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*Square Project: V.M. Agate (Palmanova, UD), A.M. Anelli (Pistoia), L. Apicella (Salerno), D. Avino (Vairano Scalo, CE), Y. Battaglia (Ferrara), P.L. Bedani (Ferrara), G. Bellinghieri (Messina), A.M. Bernardi (Rovigo), C. Bonifati (Corato, BA), M. Borzumati (Verbania), P.L. Botti (Mantova), M. Brigante (Campobasso), F. Brighina (San Nicola La Strada, CE), P. Calzavara (Conegliano, TV), C. Caputo (Albenga, SV), F. Cardone (Lavello, PZ), F. Cavatorta (Imperia), N. Confessore (Scafati, SA), M. Cossu (Sassari), E. Costantino (Gavardo, BS), G. Costantino (Messina), L. D’Apice (Caserta), R. D’Arcangelo (Arzano, NA), F. Dagostino (Corato, BA), A. Dal Canton (Pavia), G. Delgado (Teano, CE), R. Di Pietro (Napoli), C. Esposito (Pavia), P. Esposito (Pavia), E. Fasianos (Altamura, BA), F. Fiorini (Rovigo), P. Galeotti (Viterbo), G. Garibotto (Genova), A. Gemelli (Rovigo), M.C. Gregorini (Reggio Emilia), P. Iuliano (Benevento), L. La Peccerella (Benevento), M. Liuzzi (Benevento), M. Merola (Sorrento, NA), C. Montesano (Imperia), L.F. Morrone (Benevento), M. Napoli (Galatina, LE), F. Napolitano (Corato, BA), S. Paglia (Lavello, PZ), M. Parravano (Sora, FR), S. Pasquali (Reggio Emilia), A. Rosa (Benevento), M.L. Sambati (Taranto), P. Schiavone (Brindisi), E. Sozzo (Galatina, LE), A. Storari (Ferrara), R. Tarchini (Mantova), G. Tirino (Montesarchio, BN), L. Turchetta (Sora, FR). School Project: G. Bellinghieri (Messina), G. Beltrame (Torino), M. Bozzi (Bari), E. Casolino (Rionero in Vulture, PZ), E. Centrone (Ruvo di Puglia, BA), M. Ciccarelli (Reggio Calabria), T. Cicchetti (Rossano, CS), C. Colturi (Sondrio), E. Costantino (Gavardo, BS), G. Costantino (Messina), F. Dagostino (Corato, BA), E. Fasianos (Altamura, BA), P. Galeotti (Viterbo), Garibotto (Genova), F. Indraccolo (S. Fermo della Battaglia, CO), M. Lombardi (Borgo S. Lorenzo, FI), C. Minoretti (S. Fermo della Battaglia, CO), F. Napolitano (Corato, BA), R. Parsi (Alcamo, TP), F. Petrarulo (Bari), E. Prati (Desenzano del Garda, BS), F. Quarello (Torino), V. Rondanini (Palmi, RC), D. Russo (Barletta), P. Tira (Manerbio, BS), S. Turina (Manerbio, BS), W.D. Valentini (Rieti) WORLD KIDNEY DAY

BACKGROUND: Human immunodeficiency virus (HIV)- specific CD8-positive cytotoxic T-lymphocytes (CTL) play a key role in controlling HIV infection. Monitoring CTL response could be clinically relevant during structured therapy interruption (STI), HIV exposure, and vaccine trials. However, HLA patients' restriction and HIV variability limited the development of a CTL assay with broad specificity. MATERIALS AND METHODS: We designed an HLA-class I/HIV-1 clade independent assay for assessing HIV- specific CTL by using a computer-assisted selection ofthe CTL epitopes. Twenty-eight 15-mers were selected by peptide-binding motifs analysis using different databases (HIV-Immunology Database, SYFPEITHI, BIMAS). Altogether they putatively bind to more than 90% of HLA haplotypes in different populations, with an overall HIV-1 variability below 9%. The peptide pool was used as an antigen in an intracellular cytokine staining (ICS) assay for quantifying HIV-specific CTL response. RESULTS: The test can be performed using both fresh and cryopreserved peripheral blood mononuclear cells (PBMC), whereas GAG protein as antigen works only on fresh PBMC. A significantly higher CTL response with respect to HIV-negative controls was detected in all HIV-1 infected subjects of two groups of patients with different ethnicities (Caucasians and Africans) and coming from areas with different HIV-1 clade prevalences (clade B and A/G, respectively). In Caucasian patients, after month of STI, the number of HIV-1 specific CTL (2,896 +/- 2,780 IFN-gamma specific CD8 cells/ml) was significantly higher than that found at enrolment (2,125 +/- 4,426 IFN-gamma specific CD8 cells/ml, p< 0.05). CONCLUSIONS: These data indicate that this CTL assay is broadly specific and could represent a useful clinical tool for HIV immunodiagnostic independent of HLA-haplotype and HIV-clade variabilities.

The pandemic situation the world is facing caused by the new SARS-Cov-2 continues to evolve and still represent a real problem. With more than eight thousand reported cases infection, Cameroon stands as the seventh most affected country in Africa. Prevention remains the best way to fight against this zoonosis. However, the limited information available about this infection is a great barrier to stopping the propagation of the virus within the population, especially in rural and semi-rural areas, where the lack of financial and material resources is a reality. This study aimed to assessing Awareness and attitudes of the population of the Menoua Division on COVID-19 infection.A cross-sectional study was conducted from March 9 to April 15 2020 amongst the populations of rural and semi-rural areas of the Menoua Division. Data were collected using a questionnaire administered face to face to each participant. The analysis was carried out using the Statistical Analysis System software (SAS version 9.4). The significance threshold was set at a P value of less than 0.05.A total of 434 participants of which male majority (sex ratio 1.07) were included in this study. The most represented age group was [21 - 40] years old representing 40.29% of the participants. Approximately all participants (98.57%) were aware of the world emergency state due to Coronavirus. 75.56%, 91% and 90.93% of the participants knew respectively that having close contacts, kissing and touching the face with the hands could favor the transmission of the virus. However, nearly 91.14% were not aware of the clinical symptoms of the disease. Moreover, 85.02% responded that they would not be able to comply with the confinement measures if they were applied at national level. The level of awareness varied significantly according to the occupation (p=0.038) and the educational level (plt;0.001)of the participants.The average level of awareness of the population of the Menoua Division on COVID-19 infection was relatively low. Overcoming this pandemic disease means ensuring the flow of the correct information towards the population. Community outreach activities focus on clinical manifestations and what to do in case of COVID-19 infection as well as material and financial support should be help the population to protect themselves effectively against pandemic, particularly in rural areas and surrounding.

A new strategy that takes advantage of the synergism between NMR and LC-HRMS (SYNHMET), and that allows to obtain a unique list of the absolute concentrations of 164 metabolites in urine, is presented. Metabolite identification and quantification by this method in one of the most difficult biofluids to characterize, due to complexity and variability, is more accurate than what can be obtained using the two techniques separately. This result is achieved without the need for chemical reactions to cross-check the data between the two types of spectra, nor the use of analytical standards and calibration curves. The fact that the absolute rather than relative concentration is obtained allows the final dataset to be used to determine a patient's personalized profile. The number of quantifiable metabolites by the application of this method can be expanded in the future with further analysis. We will illustrate the use of SYNHMET in the study of urine samples from healthy subjects, patients with chronic cystitis and bladder cancer.

Objective During HIV infection, a down-modulation of CD3ζ was found on T cells, contributing to T-cell anergy. In this work, we studied the correlation between myeloid-derived suppressor cells (MDSC) frequency and T-cell CD3ζ expression. Moreover, we investigated the mechanisms of CD3ζ decrease exploited by MDSC. Design and method CD3ζ expression and MDSC frequency were evaluated by flow cytometry on peripheral blood mononuclear cells from 105 HIV-positive (HIV+) patients. The role of MDSC in the modulation of the HIV-specific T-cell response was evaluated. The level of CD3ζ mRNA and ELF-1 protein were analysed by real-time-PCR and western blot, respectively. Results We found that granulocytic-MDSC (Gr-MDSC) were expanded in HIV+ patients compared with healthy donors; in particular, in cART-treated individuals a higher Gr-MDSC frequency was observed in patients with a CD4 T-cell count below 400 cells/μl. We found an inverse correlation between the percentage of Gr-MDSC and CD3ζ level. Moreover, in-vitro MDSC depletion induced the up-regulation of CD3ζ in T cells, restoring the functionality of αβ, but not γδ T cells. The in-vitro effect of isolated MDSC on CD3ζ expression was found cell contact-dependent, and was not mediated by previously described molecules. CD3ζ down-modulation corresponds to the decrease of its mRNA induced by silencing the transcription factor ELF-1. Conclusion Our data provide new knowledge on mechanisms used by Gr-MDSC in immune-modulation and on their role in the immune reconstitution during antiviral treatments.

Antigen-specific γδ T cells represent an early innate defense known to play an important role in anti-mycobacterial immunity. We have investigated the immune functions of Vγ9Vδ2 T cells from Broncho-Alveolar lavages (BAC) samples of active TB patients. We observed that BAC Vγ9Vδ2 T cells presented a strong down-modulation of CD3 expression compared with Vγ9Vδ2 T cells from peripheral blood. Furthermore, Vγ9Vδ2 T cells mainly showed a central memory phenotype, expressed high levels of NK inhibitory receptors and TEMRA cells showed low expression of CD16 compared to circulating Vγ9Vδ2 T cells. Interestingly, the ability of BAC Vγ9Vδ2 T cells to respond to antigen stimulation was dramatically reduced, differently from blood counterpart. These observations indicate that γδ T cell functions are specifically impaired in situ by active TB, suggesting that the alveolar ambient during tuberculosis may affect resident γδ T cells in comparison to circulating cells. © 2013 Elsevier Inc.