Immunomodulatory Effect of Pleuran (β-glucan From Pleurotus Ostreatus) in Children With Recurrent Respiratory Tract Infections
Phase: NA
Conditions: Respiratory Tract Infection
Sponsor: Pleuran, s.r.o.

IPRRTI study Version 1 – 30.6.2023 
 
Immunomodulatory effect of pleuran (β-glucan from Pleurotus ostreatus) in 
children with recurrent respiratory tract infections 
IPRRTI study 
Study Protocol 
NCT number: NA 
Basic information about the study 
Timeline 
Pre-screening of patients: July-September 2023 
Date of enrolment of the first patient: 15.09.2023 
Date of enrolment of the last patient: 31.12.2023 
Study completion date: 31.03.2024 
Title of the study: 
Immunomodulatory effect of pleuran (β-glucan from Pleurotus ostreatus) in 
children with recurrent respiratory tract infections (RRTI). 
Study abbreviation: 
IPRRTI 
Principal 
investigator and 
site: 
 
Co-investigators 
and sites: 
 
Study design: 
Multicenter, prospective, randomized, double-blind, placebo-controlled study 
with a food supplement. 
Study population: 
250 patients aged 3-18 years enrolled by investigators at 36 centers, 
randomized to the active and placebo groups (1:1 ratio) 
Primary endpoints: 
To evaluate the effect of pleuran administration (Imunoglukan P4H® chewable 
tablets) compared to the placebo group on: 
• 
reduction in the number of RTIs episodes (total number) 
Secondary 
endpoints: 
To evaluate the effect of pleuran administration (Imunoglukan P4H® chewable 
tablets) compared to the placebo group on: 
• 
reduction in the number of episodes of RTI subtypes 
• 
reduction in the duration of RTI episodes (total duration, RTI subtypes) 
• 
reduction of the need for antibiotic (ATB) therapy 
• 
reduction of the number of missed days at school/nursery due to RTI 
• 
reduction of the number of missed working days due to RTIs 
• 
reduction of the number of emergency department visits due to RTI 
• 
reduction of the number of physician visits due to RTI 
• 
tolerability and safety 

Immunomodulatory effect of pleuran (β-glucan from Pleurotus ostreatus) in children with recurrent respiratory tract infections. 
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Study duration and visit plan: 3 months, 4 visits in total: V0 – initial visit, V1 – control visit after 30±7 
days from enrolment, V2 – control visit after 60±7 days from enrolment and V3 – control visit after 
90±14 days from enrolment (completion).  
Introduction 
Respiratory tract infections (RTIs) are the most common cause of morbidity at every age category, 
especially in childhood. At the same time, they are also the most common cause of consultations and 
visits in clinical practice of every pediatrician. Upper respiratory tract infections have a higher 
prevalence, and about 10-30% of patients develop lower respiratory tract infections. The most 
common cause are viruses (90-95%). 
Respiratory infections are characterized by both microbial load, respiratory epithelium response, and 
activation of inflammation and immune responses. These aspects lead to structural and functional 
changes in the respiratory tract. There is an overgrowth of goblet-shaped glands, which leads to 
excessive production of hyperviscous mucus. There is a weakening of mucociliary clearance, either due 
to the direct toxic influence of microorganisms on cilia and their oscillation, but also indirectly through 
an increase in the viscosity of mucus in the lumen of the respiratory tract. Respiratory epithelial cells 
form many pro-inflammatory and chemotactic compounds that lead to the development of 
inflammatory infiltrate in the airways. This is accompanied by oxidative stress, which impairs both the 
structure and function of biomolecules. Inflammation subsequently increases the reactivity of cough 
receptors. All these factors cause clinical signs of respiratory infections, whether from the respiratory 
tract (cough, expectoration, obstruction with wheezing, bronchial hyperreactivity, congestion and 
runny nose) or at the level of the whole organism (increased body temperature, weakness, myalgia 
and fatigue). 
Recurrent respiratory tract infections (RRTIs) 
RRTIs are a separate category of RTIs and represent a typical medical problem of preschool and school 
age. They can be defined on the basis of several aspects as infectious respiratory diseases, in which 
other more serious causes have been excluded (cystic fibrosis, primary immune disorders, congenital 
ciliary dyskinesias, respiratory malformations). Most children with RRTIs do not suffer from severe 
immunodeficiency and the high frequency of respiratory infections is due to a combination of various 
factors (immaturity of the immune system, high exposure to infectious agents in the family or 
children's collective, environmental factors, adenoid hypertrophy, etc.). In addition, allergic 
inflammation in the airways may also contribute to increased susceptibility to RTIs. 
According to data from epidemiological studies, approximately 6-10% of children between 2-6 years 
of age suffer from RRTIs. Due to their high incidence, RRTIs have an important socio-economic impact 
(affecting quality of life, missed school days, missed days at work, repeated medical examinations, 
hospitalization, as well as increased antibiotic prescription).  
However, it is equally important to define what the term RRTIs means, as the parent’s opinion and a 
doctor's point of view can differ significantly. When assessing RRTIs, in addition to their frequency, 
their course, duration, response to pharmacological treatment, cultivation of the pathogen as well as 
the development of complications are much more important. At the same time, the so-called 
"physiological morbidity of the child" must also be taken into account, which is considered to be: 
• 
Children 3-5 years: 6 – 8 RTIs with mild course, good response to treatment, predominantly 
during the autumn and winter months 
• 
Children 6-12 years: 2 – 4 RTIs with mild course, good response to treatment, predominantly 
during the autumn and winter months  

Immunomodulatory effect of pleuran (β-glucan from Pleurotus ostreatus) in children with recurrent respiratory tract infections. 
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The treatment of RRTIs consists not only from the treatment of each infection, but should be mainly 
aimed at their prevention. Therapeutic and preventive strategies should aim to support weakened 
immune system functions, e.g. by immunomodulation. Immunomodulation can be defined as 
therapeutic or preventive intervention in the activity of the immune system in order to achieve 
homeostasis.  
In practice, various preparations of natural origin are used to prevent RRTIs, but only few of them have 
scientific evidence proving efficacy with a sufficient number of relevant clinical studies and 
observations that elucidate individual aspects of the mechanisms of action as well as proven efficacy 
and safety in the management of individual diseases. Vitamins, minerals, as well as many natural 
immunomodulatory 
preparations 
(phytotherapy, 
probiotics, 
prebiotics, 
biologically 
active 
polysaccharides) are most commonly used, while individual biological effects are important – anti-
infectious, immunomodulatory, anti-inflammatory, antioxidant, etc. By using these preparations or 
their combination, a synergy of effects can be obtained in the treatment of the selected condition. 
Biologically active polysaccharides (BAP e.g., β-glucans - polymers of glucose commonly found in 
fungi, yeast, bacteria and cereals) represent one of the most studied natural immunomodulators with 
pluripotent biological effects (anti-infectious, antiviral, immunomodulatory). Selected molecules have 
an anti-inflammatory effect, inhibit the production of pro-inflammatory cytokines, and their 
preventing effect on lung damage was described in an animal model. By promoting the production of 
anti-inflammatory and regulatory substances, they restore balance in the cytokine network. The 
antiviral effect of BAP can be mediated either directly, through inhibition and/or disruption of viral 
particle integrity, or indirectly, by enhancing host antiviral defenses by supporting the body's immune 
processes.  Other significant effects of β-glucans include their significant immunomodulatory effect 
(increasing the number of NK cells, stimulating phagocytic activity, promoting the development of 
antibody production, improving the post-vaccination response) and modulating effect on T-
lymphocytes. The mechanism of action of β-glucans is mediated by several receptors, the most 
important is the Dectin-1 receptor.  
Zinc application may also have a possible positive effect on the immune system, as several studies have 
suggested its effect in coronavirus infections. In vitro, zinc inhibits RNA-polymerase activity as well as 
virion replication. Vitamin D is also important for the functioning of the immune system, its positive 
effect has been observed in protection against infections (antibacterial effect, anti-inflammatory 
effect, differentiation of monocytes into macrophages), tumor diseases (anti-proliferative and 
antidifferentiation effect), and its deficiency may be a risk factor in the development of 
immunopathological reactions (autoimmune diseases). 
The efficacy and safety of food supplement Imunoglukan P4H® (pleuran, β-glucan from Pleurotus 
ostreatus, enriched with vitamin C) has been proven in numerous clinical studies and observations in 
children and adults, and it is therefore a suitable choice for supporting weakened immunity for both 
short-term and long-term use. Open-label and double-blind, placebo-controlled studies in children 
with recurrent respiratory infections confirmed the preventive effect of Imunoglukan P4H® on the 
incidence and intensity of recurrent bacterial and viral infections in children from 1 year of age. 
Imunoglukan P4H® has also shown the ability to induce changes in specific and non-specific cellular 
immunity parameters, which have reflected into a reduced incidence of respiratory infections in 
children and adults, prevention of immunosuppression induced by intense physical stress, and 
reduction of the incidence of infectious exacerbations in patients with chronic inflammatory diseases.  
Supplementation with Imunoglukan P4H® was also able to achieve significant changes in markers of 
allergic inflammation in children (total IgE, blood eosinophil count), thus confirming previous results 
of in vitro tests, which indicated the anti-allergic potential of this active substance. Taking Imunoglukan 
P4H® supports physiological functions and maturation of the immune system in children and does not 

Immunomodulatory effect of pleuran (β-glucan from Pleurotus ostreatus) in children with recurrent respiratory tract infections. 
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cause its overstimulation. Imunoglukan P4H® ACUTE! (pleuran + vitamin C + zinc) confirmed its 
immunomodulatory effect in shortening the duration of symptoms in patients with herpes simplex 
virus type 1 (HSV-1) infection. 
The results of clinical studies have confirmed the complex immunomodulatory effect of this 
preparation without any significant side effects. 
Study design 
It is a multicenter, prospective, randomized, double-blind, placebo-controlled study with a food 
supplement at an outpatient healthcare provider.  
The investigators will enroll a total of 250 patients who meet the inclusion criteria. The proposed study 
does not imply any change in pharmacological/non-pharmacological treatment, diagnostic 
interventions administered to enrolled subjects. In case the respiratory tract infections occur during 
the study period, the patient enrolled in the study will be treated according to standard management 
plan.  
Patients will be randomized into 2 arms (active group and placebo group): 
• 
Active group: Imunoglukan P4H® chewable tablets (IMG® 50 mg + Zinc 5 mg + Vitamin D                 
10 μg in 1 tablet): 
o up to 25 kg of body weight 1 tablet once a day for 3 months 
o over 25 kg 2 tablets once a day for 3 months  
• 
Placebo group: Placebo chewable tablets (Zinc 5 mg + Vitamin D 10 μg in 1 tablet)  
o up to 25 kg of body weight 1 tablet once a day for 3 months 
o over 25 kg 2 tablets once a day for 3 months  
Discussion about design 
Multicenter, prospective, randomized, double-blind, placebo-controlled study design is a perpective 
for obtaining clinically, scientifically and statistically significant results to elucidate the impact of the 
investigational product on primary and secondary endpoints.  The 3-month duration of the study is 
justified in evaluating the modulation of the immune response and its clinical impact on such events 
as a reduction in the number of respiratory infections, shortening their duration, and reducing the 
need for ATB treatment. In addition to confirming the underlying hypothesis, the pilot study should 
also provide data on tolerability, safety and possible adverse effects.  
Subjects enrolled in the study complete an anamnestic data collection and fill out relevant 
questionnaires during control visits. 
Study endpoints 
Primary endpoints – to evaluate the effect of pleuran administration (Imunoglukan P4H® chewable 
tablets) compared to the placebo group on reduction in the number of RTIs episodes (total number) 
Secondary endpoints – to evaluate the effect of pleuran administration (Imunoglukan P4H® chewable 
tablets) compared to the placebo group on: 

Immunomodulatory effect of pleuran (β-glucan from Pleurotus ostreatus) in children with recurrent respiratory tract infections. 
IPRRTI study 
IPRRTI study Version 1 – 30.6.2023 
Inclusion criteria 
• 
signed informed consent 
• 
age 3-18 years 
• 
history of recurrent respiratory infections: 
o Age 3 to 5 years (< 6 years): 6 or more upper or lower respiratory tract infections from 
October 2022 until the end of March 2023  
o Ages 6 to 18 years (≥ 6 years): more than 3 upper or lower respiratory tract infections 
from October 2022 until the end of March 2023 
Exclusion criteria 
• 
refused informed consent 
• 
bronchopulmonary dysplasia 
• 
primary immunodeficiency 
• 
cystic fibrosis 
• 
chronic diarrhoea 
• 
chronic diseases of the lungs, kidneys and liver 
• 
malformations of the cardiovascular system 
• 
oncological disease 
• 
malnutrition 
• 
intolerance to any of the ingredients in the study product 
• 
taking preventive immunomodulatory therapy (beta-glucans, bacterial lysates, isoprinosine) 
less than 14 days prior enrolling the patient into the study 
List of data collected from study subjects 
 
Basic identification data 
o Patient ID 
o Patient Initials 
o Patient’s Date of birth 
o Date of completion of CRF / Date of visit 
Medical history and 
demographics 
o Age 
o Sex 
o Current comorbidities  
o Current treatment of comorbidities   
Diagnostic procedures 
o Patient diary (Respiratory infections) 
Primary endpoints 
o RTIs incidence (total)  
Secondary endpoints 
o RTIs subtypes incidence 
o Duration of RTIs (total duration, RTIs subtypes) 
o The need for ATB therapy for the treatment of RTIs  
• 
reduction in the number of episodes of RTI subtypes 
• 
reduction in the duration of RTI episodes (total duration, RTI subtypes) 
• 
reduction of the need for antibiotic (ATB) therapy 
• 
reduction of the number of missed days at school/nursery due to RTI 
• 
reduction of the number of missed working days due to RTIs 
• 
reduction of the number of emergency department visits due to RTI 
• 
reduction of the number of physician visits due to RTI 
• 
tolerability and safety 

Immunomodulatory effect of pleuran (β-glucan from Pleurotus ostreatus) in children with recurrent respiratory tract infections. 
IPRRTI study 
IPRRTI study Version 1 – 30.6.2023 
o Number of missed days in school/nursery due to RTIs  
o Number of missed working days due to RTIs of the child 
o Number of Emergency department visits due to RTIs  
o Number of Physician visits due to RTIs 
o Tolerability  
o Safety (occurrence of adverse events) 
Other data  
o Compliance 
 
 
Study visit and evaluation plan 
 
Required diagnostic procedures 
Patient diary "Respiratory tract infections" 
Parents or legal representatives of study subjects will record up-to-date data (incidence of infectious 
morbidity, symptoms, need for ATB therapy, physician or emergency department visits, number of 
missed days at school/nursery and work due to RTIs) throughout the study period. The filled "Patient 
diary" will be handed over to the treating physician at each visit. 
Adverse event form 
Visit number 
V0 
V1 
V2 
V3 
Day of visit 
Initial 
examination 
after 1st month 
(30 ±7 days) 
after 2nd month 
(60 ±7 days) 
after 3rd month 
(90 ±14 days) 
Informed consent 
 
Inclusion / exclusion 
criteria 
Demographics 
 
Medical history 
 
Comorbidities 
 
Dispensing of study 
product  
 
Incidence of respiratory 
tract infections (Patient 
diary) 
 
Adverse events 
 
Returning the study 
product, product 
accountability and patient 
compliance 
X 
 
X 
 
X 
 
X 
 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
X 
 
 
 
 
 
 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
X 
 
 
 
 
 
 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
X 
 
 
X 

Immunomodulatory effect of pleuran (β-glucan from Pleurotus ostreatus) in children with recurrent respiratory tract infections. 
IPRRTI study 
IPRRTI study Version 1 – 30.6.2023 
The physician fills out the Adverse event form when adverse events occur and records the occurrence 
of adverse events also in the CRF. 
Compliance  
Study subjects will return the container with remaining study medication at V3 (visit after 3 months) 
to their treating physician, who will count the number of remaining tablets. The physician will record 
the result in CRF. Patients with <75% compliance will be excluded from the study due to lack of 
collaboration. 
 
 
Appendices to the study protocol, other changes in the conduct of study 
Any amendments to the Protocol will be made in the form of an Appendix to the Protocol. Changes to 
the study are not permitted. Any unexpected and sudden changes in the conduct of study will be 
recorded in the clinical report.   
Privacy Policy 
All patients in this study will be identified with a unique ID. Identification of the patient through this ID 
will not be possible. Only the monitor can validate the source data. The unique ID of a particular patient 
will only be known to the investigator or monitor. The personal identification data entered into this 
study will be: date of birth, gender and initials. The nature of personal data identifiers is governed by 
all relevant legislative requirements.