CD4+T cells from HIV-1-infected patients recognize wild-type and mutant human immunodeficiency virus-1 protease epitopes

P>Human immunodeficiency virus (HIV)-1 protease is a known target of CD8+ T cell responses, but it is the only HIV-1 protein in which no fully characterized HIV-1 protease CD4 epitopes have been identified to date. We investigated the recognition of HIV-1 protease by CD4+ T cells from 75 HIV-1-infected, protease inhibitor (PI)-treated patients, using the 5,6-carboxyfluorescein diacetate succinimidyl ester-based proliferation assay. In order to identify putative promiscuous CD4+ T cell epitopes, we used the TEPITOPE algorithm to scan the sequence of the HXB2 HIV-1 protease. Protease regions 4-23, 45-64 and 73-95 were identified; 32 sequence variants of the mentioned regions, encoding frequent PI-induced mutations and polymorphisms, were also tested. On average, each peptide bound to five of 15 tested common human leucocyte antigen D-related (HLA-DR) molecules. More than 80% of the patients displayed CD4+ as well as CD8+ T cell recognition of at least one of the protease peptides. All 35 peptides were recognized. The response was not associated with particular HLA-DR or -DQ alleles. Our results thus indicate that protease is a frequent target of CD4+ along with CD8+ proliferative T cell responses by the majority of HIV-1-infected patients under PI therapy. The frequent finding of matching CD4+ and CD8+ T cell responses to the same peptides may indicate that CD4+ T cells provide cognate T cell help for the maintenance of long-living protease-specific functional CD8+ T cells.

Test-retest reliability of thermal quantitative sensory testing on two sites within the L5 dermatome of the lumbar spine and lower extremity.

INTRODUCTION: Quantitative sensory testing (QST) is widely used in human research to investigate the integrity of the sensory function in patients with pain of neuropathic origin, or other causes such as low back pain. Reliability of QST has been evaluated on both sides of the face, hands and feet as well as on the trunk (Th3-L3). In order to apply these tests on other body-parts such as the lower lumbar spine, it is important first to establish reliability on healthy individuals. The aim of this study was to investigate intra-rater reliability of thermal QST in healthy adults, on two sites within the L5 dermatome of the lumbar spine and lower extremity. METHODS: Test-retest reliability of thermal QST was determined at the L5-level of the lumbar spine and in the same dermatome on the lower extremity in 30 healthy persons under 40 years of age. Results were analyzed using descriptive statistics and intraclass correlation coefficient (ICC). Values were compared to normative data, using Z-transformation. RESULTS: Mean intraindividual differences were small for cold and warm detection thresholds but larger for pain thresholds. ICC values showed excellent reliability for warm detection and heat pain threshold, good-to-excellent reliability for cold pain threshold and fair-to-excellent reliability for cold detection threshold. ICC had large ranges of confidence interval (95%). CONCLUSION: In healthy adults, thermal QST on the lumbar spine and lower extremity demonstrated fair-to-excellent test-retest reliability.

Co-infection by Tritrichomonas foetus and Pentatrichomonas hominis in asymptomatic cats

Abstract: Tritrichomonas foetus, a parasite well known for its significance as a venereally transmitted pathogen in cattle, has been identified as a cause of chronic large bowel diarrhea in domestic cats in many countries of the world. In Brazil, several studies on the diagnosis of bovine trichomoniasis have been performed, but until now, no study was made regarding feline trichomoniasis. Thus, this is the first study to report the occurrence of T. foetus and Pentatrichomonas hominis in cats using morphological and molecular analysis. Feces from 77 cats were examined, four of which (5.2%) were positive for the presence of parabasalids. Morphological analysis of stained smears revealed piriform trophozoites showing the three anterior flagella, elongated nucleus and axostyle ending abruptly in fillet, characteristic of T. foetus. In scanning and transmission electron microscopy, identification characters similar to those previously reported for T. foetus were observed. The cultures containing trophozoites were submitted for molecular analysis, which resulted positive for T. foetus DNA using specific primers (TFR3 and TFR4), and all samples were positive and subjected to sequencing in which they showed 99.7-100% similarity with another isolate sequencing of T. foetus (JX960422). Although no trophozoite with consistent morphology of P. hominis has been visualized in the samples, differential diagnosis was performed using specific primers for P. hominis (TH3 and TH5) amplicon. In three of the four samples (3.89%) sequencing revealed 100% similarity when compared with another sequence of P. hominis deposited in Genbank (KC623939). Therefore, the present study revealed through the diagnostic techniques employed the simultaneous infection by T. foetus and P. hominis in the feces of cats. However, it was necessary to use more than one technique for the diagnosis of the co-infection. These results demonstrate the importance of a correct diagnosis to allow an appropriate treatment by the veterinarian.

T cell-mediated immunoregulation in the gastrointestinal tract

In the intestinal tract, only a single layer of epithelial cells separates innate and adaptive immune effector cells from a vast amount of antigens. Here, the immune system faces a considerable challenge in tolerating commensal flora and dietary antigens while preventing the dissemination of potential pathogens. Failure to tightly control immune reactions may result in detrimental inflammation. In this respect, 'conventional' regulatory CD4(+) T cells, including naturally occurring and adaptive CD4(+) CD25(+) Foxp3(+) T cells, Th3 and Tr1 cells, have recently been the focus of considerable attention. However, regulatory mechanisms in the intestinal mucosa are highly complex, including adaptations of nonhaematopoietic cells and innate immune cells as well as the presence of unconventional T cells with regulatory properties such as resident TCRgammadelta or TCRalphabeta CD8(+) intraepithelial lymphocytes. This review aims to summarize the currently available knowledge on conventional and unconventional regulatory T cell subsets (Tregs), with special emphasis on clinical data and the potential role or malfunctioning of Tregs in four major human gastrointestinal diseases, i.e. inflammatory bowel diseases, coeliac disease, food allergy and colorectal cancer. We conclude that the clinical data confirms some but not all of the findings derived from experimental animal models.

Comparative study of vertebral fractures and luxations in dogs and cats

The purpose of this retrospective study was to compare patterns of vertebral fractures and luxations in 42 cats and 47 dogs, and to evaluate the impact of species-related differences on clinical outcome. Data regarding aetiology, neurological status, radiographic appearance and follow-up were compared between the groups. The thoracolumbar (Th3-L3) area was the most commonly affected location in both cats (49%) and dogs (58%). No lesions were observed in the cervical vertebral segments in cats, and none of the cats showed any signs of a Schiff-Sherrington syndrome. Vertebral luxations were significantly more frequent in dogs (20%) than in cats (6%), whereas combined fracture-luxations occurred significantly more often in cats (65%) than in dogs (37%). Caudal vertebral segment displacement was mostly dorsal in cats and ventral in dogs, with a significant difference in direction between cats and large dogs. The clinical outcome did not differ significantly between the two populations, and was poor in most cases (cats: 61%; dogs: 56%). The degree of dislocation and axis deviation were both significantly associated with a worse outcome in dogs, but not in cats. Although several differences in vertebral fractures and luxation patterns exist between cats and dogs, these generally do not seem to affect outcome.