Preliminary results of the project A.I.D.A. (Auto Immunity: Diagnosis Assisted by computer)

In this paper, are presented the preliminary results of the A.I.D.A. (Auto Immunity: Diagnosis Assisted by computer) project which is developed in the frame of the cross-border cooperation Italy-Tunisia. According to the main objectives of this project, a database of interpreted Indirect ImmunoFluorescence (IIF) images on HEp 2 cells is being collected thanks to the contribution of Italian and Tunisian experts involved in routine diagnosis of autoimmune diseases. Through exchanging images and double reporting; a Gold Standard database, containing around 1000 double reported IIF images with different patterns including negative tests, has been settled. This Gold Standard database has been used for optimization of a computing solution (CADComputer Aided Detection) and for assessment of its added value in order to be used along with an immunologist as a second reader in detection of auto antibodies for autoimmune disease diagnosis. From the preliminary results obtained, the CAD appeared more powerful than junior immunologists used as second readers and may significantly improve their efficacy

Computer-Assisted Classification Patterns in Autoimmune Diagnostics: The AIDA Project

Antinuclear antibodies (ANAs) are significant biomarkers in the diagnosis of autoimmune diseases in humans, done by mean of Indirect ImmunoFluorescence (IIF)method, and performed by analyzing patterns and fluorescence intensity. This paper introduces the AIDA Project (autoimmunity: diagnosis assisted by computer) developed in the framework of an Italy-Tunisia cross-border cooperation and its preliminary results. A database of interpreted IIF images is being collected through the exchange of images and double reporting and a Gold Standard database, containing around 1000 double reported images, has been settled. The Gold Standard database is used for optimization of aCAD(Computer AidedDetection) solution and for the assessment of its added value, in order to be applied along with an Immunologist as a second Reader in detection of autoantibodies. This CAD system is able to identify on IIF images the fluorescence intensity and the fluorescence pattern. Preliminary results show that CAD, used as second Reader, appeared to perform better than Junior Immunologists and hence may significantly improve their efficacy; compared with two Junior Immunologists, the CAD system showed higher Intensity Accuracy (85,5% versus 66,0% and 66,0%), higher Patterns Accuracy (79,3% versus 48,0% and 66,2%), and higher Mean Class Accuracy (79,4% versus 56,7% and 64.2%)

Long-term pertussis-specific immune responses to a combined diphtheria, tetanus, tricomponent acellular pertussis and hepatitis B vaccine in pre-term infants

Immunoglobulin G (IgG) antibody titres against pertussis antigens, Bordetella pertussis-specific proliferation and cytokine production by peripheral blood mononuclear cells (PBMCs) were evaluated at the age of 5-6 years in 13 children who had been pre-term infants with a gestational age (GA) of < or = 31 weeks, 10 who had been pre-term infants with a GA of 32-37 weeks, and 15 who had been term infants with a GA of 38-42 weeks. All of the infants had been immunised with a combined diphtheria, tetanus, tricomponent acellular pertussis and hepatitis B vaccine (DTaP-HBV) at 3, 5, and 11 months of post-natal age. Our results show that the long-term immune responses induced by primary pertussis vaccination in the pre-term infants (especially those with a GA of < or = 31 weeks) were qualitatively and quantitatively lower than those observed in the term infants. In order to maintain an adequate pertussis-specific immune response, in term children a booster at 5-6 years of age may be suggested, whereas in pre-term an earlier booster should be useful

Differential regulation of Th1 and Th2 cells by p91–110 and p21–40 peptides of the 16-kD α-crystallin antigen of Mycobacterium tuberculosis

Permissively recognized peptides which can activate lymphocytes from subjects with a variety of class II HLA types are interesting diagnostic and vaccine candidates. In this study we generated T helper clones reactive to the permissively recognized p21–40 and p91–110 peptides of the 16-kD heat shock protein of Mycobacterium tuberculosis. All the clones specific for p91–110 secreted interferon-gamma (IFN-γ) and were of the Th1 phenotype. By contrast, the p21–40 peptide favoured the generation of IL-4-producing clones. Antibody blockade established that the peptide-specific Th clones could either be DR-, DP- or DQ-restricted. Thus, two permissively recognized sequences p21–40 and p91–110 from the same mycobacterial antigen can drive the differentiation of functionally distinct T helper subsets. Attempts to immunize against tuberculosis should bear in mind epitope specificity if a favourable Th subtype response is to be generated

T-Cell Recognition of Mycobacterial GroES Peptides in Thai Leprosy Patients and Contacts

We report here the mapping of T-cell-stimulatory determinants of the GroES 10-kDa heat shock protein homologues from Mycobacterium leprae and Mycobacterium tuberculosis, which are known as major immunogens in mycobacterial infections. Peripheral blood mononuclear cells (PBMC) from treated tuberculoid leprosy or lepromatous leprosy patients and from healthy household or hospital staff contacts of the patients were cultured with 20 16-mer peptides covering the entire sequences of both M. leprae and M. tuberculosis GroES. The total number of recognized peptides was found to be the largest in family contacts, while responder frequencies to the individual tested peptides varied (5 to 80%) with specificity between the patient and contact groups. Proliferative responses to some peptides showed positive or negative associations of low statistical significance with DR and DQ alleles, though responses to most GroES peptides were genetically permissive. Notably, the sequence of the 25–40 peptide of M. leprae, but not that of M. tuberculosis, was more frequently stimulatory in tuberculoid leprosy patients than in either group of sensitized healthy contacts. This peptide bound to a number of HLA-DR molecules, of which HLA-DRB5*0101 had the strongest affinity. The epitope core binding to this allele was localized to the 29-to-37 sequence, and its key residue was localized to the M. leprae-specific glutamic acid at position 32. This epitope may be of interest for the development of a blood test- or skin test-based diagnostic reagent for tuberculoid leprosy, subject to further clinical evaluation in untreated patients