Immunomodulatory Effects of Risperidone and Methylphenidate on Dendritic Cells in Methamphetamine Addicts: Insights from a Detoxification Study

Immunomodulatory Effects of Risperidone and Methylphenidate on Dendritic Cells in Methamphetamine Addicts: Insights from a Detoxification Study | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Immunomodulatory Effects of Risperidone and Methylphenidate on Dendritic Cells in Methamphetamine Addicts: Insights from a Detoxification Study Ghasem Mosayebi, Seyed Mohammad Moazzeni, Hadiseh Farahani, Hassan Solhi, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5381470/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background and aim: Drug abuse can impact the function of immune cells, leading to a compromised immune system response. This study aimed to investigate the immunomodulatory effects of methamphetamine and its detoxification agents on peripheral blood dendritic cells. Methods: Thirty individuals with methamphetamine addiction were divided into two groups. One group received treatment with Risperidone, while the second group was treated with a combination of Risperidone and Methylphenidate. The percentages of myeloid and plasmacytoid dendritic cells, as well as their surface markers, were assessed before and after the detoxification program. Results: The percentages of both CD11c+ and CD123+ dendritic cells in peripheral blood were significantly lower in methamphetamine addicts compared to the control group. Detoxification with Risperidone corrected this reduction, while the combination of Risperidone and Methylphenidate produced adverse effects. The expression of HLA-DR, CD11c, and CD123 markers was downregulated in the dendritic cells of methamphetamine addicts. Treatment with Risperidone restored these markers, whereas the combination therapy further exacerbated the downregulation of these markers. Conclusion: The findings suggest that detoxification with Risperidone may help ameliorate the immunological disorders associated with methamphetamine use. Dendritic cells Methamphetamine Risperidone Methylphenidate Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Narcotics exert a profound influence on various body systems, including the immune system (1). This effect is observed irrespective of the duration of use, whether short-term or long-term. The underlying mechanism of action primarily involves three key receptors: µ (MOR), κ (KOR), and δ (DOR) opioid receptors (2). Narcotics are known to suppress immune function, which is a significant factor contributing to the increased prevalence of infectious diseases and cancers among individuals with substance use disorders (3–5). Additionally, the immunosuppressive effects of these substances can exacerbate the severity and prolong the duration of infections (6). Conversely, some studies suggest that short-term use of narcotics as analgesics may stimulate certain aspects of the immune system (7). Methamphetamine (METH) is a highly addictive central nervous system stimulant (8). Its widespread use presents substantial public health and economic challenges globally (9). The drug induces feelings of euphoria, excitement, and weight loss, effects that are comparable to those produced by cocaine. Methamphetamine acts as an agonist at dopamine receptors, leading to increased dopamine release (10). Detoxification is the initial step for individuals entering rehabilitation programs for substance use disorders. During this process, antipsychotic medications such as risperidone—an antagonist of opioid receptors and dopamine D2 receptors—or a combination of antipsychotics and methylphenidate (Ritalin), a stimulant, may be administered to alleviate withdrawal symptoms associated with methamphetamine use (11–13). Historically, lymphocytes were the primary focus in studies examining the immunomodulatory effects of various substances. However, recent findings indicate that immunomodulatory agents predominantly exert their effects by suppressing the differentiation, maturation, and activation of dendritic cells (DCs) (14). Dendritic cells play a pivotal role in presenting antigens to T lymphocytes and are essential for both innate and adaptive immune responses (15). They exhibit unique responses to various pathogens due to their diverse subgroups and strategic distribution throughout the body. In humans, dendritic cells are classified into two major subgroups: CD11c + myeloid dendritic cells (mDCs) and CD123 + lymphoid dendritic cells (plasmacytoid DCs, pDCs) (16). While extensive research has been conducted on the effects of methamphetamine in murine models (17), there is a growing interest in understanding its impact on human subjects. However, our comprehension of methamphetamine's effects on the human immune system remains limited (14). The objective of this study was to investigate the immunomodulatory effects of methamphetamine and its detoxification agents on dendritic cells. Material and methods Participants : A total of 30 individuals with methamphetamine addiction and 30 healthy control subjects were enrolled in the study. The control group was closely matched to the addicted group in terms of age, sex, weight, height, and body mass index (BMI). Control subjects were selected from a population of university staff and students who had never used psychotropic drugs. The general health of the control group was confirmed through physical examinations and routine biochemical and hematological tests. The study was approved by the Ethics Committee, and all participants provided informed consent. All volunteers were free from infectious diseases, active inflammatory conditions, and other disorders affecting the immune system. Urine samples collected from the control group tested negative for morphine and methamphetamine. The addicted group was selected based on specific criteria: individuals who consumed more than 500 mg of methamphetamine daily for over one year, had a history of addiction to other drugs for no longer than three months in the past, and exhibited positive urine test results for methamphetamine above the detection threshold. The group of 30 addicted volunteers was divided into two subgroups of 15 individuals each. One subgroup received risperidone, while the other subgroup was administered both risperidone and methylphenidate concurrently. The initial dose of risperidone was 5 mg, which was increased to 7.5 mg by the end of the study. Methylphenidate was administered in a tapering manner over ten days (10 mg every 8 hours). The immune response of the addicted group was evaluated at three time points: at the beginning of the trial, at the end of detoxification, and up to one month following detoxification. Flow cytometry: Mononuclear cells were isolated from peripheral blood samples using Ficoll density gradient centrifugation. A concentration of 25 × 10⁶ cells was obtained, and 20 µl (equivalent to 5 × 10⁵ cells) was transferred to FACS tubes. Each tube received 15 µl of a cocktail containing anti-CD3, CD14, CD16, CD19, CD20, and CD56 antibodies labeled with FITC (Lin-1), along with 5 µl of anti-CD34 antibody labeled with FITC (to exclude CD34 + HLA-DR + precursors) and 5 µl of HLA-DR antibody labeled with PerCP. For three-color flow cytometry analysis, 5 µl of PE-labeled CD123 antibody was added to identify Lin-/HLA-DR+/CD123 + plasmacytoid dendritic cells (pDCs), while another tube received 5 µl of PE-labeled CD11c antibodies to identify Lin-/HLA-DR+/CD11c + myeloid dendritic cells (mDCs). Samples were analyzed using a FACScan flow cytometer (BD Biosciences, San Jose, CA), and data were processed using FlowJo software (18). Statistical analysis: One-way analysis of variance (ANOVA) was employed to assess statistical significance between experimental groups. All data are presented as mean ± SEM. P-values < 0.05 were considered statistically significant (*p < 0.05, **p < 0.01). Results 1-Selection of volunteers: Based on the selection criteria, 30 healthy individuals were chosen as the control group, while 30 individuals with methamphetamine addiction were selected as the experimental group (Table 1 ). There were no statistically significant differences in age, height, weight, or body mass index (BMI) between the two groups (control and experimental). The general health of the control group was confirmed through physical examinations and routine hematological and biochemical tests (results not shown). Table 1 Demographic characteristics of volunteers included in the study Control group Experiment group Sex Male Male (Mean of years ± SD) Age 30.57 ± 3.21 30.17 ± 5.47 (Mean of cm ± SD) Height 172.29 ± 6.07 171.13 ± 6.04 (Mean of kg ± SD) Weight 73.80 ± 5.63 69.85 ± 12.26 Body Mass Index (Mean ± SD) 24.87 ± 1.57 23.912 ± 4.11 2- Determination of the frequency of peripheral blood dendritic cells in volunteers This study identified two major dendritic cell (DC) subsets in peripheral blood mononuclear cells (PBMCs): Lin − HLA-DR + CD11c + myeloid dendritic cells (mDCs) and Lin − HLA-DR + CD123 + plasmacytoid dendritic cells (pDCs) (Fig. 1 ). To quantify the number of dendritic cells, 100,000 cells were counted within the gate of mononuclear cells. The percentage of peripheral blood dendritic cells in methamphetamine addicts was significantly lower compared to the control group (Fig. 2 A). The percentage of CD11c + DCs in the peripheral blood of methamphetamine addicts also showed a significant decrease compared to controls (Fig. 2 B). Similarly, the percentage of CD123 + DCs in the peripheral blood of methamphetamine addicts was lower than that of the controls (Fig. 2 C). Immune responses in methamphetamine addicts were assessed at three distinct time points: at the beginning of the trial, at the end of detoxification, and up to one month after detoxification (prior to the onset of withdrawal symptoms). Detoxification with risperidone restored the percentage of peripheral blood dendritic cells in methamphetamine addicts compared to the control group. However, detoxification with both risperidone and methylphenidate did not reverse this reduction (Fig. 3 A). As illustrated in Fig. 3 B, detoxification with risperidone corrected the percentage of CD11c + DCs in the peripheral blood of methamphetamine addicts. In contrast, detoxification with risperidone and methylphenidate did not restore the reduced percentage of CD11c + DCs in these individuals. Risperidone treatment led to a significant increase in the previously reduced percentage of CD123 + dendritic cells in the peripheral blood of methamphetamine addicts. However, when compared to the control group, detoxification with both risperidone and methylphenidate resulted in a significant decrease in the percentage of CD123 + DCs in the peripheral blood of addicts (Fig. 3 C). 3-The expression of HLA-DR, CD11c and CD123 markers on CD11c + and CD123 + dendritic cells in volunteers This study evaluated the mean fluorescent intensity (MFI), which represents the expression level of cell surface markers, for HLA-DR, CD11c, and CD123 on dendritic cells in the peripheral blood of volunteers. As depicted in Fig. 4 A, the expression level of HLA-DR on the surface of peripheral blood dendritic cells from methamphetamine addicts was significantly decreased compared to that in the control group. Furthermore, the expression level of CD123 on the surface of peripheral blood CD123 + DCs from methamphetamine addicts was significantly lower than that observed in the control group (Fig. 4 C). As shown in Fig. 5A, detoxification with risperidone significantly altered the reduced expression of the HLA-DR molecule on the surface of peripheral blood dendritic cells in methamphetamine addicts. However, the combination of risperidone and methylphenidate did not restore the decreased expression of HLA-DR on peripheral blood dendritic cells in this group. Detoxification with risperidone resulted in a significant increase in the expression of the CD11c molecule on CD11c + dendritic cells in the peripheral blood of methamphetamine addicts. In contrast, the combination of risperidone and methylphenidate did not improve the reduced expression of CD11c on the surface of these CD11c + dendritic cells (Fig. 5B). Neither detoxification regimen was able to correct the diminished expression of the CD123 molecule on the surface of CD123 + dendritic cells in the peripheral blood of methamphetamine addicts when compared to the control group (Fig. 5C). Discussion Dendritic cells play a crucial role in processing and presenting antigens, thereby stimulating naïve T cells and regulating the quality of subsequent immune responses. In the bone marrow, newly generated dendritic cells migrate through the bloodstream to various peripheral tissues (19). Consequently, the presence of circulating blood dendritic cells can provide insights into the overall status of the immune system. For example, a study by Lissoni et al. found that decreased levels of circulating blood dendritic cells were strongly associated with suppressed immune function in patients with advanced malignant tumors (20). One contributing factor to the increased incidence of infections and cancers among substance abusers is the immunosuppressive effects of drugs (21, 22). In the current study, we assessed the overall percentages of two main subgroups of dendritic cells in the peripheral blood of methamphetamine addicts and examined the surface markers of these cells. Our data indicate that the methamphetamine-addicted group exhibited significantly lower total dendritic cell counts, as well as reduced numbers of CD11c + and CD123 + dendritic cells compared to healthy controls. This finding suggests a potential defect in their immune system. Several factors may contribute to the decrease in dendritic cell percentages, including reduced production rates in the bone marrow, disruptions in monocyte responses to differentiation signals, inhibition of factors that promote dendritic cell differentiation, or the induction of apoptosis in dendritic cells (23). Although further studies are needed to confirm these possibilities, our findings suggest that dendritic cells are involved in the immunopathogenesis of disorders associated with methamphetamine use. In methamphetamine addicts, detoxification with risperidone restored the total percentage of dendritic cells as well as the numbers of CD11c + and CD123 + dendritic cells to levels comparable to those of the control group. In contrast, treatment with a combination of risperidone and methylphenidate resulted in adverse effects. HLA-DR is a critical molecule that stimulates T cells and serves as an indicator of dendritic cell activity. It is expressed on the surface of antigen-presenting cells, including dendritic cells, monocytes, macrophages, and B cells. HLA-DR presents processed extracellular antigens to T helper cells, and its expression level significantly influences the initiation of specific immune responses. A reduction or absence of HLA-DR can impair antigen-presenting cell function, leading to inadequate antigen presentation and diminished cytokine secretion (23). Cytokines regulate the expression of HLA-DR in an antagonistic manner. For instance, IFN-γ enhances HLA-DR expression and bolsters cellular immune responses, while IL-10 inhibits its expression (24). Our data indicate that the expression of HLA-DR is decreased in the dendritic cells of individuals addicted to methamphetamine. HLA-DR is a critical surface marker on dendritic cells responsible for activating T cells; thus, a reduction in its expression may signify immunoparalysis. The diminished expression of HLA-DR not only impairs dendritic cell function but also increases the susceptibility of methamphetamine addicts to infectious diseases (25). Our previous study demonstrated elevated levels of IL-10 in the serum of these individuals (26). However, further research is necessary to fully elucidate the mechanisms underlying this reduction. The decreased expression of HLA-DR in methamphetamine addicts was restored following treatment with risperidone. In contrast, co-administration of risperidone and methylphenidate exacerbated the downregulation of HLA-DR expression. CD11c, a member of the integrin family, plays a vital role in cellular attachment and is abundantly expressed on the surface of myeloid dendritic cells (27). This study revealed that methamphetamine significantly reduced the expression of the CD11c molecule in peripheral blood CD11c + dendritic cells among addicts. Risperidone treatment restored CD11c expression to levels comparable to those in the control group; however, co-administration with methylphenidate exacerbated this reduction. Lymphoid dendritic cells require IL-3 for differentiation, leading to high expression of the IL-3 receptor alpha chain (CD123) (28). Methamphetamine notably decreased the expression of CD123. Detoxification with risperidone increased CD123 expression; however, when combined with methylphenidate, this expression was reduced, although these changes were not statistically significant. Overall, these data suggest that long-term consumption of methamphetamine has an immunosuppressive effect. Risperidone acts as an antagonist to methamphetamine, and its use as a detoxifying agent reverses this immunosuppressive state. Conversely, methylphenidate, which is a stimulant similar to methamphetamine, exacerbates the immunosuppressive condition of the immune system. Conclusion Dendritic cells in individuals addicted to methamphetamine may lose their ability to respond effectively to invading pathogens, potentially increasing their susceptibility to infections. Impairments in dendritic cell function could also diminish the effectiveness of vaccines in this population. During detoxification from methamphetamine, the administration of risperidone was found to mitigate some of the adverse effects associated with methamphetamine use. However, the combination of risperidone and methylphenidate exacerbated these negative effects. Although this study is grounded in natural science experimentation, its findings may have practical implications for clinical practice. Declarations Acknowledgments This work was supported by the Research Council of Arak University of Medical Sciences. Funding This study was funded by the Council of Arak University of Medical Sciences (grant number 658). Availability of data and materials The datasets generated and/or analyzed during the current study are available from the corresponding author upon reasonable request. Conflict of interest The authors declare that there is no conflict of interest. Ethics approval and consent to participate The Ethics Committee of Arak University of Medical Sciences approved this study (Ethical Code: IR.ARAKMU.REC.1390.13.9). References Ghazavi A, Solhi H, Moazzeni SM, Rafiei M, Mosayebi G. Cytokine profiles in long-term smokers of opium (Taryak). Journal of addiction medicine. 2013;7(3):200-3. Eisenstein TK. The Role of Opioid Receptors in Immune System Function. Frontiers in immunology. 2019;10. Al-Hashimi M, Scott S, Thompson J, Lambert D. Opioids and immune modulation: more questions than answers. British journal of anaesthesia. 2013;111(1):80-8. Roy S, Ninkovic J, Banerjee S, Charboneau RG, Das S, Dutta R, et al. 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Lissoni P, Vigore L, Ferranti R, Bukovec R, Meregalli S, Mandala M, et al. Circulating dendritic cells in early and advanced cancer patients: diminished percent in the metastatic disease. Journal of biological regulators and homeostatic agents. 1999;13(4):216-9. Lei L, Gong X, Wen C, Zeng S, Lei Q. Research progress on the effects of opioids on the immune system. Trends in Anaesthesia and Critical Care. 2024;57:101372. Sun Q, Li Z, Wang Z, Wang Q, Qin F, Pan H, et al. Immunosuppression by opioids: Mechanisms of action on innate and adaptive immunity. Biochemical Pharmacology. 2023;209:115417. Van Leeuwen-Kerkhoff N, Westers TM, Poddighe PJ, Povoleri GAM, Timms JA, Kordasti S, et al. Reduced frequencies and functional impairment of dendritic cell subsets and non-classical monocytes in myelodysplastic syndromes. Haematologica. 2022;107(3):655-67. Ozdemir AT, Oztatlici M, Ozgul Ozdemir R, Cakir B, Ozbilgin K, Dariverenli E. The effects of preconditioning with IFN-γ, IL-4, and IL-10 on costimulatory ligand expressions of mesenchymal stem cells. Int J Med Biochem. 2021;4(2):121-30. Morel AS, Coulton G, Londei M. Regulation of major histocompatibility complex class II synthesis by interleukin-10. Immunology. 2002;106(2):229-36. Akbari A, Mosayebi G, Samiei AR, Ghazavi A. Methadone therapy modulate the dendritic cells of heroin addicts. International immunopharmacology. 2019;66:330-5. Wu J, Wu H, An J, Ballantyne CM, Cyster JG. Critical role of integrin CD11c in splenic dendritic cell capture of missing-self CD47 cells to induce adaptive immunity. Proceedings of the National Academy of Sciences of the United States of America. 2018;115(26):6786-91. Podolska MJ, Grützmann R, Pilarsky C, Bénard A. IL-3: key orchestrator of inflammation. Frontiers in immunology. 2024;15:1411047. Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5381470","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":382104500,"identity":"07ab0c1b-a7b2-4253-aab0-6bff868fbec9","order_by":0,"name":"Ghasem Mosayebi","email":"","orcid":"","institution":"Arak University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Ghasem","middleName":"","lastName":"Mosayebi","suffix":""},{"id":382104501,"identity":"ca78a0a3-609c-4b58-b721-e2f9d8373a4b","order_by":1,"name":"Seyed Mohammad Moazzeni","email":"","orcid":"","institution":"Tarbiat Modares University","correspondingAuthor":false,"prefix":"","firstName":"Seyed","middleName":"Mohammad","lastName":"Moazzeni","suffix":""},{"id":382104502,"identity":"bed53e56-883d-4bfd-a085-6824e39b7441","order_by":2,"name":"Hadiseh Farahani","email":"","orcid":"","institution":"Arak University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Hadiseh","middleName":"","lastName":"Farahani","suffix":""},{"id":382104503,"identity":"6a11341c-bf0c-4450-be21-0abe0bca31be","order_by":3,"name":"Hassan Solhi","email":"","orcid":"","institution":"Arak University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Hassan","middleName":"","lastName":"Solhi","suffix":""},{"id":382104504,"identity":"e0d58b98-c9fc-49dd-9163-69d4fc37fb17","order_by":4,"name":"Mohammad Rafiei","email":"","orcid":"","institution":"Arak University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Mohammad","middleName":"","lastName":"Rafiei","suffix":""},{"id":382104505,"identity":"d698a02f-ac9f-4ac3-b769-acc384bfa799","order_by":5,"name":"Ali Ghazavi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+UlEQVRIiWNgGAWjYDACCSjJx8x8AMhIgAobEKGFjZktgSQtDAxsDDwGSFrwAPno5scffrZZyLOx83z8XFGTFm1wgPnhB4aCezi1GN45ZmDY2yZh2MbMu1nyzLGc3A0H2IwlGAyKcWuZkWCQwHNGghGoZYNkA1sFUAuDGdAvuF1oOCP9w8E/ZyTs25h5Hv9s+AfSwv4NrxZ5iRzDZp4KiUSgFjbJxjaQw3jw22IgkVPMLFMhkdzGzGZm2diXljvzME+xRAI+W2akb/74xqDOtp//8OObDd+Sc/uOt2/88OEPHlsOYAgxM+CPHvkGPJKjYBSMglEwCsAAAFzpTwHw1PboAAAAAElFTkSuQmCC","orcid":"","institution":"Arak University of Medical Sciences","correspondingAuthor":true,"prefix":"","firstName":"Ali","middleName":"","lastName":"Ghazavi","suffix":""}],"badges":[],"createdAt":"2024-11-03 09:53:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5381470/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5381470/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":70197354,"identity":"908f0837-9d00-4d67-ac86-88dd89b313a5","added_by":"auto","created_at":"2024-11-29 11:45:59","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":190531,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eExample of flow cytometry analysis and gating strategy for identifying and quantifying blood dendritic cell subsets\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003eThe PBMC (peripheral blood mononuclear cell) population is identified by a combination of forward/side scatter characteristics (A), and DC was identified within the lineage (CD3, CD14, CD16, CD19, CD20, CD56, and CD34)-negative (Lin−) HLA-DR+ (B). One representative experiment demonstrates the gating strategy for identifying DC subsets (C and D). mDCs: myeloid DCs (CD11c+), pDCs: plasmacytoid DCs (CD123+).\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5381470/v1/c221425b1ff7c06b560ab874.png"},{"id":70197358,"identity":"7ec7f5c1-510d-47db-a9ff-cc8fc4868069","added_by":"auto","created_at":"2024-11-29 11:45:59","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":57263,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePercentage of peripheral blood dendritic cells, CD11c, and CD123 dendritic cells in the control group and methamphetamine addicts.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePercentage of peripheral blood dendritic cells (A), peripheral blood CD11c+ (B), and peripheral blood CD123+ dendritic cells (C) in the control group and methamphetamine addicts. DC= Dendritic cell, Meth= methamphetamine. Data are presented as mean ± standard error. The results were extracted from the analysis of 100 thousand mononuclear cells. The sign * indicates P\u0026lt;0.05 compared to the control.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5381470/v1/ea01d5a7a851640378fdbdcb.png"},{"id":70197592,"identity":"e78ee166-bcad-4a27-a756-0b355b4f67b3","added_by":"auto","created_at":"2024-11-29 11:54:15","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":90574,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe percentage of dendritic cells, CD11c+ dendritic cells, and CD123+ dendritic cells in the peripheral blood of methamphetamine addicts during the detoxification period.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe percentage of dendritic cells (A), CD11c+ dendritic cells (B), and CD123+ dendritic cells (C) in the peripheral blood of methamphetamine addicts during the detoxification period.\u003c/p\u003e\n\u003cp\u003eDC = Dendritic cell, RisMeth = Methamphetamine addicts treated with risperidone, RisMpMeth = Methamphetamine addicts treated with risperidone and methylphenidate, a =beginning of the trial, B = End of detoxification, c = no more than one month after detoxification. Data is presented as mean ± standard deviation. The results are obtained from the analysis of 100,000 mononuclear cells. The signs *, **, and *** represented P\u0026lt;0.05, P \u0026lt; 0.005, and P\u0026lt;0.0005, respectively, and were used to compare groups.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5381470/v1/b3a07622d9520d3aa67d7efe.png"},{"id":70197357,"identity":"0b5c2a65-9d8c-4faa-804b-d5b6b4c4d139","added_by":"auto","created_at":"2024-11-29 11:45:59","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":68885,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMean fluorescence intensity (MFI) of HLA-DR, CD11c, and CD123 peripheral blood dendritic cells.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMean fluorescence intensity (MFI) of the HLA-DR peripheral blood dendritic cell molecule on the surface of peripheral blood dendritic cells (A), the CD11c molecule on the surface of CD11c+ dendritic cells (B), and the CD123 molecule on the surface of CD123+ dendritic cells (C) of methamphetamine addicts.\u003c/p\u003e\n\u003cp\u003eDC= Dendritic cell, Meth= methamphetamine. Data are presented as mean ± standard error. The results were extracted from the analysis of 100 thousand mononuclear cells. The sign * indicates P\u0026lt;0.05 compared to the control.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5381470/v1/42f2f6344a42a72c5340c76d.png"},{"id":70197356,"identity":"7cee3aaa-4880-41e1-bcb0-e6dd23d12a6a","added_by":"auto","created_at":"2024-11-29 11:45:59","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":85870,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMean fluorescence intensity (MFI) of the HLA-DR, CD11c, and CD123 molecules on the surface of peripheral blood dendritic cells.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe level of HLA-DR molecule expression on the surface of dendritic cells (A), CD11c on the surface of CD11c+ dendritic cells (B), and CD123 molecule on the surface of CD123+ dendritic cells (C) in the peripheral blood of methamphetamine addicts during the detoxification period.\u003c/p\u003e\n\u003cp\u003eDC = Dendritic cell, RisMeth = Methamphetamine addicts treated with risperidone, RisMpMeth = Methamphetamine addicts treated with risperidone and methylphenidate, a =beginning of the trial, B = End of detoxification, c = no more than one month after detoxification. Data is presented as mean ± standard deviation. The results are obtained from the analysis of 100,000 mononuclear cells. The signs *, **, and *** represented P\u0026lt;0.05, P \u0026lt; 0.005, and P\u0026lt;0.0005, respectively, and were used to compare groups.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-5381470/v1/7f89809954619bb7c48fe184.png"},{"id":72579458,"identity":"34d7ff4c-622c-483e-b51b-b55a2d0e1d0e","added_by":"auto","created_at":"2024-12-30 05:01:56","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1033139,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5381470/v1/1a7bebea-07fb-408c-b0f9-f18d1a5e030a.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Immunomodulatory Effects of Risperidone and Methylphenidate on Dendritic Cells in Methamphetamine Addicts: Insights from a Detoxification Study","fulltext":[{"header":"Introduction","content":"\u003cp\u003eNarcotics exert a profound influence on various body systems, including the immune system (1). This effect is observed irrespective of the duration of use, whether short-term or long-term. The underlying mechanism of action primarily involves three key receptors: \u0026micro; (MOR), κ (KOR), and δ (DOR) opioid receptors (2).\u003c/p\u003e \u003cp\u003eNarcotics are known to suppress immune function, which is a significant factor contributing to the increased prevalence of infectious diseases and cancers among individuals with substance use disorders (3\u0026ndash;5). Additionally, the immunosuppressive effects of these substances can exacerbate the severity and prolong the duration of infections (6). Conversely, some studies suggest that short-term use of narcotics as analgesics may stimulate certain aspects of the immune system (7).\u003c/p\u003e \u003cp\u003eMethamphetamine (METH) is a highly addictive central nervous system stimulant (8). Its widespread use presents substantial public health and economic challenges globally (9). The drug induces feelings of euphoria, excitement, and weight loss, effects that are comparable to those produced by cocaine. Methamphetamine acts as an agonist at dopamine receptors, leading to increased dopamine release (10).\u003c/p\u003e \u003cp\u003eDetoxification is the initial step for individuals entering rehabilitation programs for substance use disorders. During this process, antipsychotic medications such as risperidone\u0026mdash;an antagonist of opioid receptors and dopamine D2 receptors\u0026mdash;or a combination of antipsychotics and methylphenidate (Ritalin), a stimulant, may be administered to alleviate withdrawal symptoms associated with methamphetamine use (11\u0026ndash;13).\u003c/p\u003e \u003cp\u003eHistorically, lymphocytes were the primary focus in studies examining the immunomodulatory effects of various substances. However, recent findings indicate that immunomodulatory agents predominantly exert their effects by suppressing the differentiation, maturation, and activation of dendritic cells (DCs) (14).\u003c/p\u003e \u003cp\u003eDendritic cells play a pivotal role in presenting antigens to T lymphocytes and are essential for both innate and adaptive immune responses (15). They exhibit unique responses to various pathogens due to their diverse subgroups and strategic distribution throughout the body. In humans, dendritic cells are classified into two major subgroups: CD11c\u0026thinsp;+\u0026thinsp;myeloid dendritic cells (mDCs) and CD123\u0026thinsp;+\u0026thinsp;lymphoid dendritic cells (plasmacytoid DCs, pDCs) (16).\u003c/p\u003e \u003cp\u003eWhile extensive research has been conducted on the effects of methamphetamine in murine models (17), there is a growing interest in understanding its impact on human subjects. However, our comprehension of methamphetamine's effects on the human immune system remains limited (14).\u003c/p\u003e \u003cp\u003eThe objective of this study was to investigate the immunomodulatory effects of methamphetamine and its detoxification agents on dendritic cells.\u003c/p\u003e"},{"header":"Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003eParticipants\u003c/b\u003e:\u003c/h2\u003e \u003cp\u003eA total of 30 individuals with methamphetamine addiction and 30 healthy control subjects were enrolled in the study. The control group was closely matched to the addicted group in terms of age, sex, weight, height, and body mass index (BMI). Control subjects were selected from a population of university staff and students who had never used psychotropic drugs. The general health of the control group was confirmed through physical examinations and routine biochemical and hematological tests. The study was approved by the Ethics Committee, and all participants provided informed consent.\u003c/p\u003e \u003cp\u003eAll volunteers were free from infectious diseases, active inflammatory conditions, and other disorders affecting the immune system. Urine samples collected from the control group tested negative for morphine and methamphetamine.\u003c/p\u003e \u003cp\u003eThe addicted group was selected based on specific criteria: individuals who consumed more than 500 mg of methamphetamine daily for over one year, had a history of addiction to other drugs for no longer than three months in the past, and exhibited positive urine test results for methamphetamine above the detection threshold.\u003c/p\u003e \u003cp\u003eThe group of 30 addicted volunteers was divided into two subgroups of 15 individuals each. One subgroup received risperidone, while the other subgroup was administered both risperidone and methylphenidate concurrently. The initial dose of risperidone was 5 mg, which was increased to 7.5 mg by the end of the study. Methylphenidate was administered in a tapering manner over ten days (10 mg every 8 hours). The immune response of the addicted group was evaluated at three time points: at the beginning of the trial, at the end of detoxification, and up to one month following detoxification.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eFlow cytometry:\u003c/h3\u003e\n\u003cp\u003eMononuclear cells were isolated from peripheral blood samples using Ficoll density gradient centrifugation. A concentration of 25 \u0026times; 10⁶ cells was obtained, and 20 \u0026micro;l (equivalent to 5 \u0026times; 10⁵ cells) was transferred to FACS tubes. Each tube received 15 \u0026micro;l of a cocktail containing anti-CD3, CD14, CD16, CD19, CD20, and CD56 antibodies labeled with FITC (Lin-1), along with 5 \u0026micro;l of anti-CD34 antibody labeled with FITC (to exclude CD34\u0026thinsp;+\u0026thinsp;HLA-DR\u0026thinsp;+\u0026thinsp;precursors) and 5 \u0026micro;l of HLA-DR antibody labeled with PerCP.\u003c/p\u003e \u003cp\u003eFor three-color flow cytometry analysis, 5 \u0026micro;l of PE-labeled CD123 antibody was added to identify Lin-/HLA-DR+/CD123\u0026thinsp;+\u0026thinsp;plasmacytoid dendritic cells (pDCs), while another tube received 5 \u0026micro;l of PE-labeled CD11c antibodies to identify Lin-/HLA-DR+/CD11c\u0026thinsp;+\u0026thinsp;myeloid dendritic cells (mDCs).\u003c/p\u003e \u003cp\u003eSamples were analyzed using a FACScan flow cytometer (BD Biosciences, San Jose, CA), and data were processed using FlowJo software (18).\u003c/p\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis:\u003c/h2\u003e \u003cp\u003eOne-way analysis of variance (ANOVA) was employed to assess statistical significance between experimental groups. All data are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM. P-values\u0026thinsp;\u0026lt;\u0026thinsp;0.05 were considered statistically significant (*p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, **p\u0026thinsp;\u0026lt;\u0026thinsp;0.01).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e1-Selection of volunteers:\u003c/h2\u003e \u003cp\u003eBased on the selection criteria, 30 healthy individuals were chosen as the control group, while 30 individuals with methamphetamine addiction were selected as the experimental group (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). There were no statistically significant differences in age, height, weight, or body mass index (BMI) between the two groups (control and experimental). The general health of the control group was confirmed through physical examinations and routine hematological and biochemical tests (results not shown).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDemographic characteristics of volunteers included in the study\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl group\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eExperiment group\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSex\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e(Mean of years\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) Age\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e30.57\u0026thinsp;\u0026plusmn;\u0026thinsp;3.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30.17\u0026thinsp;\u0026plusmn;\u0026thinsp;5.47\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e(Mean of cm\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) Height\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e172.29\u0026thinsp;\u0026plusmn;\u0026thinsp;6.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e171.13\u0026thinsp;\u0026plusmn;\u0026thinsp;6.04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e(Mean of kg\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) Weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e73.80\u0026thinsp;\u0026plusmn;\u0026thinsp;5.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e69.85\u0026thinsp;\u0026plusmn;\u0026thinsp;12.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBody Mass Index (Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e24.87\u0026thinsp;\u0026plusmn;\u0026thinsp;1.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.912\u0026thinsp;\u0026plusmn;\u0026thinsp;4.11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2- Determination of the frequency of peripheral blood dendritic cells in volunteers\u003c/h2\u003e \u003cp\u003eThis study identified two major dendritic cell (DC) subsets in peripheral blood mononuclear cells (PBMCs): Lin\u0026thinsp;\u0026minus;\u0026thinsp;HLA-DR\u0026thinsp;+\u0026thinsp;CD11c\u0026thinsp;+\u0026thinsp;myeloid dendritic cells (mDCs) and Lin\u0026thinsp;\u0026minus;\u0026thinsp;HLA-DR\u0026thinsp;+\u0026thinsp;CD123\u0026thinsp;+\u0026thinsp;plasmacytoid dendritic cells (pDCs) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTo quantify the number of dendritic cells, 100,000 cells were counted within the gate of mononuclear cells. The percentage of peripheral blood dendritic cells in methamphetamine addicts was significantly lower compared to the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). The percentage of CD11c\u0026thinsp;+\u0026thinsp;DCs in the peripheral blood of methamphetamine addicts also showed a significant decrease compared to controls (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). Similarly, the percentage of CD123\u0026thinsp;+\u0026thinsp;DCs in the peripheral blood of methamphetamine addicts was lower than that of the controls (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003eImmune responses in methamphetamine addicts were assessed at three distinct time points: at the beginning of the trial, at the end of detoxification, and up to one month after detoxification (prior to the onset of withdrawal symptoms). Detoxification with risperidone restored the percentage of peripheral blood dendritic cells in methamphetamine addicts compared to the control group. However, detoxification with both risperidone and methylphenidate did not reverse this reduction (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eAs illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB, detoxification with risperidone corrected the percentage of CD11c\u0026thinsp;+\u0026thinsp;DCs in the peripheral blood of methamphetamine addicts. In contrast, detoxification with risperidone and methylphenidate did not restore the reduced percentage of CD11c\u0026thinsp;+\u0026thinsp;DCs in these individuals.\u003c/p\u003e \u003cp\u003eRisperidone treatment led to a significant increase in the previously reduced percentage of CD123\u0026thinsp;+\u0026thinsp;dendritic cells in the peripheral blood of methamphetamine addicts. However, when compared to the control group, detoxification with both risperidone and methylphenidate resulted in a significant decrease in the percentage of CD123\u0026thinsp;+\u0026thinsp;DCs in the peripheral blood of addicts (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003e \u003cb\u003e3-The expression of HLA-DR, CD11c and CD123 markers on CD11c\u0026thinsp;+\u0026thinsp;and CD123\u0026thinsp;+\u0026thinsp;dendritic cells in volunteers\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThis study evaluated the mean fluorescent intensity (MFI), which represents the expression level of cell surface markers, for HLA-DR, CD11c, and CD123 on dendritic cells in the peripheral blood of volunteers. As depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA, the expression level of HLA-DR on the surface of peripheral blood dendritic cells from methamphetamine addicts was significantly decreased compared to that in the control group.\u003c/p\u003e \u003cp\u003eFurthermore, the expression level of CD123 on the surface of peripheral blood CD123\u0026thinsp;+\u0026thinsp;DCs from methamphetamine addicts was significantly lower than that observed in the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003eAs shown in Fig.\u0026nbsp;5A, detoxification with risperidone significantly altered the reduced expression of the HLA-DR molecule on the surface of peripheral blood dendritic cells in methamphetamine addicts. However, the combination of risperidone and methylphenidate did not restore the decreased expression of HLA-DR on peripheral blood dendritic cells in this group.\u003c/p\u003e \u003cp\u003eDetoxification with risperidone resulted in a significant increase in the expression of the CD11c molecule on CD11c\u0026thinsp;+\u0026thinsp;dendritic cells in the peripheral blood of methamphetamine addicts. In contrast, the combination of risperidone and methylphenidate did not improve the reduced expression of CD11c on the surface of these CD11c\u0026thinsp;+\u0026thinsp;dendritic cells (Fig.\u0026nbsp;5B).\u003c/p\u003e \u003cp\u003eNeither detoxification regimen was able to correct the diminished expression of the CD123 molecule on the surface of CD123\u0026thinsp;+\u0026thinsp;dendritic cells in the peripheral blood of methamphetamine addicts when compared to the control group (Fig.\u0026nbsp;5C).\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eDendritic cells play a crucial role in processing and presenting antigens, thereby stimulating na\u0026iuml;ve T cells and regulating the quality of subsequent immune responses. In the bone marrow, newly generated dendritic cells migrate through the bloodstream to various peripheral tissues (19). Consequently, the presence of circulating blood dendritic cells can provide insights into the overall status of the immune system. For example, a study by Lissoni et al. found that decreased levels of circulating blood dendritic cells were strongly associated with suppressed immune function in patients with advanced malignant tumors (20).\u003c/p\u003e \u003cp\u003eOne contributing factor to the increased incidence of infections and cancers among substance abusers is the immunosuppressive effects of drugs (21, 22). In the current study, we assessed the overall percentages of two main subgroups of dendritic cells in the peripheral blood of methamphetamine addicts and examined the surface markers of these cells.\u003c/p\u003e \u003cp\u003eOur data indicate that the methamphetamine-addicted group exhibited significantly lower total dendritic cell counts, as well as reduced numbers of CD11c\u0026thinsp;+\u0026thinsp;and CD123\u0026thinsp;+\u0026thinsp;dendritic cells compared to healthy controls. This finding suggests a potential defect in their immune system.\u003c/p\u003e \u003cp\u003eSeveral factors may contribute to the decrease in dendritic cell percentages, including reduced production rates in the bone marrow, disruptions in monocyte responses to differentiation signals, inhibition of factors that promote dendritic cell differentiation, or the induction of apoptosis in dendritic cells (23). Although further studies are needed to confirm these possibilities, our findings suggest that dendritic cells are involved in the immunopathogenesis of disorders associated with methamphetamine use.\u003c/p\u003e \u003cp\u003eIn methamphetamine addicts, detoxification with risperidone restored the total percentage of dendritic cells as well as the numbers of CD11c\u0026thinsp;+\u0026thinsp;and CD123\u0026thinsp;+\u0026thinsp;dendritic cells to levels comparable to those of the control group. In contrast, treatment with a combination of risperidone and methylphenidate resulted in adverse effects.\u003c/p\u003e \u003cp\u003eHLA-DR is a critical molecule that stimulates T cells and serves as an indicator of dendritic cell activity. It is expressed on the surface of antigen-presenting cells, including dendritic cells, monocytes, macrophages, and B cells. HLA-DR presents processed extracellular antigens to T helper cells, and its expression level significantly influences the initiation of specific immune responses. A reduction or absence of HLA-DR can impair antigen-presenting cell function, leading to inadequate antigen presentation and diminished cytokine secretion (23).\u003c/p\u003e \u003cp\u003eCytokines regulate the expression of HLA-DR in an antagonistic manner. For instance, IFN-γ enhances HLA-DR expression and bolsters cellular immune responses, while IL-10 inhibits its expression (24).\u003c/p\u003e \u003cp\u003eOur data indicate that the expression of HLA-DR is decreased in the dendritic cells of individuals addicted to methamphetamine. HLA-DR is a critical surface marker on dendritic cells responsible for activating T cells; thus, a reduction in its expression may signify immunoparalysis. The diminished expression of HLA-DR not only impairs dendritic cell function but also increases the susceptibility of methamphetamine addicts to infectious diseases (25). Our previous study demonstrated elevated levels of IL-10 in the serum of these individuals (26). However, further research is necessary to fully elucidate the mechanisms underlying this reduction.\u003c/p\u003e \u003cp\u003eThe decreased expression of HLA-DR in methamphetamine addicts was restored following treatment with risperidone. In contrast, co-administration of risperidone and methylphenidate exacerbated the downregulation of HLA-DR expression.\u003c/p\u003e \u003cp\u003eCD11c, a member of the integrin family, plays a vital role in cellular attachment and is abundantly expressed on the surface of myeloid dendritic cells (27). This study revealed that methamphetamine significantly reduced the expression of the CD11c molecule in peripheral blood CD11c\u0026thinsp;+\u0026thinsp;dendritic cells among addicts. Risperidone treatment restored CD11c expression to levels comparable to those in the control group; however, co-administration with methylphenidate exacerbated this reduction.\u003c/p\u003e \u003cp\u003eLymphoid dendritic cells require IL-3 for differentiation, leading to high expression of the IL-3 receptor alpha chain (CD123) (28). Methamphetamine notably decreased the expression of CD123. Detoxification with risperidone increased CD123 expression; however, when combined with methylphenidate, this expression was reduced, although these changes were not statistically significant.\u003c/p\u003e \u003cp\u003eOverall, these data suggest that long-term consumption of methamphetamine has an immunosuppressive effect. Risperidone acts as an antagonist to methamphetamine, and its use as a detoxifying agent reverses this immunosuppressive state. Conversely, methylphenidate, which is a stimulant similar to methamphetamine, exacerbates the immunosuppressive condition of the immune system.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eDendritic cells in individuals addicted to methamphetamine may lose their ability to respond effectively to invading pathogens, potentially increasing their susceptibility to infections. Impairments in dendritic cell function could also diminish the effectiveness of vaccines in this population. During detoxification from methamphetamine, the administration of risperidone was found to mitigate some of the adverse effects associated with methamphetamine use. However, the combination of risperidone and methylphenidate exacerbated these negative effects. Although this study is grounded in natural science experimentation, its findings may have practical implications for clinical practice.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Research Council of Arak University of Medical Sciences.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was funded by the Council of Arak University of Medical Sciences (grant number 658).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and/or analyzed during the current study are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that there is no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Ethics Committee of Arak University of Medical Sciences approved this study (Ethical Code: IR.ARAKMU.REC.1390.13.9).\u0026nbsp;\u003c/p\u003e"},{"header":" References","content":"\u003col\u003e\n \u003cli\u003eGhazavi A, Solhi H, Moazzeni SM, Rafiei M, Mosayebi G. 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Reduced frequencies and functional impairment of dendritic cell subsets and non-classical monocytes in myelodysplastic syndromes. Haematologica. 2022;107(3):655-67.\u003c/li\u003e\n \u003cli\u003eOzdemir AT, Oztatlici M, Ozgul Ozdemir R, Cakir B, Ozbilgin K, Dariverenli E. The effects of preconditioning with IFN-\u0026gamma;, IL-4, and IL-10 on costimulatory ligand expressions of mesenchymal stem cells. Int J Med Biochem. 2021;4(2):121-30.\u003c/li\u003e\n \u003cli\u003eMorel AS, Coulton G, Londei M. Regulation of major histocompatibility complex class II synthesis by interleukin-10. Immunology. 2002;106(2):229-36.\u003c/li\u003e\n \u003cli\u003eAkbari A, Mosayebi G, Samiei AR, Ghazavi A. Methadone therapy modulate the dendritic cells of heroin addicts. International immunopharmacology. 2019;66:330-5.\u003c/li\u003e\n \u003cli\u003eWu J, Wu H, An J, Ballantyne CM, Cyster JG. Critical role of integrin CD11c in splenic dendritic cell capture of missing-self CD47 cells to induce adaptive immunity. Proceedings of the National Academy of Sciences of the United States of America. 2018;115(26):6786-91.\u003c/li\u003e\n \u003cli\u003ePodolska MJ, Gr\u0026uuml;tzmann R, Pilarsky C, B\u0026eacute;nard A. IL-3: key orchestrator of inflammation. Frontiers in immunology. 2024;15:1411047.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Dendritic cells, Methamphetamine, Risperidone, Methylphenidate","lastPublishedDoi":"10.21203/rs.3.rs-5381470/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5381470/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground and aim:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDrug abuse can impact the function of immune cells, leading to a compromised immune system response. This study aimed to investigate the immunomodulatory effects of methamphetamine and its detoxification agents on peripheral blood dendritic cells.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThirty individuals with methamphetamine addiction were divided into two groups. One group received treatment with Risperidone, while the second group was treated with a combination of Risperidone and Methylphenidate. The percentages of myeloid and plasmacytoid dendritic cells, as well as their surface markers, were assessed before and after the detoxification program.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe percentages of both CD11c+ and CD123+ dendritic cells in peripheral blood were significantly lower in methamphetamine addicts compared to the control group. Detoxification with Risperidone corrected this reduction, while the combination of Risperidone and Methylphenidate produced adverse effects. The expression of HLA-DR, CD11c, and CD123 markers was downregulated in the dendritic cells of methamphetamine addicts. Treatment with Risperidone restored these markers, whereas the combination therapy further exacerbated the downregulation of these markers.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe findings suggest that detoxification with Risperidone may help ameliorate the immunological disorders associated with methamphetamine use.\u003c/p\u003e","manuscriptTitle":"Immunomodulatory Effects of Risperidone and Methylphenidate on Dendritic Cells in Methamphetamine Addicts: Insights from a Detoxification Study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-29 11:45:54","doi":"10.21203/rs.3.rs-5381470/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"fde827cd-9743-451d-80a2-f0b152c2b63c","owner":[],"postedDate":"November 29th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-30T04:53:46+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-29 11:45:54","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5381470","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5381470","identity":"rs-5381470","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}