Decreased anti-donor major histocompatibility complex class I and increased class II alloantibody response in allograft tolerance in adult rats

Permanent tolerance to allografts can be induced in adult rats by donor-specific transfusions (DST) prior to transplantation. We have previously reported, in a model of heart allograft, the presence of a heavy leukocyte infiltrate, in the allograft which displayed a strong allospecific cytotoxicity when tested in vitro against donor cells, and a strong accumulation of mRNA for granzyme A and perforin in vivo. In contrast, there was a major decrease in the accumulation of mRNA for interleukin-2 and interferon-γ. These results suggested that the DST-induced tolerance was associated with a decrease in type-1 T helper (Th1) cell function. The major role of preformed antibodies in xeno and allorejection is clearly established. Nevertheless, the consequences of alloantibody production in acute rejection and tolerance induction remains to be elucidated. We here analyze the alloantibody response in rejecting and DST-treated recipients. We show that, after transplantation, tolerant recipients, in contrast to rejecting ones, mount a low IgM alloresponse that switches to low IgG production. Detailed analysis of IgG alloantibodies in DST-treated recipients revealed that their production decrease was not equally distributed. Whereas rejecting animals mounted a strong anti-class I and II IgG alloantibody response, DST-treated recipients produced anti-class II and low titers of anti-class I IgG alloantibodies. Furthermore, among IgG subclasses, tolerant recipients predominantly produced IgG2a, a profile which, in the rat, is compatible with a Th2-controlled response. Finally, the passive transfer of immune serum from rejecting animals to DST-treated recipients could abrogate the tolerance. We suggest that the absence of anti-class I alloantibodies combined with preserved and/or increased anti-class II production plays a major role in graft tolerance in this model. These results reinforced the role of alloantibodies in rejection and in induction of tolerance.     

Computed tomography and histological evaluation of xenogenic and biomimetic bone grafts in three-wall alveolar defects in minipigs

Objectives

This study aimed to compare the performance of a xenograft (XG) and a biomimetic synthetic graft (SG) in three-wall alveolar defects in minipigs by means of 3D computerised tomography and histology.

Materials and methods

Eight minipigs were used. A total of eight defects were created in the jaw of each animal, three of which were grafted with XGs, three with SGs, and two were left empty as a negative control. The allocation of the different grafts was randomised. Four animals were euthanised at 6 weeks and four at 12 weeks. The grafted volume was then measured by spiral computed tomography to assess volume preservation. Additionally, a histological analysis was performed in undecalcified samples by backscattered scanning electron microscopy and optical microscopy after Masson’s trichrome staining.

Results

A linear mixed-effects model was applied considering four fixed factors (bone graft type, regeneration time, anatomic position, and maxilla/mandible) and one random factor (animal). The SG exhibited significantly larger grafted volume (19%) than the XG. The anterior sites preserved better the grafted volume than the posterior ones. Finally, regeneration time had a positive effect on the grafted volume. Histological observations revealed excellent osseointegration and osteoconductive properties for both biomaterials. Some concavities found in the spheroidal morphologies of SGs were associated with osteoclastic resorption.

Conclusions

Both biomaterials met the requirements for bone grafting, i.e. biocompatibility, osseointegration, and osteoconduction. Granule morphology was identified as an important factor to ensure a good volume preservation.

Clinical relevance

Whereas both biomaterials showed excellent osteoconduction, SGs resulted in better volume preservation.

     

Glucose homeostasis in acromegaly: effects of long-acting somatostatin analogues treatment

OBJECTIVE: Acromegaly is a syndrome with a high risk of impaired glucose tolerance (IGT) and diabetes mellitus (DM). Somatostatin analogues, which are used for medical treatment of acromegaly, may exert different hormonal effects on glucose homeostasis. Twenty-four active acromegalic patients were studied in order to determine the long-term effects of octreotide-LAR and SR-lanreotide on insulin sensitivity and carbohydrate metabolism. DESIGN: Prospective study. PATIENTS: We studied 24 patients with active acromegaly, 11 males and 13 females, aged 50.7 +/- 12.7 years, body mass index (BMI) 30.1 +/- 4.8 (kg/m2). MEASUREMENTS: All patients underwent an oral glucose tolerance test (OGTT) and 12 also had an euglycaemic hyperinsulinaemic clamp. All patients were evaluated at baseline and after 6 months of somatostatin analogues therapy. RESULTS: Acromegalic patients showed low M-values in respect to the control group at baseline (P<0.05), followed by a significant improvement after 6 months of therapy (P<0.005 vs. baseline). Serum glucose levels at 120 min during OGTT worsened (P<0.05) during somatostatin analogs therapy in patients with normal glucose tolerance, but not in those with impaired glucose tolerance or diabetes mellitus. This was associated with a reduced (P<0.05) and 30 min delayed insulin secretion during OGTT. Also, HbA1c significantly deteriorated in all subjects after treatment (4.7 +/- 0.6% and 5.1 +/- 0.5%, basal and after six months, respectively, P<0.005). CONCLUSION: In acromegalic patients, somatostatin analogues treatment reduces insulin resistance, and also impairs insulin secretion. This may suggest that the use of oral secretagogue hypoglycaemic agents and/or insulin therapy should be considered rather than insulin sensitizers, as the treatment of choice in acromegalic patients who develop frank hyperglycaemia during somatostatin analogues therapy.     

Unsuccessful treatment with voriconazole of a brain abscess due to Cladophialophora bantiana

Cladophialophora bantiana is a dematiaceous fungus, associated with brain abscess in normal or immunosuppressed patients. We report a case of CNS infection in this agent unsuccessfully treated by surgery and various antifungal compounds including high doses of voriconazole (6 mg/kg bid). No adverse effects related to this compound were observed.     

Donor-specific allograft tolerance by administration of recipient-derived immature dendritic cells and suboptimal immunosuppression

We recently showed that injection of recipient-type immature bone marrow-derived dendritic cells (iBMDCs) the day before transplantation induced a significant prolongation of allograft survival. This study aimed at improving the administration protocol to induce allograft tolerance. Various amounts of iBMDCs were administered to syngeneic LEW.1A rats before and after transplantation of an allogeneic LEW.1W heart, with or without additional suboptimal immunosuppression. Allograft survival was not improved by repeated injections of syngeneic iBMDCs or by additional treatment with low-dose rapamycin. Combining injections of iBMDCs and LF 15-0195 showed a striking synergistic effect and induced definitive allograft acceptance in 92% of recipients. Tolerant recipients accepted donor-type, but not third-party type skin grafts, suggesting the development of regulatory mechanisms capable of maintaining donor-specific tolerance. The reported findings may contribute to the development of new therapeutic strategies to induce transplantation tolerance in humans.     

PTEN permits acute increases in D3-phosphoinositide levels following TCR stimulation but inhibits distal signaling events by reducing the basal activity of Akt

Phosphoinositide 3-kinase (PI3K) is important in TCR signaling. PI3K generates phosphatidylinositol 3, 4, 5-trisphosphate (PI-3,4,5-P3), which regulates membrane localization and/or activity of multiple signaling proteins. PTEN (phosphatase and tensin homologue deleted on chromosome 10) opposes PI3K, reversing this reaction. Maintaining the balance between these two enzymes is important for normal T cell function. Here we use the PTEN-null Jurkat T cell line to address the role of PTEN in modulating proximal and distal TCR-signaling events. PTEN expression at levels that restored low basal Akt phosphorylation (an indicator of PI-3,4,5-P3 levels), but which were not themselves cytotoxic, had minimal effect on TCR-stimulated activation of phospholipase Cgamma1 and Ca2+ flux, but reduced the duration of extracellular signal-regulated kinase (Erk) activation. Distal signaling events, including nuclear factor of activated T cells (NFAT) activation, CD69 expression and IL-2 production, were all inhibited by PTEN expression. Notably, PTEN did not block TCR-stimulated PI-3,4,5-P3 accumulation. The effect of PTEN on distal TCR signaling events was strongly correlated with the loss of the constitutive Akt activation and glycogen synthase kinase-3 (GSK3) inhibition that is typical of Jurkat cells, and could be reversed by expression of activated Akt or pharmacologic inhibition of GSK3. These results suggest that PTEN acts in T cells primarily to control basal PI-3,4,5-P3 levels, rather than opposing PI3K acutely during TCR stimulation.     

Appropriateness of a pneumococcal conjugate vaccine in Brazil: potential impact of age and clinical diagnosis,with emphasis on meningitis

The distribution of pneumococcal serotypes in Brazil was analyzed by age group and clinical diagnosis, using data obtained during 20 years of national surveillance. Serotypes 1 and 5 remained among the main serotypes in all age groups, increasing in frequency with age. Serotype 14 was prevalent among children, whereas serotypes 3 and 4 were most prevalent among the adult population. The potential impact of the 7- and 9-valent conjugate vaccines on children up to age 5 years with severe pneumococcal diseases was 58.2% and 73%, respectively; the highest coverage of the 7-valent vaccine for pneumonia was achieved for children aged 7 months to 2 years (70%), whereas, for meningitis, it was observed for children aged 7 months to 5 years (58.6%). The use of conjugate vaccine may be of potential benefit by reducing the childhood sequelae and mortality of pneumococcal infection in Brazil.     

The Ocular Hypertension Treatment Study: reproducibility of cup/disk ratio measurements over time at an optic disc reading center.

PURPOSE: To determine the reproducibility over time of visual estimates of the horizontal cup/disk ratio by trained technicians from optic disk stereophotographs. METHODS: Baseline optic disk stereophotographs are graded at entry and regraded annually in a masked fashion. The 1,636 participants in the Ocular Hypertension Treatment Study (OHTS) undergo stereoscopic optic disk photography at study entry and annually thereafter. Stereophotographs are graded independently by two technicians at the Optic Disc Reading Center. If the readers' estimates of horizontal cup/disk ratio differ by more than 0.2 disk diameters (DD), they attempt to reach a consensus; if they cannot, the horizontal cup/disk ratio is adjudicated by a glaucoma specialist. RESULTS: The percent of regradings differing by 0.2 DD or more from the estimate of horizontal cup/disk ratio made at entry was 4%, 6%, and 7%, respectively at years 1, 2, and 3. The percent differing by more than 0.2 DD was 1% or less at all years. Intraclass correlation coefficients were 0.93, 0.92, and 0.92, respectively. Estimates of horizontal cup/disk ratio from sequential full-frame photographs and simultaneous split-frame photographs appeared comparable and equally reproducible. Gradings by technicians were comparable to gradings by glaucoma specialists. CONCLUSIONS: High reproducibility between repeated gradings of baseline horizontal cup/disk ratio was achieved by trained technicians adhering to a rigorous protocol. Horizontal cup/disk ratio measurements in OHTS are sufficiently reproducible to provide information about the relationship of cup/disk ratio to the prognosis of individuals with ocular hypertension.