Collection Citation: Katrina M Pollock, Calliope Dendrou 2023. An experimental medicine study of seasonal influenza vaccination responses in Lymph nodE single-cell Genomics in AnCestrY (LEGACY01). protocols.io https://dx.doi.org/10.17504/protocols.io.n92ldpw5nl5b/v1
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Abstract
This is a collection of protocols of an experimental medicine study of seasonal influenza vaccination responses in Lymph nodE single-cell Genomics in AnCestrY (LEGACY01)
This protocol was constructed using the Imperial College Research Governance and Integrity Team template (Template Ref: RGIT_TEMP_027, Template V6.0 04Nov2021). The authors wish to credit the MRC CTU at UCL for use of their Protocol Template version 8.0 in drafting of this protocol, which describes the LEGACY01 study and provides information about procedures for entering participants.Every care was taken in its drafting, but corrections or amendments may be necessary. These will be circulated to investigators in the study. Problems relating to this study should be referred, in the first instance, to the Chief Investigator.
COMPLIANCE
The study will be conducted in compliance with the approved protocol, the Declaration of Helsinki 1996, the principles of Good Clinical Practice (GCP) ICH topic E6 (R2), and revision E6 (R3) EWG, General Data Protection Regulation and the UK Data Protection Act 2018, and the UK Policy Framework for Health and Social Care Research.
PROTOCOL DEVELOPMENT TEAM
A
B
C
Name
Address
Email and telephone
Mark Coles (MC)
Kennedy Institute of Rheumatology, University of Oxford Oxford, OX3 7FY
mark.coles2@kennedy.ox.ac.uk
(44) 1865 612675
Calli Dendrou (CD)
Wellcome Centre for Human Genetics, University of
Oxford, Oxford OX3 7BN
Clinical queries should be directed to Katrina Pollock who will direct the query to the appropriate person.
SPONSOR
Imperial College London is the main research Sponsor for this study.For further information regarding the sponsorship conditions, please contact the Head of Regulatory Compliance at:
Research Governance and Integrity Team
Imperial College London and Imperial College Healthcare NHS Trust
An experimental medicine study of seasonal influenza vaccination responses in Lymph nodE single-cell Genomics in AnCestrY
ACRONYM
LEGACY01
IRAS ID
314444
SPONSOR
Imperial College London
DESIGN
Experimental medicine study; single arm, non-randomised, open label
SETTING
Secondary care (NHS) and academic research facilities
AIM
To investigate human immune responses in lymph node cells before and after immunisation with a seasonal influenza vaccine
OBJECTIVES
Primary objective: To generate a single cell atlas of lymph node cells before and after immunisation with seasonal influenza vaccine.
Secondary objective: To compare serum antibody responses before and after immunisation.
Exploratory: To compare cellular immune responses in various immune compartments e.g. blood and lymph nodes, against antigens including influenza before and after immunisation with seasonal influenza vaccine.
Exploratory: To compare immune responses in various immune
compartments (e.g., blood and lymph nodes) against antigens including influenza before and after immunisation to help inform vaccine development and testing across different ethnicities.
Capacity building and training: To build capacity with respect to staff expertise and resource between the three partner institutions to support this project and future similar research.
OUTCOME MEASURES
Outcome measures may include but are not limited to the following assays
1. Single cell RNA sequencing analysis of LNC and matched paired
PBMC
2. Binding ELISA specific for influenza/A antigens e.g.
haemagglutinin
3. Intracellular cytokine secretion or activation induced marker
assay of PBMC and LNC
4. Genotypic assays of areas of the genome of immunological
relevance may include tests such as HLA-testing.
POPULATION
Healthy adults aged 18 – 55 years n=30
Cohort 1 in influenza season 2022 to 2023
Cohort 2 in influenza season 2023 to 2024
ELIGIBILITY
Individuals with African or Asian ancestry
DURATION
Three years
FLOW DIAGRAM AND STUDY SCHEDULE
Figure 3.Timeline of study sampling and vaccination events for enrolled participants including lymph node FNA and influenza vaccination. Participants enrol at Visit 2 and undergo paired peripheral blood and lymph node sampling, vaccination at Visit 3, repeat paired lymph node sampling at Visit 4 and then phlebotomy at Visit 5.
Blood draw
Vaccine
FNA
Table 1. Schedule of investigations, treatments, and assessments
A
B
C
D
E
F
G
Study visit
V1
V2
V2a
V3
V4
V5
Visit location
Site
Site
Remote (by phone)
Site
Site
Site
Visit type
Screening
Enrolment: FNA1
Follow up
Vaccination
Follow-up: FNA2
Follow up
Study week
minus 24 to minus 1
0
0
1
2
5
Study day4
minus 168 to minus 1
0
5
7
12
35
Window (days)
NA
NA
minus 1 to plus 1
0 to plus 161
minus 2 to plus 2*
minus 2 to plus 14
Informed consent
x
Demographics
x
Medical history
x
Weight and height, calculate BMI
x
Blood borne virus screen (approx. 6 mL)1
x
Laboratory safety tests (approx. 10 mL)2
x
Concomitant medication3
x
x
x
x
x
x
COVID-19 symptoms and trigger COVID-19 test3
x
x
-
x
x
x
Urinary pregnancy test3
x
x
Symptom directed physical examination3
x
x
x
x
x
Inspection of the FNA site3
x
x
x
x
x
Vital signs3
x
x
x
x
x
Ultrasound scan3
x
x
Lymph node fine needle aspiration
x
x
Vaccination
x
Blood for serum immunoassays (6mL)3
x
x
x
x
Blood for cellular and plasma immunoassays (42mL)3
x
x
x
x
Blood for RNA PAXgene tube (2.5mL)3
x
x
Blood for HLA testing (3-4 mL)3
x
Adverse events check3
x
x
x
x
x
Blood volume (approx.) (mL)
16
52
-
50.5
50.5
48
1. Detection of antibodies and/or antigen for HIV, hepatitis B and hepatitis C
2. Full blood count, liver function, renal function, non-fasting glucose
3. At visits which include FNA or vaccination, there will be an AE check and vital signs pre-FNA/pre-vaccination, and again at least 30 min after. At visits which include FNA, there will be an inspection of the FNA site pre-FNA, and again at least 30 min after. All other procedures/assessments at these visits will be pre-FNA/pre-vaccination only.
4. The timings of V2a and V3 are set according to that of V2; the timings of V4 and V5 are set according to that of V3.
* This is the preferred window. However, if the FNA2 visit cannot be scheduled within the preferred window, it can take place up to 21 days after the vaccination without being a protocol deviation. Every effort should be made to schedule the FNA2 visit as close to 5 days post-vaccination as possible.
INDEMNITY
Imperial College London holds negligent harm and non-negligent harm insurance policies which apply to this study. Imperial College Healthcare NHS Trust holds standard NHS Hospital Indemnity and insurance cover with NHS Resolution for NHS Trusts in England, which apply to this study.
SPONSOR
Imperial College London will act as the main Sponsor for this study.Delegated responsibilities will be assigned to Imperial College Healthcare NHS Trust.
FUNDING
The Chan Zuckerberg Initiative is funding this study. Participants will be paid for each visit they complete, for their inconvenience and travel, at the end of their participation in the study, as follows:
Screening (V1): £10
Visits which include an FNA (V2 and V4): £120
Vaccination visit (V3): £80
Follow-up visit V5: £60
PATIENT AND PUBLIC INVOLVEMENT
Patient and Public Involvement and Engagement (PPIE) in research is research being carried out ‘with’ or ‘by’ members of the public rather than ‘to’, ‘about’ or ‘for’ them. The term “patient and public” includes patients, participants, carers and people who use health and social care services as well as people from specific communities and from organisations that represent people who use services.
The protocol has been reviewed and approved by the LEGACY01 PPIE committee.
PUBLICATION POLICY
The preparation of a manuscript for publication in a peer-reviewed professional journal or an abstract for presentation, oral or written, to a learned society or symposium will be discussed on the study calls and with the PPIE Advisory Group. Details of dissemination can be found in the study specific communication plan.
Authorship will reflect work done by the investigators and other personnel involved in the analysis and interpretation of the data, in accordance with generally recognised principles of scientific collaboration.
PROTOCOL AMENDMENTS
The protocol v2.0 will be the first version approved for use.
REFERENCES
Huang C, Wang Y, Li X, et al. (2020) Clinical Features of Patients Infected With 2019 Novel Coronavirus in Wuhan, China. Lancet 395:497-506.
Petersen LR, Jamieson DJ, Powers AM, Honein MA (2016) Zika virus. N Engl J Med 374:1552-1563.
WHO Ebola Response Team (2014) Ebola virus disease in West Africa—the first 9 months of the epidemic and forward projections. N Engl J Med 371:1481-1495.
Ball P (2021) The lightning-fast quest for COVID vaccines – and what it means for other diseases. Nature 589:16-18.
Christy C, Pichichero ME, Reed GF, et al. (1995) Effect of gender, race, and parental education on immunogenicity and reported reactogenicity of acellular and whole-cell pertussis vaccines. Pediatrics 96:584-587.
Kurupati R, Kossenkov A, Haut L, et al. (2016) Race-related differences in antibody responses to the inactivated influenza vaccine are linked to distinct pre-vaccination gene expression profiles in blood. Oncotarget 7:62898-628911.
Haralambieva IH, Salk HM, Lambert ND, et al. (2014) Associations between race, sex and immune response variations to rubella vaccination in two independent cohorts. Vaccine 32:1946-1953.
Sharma S, Hagbom M, Svensson L, Nordgren J (2020) The impact of human genetic polymorphisms on rotavirus susceptibility, epidemiology, and vaccine take. Viruses 12:324.
D’Souza RS, Wolfe I (2021) COVID-19 vaccines in high-risk ethnic groups. Lancet 397:1348.
Peng K, Safonova Y, Shugay M (2021) Diversity in immunogenomics: the value and the challenge. Nat Methods doi: 10.1038/s41592-021-01169-5.
Razai MS, Osama T, McKechnie DGJ, Majeed A (2021) COVID-19 vaccine hesitancy among ethnic minority groups. BMJ 372:n513.
Liang F, Lindgren G, Sandgren KJ et al. (2017) Vaccine priming is restricted to draining lymph nodes and controlled by adjuvant-mediated antigen uptake. Sci Transl Med 9:eaal2094.
Cirelli KM, Carnathan DG, Nogal B et al. (2019) Slow delivery immunization enhances HIV neutralizing antibody and germinal center responses via modulation of immunodominance. Cell 177:1153-1171.
Havenar-Daughton C, Carnathan DG, Torrents de la Peña A et al. (2016) Direct probing of germinal center responses reveals immunological features and bottlenecks for neutralizing antibody responses to HIV Env trimer. Cell Rep 17:2195-2209.
Havenar-Daughton C, Newton IG, Zare SY et al. (2020) Normal human lymph node T follicular helper cells and germinal center B cells accessed via fine needle aspirations. J. Immunol. Methods 479:112746.
Turner JS, Zhou JQ, Han J (2020) Human germinal centres engage memory and naïve B cells after influenza vaccination. Nature 586:127-132.
Corridoni D, Antanaviciute A, Gupta T, et al. (2020) Single-cell atlas of colonic CD8+ T cells in ulcerative colitis. Nat Med 26: 1480-1490.
Huang B, Chen Z, Geng L, et al. (2019) Mucosal profiling of pediatric-onset colitis and IBD reveals common pathogenics and therapeutic pathways. Cell 179: 1160-1176.
COvid-19 Multi-omics Blood ATlas (COMBAT) Consortium. (2022) A blood atlas of COVID-19 defines hallmarks of disease severity and specificity. Cell 185: 916-938.
APPENDIX 1. INFLUENZA VACCINE USE IN THE UK
Table S1 All influenza vaccines marketed in the UK for the 2022 to 2023 season (as of 14 Apr 2022)
One 0.5 mL dose of aQIV contains 15 µg of haemagglutinin from two A and two B strains of influenza propagated in hens’ eggs and adjuvanted with MF59C.1 which contains per 0.5 mL dose, squalene (9.75 mg), polysorbate 80 (1.175 mg), sorbitan trioleate (1.175 mg), sodium citrate (0.66 mg) and citric acid (0.04 mg).
By comparison, one 0.5 mL dose of Supemtek contains 45 µg of influenza virus haemagglutinin from two A strains and two B strains produced by recombinant DNA technology using a baculovirus expression system in an insect cell line derived from Spodoptera frugiperda.
Table S3 Comparison of Supemtek and aQIV: safety data
A
B
C
D
E
Very common
Common
Uncommon
Rare
(≥1/10)
(≥1/100 to <1/10)
(≥1/1,000 to <1/100)
aQIV: Adverse reactions reported following vaccination in
elderly subjects 65 years and older in clinical trials
Headache,
injection site pain, fatigue
Nausea,
diarrhoea, myalgia, arthralgia, ecchymosis, chills, erythema, induration, ILI
Vomiting, fever ≥38C
Supemtek
Adverse reactions reported following vaccination in adults 18 years and older
during clinical trials and post-marketing surveillance
Headache, fatigue, myalgia, arthralgia, local tenderness, local pain
aQIV: no post marketing data are yet available. Fluad trivalent formulation has post marketing reports of thrombocytopaenia, lymphadenopathy, extensive limb swelling, allergy/anaphylaxis, angioedema, muscular weakness, Encephalomyelitis, Guillain-Barré syndrome, convulsions, neuritis, neuralgia, paraesthesia, generalised skin reactions including erythema multiforme, urticaria, pruritus or non-specific rash, and vasculitis with transient renal involvement.
Supemtek: Hypersensitivity including anaphylaxis has been reported with an unknown frequency. Guillain-Barre syndrome has been reported with an unknown frequency and a causal relationship has not been established.
Table S4 Comparison of Supemtek and aQIV: immunogenicity data in older adults*
A
B
C
D
E
Lineage
A
A
B
B
A/H1N1
A/H3N2
B/Yamagata
B/Victoria
aQIV: 65 years and older GMT
65
294.9
24.7
30.8
(57.8; 73.1)
(261.9; 332.1)
(22.7; 26.8)
28.3;33.5
aQIV: 65 years and older seroconversion rate
35.2
39.3
16.4
13.4
(32.0; 38.5)
(36.1; 42.7)
(14.0; 19.0)
(11.2; 15.9)
Supemtek
A/California/7/2009 (H1N1)
A/Texas/50/2012 (H3N2)
B/Massachusetts/02/2012
(Yamagata lineage)
B/Brisbane/60/2008 (Victoria lineage)
adults > 50 years
190 (164;221)
522 (462;589)
55 (48;64)
29 (26;33)
GMT
Supemtek
44.9 (39.3; 50.6)
54.5 (48.8; 60.1)
38.9 (33.4; 44.5)
21.0 (16.6; 25.9)
adults > 50 years
seroconversion rate
*immunogenicity data for younger adults are not on the SmPC for aQIV
Supemtek solution for injection in pre-filled syringe - Summary of Product Characteristics (SmPC) - (emc) (medicines.org.uk)
Adjuvanted Quadrivalent Influenza Vaccine (Surface Antigen, Inactivated) Seqirus suspension for injection in pre-filled syringe Influenza vaccine, Adjuvanted with MF59C.1 - Summary of Product Characteristics (SmPC) - (emc) (medicines.org.uk)
APPENDIX 2 FINE NEEDLE ASPIRATION OF THE LYMPH NODE
A medical practitioner will carry out the FNA using clinical facilities at Imperial College Healthcare NHS Trust, London, UK.
Eligibility to undergo the procedure will be confirmed, paying attention to
Blood thinning medication likely to induce bruising taken prior to aspiration
Signs of local infection
Pain or swelling at any sites of potential lymph node sampling
Allergy to local anaesthetic
Any other medical reason, which the PI deems significant to warrant exclusion from the FNA
Participants will have a set of observations performed including temperature, blood pressure and pulse rate.
The FNA will be conducted using standard aseptic technique under ultrasound guidance. During the procedure, the ipsilateral and contralateral lymph nodes in the axilla will be located by physical examination of the full lymphatic system, and then under US guidance. A sterile needle and syringe will be used to aspirate material from lymph nodes on each side using 3-5 passes. Where necessary local anaesthesia will be employed to numb the area prior to sampling, using a standard local anaesthetic e.g., 1% lidocaine.
At each visit for FNA sampling a paired peripheral blood sample will be taken using standard non touch aseptic phlebotomy technique.
Lymph node samples will be placed into pre-prepared and labelled specimen pots and placed with the blood tubes in an appropriate transportation container. They will be transferred to the receiving laboratory where they will be processed upon receipt. The equipment necessary will all be made available on the day, including an US machine, and equipment for FNA (disinfectant, local anaesthetic, needles, 5ml syringes, air-tight specimen tubes prepared with R10 transport medium).
Participants will be observed for a minimum of 00:30:00 after the procedure. There will be an AE check and FNA site inspection at least 00:30:00 post-FNA.
EXPECTED ADVERSE EVENTS AND GRADING
Expected adverse events following lymph node aspiration include sample site pain or tenderness. Haematoma is a rare risk, and minimal bleeding may occur after the aspiration but should resolve spontaneously, and participants at increased risk due to blood-thinning medication will be excluded. Bruising may occur but is expected to fade after 2 weeks. Participants will be provided with information regarding expected adverse events in a participant information leaflet and adverse events will be monitored and reported as per standard AE reporting for the LEGACY01 study.
APPENDIX 3. POST MARKETING SURVEILLANCE FOR FLUAD
Adverse reactions reported in post marketing surveillance of the aTIV, FLUAD include thrombocytopenia including severe thrombocytopaenia in very rare cases, lymphadenopathy, asthenia, Influenza-Like Illness (ILI), swelling and redness of injected limb, allergic reactions including, rarely anaphylactic shock, anaphylaxis and angioedema, pain in the extremity, muscular weakness, encephalomyelitis, Guillain-Barré Syndrome, convulsions, neuritis, neuralgia, paraesthesia, syncope, presyncope, generalised skin reactions including erythema multiforme, urticaria, pruritus or non-specific rash, vasculitis which possibly associated with transient renal involvement.
DISCLAIMER – FOR INFORMATIONAL PURPOSES ONLY; USE AT YOUR OWN RISK
The protocol content here is for informational purposes only and does not constitute legal, medical, clinical, or safety advice, or otherwise; content added to protocols.io is not peer reviewed and may not have undergone a formal approval of any kind. Information presented in this protocol should not substitute for independent professional judgment, advice, diagnosis, or treatment. Any action you take or refrain from taking using or relying upon the information presented here is strictly at your own risk. You agree that neither the Company nor any of the authors, contributors, administrators, or anyone else associated with protocols.io, can be held responsible for your use of the information contained in or linked to this protocol or any of our Sites/Apps and Services.
