|Job Title||Scientific Officer|
|Date of Scholarship||7th May to 25th June 2012|
|Host Country||London School of Hygiene and Tropical Medicine (LSTHM), London, UK|
|Scholarship Title||Optimisation of immunological and molecular assays for activated NK responses in HIV infection|
My work-visit training attachment was at the Department of Clinical Research and Immunology of Infectious Diseases Laboratory, London School of Hygiene and Tropical Medicine (LSHTM) in UK. The aim of my work was to learn techniques that will define the immune and molecular characteristics of activated NK cells from blood samples of HIV infected individuals.
At the molecular biology laboratory Unit, I was trained on: 1) DNA extraction from buccal brushes samples, using a manual method; 2) DNA extraction from whole blood and PBMCs using Qiagen kit; 3) Whole genome amplification; 4) KIR gene amplification; 5) Separation of nucleic acid fragments from PCR product using agarose gel electrophoresis. I then used these methods to carry out KIR typing with 16-PCR-SSP which detects the presence or absence of KIR genes in sixteen separate multiplex PCR reactions. I was taught how to interpret the results of such genotyping.
At the immunology laboratory I was trained on: 1) PBMC isolation and stimulation methods using specific antigens; 2) characterization of NK phenotypes and NK functional assays with multiparametric flow cytometry; 3) optimisation of immunological methods includes i) NK cell staining on fixed and frozen stimulated PBMCs, ii) two-step-staining versus one-step-staining, iii) extracellular staining before and after cytofixation, and iiii) permeabilisation using BD fix/perm kit versus PFA/saponin
This HIV Research Trust Scholarship has allowed me to gain new techniques that I am able to currently apply in an EDCTP-funded work that seek to assess the relevance of activated NK responses in individuals who received candidate HIV vaccines in a phase I clinical trial in West Africa. I am a lead technical support in this project which is part of a West Africa collaborative study that involved a regional HIV network between Senegal, Gambia and Guinea Bissau. As the Senior Scientific Officer, I am applying the techniques also in the immunology work in HIV sero-discordant couples and HIV sex-workers cohort. I was proud to receive a prize award for one of the best 5 abstracts presented at the Africa HIV meeting, ICASA October 2013 in Cape Town, South Africa, which was based on the above described work. These data will help my plan to write a post-doctoral fellowship grant on cellular immune responses in HIV and HIV-coinfections in order to further my research career.
The knowledge and skills I gained has strengthened the capacity for Immunological analyses at the Immunology Unit of my Institution, Laboaratory of Bacteriology and Virology of Cheikh Anta Diop University (LBV_UCAD) in Senegal. LBV-UCAD has recently acquired a 9-colour Cyan Flow Cytometer and I am able to train key staff and users as well as manage the upkeep of the equipment for optimal use in our research programs.
The HIV Trust training opportunity has complemented the training agenda of our regional HIV research consortium to build critical mass of research leadership. This network, called West Africa Platform for HIV Intervention Research (WAPHIR), was created in 2010 by my lead supervisors and supported by the Canadian Global Health Research Initiative (GHRI) in order to harness our regional HIV cohort and database resources for collaborative research and clinical trials. I am now able to integrate my technical applications and research ideas within the joint studies between LBV-UCAD of Senegal, MRC Unit of the Gambia and INSA-BHP of Guinea Bissau.