IgM are distinct, however. Unlike cattle, a codon deletion (GTG encoding valine at position 507 in cattle) and an insertion (GGC encoding glycine at position 532) occur in buffalo Cµ4. Three N-linked glycosylation (Asn-X-Thr/Ser) sites (one at position 325–327 in Cµ2; two at positions 372–374 and 394–396 in Cµ3) differentiate buffalo IgM from cattle and sheep. Similar to cattle, buffalo IgM has fewer prolines in Cµ2, which acts as hinge, which restricts Fab arm flexibility. Increased structural flexibility of the C1q-binding site in Cµ3 compensates for the rigid buffalo Cµ2 domain. Secondary structure of C1q-binding site is distinct in buffalo and cattle IgM where long alpha-helical structure is predominant that may be relevant to complement fixation function. Conserved protein motif “Thr-Cys-Thr-Val-Ala-His” provides protein signatures of C1q-binding region of ruminant species. The distinct structural features of C1q-binding site of buffalo and cattle IgM seem to be of functional significance and, therefore, useful in designing antibody based therapeutics.

Synergy in B-Cell Activation between Toll-Like Receptor 9 and Transmembrane Activator and Calcium-Modulating Cyclophilin Ligand Interactor (TACI) in A181E/C104R Compound Heterozygous Siblings

Purpose. Approximately 9% of common variable immunodeficiency (CVID) patients harbor variants in the transmembrane activator and CAML interactor gene, TACI, which contribute to CVID development. We found identical compound heterozygous TACI variants (C104R and A181E) in kindred of which one sibling had severe CVID with refractory auto immunity, and a second sibling remained asymptomatic. This study investigated possible differences in B-cell phenotype and function that could explain this divergent clinical expression. Methods. C104R and A181E TACI variants were identified through Sanger sequencing. Phenotypic evaluation of the lymphocyte compartment was performed by flow cytometry analyses. Lymphoblastoid cell lines (LCL) from the index patient, asymptomatic sibling, and controls were generated. Intracellular TACI expression was determined, and activation-associated calcium flux capacity was measured. In vitro stimulation assays and RT PCR were performed. Results. Both intracellular levels and surface expressed TACI protein were higher in the asymptomatic sibling than the CVID patient as were TACI-triggering-induced mRNA expression AID and production of Ig class-switched antibodies. In analogy, the asymptomatic sibling displayed enhanced Toll-like receptor 9 expression and signaling, suggesting a compensatory immune mechanism. Conclusions. Posttranscriptional regulation of TACI protein and cross-talk with TLR9 signaling may contribute to phenotypic diversity between individuals with TACI variants.