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Official websites use. Share sensitive information only on official, secure websites. An accumulating body of evidence suggests that the endocannabinoid system plays a significant role in pathophysiological processes and impacts disease severity. Here we investigate the possible role of a cannabinoid receptor type 2 CB2 functional variant in determining disease severity and the potential pharmacological therapeutic effects of CB2 activation in viral respiratory infection. The frequency distribution of respiratory syncytial virus RSV -the main cause of severe cases of bronchiolitis and pneumonia in children-was studied in all collected samples. Of all the children enrolled in the study, 83 patients Collectively, CB2 is associated with RSV severity during infancy and may serve as a therapeutic target in RSV infection through the alleviation of virus-associated immunopathology. Acute respiratory tract infections ARTI are one of the most important causes of morbidity and mortality in children; they are estimated to be responsible for approximately two million childhood deaths globally. Among the commonly encountered respiratory viruses, respiratory syncytial virus RSV has a tendency to cause severe respiratory tract manifestations, primarily bronchiolitis and pneumonia, among infants and young children. Bronchiolitis and pneumonia are the most common lower respiratory tract infections in children and the leading cause of hospital admission under six months of age. RSV infection leads to infiltration of various immune cell types; if the lung recruitment of immune cells is dysregulated, the balance of viral control versus tissue damage is lost, resulting in pathology and severe disease. Cannabinoids refer to a group of diverse components that include substances of the plant Cannabis sativa , endocannabinoids, and synthetic ingredients. High-level CB2 expressions by immune cells and the inducible expressions of these receptors in inflammatory condition suggest that the anti-inflammatory and immunomodulatory action of cannabinoids are CB2-dependent. The receptor carrying R showed a reduced immune modulation function when activated by cannabinoids. Although the effects of the endocannabinoid system on immunity have received considerable attention, their impact on respiratory viral immunopathology are still unclear. It may be beneficial to explore the implication of the cannabinoid system and the virus may to maintain immune homeostasis in viral infections. In this regard, we studied the possible role of the CB2 Q63R functional variant in respiratory disease severity in children and the frequency distribution of RSV infection in children. The results reported here provide evidence that CB2 receptors play an important role in RSV disease severity. Between December and April , we admitted patients 90 inpatients and 90 outpatients to the Bahrami Children Hospital. The patients' ages ranged from one month to 22 months with median and mean ages of three and 4. The common clinical symptoms were sneezing, runny nose, cough, dyspnea, and fever. The details of demographic, age, gender, month, and clinical data according to the history of inpatients and outpatients are shown in Table 1. Males The highest rates of RSV infection were detected during winter. We found a significant difference between genotypic and allelic distributions of the Q63R polymorphism between the inpatients and outpatients. The relative odds ratio suggested that the risk of hospitalization was more than three-fold in children with the QQ genotype. The same result was observed in RSV-positive inpatients and outpatients. Thus, the common allele is associated with a reduced risk of hospitalization in RSV-positive patients. We found no significant difference in the amount of the CB2 expression between the lung cells of infected and uninfected mice Fig. Primary RSV infection elicits an inflammatory response comprising a mixed population of leukocytes that infiltrates the pulmonary airways. Experimental design. The secreted inflammatory and anti-inflammatory mediators were measured as indicators of immune cell function in response to RSV challenge. In agreement with the influx of BAL immune cells, the CB2 blockade impacts the accumulation of immune cells in the peribronchial and perivascular spaces of the lungs, although the differences did not reach statistical significance. In this experiment, we followed weight loss to determine whether the CB2 blockade and activation have any impact on disease severity. Weight loss is the hallmark of disease severity during RSV infection in the mice model. Furthermore, our results did not show that the CB2 blockade and activation impacted viral replication Fig. CB2 receptors were blockade via using AM or activated through JWH daily and the lung pathology were determined on day 5 after infection. A Representative slides of hematoxylin and eosin-stained lungs were analyzed and scored on day 5 after infection. B Pathology scores percentage for each group are shown. A The graph shows changes in body weight 5 days after the primary RSV or mock infection. The detection of specific viral causes of infection provides a useful starting point for understanding illnesses attributable to ARTIs and will guide future research studies. Epidemiological studies on the prevalence of RSV infection are essential for developing diagnostic methods, efficient drugs, and vaccine design. The frequency distribution of RSV infection in our study was The RSV positivity rate was higher among inpatient children. The majority of RSV-positive children Significant peaks of RSV prevalence were detected in the winter. This finding underlined the seasonal characteristics of RSV infections in our study group—this was consistent with the age patterns and seasonal RSV prevalence reported in other studies. The present study primarily focuses on RSV infection as the main cause of severe respiratory tract manifestations in children, while the possibility of other viral respiratory infections, such as rhinovirus, metapenumovirus, parainfluenza virus, and boca virus, needs further studies. Our knowledge of RSV pathogenesis and disease severity has increased over the last few years. It is crucial to understand how RSV interacts with its host to facilitate the development of safe and effective therapeutic interventions. The data from our study, for the first time, suggests that those with the CB RR variant is less prone to developing severe respiratory tract infections and those with the CB QQ variant is associated with more severe respiratory tract diseases and the risk of hospitalization. Although the study does not consider all respiratory viruses, the same result was observed in RSV-positive infants carrying the Q allele. An explanation for the association between the CB2 Q63R variant and the risk of disease severity and hospitalization in children with acute respiratory tract infections may be that the immune response with QQ variants were more inhibited when activated by an endogenous cannabinoid—i. Sip et al. In the animal study, we found that RSV infection can affect the CB2 expression in BAL cells—this is in line with its high-level expression by immune cells and the inducible expression in inflammatory condition. The results of our study are in line with previous reports on the role of cannabinoids in influenza infection outcomes. However, to replicate the RSV infection seen in children with different disease severity in a better way, various RSV challenge doses should be tested in future studies. Since the activation of CB2 triggers anti-inflammatory action, targeting these receptors may be a novel and effective approach for the treatment of RSV-associated immunopathology. These effects result in the control of RSV-induced lung pathology and weight loss. This observation was consistent with the immunosuppressive effects of CB2 activation reported by others. Neutrophils play an important role in the immunopathogenesis of RSV infection. However, it is important to ensure that the JWH effects are attenuated with AM in future studies. In conclusion, given the breadth of cannabinoids-mediated regulation of the immunity function and the complicated immunopathology associated with RSV infection, our results indicate that: i the CB2 Q63R variant is associated with the clinical course of acute respiratory viral infections ii experimental RSV-induced immunopathogenesis can be modulated in part by endocannabinoids, as observed by blocking the CB2 signaling, and iii RSV-induced immunopathogenesis can be modulated by CB2 activation and may be a novel approach for the treatment of RSV bronchiolitis in children. We recruited Iranian children under two years of age with clinically suspected acute respiratory viral infection, including 90 hospitalized patients severe ARTI and 90 outpatients mild ARTI during the winter season of at Bahrami Children's Hospital, Tehran, Iran. According to the Bahrami Children's Hospital protocol used for treatment of severe ARTI; the hospitalization criteria contain respiratory distress including tachypnea, nasal flaring, chest retractions, or grunting. A clinical questionnaire was used to collect data from all patients including age, gender, and symptoms like bronchiolitis, pneumonia, fever, sore throat, cough, dyspnea, runny nose, nasal congestion, and sneezing. The current human study was approved by science and bioethics committee of Tehran University of Medical Sciences. All samples were genotyped for the CNR2 rs variant, which changes the second and third adenosine at codon 63 CAA to guanosine CGG leading to the missense variant Q63R in the first intracellular signaling loop of the encoded CB2 protein. Viral RNA was extracted from nasopharyngeal swab samples using viral high pure nucleic acid extraction kit following manufacturers' instructions Roche, Germany. The animals were transferred and maintained in their home cages one week before the beginning of the experiments. All animal experiments were approved by the animal ethics committee of the Tehran University of Medical Sciences. Drugs were dissolved in dimethyl sulfoxide and diluted in normal saline 0. Virus stuck was propagated on HEp-2 cells and purification was performed using polyethylene glycol precipitation as described previously by our group. The relative level of gene expression was determined by the comparative threshold cycle method as described by the manufacturer. BALF was obtained 5 days after infection as described previously by our group. Concentrations of cytokines in the samples were calculated by interpolation from the standard curve. Five day after challenge, mice were sacrificed and their lung was obtained, and histology slides were prepared as described previously by our group. Nucleic acid was prepared from supernatants of BALF using viral high pure nucleic acid extraction kit following manufacturers' instructions Roche, Germany. The OR was adjusted by age and sex in the logistic regression model. A linear logistic regression was performed to analyze clinical data with respect to the CB2 Q63R variant. Graph preparation and statistical analyses in animal study were performed using GraphPad Prism v6. The normality of date was performed using Kolmogorov—Smirnov test. Differences between two groups were carried out using Student's t test for unpaired data. This project was extracted from a PhD thesis. As a library, NLM provides access to scientific literature. Find articles by Alireza Tahamtan. Find articles by Yazdan Samieipoor. Find articles by Fatemeh Sadat Nayeri. Find articles by Ali Akbar Rahbarimanesh. Find articles by Anahita Izadi. Find articles by Ali Rashidi-Nezhad. Find articles by Masoumeh Tavakoli-Yaraki. Find articles by Mohammad Farahmand. Find articles by Louis Bont. Find articles by Fazel Shokri. Find articles by Talat Mokhatri-Azad. Find articles by Vahid Salimi. Demographic, gender, age, month, and clinical data according to inpatients and outpatients. Open in a new tab. Genotype and allele frequencies of CNR2 gene in patients with acute respiratory infection. Demographic, gender, age, and clinical data according to CNR2 variants. Similar articles. Add to Collections. Create a new collection. Add to an existing collection. Choose a collection Unable to load your collection due to an error Please try again. Add Cancel.
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Official websites use. Share sensitive information only on official, secure websites. An accumulating body of evidence suggests that the endocannabinoid system plays a significant role in pathophysiological processes and impacts disease severity. Here we investigate the possible role of a cannabinoid receptor type 2 CB2 functional variant in determining disease severity and the potential pharmacological therapeutic effects of CB2 activation in viral respiratory infection. The frequency distribution of respiratory syncytial virus RSV -the main cause of severe cases of bronchiolitis and pneumonia in children-was studied in all collected samples. Of all the children enrolled in the study, 83 patients Collectively, CB2 is associated with RSV severity during infancy and may serve as a therapeutic target in RSV infection through the alleviation of virus-associated immunopathology. Acute respiratory tract infections ARTI are one of the most important causes of morbidity and mortality in children; they are estimated to be responsible for approximately two million childhood deaths globally. Among the commonly encountered respiratory viruses, respiratory syncytial virus RSV has a tendency to cause severe respiratory tract manifestations, primarily bronchiolitis and pneumonia, among infants and young children. Bronchiolitis and pneumonia are the most common lower respiratory tract infections in children and the leading cause of hospital admission under six months of age. RSV infection leads to infiltration of various immune cell types; if the lung recruitment of immune cells is dysregulated, the balance of viral control versus tissue damage is lost, resulting in pathology and severe disease. Cannabinoids refer to a group of diverse components that include substances of the plant Cannabis sativa , endocannabinoids, and synthetic ingredients. High-level CB2 expressions by immune cells and the inducible expressions of these receptors in inflammatory condition suggest that the anti-inflammatory and immunomodulatory action of cannabinoids are CB2-dependent. The receptor carrying R showed a reduced immune modulation function when activated by cannabinoids. Although the effects of the endocannabinoid system on immunity have received considerable attention, their impact on respiratory viral immunopathology are still unclear. It may be beneficial to explore the implication of the cannabinoid system and the virus may to maintain immune homeostasis in viral infections. In this regard, we studied the possible role of the CB2 Q63R functional variant in respiratory disease severity in children and the frequency distribution of RSV infection in children. The results reported here provide evidence that CB2 receptors play an important role in RSV disease severity. Between December and April , we admitted patients 90 inpatients and 90 outpatients to the Bahrami Children Hospital. The patients' ages ranged from one month to 22 months with median and mean ages of three and 4. The common clinical symptoms were sneezing, runny nose, cough, dyspnea, and fever. The details of demographic, age, gender, month, and clinical data according to the history of inpatients and outpatients are shown in Table 1. Males The highest rates of RSV infection were detected during winter. We found a significant difference between genotypic and allelic distributions of the Q63R polymorphism between the inpatients and outpatients. The relative odds ratio suggested that the risk of hospitalization was more than three-fold in children with the QQ genotype. The same result was observed in RSV-positive inpatients and outpatients. Thus, the common allele is associated with a reduced risk of hospitalization in RSV-positive patients. We found no significant difference in the amount of the CB2 expression between the lung cells of infected and uninfected mice Fig. Primary RSV infection elicits an inflammatory response comprising a mixed population of leukocytes that infiltrates the pulmonary airways. Experimental design. The secreted inflammatory and anti-inflammatory mediators were measured as indicators of immune cell function in response to RSV challenge. In agreement with the influx of BAL immune cells, the CB2 blockade impacts the accumulation of immune cells in the peribronchial and perivascular spaces of the lungs, although the differences did not reach statistical significance. In this experiment, we followed weight loss to determine whether the CB2 blockade and activation have any impact on disease severity. Weight loss is the hallmark of disease severity during RSV infection in the mice model. Furthermore, our results did not show that the CB2 blockade and activation impacted viral replication Fig. CB2 receptors were blockade via using AM or activated through JWH daily and the lung pathology were determined on day 5 after infection. A Representative slides of hematoxylin and eosin-stained lungs were analyzed and scored on day 5 after infection. B Pathology scores percentage for each group are shown. A The graph shows changes in body weight 5 days after the primary RSV or mock infection. The detection of specific viral causes of infection provides a useful starting point for understanding illnesses attributable to ARTIs and will guide future research studies. Epidemiological studies on the prevalence of RSV infection are essential for developing diagnostic methods, efficient drugs, and vaccine design. The frequency distribution of RSV infection in our study was The RSV positivity rate was higher among inpatient children. The majority of RSV-positive children Significant peaks of RSV prevalence were detected in the winter. This finding underlined the seasonal characteristics of RSV infections in our study group—this was consistent with the age patterns and seasonal RSV prevalence reported in other studies. The present study primarily focuses on RSV infection as the main cause of severe respiratory tract manifestations in children, while the possibility of other viral respiratory infections, such as rhinovirus, metapenumovirus, parainfluenza virus, and boca virus, needs further studies. Our knowledge of RSV pathogenesis and disease severity has increased over the last few years. It is crucial to understand how RSV interacts with its host to facilitate the development of safe and effective therapeutic interventions. The data from our study, for the first time, suggests that those with the CB RR variant is less prone to developing severe respiratory tract infections and those with the CB QQ variant is associated with more severe respiratory tract diseases and the risk of hospitalization. Although the study does not consider all respiratory viruses, the same result was observed in RSV-positive infants carrying the Q allele. An explanation for the association between the CB2 Q63R variant and the risk of disease severity and hospitalization in children with acute respiratory tract infections may be that the immune response with QQ variants were more inhibited when activated by an endogenous cannabinoid—i. Sip et al. In the animal study, we found that RSV infection can affect the CB2 expression in BAL cells—this is in line with its high-level expression by immune cells and the inducible expression in inflammatory condition. The results of our study are in line with previous reports on the role of cannabinoids in influenza infection outcomes. However, to replicate the RSV infection seen in children with different disease severity in a better way, various RSV challenge doses should be tested in future studies. Since the activation of CB2 triggers anti-inflammatory action, targeting these receptors may be a novel and effective approach for the treatment of RSV-associated immunopathology. These effects result in the control of RSV-induced lung pathology and weight loss. This observation was consistent with the immunosuppressive effects of CB2 activation reported by others. Neutrophils play an important role in the immunopathogenesis of RSV infection. However, it is important to ensure that the JWH effects are attenuated with AM in future studies. In conclusion, given the breadth of cannabinoids-mediated regulation of the immunity function and the complicated immunopathology associated with RSV infection, our results indicate that: i the CB2 Q63R variant is associated with the clinical course of acute respiratory viral infections ii experimental RSV-induced immunopathogenesis can be modulated in part by endocannabinoids, as observed by blocking the CB2 signaling, and iii RSV-induced immunopathogenesis can be modulated by CB2 activation and may be a novel approach for the treatment of RSV bronchiolitis in children. We recruited Iranian children under two years of age with clinically suspected acute respiratory viral infection, including 90 hospitalized patients severe ARTI and 90 outpatients mild ARTI during the winter season of at Bahrami Children's Hospital, Tehran, Iran. According to the Bahrami Children's Hospital protocol used for treatment of severe ARTI; the hospitalization criteria contain respiratory distress including tachypnea, nasal flaring, chest retractions, or grunting. A clinical questionnaire was used to collect data from all patients including age, gender, and symptoms like bronchiolitis, pneumonia, fever, sore throat, cough, dyspnea, runny nose, nasal congestion, and sneezing. The current human study was approved by science and bioethics committee of Tehran University of Medical Sciences. All samples were genotyped for the CNR2 rs variant, which changes the second and third adenosine at codon 63 CAA to guanosine CGG leading to the missense variant Q63R in the first intracellular signaling loop of the encoded CB2 protein. Viral RNA was extracted from nasopharyngeal swab samples using viral high pure nucleic acid extraction kit following manufacturers' instructions Roche, Germany. The animals were transferred and maintained in their home cages one week before the beginning of the experiments. All animal experiments were approved by the animal ethics committee of the Tehran University of Medical Sciences. Drugs were dissolved in dimethyl sulfoxide and diluted in normal saline 0. Virus stuck was propagated on HEp-2 cells and purification was performed using polyethylene glycol precipitation as described previously by our group. The relative level of gene expression was determined by the comparative threshold cycle method as described by the manufacturer. BALF was obtained 5 days after infection as described previously by our group. Concentrations of cytokines in the samples were calculated by interpolation from the standard curve. Five day after challenge, mice were sacrificed and their lung was obtained, and histology slides were prepared as described previously by our group. Nucleic acid was prepared from supernatants of BALF using viral high pure nucleic acid extraction kit following manufacturers' instructions Roche, Germany. The OR was adjusted by age and sex in the logistic regression model. A linear logistic regression was performed to analyze clinical data with respect to the CB2 Q63R variant. Graph preparation and statistical analyses in animal study were performed using GraphPad Prism v6. The normality of date was performed using Kolmogorov—Smirnov test. Differences between two groups were carried out using Student's t test for unpaired data. This project was extracted from a PhD thesis. As a library, NLM provides access to scientific literature. Find articles by Alireza Tahamtan. Find articles by Yazdan Samieipoor. Find articles by Fatemeh Sadat Nayeri. Find articles by Ali Akbar Rahbarimanesh. Find articles by Anahita Izadi. Find articles by Ali Rashidi-Nezhad. Find articles by Masoumeh Tavakoli-Yaraki. Find articles by Mohammad Farahmand. Find articles by Louis Bont. Find articles by Fazel Shokri. Find articles by Talat Mokhatri-Azad. Find articles by Vahid Salimi. Demographic, gender, age, month, and clinical data according to inpatients and outpatients. Open in a new tab. Genotype and allele frequencies of CNR2 gene in patients with acute respiratory infection. Demographic, gender, age, and clinical data according to CNR2 variants. Similar articles. Add to Collections. Create a new collection. Add to an existing collection. Choose a collection Unable to load your collection due to an error Please try again. Add Cancel.
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