Immunological consequences of “immune reconstitution therapy” in multiple sclerosis: A systematic review

Immune reconstitution therapy (IRT) is an emerging concept for the treatment of multiple sclerosis (MS) that is given intermittently and can induce long-term remission of MS that is sustained in treatment-free periods. A systematic literature review was performed to identify and summarize current knowledge regarding the short- and long-term immunological consequences of different IRTs and CD20 depleting therapies on the cellular level in patients with MS. A total of 586 articles published between January 2010 and September 2019 were identified and screened; 44 studies met inclusion criteria for the review. All the treatments considered appeared to produce both qualitative and quantitative changes in the immune cell populations of patients with MS that resulted in a more anti-inflammatory immune profile. Autologous hematopoietic stem cell transplantation produced the longest-lasting and greatest effects on a wide range of immune cells. Many patients achieved prolonged depletion of the adaptive immune system when alemtuzumab and cladribine tablets were administered as short courses of therapy; however, a proportion of patients required retreatment to maintain these effects. Alemtuzumab may produce greater depletion of both CD4+ and CD8+ T cells than cladribine tablets, although both treatments similarly deplete B cells. Recovery of B cells before T cell recovery and hyperpopulation of B cells after alemtuzumab may contribute to secondary autoimmunity. Cladribine tablets had a greater effect on B cells than T cells, and no hyperpopulation of B cells was observed after treatment with cladribine tablets. Ocrelizumab and rituximab require regular repeated treatment every 6 months to maintain depletion of B and T cells. Effects of the drug treatments on the innate immune system were minor compared with those on the adaptive immune system. Additional characterization of the cellular changes occurring during IRT and CD20 depletion may lead to further improvement in the understanding of the pathogenesis of MS and the future development of therapies with even longer lasting effects. Although the treatments considered in this review improve quality of life and outcomes for patients with MS, a cure for this debilitating disease is not yet in sight.     

A “Geographic Information Systems” Based Technique for the Study of Microvascular Networks

An automated system (ANET) has been developed to construct interactive maps of microvascular networks, calculate blood flow parameters, and simulate microvascular network blood flow using the geographic information systems (GIS) technology. ANET enables us to automatically collect and display topological, structural, and functional parameters and simulate blood flow in microvascular networks. The user-definable programming interface was used for the manipulation of drawings and data. Visual enhancement techniques such as color can be used to display useful information within a network. In ANET the network map becomes a graphical interface through which network information is stored and retrieved and simulations of microvascular network blood flow are carried out. We have used ANET to study the effects of ionizing radiation on normal tissue microvascular networks. Our results indicate that while vessel diameters significantly increased with age in control animals they decreased in irradiated animals. The tortuosity of irradiated vessels (16.3 ± 1.1 mean±standard error of the mean) was significantly different from control vessels (10.0 ± 1.3) only at 7 days postirradiation. Average red blood cell transit time was significantly different between control (1.6 ± 0.6 s) and irradiated (10.7 ± 5.7 s) microvascular networks at 30 days postirradiation. ANET provides an effective tool for handling the large volume of complex data that is usually obtained in microvascular network studies and for simulating blood flow in microvascular networks. © 1999 Biomedical Engineering Society. PAC99: 8764-t, 8719Tt, 0705Pj, 8750Gi     

Versatile supramolecular pDNA vehicles via “click polymerization” of β-cyclodextrin with oligoethyleneamines

Herein, we report the efficient synthesis of high molecular weight polymers (up to 331 kDa) that contain β-cyclodextrin within the polymer backbone and the examination of these structures for pDNA delivery within cultured mammalian cells. Two series of polymers were synthesized, one with variation in oligoethyleneamine stoichiometry, Cd1₄₆, Cd2₄₄, Cd3₄₉, and Cd4₄₇ (1-4 oligoethyleneamines in the repeat unit, respectively and similar degree of polymerization, n_w = 44–49) and another with variation in polymer length (four ethyleneamines in the repeat unit), Cd4₂₇, Cd4₄₇, Cd4₉₃, and Cd4₂₀₀ [n_w = 27, 47, 93, 200] via the “click reaction”. The two series of polymers revealed efficient pDNA binding and compaction through gel electrophoresis, dynamic light scattering, and transmission electron microscopy experiments. The DNase protection assay showed a decrease in pDNA degradation with an increase in the polymer amine stoichiometry, where polymer Cd3₄₉ and all of the Cd4 analogs completely protected pDNA for up to 8 h in serum. The cellular uptake and gene expression profiles were examined in HeLa cells, which similarly demonstrated that both the series of polymers had high pDNA delivery where, Cd3₄₉ and Cd4₉₃ had the most effective luciferase gene expression. In addition, the cell viability profiles were quite high with all of the structures.     

RETRACTED ARTICLE: An Experimental Study of the Neurogenic and the Immunological Contribution to “Tennis Elbow” in Rats

In the present study the content of substance P (SP)-, neurokinin A (NKA)-, calcitonin gene-related peptide (CGRP)- and neuropeptide Y (NPY)-like immunoreactivity (-LI) was measured in rats cerebrospinal fluid (CSF), plasma and perfusates (PF) from both elbow enthesis during acute inflammation. Either substance P, SP, (10–5 M, 0.01 ml) or human recombinant interleukin-1 (hrIl-1 , 0.01 ml) were injected into the right enthesis of the extensor carpi radialis brevis (ECRB). The left ECRB and both ECRBs of control rats, were injected with 0.01 ml saline. Samples of CSF, plasma and PF from both ECRBs were obtained at 2, 6, and 24 h following injection and neuropeptide-LI was analysed by specific radioimmunoassays. Neuropeptide-LI was compared with control values and between the treated groups. In both treated groups NKA- and CGRP-LI was increased in CSF and NKA-LI decreased in plasma, while CGRP- and NPY-LI were raised to a similarly significant degree in the enthesis of the ECRB. SP-LI was increased in ECRB PF in comparison with controls and NKA-LI levels were higher in the hrIl-1 group both in comparison with controls and between treated groups. In summary an unilateral injection of either SP or hrIl-1 into the enthesis of the ECRB of the rat showed a similar influence at 2, 6, and 24 h following injection. The most pronounced changes in neuropeptide-LI occurred in the ECRB PF of both treated groups.     

Blood testosterone in rats: Correlation of the level of individual anxiety and its impairment after “death threat”

Studies of identical groups of male Wistar rats after preliminary selection to give groups including extreme behavioral types with low and high rankings on the anxiety scale showed that blood testosterone concentrations in intact rats (controls) correlated negatively with anxiety ranking, i.e., minimal hormone concentrations (no greater than 5 nM) corresponded to high levels of anxiety — with a predominance of passive defensive behavioral components on testing. Short-term exposure to a “death threat” situation (sight of a boa attacking and eating two individuals from the group of rats) impaired this correlational relationship in a manner comparable to the sequelae of chronic neuroticization by unavoidable pain stimulation. Impairments were manifest as scatter in measures in low-anxiety animals (3–21 nM). This characteristic, reflecting the multitude of adaptive pathways in the population in threat situations, distinguishes this type of action from neuroticization by unavoidable pain stimulation, which leveled out individual differences and decreased the hormone level. __________ Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti imeni I. P. Pavlova, Vol. 57, No. 5, pp. 591–597, September–October, 2007.     

“Artificial Lymphatic System”: A New Approach to Reduce Interstitial Hypertension and Increase Blood Flow, pH and pO2 in Solid Tumors

A mechanical drainage system, the artificial lymphatic system (ALS), consisting of a vacuum source and drain, is evaluated for its ability to aspirate the interstitial fluids responsible for the elevated interstitial fluid pressure (IFP) observed in solid tumors. IFP, pH, and pO₂ radial profiles were measured before and after aspiration using wick-in-needle (WIN) probes, needle pH and oxygen electrodes, respectively. Laser Doppler flowmetry measured temporal changes in blood flow rate (BFR) at the tumor surface during aspiration. The WIN probe and IFP profile data were analyzed using numerical simulation and distributed mathematical models, respectively. The model parameter, p _E reflecting central tumor IFP, was reduced from 15.3 to 5.7 mm Hg in neuroblastoma and from 13.3 to 12.1 mm Hg in Walker 256, respectively, following aspiration. The simulation demonstrated that spatial averaging inherent in WIN measurements reduced the calculated magnitude of the model parameter changes. IFP was significantly lower (p < 0.05), especially in regions surrounding the drain, and BFR was significantly higher (p < 0.05) following 25 and 45 min of aspiration, respectively; pH and pO₂ profiles increased following aspiration. The experimental and mathematical findings suggest that ALS aspiration may be a viable way of reducing IFP and increasing BFR, pO₂ and pH and should enhance solid tumor chemo and radiation therapy. © 2000 Biomedical Engineering Society. PAC00: 8719Tt, 8715Vv, 8719Uv, 8780-y, 8719Xx     

Immuno- and Enzyme-histochemistry of HRP for Demonstration of Blood Vessel Permeability in Mouse Thymic Tissues by “In Vivo Cryotechnique”

It is difficult to understand the in vivo permeability of thymic blood vessels, but “in vivo cryotechnique” (IVCT) is useful to capture dynamic blood flow conditions. We injected various concentrations of horseradish peroxidase (HRP) with or without quantum dots into anesthetized mice via left ventricles to examine architectures of thymic blood vessels and their permeability at different time intervals. At 30 sec after HRP (100 mg/ml) injection, enzyme reaction products were weakly detected in interstitium around some thick blood vessels of corticomedullary boundary areas, but within capillaries of cortical areas. At 1 and 3 min, they were more widely detected in interstitium around all thick blood vessels of the boundary areas. At 10 min, they were diffusely detected throughout interstitium of cortical areas, and more densely seen in medullary areas. At 15 min, however, they were uniformly detected throughout interstitium outside blood vessels. At 30 min, phagocytosis of HRP by macrophages was scattered throughout the interstitium, which was accompanied by decrease of HRP reaction intensity in interstitial matrices. Thus, time-dependent HRP distributions in living mice indicate that molecular permeability and diffusion depend on different areas of thymic tissues, resulting from topographic variations of local interstitial flow starting from corticomedullary areas.     

Rapid Diagnosis of Cryptococcal Meningitis by Use of Lateral Flow Assay on Cerebrospinal Fluid Samples: Influence of the High-Dose “Hook” Effect

Cryptococcal meningitis is the most frequent cause of meningitis and a major cause of mortality in HIV-infected adults in Africa. This study evaluated the performance of the lateral flow assay (LFA) on cerebrospinal fluid (CSF) samples for the diagnosis of cryptococcal meningitis against that of existing diagnostic tests. LFA performed on 465 undiluted CSF samples had a sensitivity of 91%. When the LFA was paired with Gram staining, a sensitivity of 100% was achieved after implementation of a dilution step for samples with negative LFA results and the presence of yeasts on microscopy. Microscopy is essential for preventing the reporting of false-negative results due to the high-dose “hook” effect.     

“No Wash” Albumin-Dextran Dilution for Double-Unit Cord Blood Transplantation is Safe with High Rates of Sustained Donor Engraftment

Washing cord blood (CB) grafts involves product manipulation and may result in cell loss. We investigated double-unit CB transplantation (CBT) using red blood cell (RBC)–depleted units diluted with albumin-dextran in patients with hematologic malignancies. One-hundred thirty-six patients (median age, 43 years; range, 4 to 71; median weight, 69 kilograms (kg); range, 24 to 111) underwent transplantation with a 4/6 to 6/6 HLA-matched graft. Patients ≤ 20 kg were excluded, as they only received washed units. Units were diluted a median of 8 fold to a median volume of 200 mL/unit. The median infused total nucleated cell doses were 2.7 (larger unit) and 2.0 (smaller unit) x 10⁷/kg, respectively, and the median post-thaw recovery was 86%. Units were infused consecutively (median, 45 minutes/unit). While only 17 patients (13%) had no infusion reactions, reactions in the remaining 119 patients were almost exclusively mild-moderate (by CTCAE v4 criteria 12 grade 1, 43 grade 2, 63 grade 3) with only 1 patient (< 1%) having a severe (grade 4) reaction. Moreover, most were easily treated. Grade 2 to 3 hypertension was the most common in 101 (74%) patients. The cumulative incidence of sustained donor-derived neutrophil engraftment was high: 95% in myeloablative and 94% in nonmyeloablative CBT recipients. With appropriate supportive care, double-unit CBT with RBC-depleted grafts infused after albumin-dextran dilution is safe with high rates of engraftment in patients > 20 kg.