Work

Teaching advanced flow cytometry in Africa: 10 years of lessons learned

Year: 2016

Type: article

Abstract: Africa is particularly burdened by major infectious diseases and there are many groups throughout the continent whose research focuses on understanding cellular immunity to these pathogens1. There is thus a continuous need for improving and further developing scientific skills in different regions that can enable the design, conduct and interpretation of data that support the immune monitoring of clinical research studies2. To enhance technical and theoretical knowledge of African immunologists and in response to the paucity of training opportunities on the continent, the first Intracellular Cytoakine Staining and Flow Cytometry Workshop was held in 2005 in South Africa. Since then the workshop has been held biennially, with the aim of training a new generation of African researchers so that cutting edge technology can be applied and human capital can be perpetuated locally by peer-training. We deemed it appropriate to relay our experiences to the international cytometry community and to promote discussion about access to flow cytometry training with those involved in similar educational efforts on the African continent. Applicants based in Africa were selected for workshop attendance through a peer-review process based on: (I) the scientific merit of each submitted abstract; (II) flow cytometry experience; (III) training needs (at both individual and institutional levels); and (IV) potential impact on career advancement. All selected participants received a full scholarship that covered all expenses, international and local transport, accommodation and meals. Funding was secured from a range of sources including educational grants, academic funding, private foundations and vendors (reported under Acknowledgements section). Participant selection was highly competitive due to limited funding and the practical nature of the interactive workshop. On average, fewer than half of the applicants were selected (e.g., 28% in 2015), indicating the high demand for advanced flow cytometry training in Africa. Over the six workshops held between 2005–2015, 106 scientists working in 22 African countries attended (Fig. 1A), an average of 18 participants (range: 14–22) per workshop. Participants included junior faculty/postdoctoral fellows, postgraduate students, as well as laboratory managers and senior personnel. Such participant diversity translated into broad training demands specifically including optimization of instrument performance, appropriate design of new studies, analysis of existing data-sets and training/supervision of others. (A) Geographical distribution of institutional affiliations of flow cytometry workshop participants (n = 106 from 2005 to 2015). (B) Participant learning was evaluated by answering the same questions (n = 14) at the beginning (Day 1) and end (Day 5) of the workshop programme (Wilcoxon signed rank test). A fold change above 1 (dotted line) indicates improvement in questionnaire score (representative data are from the 2015 workshop). (C) Proportion of faculty members with African affiliation for each workshop. The 5-day flow cytometry workshop was designed to address such needs through a combination of lectures and hands-on tutorials focusing on the measurement of immune responses using multiparameter flow cytometry 3. The course program increased in complexity over time to keep apace with technology advances. In 2015 the workshop addressed: (I) basic principles of flow cytometry; instrument set-up, optimization and quality control 4; (II) panel design, antibody titrations and compensation 5, 6; (III) experiment design and optimization 7, 8; (IV) data visualization, analysis, interpretation and reporting 9-11; (V) controls, data validation and troubleshooting 12; and (VI) principles of assay standardization and validation for clinical trials according to Good Clinical Laboratory Practice guidelines 13, 14 (Supporting Information tables 1 and 2). In these workshops, we progressed from demonstrating principles for 4 to >6 parameter instruments in the latest workshops. Hands-on tutorials initially included sample processing, but due to time limitations and broadening of the course scope, have recently been limited to those perceived as the most difficult and critical topics: instrument optimization and quality control, sample acquisition and data analysis. Most participants were extremely appreciative about the hands-on data analysis sessions, conducted using FlowJo software (FlowJo, LLC, Ashland Oregon), which is available free-of-charge to all scientists based in Africa. Learning was evaluated by initial and final knowledge assessments (Supporting Information 3). Moreover, daily homework was assigned and discussed the following day during highly interactive sessions. Participants left the workshop with a better knowledge of current techniques. For example, an average two-fold increase in the scores achieved at workshop completion was observed when compared to the pre-workshop assessment in 2015 (Fig. 1B). To ensure that the content and format of the workshop suited participants' needs over the years, anonymous feedback provided by each attendee at the end of the workshop was heeded to tailor and adjust the program of the next workshop, thus ensuring continuous improvement. Overall, the trainees expressed overwhelming enthusiasm, exemplified by this comment: “It will help our whole team to improve the quality of our data, [and] prepare me for my dream, a PhD.” As this workshop was designed for more advanced flow cytometry users, some participants with limited experience regarded the program as too concentrated and suggested stretching the course over a longer period of time. This has not been possible so far due to resource limitations. English was the official language adopted for the workshops, and some participants coming from French-speaking countries perceived this as a barrier. French-speaking faculty members could assist when needed, but it's clear that many flow cytometry users would benefit from regional workshops conducted in languages other than English. The course also provided a network of regional and international contacts, teaching materials and technical resources for participants to return home with and share with their peers. Post-training networking and resource sharing tools have been limited to date. We are working towards a more structured approach, building on existing platforms such as the Immunopaedia internet portal (immunopaedia.org.za). From the start scientists within and outside Africa contributed to the organizing committee and faculty team. However, greater representation of faculty members affiliated to African institutions in 2013 and 2015 bears testament to the sustainable impact of the workshop (Fig. 1C). Notably, most of the recent African faculty members had attended the workshop as participants in the past. The longer-term impact of the workshop was recently evaluated by a comprehensive survey, conducted in May/June 2016 (Supporting Information 4). Sixty-five participants who attended workshops from 2009 onwards were contacted via e-mail. Among the 34 respondents (52%), all were still working in Africa, 88% reported that attending the workshop had been important to achieve their career goals and thought the workshop provided good networking opportunities. All interviewees would recommend the workshop to their peers and expressed interest in further training through e-learning. Almost 90% of the respondents had trained peers at their home institution after the workshop (Fig. 2A), often adapting teaching materials shared during the workshop (Fig. 2B). Importantly, the majority of respondents reported paucity of local training opportunities (Fig. 2C) and that their home research organizations could not afford the cost for such training (Fig. 2D). We believe that “training the trainers” is the most effective and empowering approach, especially since access to advanced flow cytometry training on the African continent has improved only modestly over the past 10 years, according to respondents and our own observations. Results from the 2016 survey. The survey was submitted to 65 participants who attended one of the workshops held between 2009 and 2015, and was completed by 34 participants (52%). Impact of the flow cytometry workshop was assessed in terms of capacity building: (A) spread of knowledge among peers and (B) utility of training materials provided. (C) Availability and (D) affordability of alternative flow cytometry training opportunities were also evaluated. In light of the increasing needs and efforts to conduct clinical research in Africa, we believe that these workshops serve not only to enhance local African scientific capacity, but also contribute to the global health agenda. In 2016 only 10 of 1,725 (0.6%) ISAC members held African affiliations (Michelle Butler, ISAC Executive Director, personal communication). There is a clear opportunity for the Society to expand and partner with African organizations and initiatives to further its reach on the African continent, to meaningfully contribute to capacity building and to broaden the diversity of its members. In conclusion, there is further need for regional face-to-face flow cytometry training initiatives, ideally tailored to the participants' experience and needs, and offered free-of-charge. Some of this unmet need for flow cytometry training in Africa could be fulfilled by open-access e-learning-based courses, such as those offered by CYTOA University (cytoau.org). The organizers thank all participants who attended the flow cytometry workshops, and in particular those who contributed to the 2016 survey. Faculty 2005–2015: John Altman, Erica Andersen-Nissen, Omu Anzala, Michael Betts, Marty Bigos, Gaelle Breton, Wendy Burgers, Stephen De Rosa, One Dintwe, Helene Dujardin, Michael Eller, Guido Ferrari, Deborah Glencross, Clive Gray, Willem Hanekom, Alexandre Harari, Jane Hughes, Kondwani Jambo, Claire Landry, Holden Maecker, John Mantovani, Agatha Masemola, Elisa Nemes, Janet Ottinger, Stephen Perfetto, John Quinn, Catherine Riou, Mario Roederer, Thomas Scriba, Rafick-Pierre Sekaly, Erica Smit, Andreia Soares, Sara Suliman, Kayla Treister, Claudio Vallan. Logistical and Administrative Support: Dhuraiyah Abdullatief, Carina Kriel, Kathryn Norman. Funders and Sponsors: African AIDS Vaccine Programme (AAVP)-World Health Organisation (WHO), Becton Dickinson (BD), Beckman Coulter, Biocom Biotech (South Africa), Canada-Africa Prevention Trials Network (CAPT), Center for AIDS Research (CFAR, Duke University Medical Center), Clinical Infectious Diseases Research Initiative (CIDRI, South Africa), Global Health Research Initiative (GHRI, Canada), HIV Vaccine Trial Network (HVTN, US), Institute of Infectious Disease and Molecular Medicine (IDM, South Africa), International Union of Immunological Societies (IUIS), National Institute for Communicable Diseases (NICD, South Africa), National Institute of Allergy and Infectious Disease (NIAID) and Office of AIDS Research (OAR), National Institute of Health (NIH, US), South African Immunology Society (SAIS), Scientific Group (South Africa), TreeStar/FlowJo, United Nations Programme on HIV/AIDS (UNAIDS). Additional Supporting Information may be found in the online version of this article. Supporting Information Supporting Information Supporting Information Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Source: Cytometry Part A

DOI: 10.1002/cyto.a.23015