Horizon 2020 – The Framework Programme for Research and Innovation Projects

Personalized in Vtro tests to detect immune responses to infectious agents – Acronym PERVIMMUNE

Horizon 2020 – The Framework Programme for Research and Innovation

Participant
Participant organisation name
Country
Gennaro De Libero: co-ordinator, immunology of infectious diseasesUniversity Hospital BaselSwitzerland
Roberto Nisini: Scientific partner; detection of immune response in patients with infectious diseasesIstituto Superiore di Sanità, RomeItaly
Peter Seeberger: Scientific partner; synthesis of oligosaccharides and lipidsMax Planck Institute of Colloids and Interfaces, BerlinGermany
Giovanni Delogu: Clinical partner; diagnosis of tuberculosis and other infectionsUniversitá Cattolica del Sacro Cuore, RomeItaly
Riccardo Manganelli: Scientific partner; recombinant bacterial proteinsUniversity of PadovaItaly
Amit Singhal: Scientific partner; development of POC devicesSingapore Immunology Network, A-STAR, SingaporeSingapore
Mircea I. Popa: Clinical partner; tuberculosis diagnosisIPNEU “Carol Davila” and ”Marius Nasta” National Institute for Pneumophtysiology, BucarestRomania
Beate Kampmann: Clinical partner; pediatric tuberculosis diagnosisImperial College LondonUK
Utkan Demirci: Scientific partner; expertise in development of POC devicesStanford UniversityUSA
Immunotools. New generation ELISASMEGermany
Koek Biotech. Microfluidic devicesSMETurkey
Mamadou DafféCNRS, ToulouseFrance
 

Summary

An important unmet medical need in current clinical practice is the use of appropriate diagnostic tools to implement early identification of diseases, proper stratification of patients and the possibility to take fast therapeutic decisions. Innovative diagnostic tools should utilize reliable technologies, be based on robust biomarkers and also possibly rely on novel physio-pathological concepts capable of improving final clinical decisions.

A diagnostic device has also to provide highly sensitive and disease-specific information, and if it works with novel technologies, these must be highly reproducible, low in cost, applicable in every clinical setting and should be effective at point-of-care (POC).

PERVIMMUNE proposes the development of a device allowing novel in vitro tests, with the aim of using them as diagnostic tools for infectious diseases.

In preliminary work we have generated a prototype chip that uses nanobeads and that is capable of performing enzyme-linked immunoassays (ELISA) to detect anti-tuberculosis responses in clinical samples in a short time (15 minutes). Other important features of our prototype are its sensitivity (already in the same range as classical plate ELISA assays and better than the currently available commercial test for TB), specificity (no crossreactivity has been found with different protein and lipid antigens) and robustness (a high degree of reproducibility has been found in repeated evaluations of the same sera against the same antigens). In addition, the prototype works without the need for complex motors or optics, and all reagents are in place within the microchip, with no need for additional pipetting.

This chip can be implemented to perform T cell activation assays and antigen detection assays in addition to ELISA assays. These new applications will rely on the utilisation of the novel technologies described below.

The aims of PERVIMMUNE are to develop an affordable device using multidisciplinary scientific and technological knowledge:

  1. to detect the antibody response to microbial protein, lipid and oligosaccharide antigens;
  2. to detect T cell responses specific for microbial antigens relevant to disease diagnosis;
  3. to detect microbial products in biofluids;
  4. to validate the clinical applications of the chip;
  5. to promote the clinical use of the chip and implement sustainability of the health-care system.

The disease that will be used as clinical proof of principle is Tuberculosis (TB) because of the following important reasons. TB, caused by Mycobacterium tuberculosis (Mtb) infection, remains one of the world’s most important infectious diseases, with an estimated 9 million cases and 1.4 million deaths per year. It is a major threat to the global health, particularly due to the emergence of multiple drug-resistant (MDR) and extensively drug-resistant (XDR) TB strains. Therefore, novel and more stringent diagnostic tests and therapeutic regimens are absolutely needed. Upon infection about 5% of individuals develop an active disease, whereas most infected individuals control the bacterial growth and become latently infected. Patients with latent disease may undergo reactivation during their life and represent an immense reservoir of infecting mycobacterial bacilli.

Diagnosis of TB infection is essential not only for treatment of the infected individual but also for controlling its spread within the human population. Treatment and management of TB remains difficult due to the low specificity of clinical diagnosis and poor performance of diagnostic methods available. Currently, the routine diagnosis of active TB is based mainly on sputum microscopy (provides answer in 2-3 days, labour intensive and has modest sensitivity of 34-80%) and culture of MTB bacilli (require 2-3 weeks and has sensitivity of >95%). Recently PCR-based tests have been developed, however their costs and requirements for appropriate laboratory settings, make their use very cumbersome. An immunological-based test would be of great value if capable of restricting the number of people who required screening with more costly tests. It might also discriminate patients with active vs. latent disease, thus facilitating the therapeutic decisions to be taken. The same test might also identify individuals who may develop active disease before clinical symptoms appear. This would be useful in clinical settings where there is a predisposition to TB reactivation following the use of immunobiologics (anti-TNF-α monoclonal antibodies) or immunodeficiencies (patients with organ transplantations or HIV-infection).

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