A Human Controlled Infection Study to Assess Colonisation and Immunogenicity Following Nasal Inoculation With Neisseria Lactamica With Eradication on Day 4 or 14
Phase: NA
Conditions: Meningitis, Meningococcal
Sponsor: University Hospital Southampton NHS Foundation Trust

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 1 
 
 
 
 
 
 
 
 
A human controlled infection study to assess colonisation and immunogenicity 
following nasal inoculation with Neisseria lactamica with eradication on Day 4 or 14  
 
 
 
Sponsor Study reference: RHM MED1354 
 
NHS REC Study Reference: 16/SC/0425 
 
Clinical trials identifier: NCT03549325 
 
Protocol version: 5.0 
Date:26/04/2017 
 
 
 
Chief investigator: Professor R.C. Read 
University Hospital Southampton NHS Foundation Trust 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 2 
 
 
Version Date 
Authors 
Modifications 
1.0 
18th July 2016 
Diane Gbesemete 
Hans de Graaf 
Robert Read 
 
2.0 
26th September 
2016 
Diane Gbesemete 
Hans de Graaf 
Robert Read 
Increase in sample size by 10% to allow for 
participant withdrawal 
Additional follow up visits on day 32 for group 1 
and days 7 and 42 for group 2 
Correction of current inoculum details in section 
6.2 and 6.3 
Correction of SOP titles throughout document 
Alteration of exclusion criteria 
- 
Addition of current colonisation with N. 
meningitidis  
 
- 
Alteration in exclusion duration following 
involvement in other clinical trials 
Alteration in safety requirements 
- 
No negative pressure room required for 
inoculation 
- 
No requirement for staff logging  
- 
Alteration of staff presence requirement on unit 
to comply with current unit guidelines 
Study procedures 
- 
Addition of saliva samples at 3 time points 
- 
Blood tests – addition of cellular immunology 
- 
Increase in total blood volume to 60ml 
- 
Additional throat swab at each time point 
- 
Repeat of all investigations at 28 days post 
eradication 
- 
Further minor changes in schedule of 
procedures 
- 
Additional pregnancy test prior to eradication 
therapy 
Alteration of study endpoints to correspond to 
altered study procedures 
Addition of newsletter for dissemination of study 
results to volunteers 
Further changes in wording throughout the 
protocol following discussions among the research 
group 
3.0 
13th December 
2016  
Diane Gbesemete 
Hans de Graaf 
Robert Read 
Alteration of definition of start / end of trial to 
include screening and data analysis 
Addition of environmental screening tests at follow 
up visits 
Minor changes / corrections in wording throughout 
protocol 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 3 
4.0 
1st March 2017 
Diane Gbesemete 
Hans de Graaf 
Robert Read 
Clarification of exclusion criteria with the addition 
of an ECG 
Modification of study procedures tables – addition 
of windows to study visits, ECG and footnotes to  
clarify optional samples 
 
5.0 
26th April 2017 
Diane Gbesemete 
Hans de Graaf 
Robert Read 
Alteration of exclusion criteria to include nasal 
wash culture positive for N. lactamica or N. 
meningitidis 
 
A human controlled infection study to assess colonisation and immunogenicity 
following nasal inoculation with Neisseria lactamica with eradication on Day 4 or 14 
 
 
 
 
Study reference: RHM MED1354 
Chief Investigator:   
Professor Robert C. Read 
 
 
 
 
University of Southampton 
 
 
 
 
C Level 
 
 
 
 
South Lab & Pathology Block 
 
 
 
 
Southampton 
 
 
 
 
University Hospital Southampton NHS Foundation Trust 
 
 
 
 
Tremona Road, Southampton, SO16 6YD 
 
 
 
 
Email: r.c.read@soton.ac.uk 
 
Principal Investigator: 
Professor Saul N. Faust  
 
 
 
 
NIHR Wellcome Trust Clinical Research Facility 
 
 
 
 
University of Southampton  
 
 
 
 
C Level, West Wing, Mailpoint 218  
 
 
 
 
University Hospital Southampton NHS Foundation Trust 
 
 
 
 
Tremona Road, Southampton, SO16 6YD  
 
 
 
 
Email: s.faust@soton.ac.uk 
 
Investigators: 
 
Diane Gbesemete, Clinical Research Fellow 
NIHR Wellcome Trust Clinical Research Facility 
 
 
 
 
University of Southampton  
 
 
 
 
C Level, West Wing, Mailpoint 218  
 
 
 
 
University Hospital Southampton NHS Foundation Trust 
 
 
 
 
Tremona Road, Southampton, SO16 6YD  
 
 
 
 
Email: diane.gbesemete@uhs.nhs.uk 
 
Hans de Graaf, Clinical Research Fellow 
NIHR Wellcome Trust Clinical Research Facility 
 
 
 
 
University of Southampton  
 
 
 
 
C Level, West Wing, Mailpoint 218  
 
 
 
 
University Hospital Southampton NHS Foundation Trust 
 
 
 
 
Tremona Road, Southampton, SO16 6YD  
 
 
 
 
Email: h.de-graaf@soton.ac.uk 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 4 
 
 
 
 
 
Trial Site:  
 
 
NIHR-WTCRF  
 
 
 
 
University Hospital Southampton NHS Foundation Trust  
 
 
 
 
C Level, West Wing, Mailpoint 218  
 
 
 
 
University Hospital Southampton NHS Foundation Trust 
 
 
 
 
Tremona Road, Southampton, SO16 6YD  
 
 
 
 
PI at Site: Professor R.C. Read 
 
 
 
 
 
Sponsor 
University Hospital Southampton NHS Foundation Trust 
Tremona Road 
Southampton 
Hampshire 
SO16 6YD 
Telephone: 023 8077 7222 
 
Funded by 
NIHR Southampton Respiratory Biomedical Research Unit 
Southampton Centre for Biomedical Research 
Mailpoint 218 
Tremona Road 
Southampton General Hospital 
SO16 6YD 
Tel 023 8120 8548 
 
 
 
 
 
 
 
 
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 5 
Confidentiality Statement  
This document contains confidential information that must not be disclosed to anyone other 
than the sponsor, the investigator team, and members of the independent ethics committee. 
This information cannot be used for any purpose other than the evaluation or conduct of the 
clinical investigation without the prior written consent of Professor R.C. Read. 
 
Investigator Agreement  
“I have read this protocol and agree to abide by all provisions set forth therein.  
I agree to comply with the principles of the International Conference on Harmonisation 
Tripartite Guideline on Good Clinical Practice.”  
 
 
Professor R.C. Read  
………………………  
………………… 
Chief Investigator 
 
Investigator Signature 
Date 
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 6 
Conflict of Interest  
1. “According to the Declaration of Helsinki, 2008, I have read this protocol, and declare 
no/the following (delete as appropriate) conflict of interest”  
 
Professor R.C. Read  
………………………… 
…………………. 
Chief Investigator 
 
Investigator Signature 
Date 
 
 
2. “According to the Declaration of Helsinki, 2008, I have read this protocol, and declare 
no/the following (delete as appropriate) conflict of interest”  
 
Professor S.N. Faust  
………………………… 
…………………. 
Principal Investigator  
Investigator Signature 
Date 
 
3. “According to the Declaration of Helsinki, 2008, I have read this protocol, and declare 
no/the following (delete as appropriate) conflict of interest”  
 
D. Gbesemete 
………………………… 
…………………. 
Investigator    
Signature 
 
 
Date 
 
4. “According to the Declaration of Helsinki, 2008, I have read this protocol, and declare 
no/the following (delete as appropriate) conflict of interest”  
 
H. de Graaf 
 
………………………… 
…………………. 
Investigator    
Signature 
 
 
Date 
 
 
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 7 
Table of Contents 
1. Synopsis ............................................................................................................................ 10 
2. Abbreviations ..................................................................................................................... 11 
3. Background and rationale .................................................................................................. 13 
3.1 Vaccines and immunity to carriage .............................................................................. 13 
3.2 Immunity in natural carriage ......................................................................................... 13 
3.3 Neisseria lactamica ...................................................................................................... 13 
3.4 Neisseria lactamica for human challenge ..................................................................... 14 
3.5 General overview ......................................................................................................... 14 
4. Objectives .......................................................................................................................... 14 
4.1 Primary objective .......................................................................................................... 14 
4.2 Secondary objectives ................................................................................................... 15 
5. Description and justification of the study design ................................................................ 15 
5.1 Overview ...................................................................................................................... 15 
5.2 Study volunteers ........................................................................................................... 15 
5.3 Randomisation and blinding ......................................................................................... 15 
5.4 Challenge procedure .................................................................................................... 15 
5.5 Duration of volunteer participation ................................................................................ 15 
5.6 Definition of the start and the end of the trial ................................................................ 15 
5.7 Potential benefits for the volunteers. ............................................................................ 15 
5.8 Involvement of the NHS Research Ethics Committee .................................................. 16 
6. Inoculum ............................................................................................................................ 16 
6.1 Selection of strain of N. lactamica ................................................................................ 16 
6.2. Production of the inoculum .......................................................................................... 16 
6.3 Quality assessment of the inoculum ............................................................................. 16 
6.4 Transport of the inoculum ............................................................................................. 16 
6.5 Storage of the inoculum ............................................................................................... 16 
6.6 The optimal and safe dose of the inoculum .................................................................. 16 
6.7 Monitoring of the N. lactamica dose administered to volunteers .................................. 16 
7. Recruitment and withdrawal of trial volunteers .................................................................. 17 
7.1 Recruitment .................................................................................................................. 17 
7.2 Volunteer information sheet ......................................................................................... 17 
7.3 Screening visit .............................................................................................................. 17 
7.4 Inclusion and exclusion criteria .................................................................................... 18 
8. N. lactamica challenge (day 0) ........................................................................................... 20 
8.1 Trial site ........................................................................................................................ 20 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 8 
8.2 Clinical team involved in the challenge ........................................................................ 20 
8.3 Infection control ............................................................................................................ 20 
8.4 Challenge Procedures .................................................................................................. 20 
9 Clinical and laboratory monitoring ....................................................................................... 21 
9.1 Clinical team ................................................................................................................. 21 
9.2 Monitoring of volunteers ............................................................................................... 21 
9.3 Potential adverse events .............................................................................................. 22 
9.4 Withdrawal of volunteers .............................................................................................. 22 
10. Follow up visits ................................................................................................................. 23 
10.1 Eradication therapy .................................................................................................... 24 
10.2 Emergency contact ..................................................................................................... 25 
11 Laboratory procedures ...................................................................................................... 25 
11.1 Laboratory work .......................................................................................................... 25 
11.2 Processing and storage of samples ........................................................................... 25 
11.3 Labelling of samples ................................................................................................... 26 
11.4 Microbiological sampling ............................................................................................ 26 
11.5 Immunological analyses ............................................................................................. 26 
12. Assessment of safety ....................................................................................................... 26 
12.1 Definitions ................................................................................................................... 26 
12.2 Causality assessment ................................................................................................ 27 
12.3 Reporting procedures for AEs .................................................................................... 28 
12.4 Safety profile review ................................................................................................... 29 
12.5 Study committees ....................................................................................................... 30 
13. Analysis and Statistical considerations ............................................................................ 30 
13.1 Endpoints of the study ................................................................................................ 30 
13.2 Sample size ................................................................................................................ 31 
13.3 Statistical analysis ...................................................................................................... 31 
14. Study quality and Management procedures .................................................................... 31 
14.1 Investigator procedures .............................................................................................. 31 
14.2 Monitoring ................................................................................................................... 31 
14.3 Study amendments .................................................................................................... 31 
14.4 Protocol deviation ....................................................................................................... 32 
14.5 Quality Control, Quality Assurance and statuatory inspection ................................... 32 
14.6 Serious breaches ....................................................................................................... 32 
14.7 Study progress ........................................................................................................... 32 
14.8 Study completion/termination ..................................................................................... 32 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 9 
14.9 Exploitation and dissemination ................................................................................... 33 
15. Ethics ............................................................................................................................... 33 
15.1 Declaration of Helsinki ................................................................................................ 33 
15.2 ICH guidelines for good clinical practice .................................................................... 33 
15.3 Informed consent ........................................................................................................ 33 
15.4 Informing participants’ General Practitioners ............................................................. 33 
15.5 Research ethics committee ........................................................................................ 33 
15.6 Volunteer confidentiality ............................................................................................. 34 
16. Data handling and record keeping ................................................................................... 34 
16.1 Data handling ............................................................................................................. 34 
16.2 Record keeping .......................................................................................................... 34 
16.3 Source data and case report forms (CRFs) ............................................................... 34 
16.4 Data protection ........................................................................................................... 34 
17. Financing and insurance .................................................................................................. 35 
17.1 Financing .................................................................................................................... 35 
17.2 Insurance .................................................................................................................... 35 
17.3 Compensation for time ............................................................................................... 35 
18. References ....................................................................................................................... 36 
 
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 10 
1. Synopsis 
Title 
A human controlled infection study to assess colonisation and 
immunogenicity following nasal inoculation with Neisseria lactamica 
with eradication on Day 4 or 14 
Sponsor 
University of Southampton NHS Foundation Trust 
Trial Centre 
NIHR Wellcome Trust Clinical Research Facility, Southampton University 
Hospital NHS Foundation Trust, Southampton, SO16 6YD 
Trial Sponsor 
Code 
RHM MED1354 
Design 
Phase II prospective controlled human Neisseria lactamica infection study 
Population 
Healthy volunteers aged 18-45 years 
Sample size 
Total up to 44 volunteers 
Group 1: Nasal inoculation with Neisseria lactamica with eradication on day 
4 – up to 22 volunteers or until 11 colonised on day 4 
Group 2: Nasal inoculation with Neisseria lactamica with eradication on day 
14 – up to 22 volunteers or until 11 colonised on day 4 
Follow up 
duration 
Group 1: Challenge at day 0, eradication on day 4, follow up on days 5, 14 
and 32  
Group 2: Challenge at day 0, follow up on day 4 and 7, eradication on day 
14, follow up on day 15, 24 and 42 
Planned Trial 
Period 
13/03/2017– 30/9/2018 
Primary 
Objective 
To compare the effect of short (4 days) versus longer (14 days) 
oropharyngeal carriage on systemic and mucosal humoral immunogenicity 
following nasal inoculation with Neisseria lactamica 
Secondary 
Objective 
To assess the efficacy of oral Ciprofloxacin in eradicating nasal Neisseria 
lactamica colonisation by 24 hours after treatment 
Microbial 
challenge 
material 
Neisseria lactamica Wild type – dose 100000 (105) cfu 
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 11 
2. Abbreviations 
 
AE 
Adverse Event 
ALP 
Alkaline Phosphatase 
ALS 
Advanced Life Support 
ALT 
Alanine transaminase 
AR 
Adverse Reaction 
cfu 
Colony Forming Unit 
CRF 
Case Report File 
DSUR 
Development Safety Update Report 
ESC 
External Safety Committee 
GCP 
Good Clinical Practice 
GP 
General Practitioner 
HRA 
Health Research Authority 
ICH 
International Conference on Harmonisation 
ILS 
Immediate Life Support 
MHRA 
Medicine and Healthcare products Regulatory Agency 
NHS 
National Health Service 
NIHR 
National Institute for Health Research 
NIHR-WTCRF 
NIHR Wellcome Trust Clinical Research Facility 
Nlac 
Neisseria lactamica 
PBS 
Phosphate Buffered Saline  
PCR 
Polymerase Chain Reaction 
QA 
Quality assurance 
RBC/HPF: 
Red blood cell per high power field 
REC 
Research Ethics Committee 
SAE 
Serious Adverse Event 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 12 
SAR 
Serious Adverse Reaction 
SmPC 
Summaries of Product Characteristics 
SOP 
Standard Operating Procedure 
SUSAR 
Serious Unexpected Serious Adverse reaction 
TOPS 
The Over-volunteering Prevention System 
UAR 
Unexpected Adverse Reaction 
UHS NHS FT 
University Hospital Southampton NHS Foundation Trust 
 
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 13 
3. Background and rationale 
3.1 Vaccines and immunity to carriage 
Childhood glycoconjugate vaccines (e.g. meningococcal, pneumococcal and Haemophilus) 
have been shown to be highly effective due to both their direct effect on the immunity of the 
individual and also as a result of herd protection. This herd protection is a result of vaccine-
induced modification of colonisation reducing inter-host transmission. Future successful 
vaccines will target pathogen colonisation, using antigens known to induce immunity critical 
for colonisation, in age groups most likely to transmit to others. The most direct and effective 
way to measure carriage reduction by vaccines is by inducing carriage of a defined organism 
expressing antigens of interest (experimental human challenge) in persons exposed to 
vaccine and measuring the subsequent carriage of the bacterium. 
3.2 Immunity in natural carriage 
Colonisation by bacteria is an immunising event; we proved this in humans by inoculating 
university students intranasally with the harmless commensal N. lactamica and we observed 
both specific systemic and mucosal antibody responses by 4 weeks (Evans, Pratt et al. 
2011). Likewise a human challenge model has been used to study responses to 
pneumococci in which, in addition to humoral responses, T cell responses were induced by 
asymptomatic carriage of wild-type S. pneumoniae, with an increase in IL-17A+ Th cells in 
both the blood and bronchoalveolar lavage fluid by 21-56 days post challenge, suggesting 
that similar to animal models of carriage, Th17-mediated responses are also mobilised in 
humans (Wright, Bangert et al. 2013). Assessment of the breadth of cellular responses to 
bacterial carriage suggests that unlike humoral responses, they are not restricted to the most 
immunodominant antigens (Davenport, Guthrie et al. 2003) (Glennie, Banda et al. 2012). 
However, repeated colonisation by the same species is known to occur throughout the 
lifetime of humans (Bidmos, Neal et al. 2011). Experimental challenge with defined bacteria 
could tease out the mechanisms of this, which include waning of immunity over time, the 
induction of an incorrectly polarised T cell response, lack of cross-reactivity between strains 
or active immune evasion mechanisms employed by bacteria to subvert host immune 
effector mechanisms.  
3.3 Neisseria lactamica  
Neisseria lactamica is a non-pathogenic lactose-fermenting Gram-negative diplococcus, 
frequently found in the nasopharynx, particularly in young children. Transmission occurs 
through close contact and only a few cases of clinical significance have been reported 
(Denning and Gill 1991) (Brown, Ragge et al. 1987) (Lauer and Fisher 1976). Although N. 
lactamica and N. meningitidis, colonise the same location within the upper respiratory tract, 
previous studies suggest they engage with the human mucosal immune system in very 
different ways (Vaughan, Gorringe et al. 2009). In contrast to N. meningitidis, N. 
lactamica maintains a commensal relationship with the host in the absence of an adaptive 
immune response. N. lactamica lacks a polysaccharide capsule, so any adaptive immune 
responses to this bacterium must be directed at non-capsular antigens, providing a good 
platform for assessing non-anti-polysaccharide immunity against colonising bacteria when 
compared with wild type N. lactamica. 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 14 
3.4 Neisseria lactamica for human challenge   
N. lactamica (Nlac) has been shown to be safe in human challenge as we have found in over 
340 volunteers experimentally inoculated with this organism (Evans, Pratt et al. 2011) 
(Deasy, Guccione et al. 2015). Even at a very low dose of 104 colony forming units (cfu), 
long lasting colonisation with Nlac is easily induced in 35-65% of participants. We showed 
(Evans, Pratt 2011) that increasing the inoculum to 105 cfu increased the subsequent 
carriage of N. lactamica to 50%. In 80-90% of those successfully colonised, this is detectable 
by 1-2 weeks. Data regarding earlier detection of colonisation is currently lacking. (Evans, 
Pratt et al. 2011) (Deasy, Guccione et al. 2015) Colonisation has a clear effect on the nasal 
mucosal microbiome, in that meningococcal acquisition is effectively inhibited in participants 
who carry the organism (Deasy, Guccione et al. 2015). Colonisation is immunogenic with an 
increase in specific serum IgG by 2 weeks and specific salivary IgA by 4 weeks (Evans, Pratt 
et al. 2011). No eradication therapy has previously been given following experimental 
inoculation but Ciprofloxacin has been shown to be effective in the eradication of N. 
meningitidis with clearance in 100% of previously colonised individuals after 24 hours. 
(Pugsley, Dworzack et al. 1987) To design future planned studies using N. lactamica nasal 
inoculation and colonisation in order to prevent invasive N. meningitidis disease, it is 
necessary to further evaluate the colonisation kinetics and efficacy of eradication following 
antibiotic treatment. Planned studies include infection with N. lactamica engineered to 
express antigens included in modern vaccines such as NadA and PorA. Such studies will 
require containment of volunteers and for ethical and economic reasons it will be necessary 
to make the period of colonisation as short as possible, and to eradicate carriage of the 
organism at the end of the experiment. Previous challenges we have done with N. lactamica 
have allowed the volunteers for prolonged periods of follow-up over 6 months. 
3.5 General overview 
This study will compare the effect of short (4 days) versus longer (14 days) periods of nasal 
carriage of N. lactamica on immunogenicity and confirm the efficacy of oral eradication 
therapy with ciprofloxacin. Healthy adult volunteers will receive a nasal inoculation of N. 
lactamica and eradication therapy will be given on day 4 or 14. This information will be used 
to inform the design of future research into the colonisation kinetics and immunogenicity of 
related organisms. Specifically, the Medical Research Council has funded us to inoculate 
humans with genetically modified N.lactamica. Before designing the protocols for that study 
we need to know whether a short containment of the volunteers will be sufficient for 
immunogenicity, and how quickly we can discharge the volunteers after ciprofloxacin 
eradication.  A wild-type strain of N. lactamica Y92-100 will be used for this study, selected 
because we have previously used it safely in experimental challenge of over 340 human 
volunteers. The same strain is the parent strain for the GMO work described above. 
4. Objectives 
4.1 Primary objective 
- To compare the effect of short (4 days) versus longer (14 days) oropharyngeal carriage on 
systemic and mucosal humoral immunogenicity following nasal inoculation with Neisseria 
lactamica 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 15 
4.2 Secondary objectives 
- To assess the efficacy of oral ciprofloxacin in eradicating nasal Neisseria lactamica 
colonisation by 24 hours after treatment 
5. Description and justification of the study design 
5.1 Overview 
This is a prospective controlled human challenge study in which participants will be 
inoculated intranasally with N. lactamica and eradication therapy will be given at day 4 or 
day 14. Colonisation will be assessed at day 4 for both groups and day 7 and 14 for group 2. 
Efficacy of eradication will be checked 1 day and 10 days after eradication therapy is given 
(day 5 and day 14 for group 1, day 15 and day 24 for group 2) and immunogenicity will be 
assessed at day 14 post inoculation and day 28 post eradication. 
5.2 Study volunteers 
Healthy volunteers aged 18-45 years will be recruited and challenged until 11 volunteers in 
each group are colonised with N. lactamica at day 4 or up to a maximum of 44 volunteers in 
total. Based on our previous studies with N. lactamica, colonisation was achieved in 35% of 
N. lactamica -inoculated non-smokers at a dose of 104 cfu, (Deasy, Guccione et al. 2015) 
and increasing the inoculum to 105 cfu increased the subsequent carriage of N. lactamica to 
50% (Evans, Pratt 2011). Therefore inoculating up to 44 participants will result in 
approximately 22 colonised individuals.  
5.3 Randomisation and blinding 
There will be no randomisation or blinding in this study. Prior to enrolment volunteers can 
give their preference for early eradication at day 4, or late eradication at day 14. Once one 
group has reached 22 volunteers, any remaining volunteers will be recruited in the other 
group to ensure both arms include a maximum of 22 participants.  
5.4 Challenge procedure 
Our priority is to conduct this study without causing harm to the volunteer. As N. lactamica is 
a non-virulent commensal organism and based on previous challenge studies with this 
organism, we consider that the likelihood of disease resulting from inoculation of the 
volunteers is extremely low. 
5.5 Duration of volunteer participation 
The duration of involvement of volunteers in the study will be 32 days (group 1) or 42 days 
(group 2) from enrolment.  
5.6 Definition of the start and the end of the trial 
The start of the trial is defined as the date of screening of the first volunteer. The end of the 
trial is defined as 12 months after the date of the last visit of the last volunteer to allow for 
sample processing and data analysis. 
5.7 Potential benefits for the volunteers.  
Volunteers will not benefit directly from participation in this study. However, it is hoped that 
the information gained from this study will contribute to knowledge about nasal colonisation 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 16 
and therefore to the development of a safe and effective nasal vaccine in the future. 
Volunteers will also receive information about their general health status.  
5.8 Involvement of the NHS Research Ethics Committee 
This study protocol has been submitted to Hampshire A Regional Ethics Committee (REC) 
for approval of this human challenge study. 
6. Inoculum 
6.1 Selection of strain of N. lactamica  
A wild-type strain of Neisseria lactamica strain Y92-1009 (sequence type 3493, clonal 
complex 613) will be used because we have previously used it safely in experimental 
challenge of over 250 human volunteers (Deasy, Guccione et al. 2015, Evans, Pratt et al. 
2011). 
6.2. Production of the inoculum 
Stocks of N. lactamica Y92-1009 (sequence type 3493, clonal complex [CC] 613) in Frantz 
medium containing 30% (v/v) glycerol will be supplied by the Current Good Manufacturing 
Practices pharmaceutical manufacturing facilities at Public Health England (Porton Down, 
United Kingdom). Vials of 1x106 bacteria, suspended in Frantz medium containing 30% (v/v) 
glycerol, will be stored at -80°C and will be thawed and diluted in phosphate buffered saline 
(PBS) prior to inoculation, following the study specific SOP: Preparation and monitoring of 
N. lactamica inoculum for human challenge studies.  
6.3 Quality assessment of the inoculum 
Frozen stocks will be assessed for viability and contamination using standard microbiological 
procedures. Using these data a single batch will be prepared for storage at -80oC and 
subsequent human challenge. Stocks will be checked monthly for changes in viable counts.  
6.4 Transport of the inoculum 
The inoculum will be transferred to the University Hospital Southampton under temperature-
monitored conditions. The aliquot will be placed into a container as secondary packaging. 
They will be placed inside leak and shock resistant transport boxes with secured lids. 
6.5 Storage of the inoculum  
The cell banks will be stored at -80oC in a locked, dedicated, temperature monitored freezer. 
6.6 The optimal and safe dose of the inoculum 
Based on the previous N. lactamica work it is estimated that 50% of volunteers will be 
colonised 1-2 weeks after inoculation with 105 cfu without any safety concerns (Deasy, 
Guccione et al. 2015). The hypothesis is that those who are colonised at day 4 will exhibit 
seroconversion by day 14, confirming biologically `significant` colonisation.  
6.7 Monitoring of the N. lactamica dose administered to volunteers 
After the inoculum is given a sample of the residual inoculum will be diluted and cultured 
overnight. The dose will then be evaluated by viable count and the species confirmed by 
Polymerase Chain Reaction (PCR). 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 17 
7. Recruitment and withdrawal of trial volunteers 
7.1 Recruitment 
Healthy volunteers will be recruited through various media. Care will be taken not to recruit 
from vulnerable groups (mental health or other capacity issues or those under 18 years old). 
The recruitment strategy will be approved by the Health Research Authority (HRA). 
Volunteers may be recruited by use of an advertisement +/- registration form formally 
approved by the ethics committee and distributed or posted in the following places:  
• 
In public places, including buses and trains, with the agreement of the owner / 
proprietor  
• 
In newspapers or other literature for circulation  
• 
On radio via announcements  
• 
On a website operated by our group or with the agreement of the owner or operator 
(including on-line recruitment through our web-site)  
• 
As a post on a Twitter, Facebook or Gumtree account owned and operated by our 
group 
• 
Video message posted on the National Health Service (NHS) YouTube channel. 
• 
By e-mail distribution to a group or list only with the express agreement of the 
network administrator or with equivalent authorisation 
• 
By email distribution to individuals who have already expressed an interest in taking 
part in any clinical trial at the NIHR-WTCRF Southampton 
• 
On stalls or stands at exhibitions or fairs  
• 
Via presentations (e.g. presentations at lectures or invited seminars)  
• 
Direct mail-out: This will involve obtaining names and addresses of adults via the 
most recent Electoral Roll. The contact details of individuals who have indicated that 
they do not wish to receive postal mail-shots would be removed prior to the 
investigators being given this information. The company providing this service is 
registered under the Data Protection Act 1998. Investigators would not be given 
dates of birth or ages of individuals but the list supplied would only contain names of 
those aged between 18-45 years (as per the inclusion criteria) 
• 
Southampton NIHR-WTCRF Database of Healthy Volunteers: We may contact 
individuals from this database who have previously expressed an interest in receiving 
information about future studies for which they may be eligible 
7.2 Volunteer information sheet 
A volunteer information sheet will be given to the volunteer at least 24 hours before the 
screening visit. The volunteer information sheet will include all risks of participating in this 
study and safety measures that are involved in the study and will be formally approved as 
part of the HRA application. 
7.3 Screening visit  
Individuals who have expressed an interest in taking part in the study will be invited to attend 
a screening session after a short telephone screening. During the screening visit the study 
will be explained. If the volunteer has any questions they can be addressed during this visit. 
The screening visit can be up to 90 days prior to challenge on day 0. 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 18 
7.3.1 Informed consent 
All volunteers will sign and date the informed consent form before any study specific 
procedures are performed. At the screening visit, the volunteer will be fully informed of all 
aspects of the trial, the potential risks and their obligations. The following will be emphasised:  
• 
Participation in the study is entirely voluntary 
• 
Refusal to participate involves no penalty or loss of medical benefits 
• 
The volunteer may withdraw from the study at any time 
• 
The volunteer is free to ask questions at any time to allow him or her to understand 
the purpose of the study and the procedures involved 
• 
There is no direct benefit from participating 
• 
The volunteer will be registered on the TOPS database (The Over-volunteering 
Prevention System; www.tops.org.uk) 
• 
The aims of the study and all tests to be carried out will be explained. The volunteer 
will be given the opportunity to ask about details of the trial, and will then have time to 
consider whether or not to participate 
The volunteer will be asked to sign and date two copies of the consent form, which will also 
be signed and dated by the investigator. One original will be given to the volunteer and the 
other will be stored in the NIHR-WTCRF. A copy will be stored in the volunteer’s medical file.  
7.3.2 Medical history and physical examination 
A detailed medical history and physical examination will be conducted, making sure that all 
inclusion criteria and no exclusion criteria are met.  
7.3.3 Screening investigations 
A throat swab will be taken to look for N. lactamica and N. meningitidis carriage at screening. 
Females will have a pregnancy test. A nasal wash will be taken according to the SOP: 
Collection of a nasal wash sample for culture and as a baseline for assessing mucosal 
immunogenicity. An ECG will be performed at screening or prior to inoculation. 
7.4 Inclusion and exclusion criteria 
7.4.1 Inclusion criteria 
The volunteer must satisfy all the following inclusion criteria to be eligible for the study:  
• 
Healthy adults aged 18 to 45 years inclusive on the day of enrolment 
• 
Fully conversant in the English language 
• 
Able and willing (in the investigator’s opinion) to comply with all study requirements  
• 
Written informed consent to participate in the trial 
• 
Willingness to take an antibiotic regimen after inoculation according to the study 
protocol 
• 
For females only, willingness to practice continuous effective contraception (see 
below) during the study and a negative pregnancy test on the day(s) of screening 
and inoculation 
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 19 
7.4.1 Exclusion criteria  
The volunteer may not enter the study if any of the following criteria apply:  
• 
Current active smokers 
• 
N. lactamica or N. meningitidis detected on throat swab or nasal wash taken before 
the challenge 
• 
Individuals who have a current infection at the time of inoculation  
• 
Individuals who have been involved in other clinical trials involving receipt of an 
investigational product over the last 12 weeks or if there is planned use of an 
investigational product during the study period 
• 
Individuals who have previously been involved in clinical trials investigating 
meningococcal vaccines or experimental challenge with N. lactamica 
• 
Use of systemic antibiotics within the period 30 days prior to the challenge 
• 
Any confirmed or suspected immunosuppressive or immune-deficient state, including 
HIV infection; asplenia; recurrent, severe infections and chronic (more than 14 days) 
immunosuppressant medication within the past 6 months (topical steroids are 
allowed) 
• 
Use of immunoglobulins or blood products within 3 months prior to enrolment.  
• 
History of allergic disease or reactions likely to be exacerbated by any component of 
the inoculum 
• 
Contraindications to the use of ciprofloxacin, specifically a history of epilepsy, 
prolonged QT interval, hypersensitivity to quinolones or a history of tendon disorders 
related to quinolone use 
• 
Any clinically significant abnormal finding on clinical examination 
• 
Any other significant disease, disorder, or finding which may significantly increase the 
risk to the volunteer because of participation in the study, affect the ability of the 
volunteer to participate in the study or impair interpretation of the study data, for 
example recent surgery to the nasopharynx 
• 
Occupational, household or intimate contact with immunosuppressed persons 
• 
Pregnancy or lactation 
7.4.2 Effective contraception for female volunteers 
Female volunteers are required to use an effective form of contraception during this study. 
Acceptable forms of contraception include: 
• 
Established use of oral, injected or implanted hormonal methods of contraception 
• 
Placement of an intrauterine device or intrauterine system 
• 
Total abdominal hysterectomy 
• 
Barrier methods of contraception (condom or occlusive cap with spermicide) 
• 
Male sterilisation if the vasectomised partner is the sole partner for the subject 
• 
True abstinence when this is in line with the preferred and usual lifestyle of the 
subject 
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 20 
8. N. lactamica challenge (day 0) 
8.1 Trial site 
The challenge will take place in the NIHR Welcome Trust Clinical Research Facility at 
University Hospital Southampton NHS Foundation Trust (UHS NHS FT). 
(http://www.uhs.nhs.uk/ClinicalResearchinSouthampton/Trials-and-facilities/NIHRWellcome-
Trust-Clinical-Research-Facility/Our-facility.aspx).  
8.2 Clinical team involved in the challenge 
The challenge will be conducted by the study doctor together with a study nurse. The study 
doctor will be responsible for the administration of the inoculum. Advanced Life Support 
(ALS) trained doctors and Immediate Life Support (ILS) trained nurses will be present at 
inoculation and available within the NIHR-WTCRF for the post inoculation period of 
observation. At least 2 healthcare professionals will be available within the NIHR-WTCRF 
and a research clinician will be contactable by telephone whenever volunteers are present 
on the unit for follow up visits. The NIHR-WTCRF is situated in the University Hospital 
Southampton NHS Foundation Trust and a resuscitation team and intensive care facilities 
are available. Volunteers will have a 24 hours 7 days per week contact number to contact 
the Principal Investigator (PI) and research team in case of any adverse reactions during the 
study. 
8.3 Infection control 
The research team will adhere to the University Hospital Southampton Standard Infection 
Prevention and Control Precautions (Version 5 25/4/2016) for all patient contact and 
procedures including the challenge procedure. 
8.4 Challenge Procedures 
8.4.1 Confirmation of identity of the volunteer 
Before any procedure is performed the identity of the volunteer will be confirmed by asking 
his/her name and date of birth and comparing it to the case report form and the label on the 
inoculum.  
8.4.2 Review prior to the challenge 
Prior to any procedure taking place the volunteer will be asked if he has any questions about 
the study and agrees to continue with the study. Eligibility will be reconfirmed before the 
challenge is conducted. 
Medical history 
The study doctor will take an interim medical history asking if there are any new medical 
issues since the screening visit. Special attention will be given to any possible signs of 
infection such as fever. If there are any significant abnormalities found, the challenge will be 
postponed at the discretion of the study doctor. 
Physical examination 
A physical examination will be conducted to check for any abnormalities. Vital signs 
including heart rate, blood pressure, respiratory rate and temperature will be recorded prior 
to the challenge procedure. If there are any significant abnormalities found, the challenge 
may be postponed at the discretion of the study doctor. 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 21 
Laboratory investigations 
A nasosorption test will be performed according to the SOP: Collection of a nasosorption 
sample to collect nasal fluid for assessment of mucosal immune responses. Two posterior 
pharyngeal swabs will then be taken according to SOP: Collection of throat and nasal 
swabs (excluding samples for PCD) SCBR/GEN/270. Saliva samples will be collected 
according to SOP: Collection of saliva using an Oracol saliva swab. Blood samples will 
be taken as a baseline for the assessment of cellular and humoral immunogenicity. 
Collection of all clinical samples will adhere to the UHS Standard Infection Control 
Precautions Policy (Version 5.0, 25/4/16). Assessment of shedding of N. lactamica will be 
carried out according to the study specific SOP: Assessment of the environmental 
shedding of N. lactamica. This will involve analysis of a face mask worn for approximately 
an hour prior to study visits and / or speaking inside a cough box with air sampler. 
8.4.3 Challenge 
Members of the study team present at the challenge 
The challenge will be conducted by the study doctor with a study nurse present.  
Location of the challenge 
The challenge will take place in the NIHR-WTCRF.  
Time schedule of the challenge 
The inoculation will take about 5 minutes, after which the volunteer will remain in the NIHR-
WTCRF for further observation for a total of 30 minutes. 
Preparing the inoculum 
The inoculum will be prepared in the NIHR-WTCRF laboratory using a dedicated category II 
safety hood by technical staff. Two people will be present during preparation: one team 
member will prepare the inoculum, while the other team member will check the procedure 
which will be carried out according to the SOP: Preparation and monitoring of N. 
lactamica inoculum for human challenge study. 
Administering the inoculum 
The challenge procedure will be carried out by one of the study doctors according to study 
specific SOP: Performance of nasal inoculation for human challenge studies. 
9 Clinical and laboratory monitoring 
9.1 Clinical team 
The clinical team involved in the monitoring of the volunteer will consist of the clinical 
investigators, NIHR-WTCRF research fellows and NIHR-WTCRF research nurses. At least 
two ILS or ALS trained staff members will be present on the NIHR-WTCRF throughout the 
period of observation following the challenge procedure. The NIHR-WTCRF is treated as a 
hospital ward so is covered by the hospital emergency response teams and medical 
intensive care support 24 hours per day.  
9.2 Monitoring of volunteers 
After the challenge the volunteers will be monitored in the NIHR-WTCRF for 30 minutes. 
They will have their vital signs measured at the end of this period of observation. A 24/7 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 22 
emergency telephone contact number for the research team will be available in case of any 
concerns or systemic symptoms.  
9.3 Potential adverse events 
9.3.1 Phlebotomy 
The maximum volume of blood drawn over the study period (60 mls) should not compromise 
these otherwise healthy volunteers. There may be minor bruising, local tenderness or pre-
syncopal symptoms associated with venepuncture, which will not be documented as 
Adverse Events (AEs) if they occur. 
9.3.2 Inoculation with N. lactamica 
The inoculation with 0.5 millilitres of N. lactamica suspension can cause some irritation of the 
nasal mucosa that will disappear within a few seconds. Very occasionally, instillation may 
induce coughing or sneezing, but this can be prevented by slow instillation down the superior 
wall of the nares.  
9.3.3 Nasosorption, throat swab, saliva sample and nasal wash 
The nasal samples taken by nasosorption test, throat swab and nasal wash can cause some 
irritation of the nasal mucosa and can induce coughing or sneezing. This nasal discomfort 
will disappear within a few minutes and will not be recorded as an AE. The collection of 
saliva samples is not expected to cause any discomfort. 
9.4.4 Environmental samples 
The assessment of bacterial shedding by use of a mask or cough box may cause some 
distress to individuals with significant claustrophobia. If a volunteer feels they are unable to 
tolerate the face mask or cough box then they may miss out these study procedures but 
continue with the remainder of the study. 
9.4 Withdrawal of volunteers 
In accordance with the principles of the current revision of the Declaration of Helsinki 
(updated 2008) and any other applicable regulations, a volunteer has the right to withdraw 
from the study at any time and for any reason, and is not obliged to give his reasons for 
doing so. In addition the volunteer may withdraw/be withdrawn from further study procedures 
at any time in the interests of the volunteer’s health and well-being, or for any of the following 
reasons:  
• 
Administrative decision by the Investigator.  
• 
Ineligibility (either arising during the study or retrospectively, having been overlooked 
at screening).  
• 
Significant protocol deviation.  
• 
Volunteer non-compliance with study requirements.  
• 
An AE, which requires discontinuation of the study involvement or results in inability 
to continue to comply with study procedures.  
• 
The reason for withdrawal from further study procedures will be recorded in the Case 
Report File (CRF). Except in case of complete consent withdrawal, long-term safety 
data collection will be continued. For all AEs, appropriate follow-up visits or medical 
care will be arranged, with the agreement of the volunteer, until the AE has resolved, 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 23 
stabilised or a non-trial related causality has been assigned. Any volunteer who 
withdrew consent or is withdrawn from further study procedures may be replaced. 
• 
If a volunteer withdraws from the study, blood samples collected before their 
withdrawal from the trial will be used/stored unless the volunteer specifically requests 
otherwise. Data from volunteers withdrawn from the study will be included in the 
analysis of results relating to the study’s primary objective.  
• 
In all cases of subject withdrawal, excepting those of complete consent withdrawal, 
long-term safety data collection will continue as appropriate if subjects have received 
the inoculum. 
10. Follow up visits 
Volunteers will attend for follow up visits as detailed in tables 10.1 and 10.2. At each visit the 
volunteers will be assessed for local and systemic adverse events and concomitant 
medication use.  Measurement of vital signs and physical examination will be performed if 
clinically indicated and laboratory samples will be taken at the time-points indicated in the 
schedule of attendances. An appointment will be made for the next follow up visit.  
 
 Group 1 
Screening 
Inoculation and follow up 
Timeline (days) 
≤ 90 
0 
4 
5 
14 
32 
TOPS confirmation 
+ 
 
 
 
 
 
Visit window 
 
 
0 
0 
+/- 2  
+/- 2  
Volunteer Information Sheet 
+ 
 
 
 
 
 
Informed consent 
+ 
 
 
 
 
 
Vital signs 
+ 
+ 
(+) 
(+) 
(+) 
(+) 
Medical history 
+ 
 
 
 
 
 
Physical examination 
+ 
(+) 
(+) 
(+) 
(+) 
(+) 
Pregnancy test (females only) 
+ 
+ 
+ 
 
 
 
Electrocardiogram 
+ 
 +a 
 
 
 
 
Review eligibility 
 
+ 
 
 
 
 
Inoculation 
 
+ 
 
 
 
 
Eradication 
 
 
+ 
 
 
 
Review of adverse events and 
concomitant medications 
 
 
+ 
+ 
+ 
+ 
Nasal wash 
+ 
 
 
 
+ 
+ 
Throat swab x 2 
  + b 
+ 
+ 
+ 
  + b 
 + b 
Nasosorption test 
 
+ 
 
 
+ 
+ 
Saliva sample 
 
+ 
 
 
+ 
+ 
Environmental samples 
 
 
+ 
  +c 
  + c 
  + c 
Immunological blood tests (ml) 
 
20 
 
 
20 
20 
Cumulative blood volume (ml) 
 
20 
 
 
40 
60 
Table 10.1: Summary of monitoring procedures during the study – Group 1; (+) if clinically 
indicated, aif screening ECG not available, bThroat swab 2 optional, cif colonised at previous 
visit 
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 24 
 
 Group 2 
Screening 
Inoculation and follow up 
Timeline (days) 
≤ 90 
0 
4 
7 
14 
15 
24 
42 
Visit window (days) 
 
 
0 
+/-2 
+/-2  
 0*  
+/-2 
+/-2 
TOPS confirmation 
+ 
 
 
 
 
 
 
 
Volunteer Information Sheet 
+ 
 
 
 
 
 
 
 
Informed consent 
+ 
 
 
 
 
 
 
 
Vital signs 
+ 
+ 
(+) 
(+) 
(+) 
(+) 
(+) 
(+) 
Medical history 
+ 
 
 
 
 
 
 
 
Physical examination 
+ 
(+) 
(+) 
(+) 
(+) 
(+) 
(+) 
(+) 
Pregnancy test (females only) 
+ 
+ 
 
 
+ 
 
 
 
Electrocardiogram 
+ 
 +a 
 
 
 
 
 
 
Review eligibility 
 
+ 
 
 
 
 
 
 
Inoculation 
 
+ 
 
 
 
 
 
 
Eradication 
 
 
 
 
+ 
 
 
 
Review of adverse events and 
concomitant medications 
 
 
+ 
+ 
+ 
+ 
+ 
+ 
Nasal wash 
+ 
 
 
 
+ 
 
 
+ 
Throat swab  x 2 
 +b 
+ 
+ 
  + b 
+ 
+ 
  + b 
  + b 
Nasosorption test 
 
+ 
 
 
+ 
 
 
+ 
Saliva sample 
 
+ 
 
 
+ 
 
 
+ 
Environmental samples 
 
 
+ 
 + c 
 + c 
 + c 
 + c 
 + c 
Immunological blood tests (ml) 
 
20 
 
 
20 
 
 
20 
Cumulative blood volume (ml) 
 
20 
 
 
40 
 
 
60 
Table 10.2: Summary of monitoring procedures during the study – Group 2; (+) if clinically 
indicated, *1 day following D14 visit, aif screening ECG not available, bThroat swab 2 
optional, cif colonised at previous visit, c1 day following D14 visit 
10.1 Eradication therapy 
Colonisation with N. lactamica will be eradicated by treatment with ciprofloxacin 500 mg 
single oral dose (BNF 2013, p350). Ciprofloxacin proved effective at eradicating N. 
meningitidis (Fraser, Gafter-Gvili et al. 2006) and should eradicate N. lactamica carriage 
within 24 hours (Pugsley, Dworzack et al. 1987). The strain used is fully sensitive to this 
antibiotic with an exceptionally low MIC on E-testing. 
Eradication therapy will be given at day 4 for group 1 and day 14 for group 2. Eradication 
efficacy will be analysed by taking a throat swab 24 hours, 10 days and 28 days after 
eradication therapy (Day 5, 14 and Day 32 for Group 1, Day 15, 24 and Day 42 for Group 2). 
Female volunteers will have a further pregnancy test prior to administering eradication 
therapy as pregnancy is a relative contraindication to its use. If a volunteer is found to be 
pregnant at this point they will not receive eradication therapy and will be withdrawn from the 
study. 
10.1.1 Compliance with the eradication therapy 
The dose will be taken under supervision of the study team (directly observed treatment)  

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 25 
 
 
10.1.2 Side effects of the eradication therapy 
The side effects of ciprofloxacin in young adults may include:  
- 
Abdominal ache, diarrhoea and nausea.  
- 
Tiredness and headaches.  
- 
Rash and itching.  
- 
Facial swelling - very rarely breathing difficulties may occur with the facial swelling. 
- 
Pain and inflammation around the joints 
Volunteers should contact the clinical study team if this occurs.  
Female volunteers using the oral contraceptive pill will be advised to use alternative forms of 
contraception for two weeks following eradication therapy as ciprofloxacin may interfere with 
the oral contraceptive pill. 
10.2 Emergency contact 
Subjects will be encouraged to contact one of the investigators on the 24 hour emergency 
mobile study telephone number if they develop symptoms between the regular follow up 
visits. The investigator will consider an extra clinical review if the volunteer has any 
symptoms that are moderate or severe. 
11 Laboratory procedures 
11.1 Laboratory work 
Standard operating procedures for all laboratory work will be followed. Investigators will 
conform to well established laboratory safety standards. 
11.2 Processing and storage of samples 
Blood samples 
Blood samples for immunological assays will be processed in the NIHR-WTCRF lab and 
frozen at -80ºC.  
Immunological assays (serology and cellular immunology specific to N. lactamica) will be 
conducted according to the procedures established in the test laboratories.  
Nasal washes, nasosorption test strips and saliva samples 
Nasal wash fluid, nasosorption test strips and saliva samples will be processed in the NIHR-
WTCRF lab and frozen at -80ºC and then analysed according to the procedures established 
in the test laboratories. 
Throat swab 1 – bacterial culture and PCR 
One throat swab at each time point will be placed into medium and centrifuged. The resulting 
solution will be spot plated and then incubated for up to 48hrs prior to the identification 
process. Positively identified Neisseria species will then be sub-cultured onto fresh plates for 
up to 24hrs. Individual colonies will be selected for storage in glycerol medium and frozen to 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 26 
-80ºC to await PCR and sequencing for exact characterisation. The remainder of the solution 
will be frozen at -80ºC for a RT PCR assay which will be conducted according to procedures 
established in the test laboratory.  
Throat swab 2 – Microbiome analysis 
The second throat swab will be applied directly to an FTA card for DNA preservation, stored 
in a pouch with desiccant pack then frozen in a -80ºC freezer for future microbiome analysis. 
Environmental samples 
Mask portions and air sample cultures will be analysed for viable count or frozen at -80ºC 
freezer for future RTPCR.  
11.3 Labelling of samples 
Samples will be clearly identified with the study code, volunteer’s unique anonymised 
identifier, sample ID and time point, and recorded in the study log. Samples will not be 
labelled with any personal identifiable information. 
11.4 Microbiological sampling 
Estimates of population density will be measured at each sampling point. Positive 
colonisation will be defined as the culture of at least one isolate from day 4 onwards after 
inoculation (Deasy, Guccione et al. 2015). We will record the duration of positive colonisation 
and the effectiveness of eradication therapy in clearing colonisation in 24 hours.  
11.5 Immunological analyses 
N. lactamica specific IgG will be measured using previously described methods. Cellular 
immunity will be assessed according to the procedures established in the test laboratories. 
Nasal fluid, collected by a nasosorption test, will be analysed for specific cytokine profiles. 
Nasal washes will be analysed for N.lactamica specific IgA and IgG. Saliva samples will be 
analysed for specific IgA. Other potentially relevant immunological assays may be performed 
on stored samples at the discretion of the investigator. 
12. Assessment of safety  
Safety of the volunteers will be assessed by analysing the frequency, incidence and nature 
of adverse events and serious adverse events arising during the study.  
12.1 Definitions  
Adverse Event (AE) 
An AE is any untoward medical occurrence in a volunteer, including a dosing error, which 
may occur during or after administration of the inoculum and does not necessarily have a 
causal relationship with the intervention. An AE can therefore be any unfavourable and 
unintended sign (including an abnormal laboratory finding), symptom or disease temporally 
associated with the study intervention, whether or not considered related to the study 
intervention.  
Adverse Reaction (AR) 
An AR is any untoward or unintended response to the inoculum. This means that a causal 
relationship between the inoculum and an AE is at least a reasonable possibility, i.e., the 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 27 
relationship cannot be ruled out. All cases judged by either the reporting medical investigator 
or the sponsors as having a reasonable suspected causal relationship to the inoculum (i.e. 
possibly, probably or definitely related to the inoculum) will qualify as adverse reactions.  
Unexpected Adverse Reaction (UAR) 
An adverse reaction, the nature or severity of which is not consistent with the applicable 
information about the inoculum in the protocol, is considered as an unexpected adverse 
reaction.  
Serious Adverse Event (SAE) 
An SAE is an AE that results in any of the following outcomes, whether or not considered 
related to the study intervention.  
• 
Death (i.e., results in death from any cause at any time)  
• 
Life-threatening event (i.e., the volunteer was, in the view of the investigator, at 
immediate risk of death from the event that occurred). This does not include an AE 
that, if it occurred in a more serious form, might have caused death.  
• 
Persistent or significant disability or incapacity (i.e. substantial disruption of one’s 
ability to carry out normal life functions).  
• 
Hospitalisation other than admission in the NIHR-WTCRF, regardless of length of 
stay, even if it is a precautionary measure for continued observation. Hospitalisation 
(including inpatient or outpatient hospitalisation for an elective procedure) for a pre-
existing condition that has not worsened unexpectedly does not constitute a serious 
AE.  
• 
An important medical event (that may not cause death, be life threatening, or require 
hospitalisation) that may, based upon appropriate medical judgment, jeopardise the 
volunteer and/or require medical or surgical intervention to prevent one of the 
outcomes listed above. Examples of such medical events include allergic reaction 
requiring intensive treatment in an emergency department or clinic, blood dyscrasias, 
or convulsions that do not result in inpatient hospitalisation.  
• 
Congenital anomaly or birth defect.  
Serious Adverse Reaction (SAR) 
An adverse event (expected or unexpected) that is both serious and, in the opinion of the 
reporting investigator or sponsors, believed to be possibly, probably or definitely due to the 
inoculum or any other study treatments, based on the information provided in the protocol.  
Suspected Unexpected Serious Adverse Reactions (SUSARs) 
A SUSAR is a SAE that is unexpected and thought to be possibly, probably or definitely 
related to the inoculum.  
12.2 Causality assessment 
For each AE, an assessment of the relationship of the AE to the study intervention(s) will be 
undertaken. The relationship of the adverse event with the study procedures will be 
categorised as unrelated, unlikely to be related, possibly related, probably related or 
definitely related (Table 12.1). An intervention-related AE refers to an AE for which there is a 
possible, probable or definite relationship to the study intervention. The investigator will use 
clinical judgment to determine the relationship. Alternative causes of the AE, such as the 
natural history of pre-existing medical conditions, concomitant therapy, other risk factors and 
the temporal relationship of the event to the challenge will be considered and investigated. 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 28 
 
0  
 
No Relationship 
No temporal relationship to the challenge and  
Alternate aetiology (clinical state, environmental or other 
interventions); and  
Does not follow known pattern of response to N.lactamica  
1  Unlikely  
Unlikely temporal relationship to the challenge and  
Alternate aetiology likely (clinical state, environmental or other 
interventions) and  
Does not follow known pattern of response to N.lactamica 
2  Possible  
Reasonable temporal relationship to the challenge; or  
Event not readily produced by clinical state, environmental or 
other interventions; or  
Follows expected pattern of response to N.lactamica 
3  Probable  
Reasonable temporal relationship to the challenge; and  
Event not readily produced by clinical state, environment, or other 
interventions or  
Follows expected pattern of response to N.lactamica 
4  Definite  
Reasonable temporal relationship to the challenge; and  
Event not readily produced by clinical state, environment, or other 
interventions; and  
Follows expected pattern of response to N.lactamica 
Table 12.1: Guidelines for assessing the relationship of an AE to inoculation with N. 
lactamica 
12.3 Reporting procedures for AEs 
AEs will be recorded in the patient file and included in the annual safety report. 
AEs that result in a volunteer’s withdrawal from the study or that are present at the end of 
the study will be followed up (if volunteer consents to this) until a satisfactory resolution or 
stabilisation occurs, or until a non-study related causality is assigned. 
12.3.1 Severity grading of clinical adverse events 
The severity of clinical and laboratory adverse events will be assessed according to the 
scales in table 12.2. 
GRADE 0  
None  
GRADE 1  
Mild: Transient or mild discomfort (< 48 hours); no medical intervention/therapy 
required 
GRADE 2  
Moderate: Mild to moderate limitation in activity - some assistance may be 
needed; no or minimal medical intervention/therapy required  
GRADE 3 
Severe: Marked limitation in activity, some assistance usually required; medical 
intervention/therapy required, hospitalisation possible  
Table 12.2: Severity grading criterion for AEs. 
12.3.2 Reporting procedures for serious AEs (SAEs) 
In order to comply with current regulations on serious adverse event reporting to regulatory 
authorities, the event will be documented accurately and notification deadlines respected. 
SAEs will be reported to the Principal Investigator immediately when the study team is aware 
of their occurrence, as described in the relevant SOP. The external safety committee will be 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 29 
notified of SAEs deemed possibly, probably or definitely related to study interventions; the 
sponsor will be notified immediately (within 24 hours) when the investigators are aware of 
their occurrence. SAEs will not normally be reported to the ethical committee(s) unless there 
is a clinically important increase in occurrence rate, an unexpected outcome, or a new event 
that is likely to affect safety of trial volunteers, at the discretion of the Chief Investigator. In 
addition to the expedited reporting above, the investigator shall include all SAEs in the 
annual Development Safety Update Report (DSUR) report.  
12.3.3 Reporting procedures for SUSARs 
The chief investigator will report all SUSARs to the ethical committee(s) within required 
timelines. The chief investigator will also inform all investigators concerned of relevant 
information about SUSARs that could adversely affect the safety of participants. In addition, 
the chief investigator will report any SUSARs relating to licensed products used in the trial 
(ciprofloxacin) to the Medicine and Healthcare products Regulatory Agency (MHRA) using 
the electronic ‘Yellow Card’ System. 
All SUSARs and deaths occurring during the study will be reported to the sponsor. For all 
deaths, any autopsy reports and relevant medical reports will be made available for reporting 
to the relevant authorities.  
12.3.4 Procedures to be followed in the event of abnormal findings 
Abnormal clinical findings from medical history, examination or blood tests, will be assessed 
as to their clinical significance. If a test is deemed clinically significant, it may be repeated, to 
ensure it is not a single occurrence. If a test remains clinically significant, the volunteer will 
be informed and appropriate medical care arranged as appropriate with the permission of 
the volunteer. Decisions to exclude the volunteer from enrolling in the trial or to withdraw a 
volunteer from the trial will be at the discretion of the Investigator.  
12.3.5 Foreseeable medical occurrences 
The following medical occurrences are foreseeable:  
• 
Local sensation effects in the nose following inoculation. 
• 
Local bruises following venesection 
• 
Adverse reactions to ciprofloxacin as detailed in the Summaries of product 
characteristics (SmPC) 
12.3.6 Adverse events of special interest 
Adverse events of special interest will be reported as SAEs. These are:  
• 
Severe hypersensitivity reactions to the inoculum (e.g. anaphylaxis) 
• 
Overdosing of the inoculum  
12.4 Safety profile review 
The safety profile will be assessed on an on-going basis by the investigators.  
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 30 
12.5 Study committees 
12.5.1 External Safety Committee (ESC) 
An external safety committee will be appointed prior to recruitment.  
The role of the ESC is to provide overall supervision for the trial and provide advice through 
its independent Chair. The ultimate decision for the continuation of the trial lies with the Chief 
investigator following advice from the ESC.   
The ESC will be responsible for reviewing and assessing this protocol prior to 
commencement of the trial, recruitment, interim monitoring of safety and effectiveness, trial 
conduct and external data. The ESC will first convene prior to trial initiation and will then 
define the frequency of subsequent meetings (at least annually).  
All correspondence between investigator and ESC will be conveyed by the investigator to 
the trial Sponsor. The study protocol and implemented safety procedures will be discussed 
with the ESC before starting the study. 
The chair of the ESC may be contacted for advice and independent review by the 
investigator or trial Sponsor in the following situations:  
• 
Following any SAE deemed to be possibly, probably, or definitely related to a study 
intervention.  
• 
Any other situation where the Investigator or trial sponsor feels independent advice 
or review is important.  
12.5.2 Ethics Committee 
The study will be reviewed by the HRA. A progress report will be submitted to the REC 12 
months after the date on which the favourable opinion on which the form was given. 
13. Analysis and Statistical considerations 
13.1 Endpoints of the study 
Primary endpoint 
• 
Rise in serological specific antibody titre comparing day 0 versus day 14 and 28 days 
post eradication and comparing early (day 4) versus late (day 14) eradication 
 
Secondary endpoints: 
• 
Assessment of colonisation – culture of N. lactamica from throat swabs at day 4 for 
group 1 and days 4, 7 and 14 for group 2 
• 
Assessment of eradication – culture of N. lactamica from throat swabs on the day of 
eradication, one day, 10 days and 28 days after eradication 
• 
Rise in mucosal specific antibody titre comparing day of screening versus day 14 and 
28 days post eradication and comparing early (day 4) versus late (day 14) 
eradication 
• 
Change in nasal cytokine profile comparing day 0 versus day 4, 14 and 28 days post 
eradication and comparing early (day 4) versus late (day 14) eradication 
• 
Difference in environmental shedding between short (group 1) and longer (group 2) 
periods of colonisation. 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 31 
Safety endpoints:  
• 
Occurrence of unsolicited adverse events within the study period 
• 
Occurrence of serious adverse events within the study period 
Safety analysis will be carried out for all volunteers that received the inoculum, regardless of 
whether or not they complete the study. 
13.2 Sample size 
The previous experimental N. lactamica challenge study showed a significant rise in 
serological antibody titre against N. lactamica over 2 weeks (Deasy, Guccione et al. 2015). 
This gave SDs on a log-10 scale of 0.11 for IgA saliva and 0.26 for serum total IgG. For this 
study, using the SD of 0.26 we will be able to confirm a 4 fold rise with 10 carriers of N. 
lactamica with 90% power using analysis of variance. 
Allowing for a drop out rate of approximately 10%, we will therefore recruit volunteers until 
we have 11 individuals colonised for each group up to a maximum of 22 volunteers for each 
group. At a dose of 104 colony forming units (cfu), long lasting colonisation with N. lactamica 
is easily induced in 35-65% of participants. We showed that increasing the inoculum to 105 
cfu increased the subsequent carriage of N. lactamica to 50% (Evans, Pratt 2011). Therefore 
approximately 44 individuals will be enrolled.  Assuming a screening failure rate of 5%, we 
will need to screen approximately 46 potential participants to achieve colonisation in 11 of 
each group.  
13.3 Statistical analysis 
Statistical analysis will be performed by the study team and Dr Nick Andrews, Public Health 
England, Colindale. Differences between groups at each time point will be assessed using 
Fisher exact test and between time points within groups by McNemar test. 
14. Study quality and Management procedures 
14.1 Investigator procedures 
Approved site-specific SOPs will be used at all clinical and laboratory sites.  
14.2 Monitoring  
Monitoring will be performed by the Sponsor according to International Conference on 
Harmonisation (ICH) Good Clinical Practice (GCP). Following written standard operating 
procedures, the monitors will verify that the clinical trial is conducted and data are generated, 
documented and reported in compliance with the protocol, GCP and the applicable 
regulatory requirements. The investigator sites will provide direct access to all trial related 
source data/documents and reports for the purpose of monitoring and auditing by the 
sponsor and inspection by local and regulatory authorities.  
14.3 Study amendments 
No amendments to this protocol will be made without consultation with, and agreement of, 
the Sponsor. Any amendments to the trial that appear necessary during the course of the 
trial must be discussed by the investigator and sponsor concurrently. If agreement is 
reached concerning the need for an amendment, it will be produced in writing by the chief 
investigator and will be made a formal part of the protocol following ethical and regulatory 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 32 
approval (NRES-REC SOPs – Version 5.1 March 2012: http://www.hra.nhs.uk/wp-
content/uploads/2013/08/NRES_SOPs_v5.1_2012.03.14.pdf).  
An administrative change to the protocol is one that modifies administrative and logistical 
aspects of a protocol but does not affect the subjects’ safety, the objectives of the trial and 
its progress. An administrative change does not require UK ethical committee or regulatory 
approval. 
Any amendments to study documents will follow established HRA and REC requirements.  
The investigator is responsible for ensuring that changes to an approved trial, during the 
period for which regulatory and ethical committee(s) approval has already been given, are 
not initiated without regulatory and ethical committee(s)’ review and approval except to 
eliminate apparent immediate hazards to the subject.  
14.4 Protocol deviation 
Any deviations from the protocol will be documented in a protocol deviation form and filed in 
the site trial master file.  
14.5 Quality Control, Quality Assurance and statuatory inspection 
The UHS R&D department QA staff will provide Quality Assurance (QA) for the trial and 
perform internal audits to check that the trial is being conducted, data recorded, analysed 
and accurately reported according to the protocol, Sponsor’s SOPs and in compliance with 
ICH GCP. The audits will also include laboratory activities according to an agreed audit 
schedule. The internal audits will supplement the sponsor’s monitoring process and will 
review processes not covered by the sponsor’s monitor.  
A Quality control (QC) plan will be established at the start of the trial, as per local SOP. 
The Sponsor, trial site and ethical committee may carry out audit to ensure compliance with 
the protocol, GCP and appropriate regulations. GCP inspections may also be undertaken by 
the regulatory authority to ensure compliance with protocol and national regulations. The 
sponsor will assist in any inspections.  
14.6 Serious breaches 
A serious breach is defined as “A breach of GCP or the trial protocol which is likely to affect 
to a significant degree – the safety or physical or mental integrity of the subjects of the trial; 
or the scientific value of the trial.”  
In the event that a serious breach is suspected the Sponsor will be informed as soon as 
possible and in turn will notify the REC and external safety committee within 7 days.  
14.7 Study progress  
The progress of the trial will be overseen by the Chief Investigator.  
14.8 Study completion/termination 
The trial will be considered complete upon the last volunteer/last visit at the site. The data 
will be sent to the sponsor in the timeframe specified in the Clinical Trial Agreement.  

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 33 
The study may be terminated early at the discretion of the Chief Investigator, Sponsor or 
External Safety Committee if there are safety concerns, concerns about compliance with 
GCP or other appropriate regulations, poor recruitment or new information becomes 
available which has an impact on the scientific validity or safety of the trial. 
14.9 Exploitation and dissemination 
The investigators will be involved in reviewing drafts of the manuscripts, abstracts, press 
releases and any other publications arising from the study. Findings will be published in peer 
reviewed journals as soon as possible, even where results prove negative. The authors will 
acknowledge that the study is funded by the NIHR Southampton Respiratory Biomedical 
Research Unit. The results of the study will be disseminated at relevant international 
scientific meetings. Volunteers will be sent a newsletter with a lay summary of the results of 
the study once the results are available. This will provide information regarding the study in 
general and not their individual results. 
15. Ethics  
15.1 Declaration of Helsinki 
The Investigator will ensure that this study is conducted according to the principles of the 
current revision of the Declaration of Helsinki 2008.  
15.2 ICH guidelines for good clinical practice  
The Investigator will ensure that this study is conducted in full conformity to the ICH 
guidelines for GCP (CPMP/ICH/135/95) July 1996.  
15.3 Informed consent  
Written informed consent will be gained from all participants and their sleeping partners 
following the provision of detailed information about the aims of the study, the level of 
involvement required, and the risks involved. Participants will be provided with an 
information sheet prior to the start of the study either in print form or via email. They will be 
encouraged to use the contact details on this form to contact the research team to get further 
information if necessary. Prior to screening the participants’ understanding of the study and 
risks involved will be explored and they will be asked to sign a consent form. 
15.4 Informing participants’ General Practitioners 
A letter describing the study and the participant’s involvement will be sent to their General 
Practitioner (GP) on the day of the screening visit. This will include contact details for the 
research team.  
15.5 Research ethics committee 
A copy of the protocol, proposed informed consent form, other written volunteer information 
and the proposed advertising material will be submitted to the REC and HRA for written 
approval, using the UK Integrated Research Application System. The investigator will submit 
and, where necessary, obtain approval from the REC for all subsequent amendments to the 
protocol and associated trial documents. A non-substantial amendment does not require UK 
ethical committee approval (NRES-REC SOPs – Version 5.1 March 2012: 
http://www.hra.nhs.uk/wp-content/uploads/2013/08/NRES_SOPs_v5.1_2012.03.14.pdf). The 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 34 
investigator will notify deviations from the protocol or SAEs occurring at the site to the 
sponsor and will notify the REC of these if necessary in accordance with procedures.  
15.6 Volunteer confidentiality 
All data will be anonymised; volunteer data will be identified by a unique study number in 
CRF and database. Separate confidential files containing identifiable information will be 
stored in secured locations. Only the sponsor representative, investigators, the clinical 
monitor, the ethical committee(s) and the regulatory authorities will have access to the 
records.  
16. Data handling and record keeping 
16.1 Data handling 
The chief investigator will be the data manager with responsibility for delegating the 
receiving, entering, cleaning, querying, analysing and storing of all data that accrues from 
the study in the site file held in the NIHR-WTCRF. The investigators will enter the data into 
the volunteers’ CRFs, which will be in a paper format. This includes safety data, laboratory 
data (both clinical and immunological) and outcome data. 
16.2 Record keeping 
The investigators will maintain and retain appropriate medical and research records and 
essential documents for this trial in compliance with ICH E6 GCP and regulatory and 
institutional requirements for the protection of confidentiality of volunteers. The chief 
investigator, co-investigators and clinical research nurses will have access to records. The 
investigators will permit authorised representatives of the sponsor, regulatory agencies and 
the monitors to examine (and when required by applicable law, to copy) clinical records for 
the purposes of quality assurance reviews, audits and evaluation of the study safety and 
progress.  
16.3 Source data and case report forms (CRFs)  
All protocol-required information will be collected in CRFs designed by the investigator. All 
source documents, excluding hospital records, will be filed in the CRF. Source documents 
are original documents, data, and records from which the volunteer’s CRF data are obtained. 
For this study these will include, but are not limited to; volunteer consent form, blood results, 
GP response letters, laboratory records and correspondence. In the majority of cases, CRF 
entries will be considered source data as the CRF is the site of the original recording (i.e. 
there is no other written or electronic record of data). In this study this will include, but is not 
limited to medical history, medication records, vital signs, physical examination records, 
urine assessments, blood results, adverse event data and details of study interventions. All 
source data and volunteer CRFs will be stored securely.  
16.4 Data protection 
The study protocol, documentation, data and all other information generated will be held in 
strict confidence. No information concerning the study or the data will be released to any 
unauthorised third party, without prior written approval of the sponsor.  

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 35 
17. Financing and insurance 
17.1 Financing  
The study will be funded by the NIHR Southampton Respiratory Biomedical Research Unit 
and will be supported by funding for Experimental Medicine by the National Institutes of 
Health Research through support from the Southampton NIHR Wellcome Trust CRF, the 
Wessex Comprehensive Research Network. 
17.2 Insurance  
 The University of Southampton has a specialist insurance policy in place, which would 
operate in the event of any participant suffering harm as a result of their involvement in the 
research. 
17.3 Compensation for time 
Volunteers will be compensated for their time and for the inconvenience caused by 
procedures as below.  
• 
Attending screening and follow-up sessions - £21 total (£15/visit plus additional £6 
travel expenses) 
• 
1 x screening 
• 
1x challenge 
• 
Up to 6 follow up visits 
 
The maximum individual volunteers will be compensated is £168 and the minimum £15 
If volunteers withdraw from the study prior to its completion they will be offered financial re-
imbursement corresponding to the number of visits attended. 
 
 

Pilot N. lactamica challenge study Protocol v5.0, 26th April 2017 
Sponsor Code RHM MED1354, IRAS ID 207453 
Page 36 
18. References 
Bidmos, F. A., K. R. Neal, et al. (2011). "Persistence, replacement, and 
rapid clonal expansion of meningococcal carriage isolates in a 
2008 university student cohort." J Clin Microbiol 49(2): 506-512. 
Brown, N. M., N. K. Ragge, et al. (1987). "Septicaemia due to Neisseria 
lactamica--initial confusion with Neisseria meningitidis." J Infect 
15(3): 243-245. 
Davenport, V., T. Guthrie, et al. (2003). "Evidence for naturally acquired 
T cell-mediated mucosal immunity to Neisseria meningitidis." J 
Immunol 171(8): 4263-4270. 
Deasy, A. M., E. Guccione, et al. (2015). "Nasal Inoculation of the 
Commensal Neisseria lactamica Inhibits Carriage of Neisseria 
meningitidis by Young Adults: A Controlled Human Infection 
Study." Clin Infect Dis 60(10): 1512-1520. 
Denning, D. W. and S. S. Gill (1991). "Neisseria lactamica meningitis 
following skull trauma." Rev Infect Dis 13(2): 216-218. 
Evans, C. M., C. B. Pratt, et al. (2011). "Nasopharyngeal colonization by 
Neisseria lactamica and induction of protective immunity against 
Neisseria meningitidis." Clin Infect Dis 52(1): 70-77. 
Fraser, A., A. Gafter-Gvili, et al. (2006). "Antibiotics for preventing 
meningococcal infections." Cochrane Database Syst Rev(4): 
CD004785. 
Glennie, S. J., D. Banda, et al. (2012). "Regulation of naturally acquired 
mucosal immunity to Streptococcus pneumoniae in healthy 
Malawian adults and children." PLoS One 7(12): e51425. 
Lauer, B. A. and C. E. Fisher (1976). "Neisseria lactamica meningitis." 
Am J Dis Child 130(2): 198-199. 
Pugsley, M. P., D. L. Dworzack, et al. (1987). "Efficacy of ciprofloxacin in 
the treatment of nasopharyngeal carriers of Neisseria 
meningitidis." J Infect Dis 156(1): 211-213. 
Vaughan, A. T., A. Gorringe, et al. (2009). "Absence of mucosal 
immunity in the human upper respiratory tract to the commensal 
bacteria Neisseria lactamica but not pathogenic Neisseria 
meningitidis during the peak age of nasopharyngeal carriage." J 
Immunol 182(4): 2231-2240. 
Wright, A. K., M. Bangert, et al. (2013). "Experimental human 
pneumococcal carriage augments IL-17A-dependent T-cell 
defence of the lung." PLoS Pathog 9(3): e1003274.