Treatment of cancer treatment-induced oral mucositis

Abstract

Compositions comprising Streptococcus salivarius and Streptococcus salivarius are provided for treating ENT diseases, particularly cancer treatment-induced oral mucositis and cancer treatment-related respiratory infections.

Description

[Technical Field] 【0001】 The present invention relates to Streptococcus salivarius for use in the treatment and prevention of diseases of the ear, nose, and throat (ENT), wherein the ENT disease is induced by or associated with cancer treatment, and preferably the ENT disease is oral mucositis or a respiratory infection. Furthermore, the present invention relates to the use of Streptococcus salivarius in maintaining healthy oral mucosa during or after cancer treatment, and to Streptococcus salivarius for use in the treatment and prevention of respiratory infections in immunocompromised patients. [Background technology] 【0002】 Cancer is a global health problem, causing the deaths of one in six people worldwide. In 2020, there were an estimated 19.3 million new cancer cases and approximately 10 million deaths from cancer globally. 【0003】 Cancer treatment modalities can be divided into conventional (traditional) categories and advanced, novel, or modern categories. Currently, more than half of ongoing medical trials worldwide focus on cancer treatment. Factors such as the type, location, and severity of the cancer guide the selection of treatment and the assessment of its course. The most widely used traditional treatment methods are surgery, chemotherapy, and radiotherapy. Chemotherapy, used alone or in combination with radiotherapy, is considered the most effective and widely used modality in cancer treatment. Modern modalities, on the other hand, include hormone therapy, anti-angiogenic therapy, stem cell therapy including hematopoietic stem cell transplantation (HSCT), immunotherapy, and dendritic cell-based immunotherapy (Debela et al., SAGE Open Med.9 (2021), doi:10.1177 / 20503121211034366). 【0004】 Chemotherapy and radiotherapy are widely used as non-surgical cancer treatments that can extend life and, in some cases, even lead to a cure. However, chemotherapy and radiotherapy cause numerous toxic side effects, including oral mucositis that severely impairs quality of life. The most widely observed disorder is oral mucositis. Approximately 40% of patients treated with chemotherapy develop mucositis, and this percentage rises to approximately 90% in head and neck cancer patients (HNC) treated with both chemotherapy and radiotherapy (Kusiak et al., Int J Environ Res Public Health.17(2020), 2464; Pulito et al., J Exp Clin Cancer Res 9(2020), 210). 【0005】 Allogeneic hyperstimulant hyperstimulant chemotherapy (HSCT) is considered the most potent post-remission antileukemia therapy for adult acute lymphoblastic leukemia. However, it frequently causes acute oral complications, including mucositis, particularly local and systemic infections of the ear, nose, and throat (ENT) region, dry mouth, and altered taste (Haverman et al., Mediators Inflamm. (2014), 378281). In particular, 85% of patients receiving intensive chemotherapy for HSCT also develop oral mucositis, and the incidence of severe oral mucositis (≥grade 3) is 100% when patients receive intensive total body irradiation (TBI) along with cyclophosphamide chemotherapy in combination with HSCT. 【0006】 Therefore, oral mucositis (OM), characterized by inflammation and damage to the oral mucosa, is the most common oral complication in patients undergoing cancer treatment. OM causes erythema, ulceration, pain, dysphagia, and malnutrition, severely impacting the patient's quality of life and sometimes leading to the discontinuation of anti-cancer treatment. Severe oral mucositis can result in unplanned hospitalization and may lead to changes in treatment plans, such as interruption of radiation therapy or reduction of chemotherapy. As a result, it can negatively affect treatment outcomes. 【0007】 To better understand omphalomyelitis (OM) and find ways to prevent and treat it, the disease mechanisms have been extensively studied. Researchers have found that OM is far more complex than initially thought. The development of mucositis has been found to be a dynamic process typically divided into five stages: initiation, primary injury response, signal amplification, ulceration, and healing. In the initial stage, cancer treatments such as chemotherapy or radiotherapy cause both DNA and non-DNA damage to basal epithelial cells, triggering a cascade of reactive oxygen species (ROS) that directly damage cells, tissues, and blood vessels. In the primary injury response stage, mucositis is driven by three pathways activated by the ROS cascade: the NF-κB pathway, the ceramide pathway, and the matrix metalloproteinase pathway. In the signal amplification stage, a range of pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6, further damage basal epithelial cells through positive feedback in the three pathways. During the ulcer formation phase, the loss of mucosal integrity promotes bacterial colonization, which in turn stimulates macrophage cells to produce more pro-inflammatory cytokines, exacerbating tissue damage. This phase causes extreme pain in the patient. Finally, during the healing phase, signals from the submucosa's extracellular matrix and mesenchyme promote cell proliferation and differentiation, re-establishing the mucosal barrier (Shu et al., Oral Oncology 102(2020), 104559). 【0008】 Although the mechanisms are well studied, controlling omandibular malformation (OM) remains challenging. Interventions including basic oral care, growth factors and cytokines, anti-inflammatory drugs, cryotherapy, and low-level laser therapy are suggested by the Multinational Society for Supportive Care of Cancer / International Society of Oral Oncology (MASCC / ISOO) clinical practice guidelines. However, only palifermin (keratinocyte growth factor-1) has been approved by the U.S. Food and Drug Administration and the European Medicines Agency for the purpose of mitigating OM in a very limited number of high-risk patient populations (Shu et al., Oral Oncology 102(2020), 104559). 【0009】 Therefore, further strategies for the treatment of oral mucositis are of paramount importance for the health and favorable treatment outcomes of patients undergoing cancer treatment. [Prior art documents] [Non-patent literature] 【0010】 [Non-Patent Document 1] Debela et al., SAGE Open Med.9 (2021), doi:10.1177 / 20503121211034366 [Non-Patent Document 2] Kusiak et al., Int J Environ Res Public Health.17(2020), 2464 [Non-Patent Document 3] Pulito et al., J Exp Clin Cancer Res 9(2020), 210 [Non-Patent Document 4] Haverman et al., Mediators Inflamm. (2014), 378281 [Non-Patent Document 5] Shu et al., Oral Oncology 102(2020), 104559 [Overview of the project] [Problems that the invention aims to solve] 【0011】 The present invention generally relates to Streptococcus salivarius for use in a method of treating or preventing ear, nose, and throat (ENT) diseases induced by or associated with cancer treatment, particularly oral mucositis (OM) induced by cancer treatment or respiratory tract infections (RTi) associated with cancer treatment, the method comprising administering to a subject needing it the probiotic microorganism Streptococcus salivarius. The present invention is based on the surprising finding that OM developed by cancer patients who have received radiotherapy and concurrent chemoradiotherapy (CCRT) respectively, or who have received HSCT, was prevented, reduced and improved in patients treated with the probiotic microorganism Streptococcus salivarius K12 before, during and / or after cancer treatment, and that the administration of the probiotic microorganism Streptococcus salivarius K12 was safe in patients undergoing cancer treatment and furthermore reduced the incidence of respiratory tract infections in said patient group. 【0012】 To date, investigations on the potential of probiotics, such as Lactobacillus brevis, Lactobacillus lactis, as well as a mixture of Bifidobacterium longum, Lactobacillus lactis, and Enterococcus faecium, to treat cancer treatment-induced OM have been reported, but the results are variable and lack reliability. See the review by Shu et al., Oral Oncology 102 (2020), 104559. Also, the mechanism of the protective effect of probiotics against various conditions generally remains unclear. 【Means for Solving the Problems】 【0013】 Experiments conducted within the scope of the present invention surprisingly revealed that the probiotic strain, namely Streptococcus salivarius K12, had a significant curative effect on the onset of OM and alleviated OM symptoms when applied to patients undergoing CCRT as shown in Example 1 or patients who had undergone HSCT as shown in Example 2. Specifically, as seen in FIG. 1 and as described in detail in Example 1, during 7 weeks of CCRT, the incidence of radiation therapy-induced oral mucositis (RIOM) with a Radiation Therapy Oncology Group (RTOG) score ≥2 decreased in patients administered oral probiotic Streptococcus salivarius K12. Furthermore, the onset of RIOM with RTOG ≥2 was delayed by 3 weeks in more than 60% of patients, and patients ingesting oropharyngeal probiotics were completely protected from severe RIOM classified as RTOG 3 and 4, in contrast to patients not ingesting probiotics. The RTOG score 4 is classified as life-threatening and includes ulcers, bleeding, or necrosis. In contrast, the RTOG score 2 is classified as moderate. 【0014】 OM that developed in patients undergoing CCRT is classified as RIOM because, in contrast to OM caused by mere chemotherapy (CIOM), radiation, in addition to its cytotoxic effect, also causes further necrosis and inflammatory effects on the oral mucosa. 【0015】 As shown in Figure 6 and described in detail in Example 2, during a 130-day study period in HSCT patients (HSCT is a treatment used for multiple malignant and non-malignant diseases, and the preceding chemotherapy is used for pre-transplant bone marrow apheresis), the incidence of OM grade ≥ 2 (WHO grading scale) was reduced in patients administered the oropharyngeal probiotic Streptococcus salivarius K12 compared to the control group that did not receive the probiotic. Notably, no ulcerative OM was observed in patients in the probiotic group during the 100-day oropharyngeal probiotic intervention. Therefore, the results of the experiments conducted according to the present invention have established for the first time a therapeutically effective probiotic-based treatment for RIOM and OM induced by HSCT and preceding chemotherapy (CIOM), respectively, in human subjects. 【0016】 During the study periods described in Examples 1 and 2, adverse events (side reactions) were also monitored, but no adverse events were reported in either study. Therefore, Streptococcus salivarius is considered safe in patients treated with radiotherapy and those who underwent HSCT, respectively. This is particularly important because patients undergoing anti-cancer treatment are immunosuppressed, and therefore, further caution and comprehensive safety evaluation are necessary before the application of probiotics. Thus, proof of concept in clinical trials is essential for the approval of probiotic therapy in humans, especially those with weakened immune function due to cancer treatment or other immunosuppressant drug treatment. 【0017】 Therefore, human trials conducted in accordance with the present invention represent an important step toward the approval of a probiotic strain, namely Streptococcus salivarius, for the treatment of cancer treatment-induced omya malformation. 【0018】 Further mouse studies were conducted to visualize the effects of chemotherapeutic agents and Streptococcus salivarius on oral epithelial cells, and therefore to monitor the effects of Streptococcus salivarius in the treatment of chemotherapy-induced omandibular malformation (OM). Anticancer drugs, in this case chemotherapeutic agents (busulfan combined with cyclophosphamide (Example 3) and 5-fluorouracil (Example 4)), were administered over 5-6 days to mimic repeated administration and the onset of OM in human subjects. This mode of administration is similar to that actually used in human patients, as chemotherapy is usually delivered over a short period, in which case damage to mucosal tissue tends to be acute (in contrast, radiotherapy, which is usually administered over several weeks, makes it difficult to establish a suitable mouse model for studying RIOM). Chemotherapy-induced OM (CIOM) usually develops within 4-7 days after the start of treatment and peaks within 2 weeks. This period can be well observed in mice. 【0019】 While CIOM and RIOM share similar cellular events, their biological pathways differ somewhat. As mentioned above, radiation, in addition to its cytotoxic effects, leads to further necrosis and inflammation of the oral mucosa. In particular, chemotherapy is administered systemically, while radiotherapy affects specific areas of the body. Furthermore, as mentioned above, there are differences in the dynamics of the treatments, which affect the clinical course. Chemotherapy can be delivered in a short time, in which case damage to mucosal tissue tends to be acute. CIOM usually develops within 4-7 days after the start of treatment and peaks within 2 weeks. Radiotherapy, in most cases, is administered in small fractions over several weeks and has a more progressive clinical course. RIOM typically develops at a cumulative dose of about 15 Gy (about 10 days later), typically reaches maximum severity at 30 Gy, and lasts for several weeks or months (Raber-Durlacher et al., Oral Oncology 46(2010)452-456). 【0020】 As described in Example 3 and shown in Figures 10, 11, and 12, Streptococcus salivarius K12 promotes the healing of CIOM in mice. In particular, chemotherapy treatment in mice resulted in significant mucosal hypoplasia and ulceration in the tongue. Furthermore, basal layer cells were only loosely aligned, and nuclear condensation, which is irreversible condensation of chromatin in the nucleus of cells undergoing necrosis or apoptosis, was observed. In addition, the tongue tissue had fewer cells in the stratum spinosum and stratum granulosum compared to healthy tissue. See Figures 10A and 10B. Treatment with Streptococcus salivarius K12 during chemotherapy restored the integrity of the tongue mucosa and partially restored the basal layer, stratum spinosum, and stratum granulosum. See Figure 10C. It was further observed that chemotherapy treatment reduced mucosal thickness by almost 50%, and when Streptococcus salivarius K12 was administered during chemotherapy, the reduction was less, i.e., about 25%. Please refer to Figures 11A-11C and Figure 12. Similar results were observed after treatment of mice with 5-Fu. As shown in Example 4 and Figure 17, the area of ​​the oral mucosa layer in mice decreased with 5-Fu treatment and significantly increased with S. salivarius K12 treatment. 【0021】 Based on these experiments, the inventors conceived the idea that inactivated Streptococcus salivarius K12 may have beneficial effects in the prevention and treatment of OM. This idea arose from the fact that inactivated probiotics have been shown to reduce systemic inflammation and preserve the cellular and molecular biological reactivity of viable probiotics (WO 2008 / 106373 A1). Furthermore, studies have been conducted to confirm that heat-inactivated Lactococcus salivarius CECT 5713 prevents the adhesion of Streptococcus mutans to hydroxyapatite, and therefore, the inactivated form can be used as a strategy to reduce the salivary concentration of this oral pathogen (Sanudo et al., Archives of Oral Biology 84(2017), 58-63). Regarding S. salivarius, it has also been shown that the cell wall contains antigenic compounds (Montague and Knox, J Gen Microbiol. 54 (1986), 237-246; Weerkamp and Jacobs, Infection and Immunity 38 (1982), 233-242). Therefore, we have developed the present invention that cell wall components may be sufficient to induce an immune response. This is in complete contradiction to the teachings of the prior art in the field of treatment of oral mucositis. For example, in Wang et al., Front. Immunol. 12 (2021), 684824, the effect of S. salivarius on the treatment of RIOM is attributed to the ability of live S. salivarius to reconstitute the oral microbiota, and therefore, based on this publication, those skilled in the art would not have attempted to use inactivated cells. 【0022】 As shown in Example 4, treatment of oral epithelial cells with heat-inactivated S. salivarius K12 actually restored the proliferative capacity of cells that had decreased during 5-Fu treatment. See Figure 16. Furthermore, the oral mucosal barrier was protected by heat-inactivated S. salivarius K12. As shown in Figure 18, the area of ​​the oral mucosal layer in mice decreased with 5-Fu treatment and significantly increased with heat-inactivated S. salivarius K12 treatment. The same effect was observed with live S. salivarius K12 (see Figure 17), which has already been shown in clinical trials to be suitable for the treatment of oral mucositis (see, for example, Example 2). Therefore, both live and heat-killed S. salivarius K12 significantly promote the proliferation of oral mucosal cells and are thus both suitable for the treatment of oral mucositis. 【0023】 Therefore, the present invention also relates to the use of inactivated S. salivarius K12 for use in the treatment of oral mucosal disorders, particularly oral mucositis, most preferably CIOM. 【0024】 Ovarian inflammatory bowel syndrome (OM) is not the only complication that can occur during cancer treatment. Due to the immunosuppressive effects of anticancer drugs, particularly chemotherapy and HSCT, respiratory infections are very common. Previous clinical trials in adults and children have already established the efficacy of daily administration of Streptococcus salivarius in reducing respiratory infections and major respiratory pathogens. 【0025】 The study described in Example 2 analyzed the effect of S. salivarius K12 on respiratory infections in HSCT patients, and therefore in patients with immune system disorders. Adjuvant treatment with S. salivarius K12 was shown to effectively reduce the prevalence of RTi in patients after HSCT, shorten the duration of respiratory symptoms, and reduce the number of days of antibiotic use. The latter finding is particularly important because the use of antibiotics in hematopoietic stem cell transplant (HSCT) patients is a subject of debate, and antibiotic use has been reported to have long-term adverse effects on overall survival after transplantation. 【0026】 Therefore, on the one hand, the safety of Streptococcus salivarius K12 has been demonstrated for the first time in immunosuppressed patients in studies conducted within the scope of the present invention, and on the other hand, its therapeutic effect on respiratory infections in HSCT patients has been demonstrated. Thus, Streptococcus salivarius can be used for the treatment of ENT diseases induced by cancer treatment, particularly ENT diseases induced by chemotherapy and / or radiotherapy, as well as for the treatment of patients who have undergone HSCT, especially immunosuppressed patients. [Brief explanation of the drawing] 【0027】 [Figure 1] The oropharyngeal probiotic Streptococcus salivarius (ENT-K12) prevents severe RIOM in nasopharyngeal cancer patients undergoing CCRT treatment. This figure shows the mean RTOG scores experienced by patients during 7 weeks of CCRT, with RIOM (RTOG=1) beginning at weeks 2 and 3 of the CCRT treatment period in the CCRT group (control group) and the CCRT-P group (probiotic-treated group), respectively. [Figure 2] Kaplan-Meier curve analysis of the first onset of RTOG level I RIOM in the CCTR group (control group) and the CCRT-P group (group administered the oropharyngeal probiotic Streptococcus salivarius (ENT-K12)). The horizontal axis is based on the start of the CCRT treatment period. The proportion of patients who did not develop RTOG level I RIOM decreased significantly faster in the CCRT group compared to the CCRT-P group (p<0.05). Y-axis: Patients who did not develop RTOG level I RIOM (%). [Figure 3]Kaplan-Meier curve analysis of the first onset of RTOG level II RIOM in the CCTR group (control group) and the CCRT-P group (group administered the oropharyngeal probiotic Streptococcus salivarius (ENT-K12)). The horizontal axis is based on the start of the CCRT treatment period. The proportion of patients who did not develop RTOG II RIOM decreased significantly faster in the CCRT group compared to the CCRT-P group (p<0.05). Y-axis: Patients who did not develop RTOG II RIOM (%). [Figure 4] Kaplan-Meier curve analysis of the first onset of RTOG level III RIOM in the CCTR group (control group) and the CCRT-P group (group administered the oropharyngeal probiotic Streptococcus salivarius (ENT-K12)). The horizontal axis is based on the start of the CCRT treatment period. The proportion of patients who did not develop RTOG III RIOM decreased significantly faster in the CCRT group compared to the CCRT-P group (p<0.05). Y-axis: Patients who did not develop RTOG III RIOM (%). [Figure 5] Kaplan-Meier curve analysis of the first onset of RTOG level IV RIOM in the CCTR group (control group) and the CCRT-P group (group administered the oropharyngeal probiotic Streptococcus salivarius (ENT-K12)). The horizontal axis is based on the start of the CCRT treatment period. There was no difference in the proportion of patients who did not develop RTOG IV RIOM between the CCRT group and the CCRT-P group (p>0.05). Only one patient in the CCRT group developed RTOG IV RIOM, while the remaining nine patients did not. Y-axis: Patients who did not develop RTOG IV RIOM (%). [Figure 6]The oropharyngeal probiotic Streptococcus salivarius (ENT-K12) prevents severe omaniosacral malformation (OM) in humans undergoing HSCT. This figure shows the mean incidence rate (OM grade ≥ 2 on the WHO grading scale) in the probiotic group (patients who underwent HSCT and were administered the oropharyngeal probiotic ENT-K12) and the control group (patients who underwent HSCT but were not administered the oropharyngeal probiotic ENT-K12). During the 130-day study period, patients who received the oropharyngeal probiotic had a reduced incidence of OM of grade 2 or higher. [Figure 7] The probability of having no RTi episodes during a 130-day study period, including 100 days of oropharyngeal probiotic intervention and a 30-day follow-up period. Kaplan-Meier analysis showed that patients in the probiotic group consistently had a higher probability of not having RTi episodes than the control group (p=0.071). [Figure 8] The cumulative duration of RTi-like symptoms during a 130-day study period, including a 100-day oropharyngeal probiotic intervention and a 30-day follow-up period. Kaplan-Meier analysis showed that the cumulative duration of RTi-like symptoms was consistently lower in the probiotic group (p=0.131). [Figure 9] The cumulative duration of RTi-like symptoms during a 130-day study period, including a 100-day oropharyngeal probiotic intervention and a 30-day follow-up period. Kaplan-Meier analysis showed that the probiotic group consistently had a shorter cumulative duration of RTi-like symptoms (p=0.131) and significantly fewer cumulative days of antibiotic use throughout the entire study period (p=0.011). [Figure 10]Representative images (80x magnification) of dorsal tongue staining showing the integrity of the tongue mucosa. A) Control group (untreated mice), B) Chemotherapy group (mice that received chemotherapy), C) Chemotherapy + ENT-K12 group (mice that received chemotherapy and were administered the oral / pharyngeal probiotic Streptococcus salivarius (ENT-K12)). In the chemotherapy group (B), significant mucosal hypoplasia and ulcer formation were observed in the tongue, and nuclear condensation, in which chromatin in the nuclei of necrotic or apoptotic cells condensed irreversibly, was observed. Basal layer cells were only loosely aligned, and the tongue tissue had fewer cells in the stratum spinosum and stratum granulosum than in the control group (A). In the chemotherapy + ENT-K12 group (C), the integrity of the tongue mucosa was restored, and the stratum basosum, stratum spinosum, and stratum granulosum were partially restored. [Figure 11] Representative images (80x magnification) of dorsal tongue staining showing mucosal thickness. A) Control group (untreated mice), B) Chemotherapy group (mice that received chemotherapy), C) Chemotherapy + ENT-K12 group (mice that received chemotherapy and were administered the oral / pharyngeal probiotic Streptococcus salivarius (ENT-K12)). In the chemotherapy group (B), mucosal thickness was reduced by almost 50% compared to the control group (A), and in the chemotherapy + ENT-K12 group (C), it was reduced by approximately 25% compared to the control group (A). [Figure 12] This figure shows the thickness of the mucosa in a longitudinal section of a mouse tongue. The thickness of the mucosa was reduced by almost 50% in chemotherapy-treated mice compared to untreated mice (healthy mucosa), while the reduction was only about 25% in mice that received chemotherapy and were also administered the oral / pharyngeal probiotic Streptococcus salivarius (ENT-K12). [Figure 13] Live-cell imaging experiments showed that the proliferative activity of oral epithelial cells (HOK cells) decreased 48 hours after 5-Fu treatment. [Figure 14] The proliferative activity of HOK cells was analyzed using the CCK-8 assay, and mitochondrial metabolic activity levels were measured as an indicator of cell proliferation capacity. Treatment with 5-Fu at concentrations exceeding 10 μg / mL for 48 hours significantly reduced the proliferative activity of HOK cells (p<0.05). [Figure 15]The proliferative activity of HOK cells was analyzed using the CCK-8 assay. Heat-killed Streptococcus salivarius ENT-K12 did not show proliferative toxicity to HOK cells at any concentration, and the cell viability of HOK cells was significantly increased by treatment with heat-killed Streptococcus salivarius ENT-K12 at concentrations >1.5 mg / mL. [Figure 16] The proliferation activity of HOK cells was analyzed using the CCK-8 assay. HOK cell proliferation was significantly reduced after treatment with 10 μg / mL of 5-Fu and significantly and dose-dependently restored after treatment with heat-killed Streptococcus salivarius ENT-K12. [Figure 17] Figures showing tissue sections (A-C) and the area of ​​the oral mucosa (D) demonstrate that 5-Fu has significant adverse effects on the integrity and permeability of the oral mucosa barrier in mice. The area of ​​the oral mucosa in mice decreased with 5-Fu treatment and significantly increased with treatment with live Streptococcus salivarius ENT-K12. [Figure 18] Figures showing tissue sections (A-C) and the area of ​​the oral mucosa (D) demonstrate that 5-Fu has significant adverse effects on the integrity and permeability of the oral mucosa barrier in mice. The area of ​​the oral mucosa in mice decreased with 5-Fu treatment and significantly increased with heat-killed Streptococcus salivarius ENT-K12 treatment. [Figure 19] Figures showing tissue sections (A-C) and mean fluorescence activity (D) indicate that Ki-67 expression was lower in the 5-Fu group than in the probiotic group (treated with live Streptococcus salivarius ENT-K12) and the control group. Therefore, treatment with live Streptococcus salivarius ENT-K12 significantly promoted the proliferation of oral mucosal cells. [Figure 20]Figures showing tissue sections (A-C) and mean fluorescence activity (D) indicate that Ki-67 expression was lower in the 5-Fu group than in the probiotic group (treated with heat-killed Streptococcus salivarius ENT-K12) and the control group. Therefore, treatment with heat-killed Streptococcus salivarius ENT-K12 significantly promoted the proliferation of oral mucosal cells. [Figure 21] Analysis of the HOK cell cycle by flow cytometry. (A) Control, (B) HOK cells treated with heat-killed Streptococcus salivarius ENT-K12, (C) HOK cells treated with 5-Fu, (D) HOK cells treated with 5-Fu and heat-killed Streptococcus salivarius ENT-K12. Compared to the control group, heat-killed Streptococcus salivarius ENT-K12 did not significantly alter the HOK cell cycle after 48 hours of treatment. 5-Fu treatment induced cell cycle arrest in the S phase and G2 / M phase, stages in which cells undergo rapid growth and protein synthesis in preparation for mitosis and cytokinesis. Heat-killed Streptococcus salivarius ENT-K12 restored the entire cell cycle after 5-Fu treatment. [Figure 22] Reactive oxygen species (ROS) levels in HOK cells were detected by flow cytometry. ROS levels in HOK cells were induced by 5-Fu treatment and significantly decreased after treatment with heat-killed S. salivarius ENT-K12. FITC-A values ​​indicate ROS levels in HOK cells that reflect DNA damage during chemotherapy (higher values ​​indicate higher ROS levels and more damage). [Modes for carrying out the invention] 【0028】 The present invention relates to the treatment of ear, nose, and throat (ENT) diseases in human subjects, wherein the ENT disease is preferably oral mucositis (OM), and the treatment comprises the administration of Streptococcus salivarius. In particular, the present invention relates to Streptococcus salivarius or a composition comprising Streptococcus salivarius for use in the treatment of ENT diseases in human subjects, wherein the ENT disease is cancer treatment-induced or cancer treatment-related ENT disease, preferably cancer treatment-induced OM or cancer treatment-related respiratory infection (RTi). With respect to respiratory infections, the present invention relates in particular to their treatment in immunosuppressed patients, such as patients who have undergone HSCT. The present invention also relates to the treatment of oral mucosal disorders, particularly oral mucositis, with inactivated Streptococcus salivarius. Furthermore, the present invention relates to the use of Streptococcus salivarius or compositions comprising Streptococcus salivarius in maintaining healthy ear, nose, and throat (ENT) regions, preferably healthy oral mucosa, in human subjects undergoing cancer treatment. Furthermore, the present invention relates to tablets, particularly lozenges, comprising Streptococcus salivarius for the indicated uses. 【0029】 Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present invention pertains. Similar or equivalent methods and materials to those described herein may be used in carrying out or testing the present invention, but exemplary methods and materials are described below. All publications, patent applications, patents, and other references referenced herein are incorporated by reference in their entirety. Materials, methods, and examples are illustrative and not intended to limit the scope. 【0030】 Other features and advantages of the present invention will become apparent from the following detailed description and claims. 【0031】 To avoid misunderstanding, expressions such as “in some embodiments,” “in certain embodiments,” “in certain cases,” “in some cases,” “in further embodiments,” and “in one embodiment” should be read with the understanding that any of the embodiments described therein combines the features of each of those embodiments, and that the disclosure is used and means to treat as if the combination of features of those embodiments were detailed in one embodiment. The same applies to any combination of embodiments and features shown in the appended claims and examples, which are also intended to be combined with features from corresponding embodiments disclosed herein, and for consistency and brevity, embodiments are characterized by dependencies, but in practice, each combination of embodiments and features that may be interpreted based on (multiple) dependencies should be considered literally disclosed and not interpreted as being selected from different options. 【0032】 The terms "human subject" and "patient" are used interchangeably in this specification. 【0033】 The term "treatment" refers to both "therapeutic treatment" and "preventive treatment." Treatment for ENT disease and treatment for OM also refer to patients who have not yet developed ENT disease and OM, respectively, but are at risk of developing such diseases and are prone to developing them. Patients at risk of developing ENT disease, particularly OM, include, for example, patients undergoing cancer treatment and otherwise immunosuppressed patients. 【0034】 The terms “cancer treatment,” “anti-cancer therapy,” “cancer treatment,” and “anti-cancer agent” are used interchangeably herein. Furthermore, cancer treatment can also be used to treat diseases other than cancer, such as autoimmune diseases. Therefore, patients receiving anti-cancer treatment do not necessarily have cancer, but instead, or in addition, may have, for example, an autoimmune disease. Thus, when referring to cancer treatment-induced omphalomyelitis (OM), OM may be induced by the treatment of cancer by cancer treatment, or by the treatment of another disease, particularly an autoimmune disease, by cancer treatment. 【0035】 More specifically, HSCT, or treatment with chemotherapy agents or antimetabolites such as methotrexate, is not only a treatment strategy for cancer patients, but HSCT is also a promising curative treatment for patients with autoimmune diseases such as bone marrow failure syndrome, congenital immunodeficiency, or multiple sclerosis. Low-dose chemotherapy and methotrexate treatment are also promising treatment approaches for autoimmune diseases such as rheumatoid arthritis, and these treatments can also cause OM in cancer patients as previously mentioned (Haverman et al., Mediators Inflamm. (2014), 378281; Gobbo et al., Photon Lasers Med 2 (2013), 71-76; Gobbo et al., Photon Lasers Med 2 (2013), 71-76). 【0036】 When referring to "cancer treatment-induced ENT disease," "cancer treatment-induced oral mucositis," "chemotherapy-induced ENT disease," "chemotherapy-induced OM (CIOM)," "radiotherapy-induced ENT disease," "radiotherapy-induced OM (RIOM)," "HSCT-induced ENT disease," "HSCT-induced OM," or similar expressions, it means that the ENT disease and OM are induced by or associated with the aforementioned treatments, respectively. 【0037】 HSCT involves the administration of healthy hematopoietic stem cells to patients with dysfunctional or depleted bone marrow. This helps to enhance bone marrow function and, depending on the disease being treated, leads to the destruction of malignant tumor cells or the production of functional cells to replace dysfunctional cells found in conditions such as immunodeficiency syndromes and hemoglobin disorders. HSCT is usually preceded by high-dose chemotherapy for pre-transplant myeloablative disruption. 【0038】 Streptococcus salivarius K12 has been marketed as a probiotic for over 15 years. S. salivarius K12 is deposited at Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstraße 7B, 38124 Braunschweig, Germany, under accession number DSM 13084, and is publicly available at the American Type Culture Collection (ATCC), POBox 1549, Manassas, VA 20108, USA, under accession number BAA-1024. S. salivarius ENT-K12 is genetically identical to S. salivarius K12 and was deposited on February 22, 2023, at Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstraße 7B, 38124 Braunschweig, Germany, with accession number DSM 34540. Streptococcus salivarius is also known as Streptococcus salivarius subsp. thermophilus ENT-K12, hence the terms "Streptococcus salivarius", "Streptococcus salivarius K12", "Streptococcus salivarius ENT-K12", "S. salivarius", "S. salivarius K12", "S. salivarius ENT-K12", and "Streptococcus thermophilus (Streptococcus "thermophilus)", "Streptococcus thermophilus K12", "Streptococcus thermophilus ENT-K12", "S. thermophilus", "S. thermophilus K12", "S. thermophilus ENT-K12", "S. salivarius subspecies thermophilus ENT-K12", "K12" and "ENT-K12" are used interchangeably in this specification. 【0039】 Experiments conducted within the scope of this invention have demonstrated that administration of Streptococcus salivarius is effective in treating cancer treatment-induced omya cystitis (OM). In particular, OM developed in cancer patients undergoing radiotherapy and cancer patients undergoing HSCT (chemotherapy-preceded omya cystitis) were shown to be prevented, reduced, and improved in patients treated with the probiotic microorganism Streptococcus salivarius concurrently with cancer treatment. Furthermore, the effect of Streptococcus salivarius in treating chemotherapy-induced OM was confirmed in mouse studies. Mouse studies also showed that inactivated Streptococcus salivarius, especially thermoinactivated Streptococcus salivarius, can be used in the treatment of OM. In addition, immune checkpoint inhibitor therapy is widely used as a treatment for cancer and autoimmune diseases, but it often causes immune-related adverse events in the oral mucosa, which can be a significant factor influencing the treatment outcomes of cancer and autoimmune diseases. 【0040】 Mucosal cells are a natural target for cancer cytotoxic regimens due to their rapid mitotic rate. Cytotoxic therapy results in epithelial damage, which usually leads to omaniosacral malformation (OM). Therefore, generally speaking, any cancer treatment, especially cancer treatment based on cytotoxic drugs, can lead to OM. Thus, patients with any cancer or any other disease requiring the administration of anticancer drugs, such as autoimmune diseases, who develop OM as a result of cancer treatment, are candidates for treatment with Streptococcus salivarius. Furthermore, OM can be associated not only with cancer treatment but also with various other factors. For example, OM is known to occur in various immunosuppressed patients (Chiappelli, Evid Based Complement Alternat Med. 2 (2005), 489-494). 【0041】 Accordingly, in one embodiment, the present invention relates to Streptococcus salivarius for use in the treatment of oral mucosal disorders, preferably omandibular malformations (OM), which are ear, nose, and throat (ENT) diseases in human subjects. In preferred embodiments, OM is induced by or associated with cancer treatment. Accordingly, in preferred embodiments, patients treated with Streptococcus salivarius are patients who have OM and cancer or autoimmune diseases and are undergoing, have completed, or are about to be treated with cancer treatment. Cancer treatments include, but are not limited to, radiotherapy, chemotherapy, chemoradiotherapy, especially concurrent chemoradiotherapy, hormone therapy, anti-angiogenic stem cell therapy including hematopoietic stem cell transplantation (HSCT), immunotherapy, dendritic cell-based immunotherapy, molecular targeted therapy, proton pump inhibitor therapy, immune checkpoint inhibitor therapy, and hormone therapy. 【0042】 In a preferred embodiment, the cancer treatment is a cancer treatment based on a cytotoxic drug. In other words, the OM preferably treated according to the present invention is an anticancer drug-induced OM, for example, a cytotoxic anticancer drug-induced OM. 【0043】 In preferred embodiments of the present invention, cancer treatments that induce OM include chemotherapy, radiotherapy, HSCT, immune checkpoint inhibitor therapy, a combination of chemotherapy and radiotherapy, a combination of HSCT and chemotherapy, a combination of HSCT and radiotherapy, or a combination of HSCT, chemotherapy, and radiotherapy. Thus, OMs treated according to the present invention are, as described above, CIOM, RIOM, HSCT-induced OM, or any combination thereof. 【0044】 Accordingly, in one embodiment, the present invention relates to Streptococcus salivarius for use in the treatment of OM in human subjects, wherein OM is induced by chemotherapy, radiotherapy, HSCT, a combination of chemotherapy and radiotherapy, a combination of HSCT and chemotherapy, a combination of HSCT and radiotherapy, or a combination of HSCT, chemotherapy and radiotherapy. In other words, patients treated with Streptococcus salivarius have OM and cancer or autoimmune disease, are undergoing or have completed cancer treatment, and cancer treatment is chemotherapy, radiotherapy, HSCT, a combination of chemotherapy and radiotherapy, a combination of HSCT and chemotherapy, a combination of HSCT and radiotherapy, or a combination of HSCT, chemotherapy and radiotherapy. 【0045】 In one embodiment, the present invention relates to Streptococcus salivarius for use in the treatment of OM, preferably in human subjects, wherein OM is CIOM. 【0046】 The present invention further relates to Streptococcus salivarius for use in the treatment of OM, preferably in human subjects, wherein the OM is RIOM. As described above, OM is referred to as RIOM when a patient has received either radiotherapy or chemoradiotherapy. 【0047】 In one embodiment, the present invention further relates to Streptococcus salivarius for use in the treatment of OM in human subjects, wherein the OM is HSCT-induced OM. 【0048】 Chemotherapeutic agents are well known to those skilled in the art and are listed in various publicly available lists, such as the one on the website of the cancer network “Chemotherapeutic Agents and Their Uses, Dosages, and Toxicities, June 1, 2016, Anne M. McDonnell, PharmD, BCOP”. In one embodiment, the chemotherapy is a combination chemotherapy of busulfan and cyclophosphamide. In another embodiment, the chemotherapy is 5-Fu chemotherapy. 【0049】 As described above, radiotherapy is usually administered over several weeks, and the appropriate dosage can be determined by those skilled in the art. As shown in Example 1, to treat NPC, radiotherapy was administered five times a week at a dose of 2 Gy per session, for a total dose of 60-70 Gy over 6-7 weeks. Therefore, in a preferred embodiment, radiotherapy is administered 1-7 times a week, preferably 2-6 times a week, more preferably 3-6 times a week, more preferably 4-6 times a week, most preferably 5 times a week, for a period of 2-10 weeks, preferably 3-10 weeks, more preferably 4-10 weeks, more preferably 5-10 weeks, more preferably 5-9 weeks, more preferably 5-8 weeks, and most preferably 6-7 weeks. Preferably, radiotherapy is administered at a dose of 2 Gy per session, for a total dose of 60-70 Gy, but naturally, the treatment regimen can vary depending on the disease being treated. In a preferred embodiment, radiotherapy is administered in conjunction with concurrent chemotherapy. This dosage form is used to treat nasopharyngeal cancer (NPC). 【0050】 As demonstrated in tests conducted within the scope of the present invention, the use of Streptococcus salivarius is efficient and safe in the treatment of omphalomyelitis (OM) in NPC patients and patients who have undergone HSCT. See Examples 1 and 2. NPC belongs to head and neck cancers. Most head and neck cancers originate from the mucosal epithelium of the oral cavity, pharynx, and larynx and are collectively known as head and neck squamous cell carcinoma (HNSCC). However, HNSCC can arise from further regions within the head and neck, including but not limited to the mucosal epithelium of the oral cavity (lips, buccal mucosa, hard palate, anterior tongue, floor of the mouth, and posterior molar triangle), nasopharynx, oropharynx (palatine tonsils, lingual tonsils, base of the tongue, soft palate, uvula, and posterior pharyngeal wall), hypopharynx (the base of the throat extending from the hyoid bone to the cricoid cartilage), and larynx. Human papillomavirus-associated HNSCC primarily arises from the palate and lingual tonsils of the oral pharynx, while tobacco-associated HNSCC primarily arises from the oral cavity, hypopharynx, and larynx. Therefore, in one embodiment, the patient treated according to the present invention has head and neck cancer, preferably NPC. Therefore, in one embodiment, OM treated according to the present invention is caused by treatment of head and neck cancer, preferably NPC, using cancer treatment. 【0051】 In a preferred embodiment, the present invention relates to Streptococcus salivarius for use in the treatment of RIOM, wherein the cancer treated by radiotherapy is head and neck cancer, preferably NPC. In a more preferred embodiment, radiotherapy is applied five times a week within 6 to 7 weeks (total dose of 60 to 70 Gy, with 2 Gy per session). 【0052】 HSCT is used to treat a variety of diseases, including cancer, leukemia, lymphoma, heart failure, neurological disorders, autoimmune diseases, immunodeficiency, metabolic or hereditary disorders. Treatment with HSCT usually begins with high-dose chemotherapy, and therefore, omphalomyelitis (OM) that develops in HSCT patients can be called comorbid omphalomyelitis (CIOM). Thus, in one embodiment, the present invention relates to Streptococcus salivarius for use in the treatment of CIOM that develops as a result of chemotherapy treatment, or as a result of chemotherapy treatment and subsequent HSCT. In a preferred embodiment, the present invention relates to Streptococcus salivarius for use in the treatment of CIOM, wherein the cancer treated with HSCT and prior chemotherapy is a hematopoietic malignancy. 【0053】 The ability to consistently and accurately assess OM is crucial for describing its incidence and severity, as well as evaluating the effectiveness of potential interventions. Three of the most commonly used criteria are the OM grading criteria (World Health Organization (WHO), Radiotherapy Oncology Group (RTOG), and Common Terminology Criteria for Adverse Events (CTCAE). A comparison is shown in Villa et al., Support Care Cancer 29 (2021), 6061-6068. All grading criteria range from Grade 0 to Grade 4, where Grade 0 is defined as no findings and Grade 4 represents the most severe form of OM. Therefore, in one embodiment, OM treated according to the present invention may be of any grade between Grade 0 and Grade 4. In Examples 1 and 2, the incidence of RIOM with RTOG ≥ 2 was reduced, and patients taking oropharyngeal probiotics were RTOG It was shown that complete protection from severe RIOM, classified as categories 3 and 4, was achieved. Furthermore, administration of oropharyngeal probiotics was shown to delay the onset of oral mucositis, reduce the risk of severe oral mucositis prevalence, dose-dependently restore reduced oral epithelial cell proliferation and cell cycle disruption after cancer treatment, prevent reduction in oral mucosal layer area after chemotherapy, and restore disruption of oral mucosal integrity. 【0054】 Therefore, in one embodiment, S. salivarius is used according to the present invention to prevent a patient from developing grade ≥ 2 OM. Therefore, in one embodiment, S. salivarius is used according to the present invention to prevent a patient from developing grade 3 or 4 OM. Therefore, in one embodiment, S. salivarius is used according to the present invention to prevent a patient from developing OM. In one embodiment, S. salivarius is used according to the present invention to delay the onset of oral mucositis and reduce the risk of the prevalence of severe oral mucositis. In one embodiment, S. salivarius is used according to the present invention to dose-dependently restore reduced proliferation and cell cycle disorders of oral epithelial cells after chemotherapy. In one embodiment, S. salivarius is used according to the present invention to prevent a decrease in oral mucosal layer area and restore disorder of oral mucosal integrity after chemotherapy. 【0055】 Furthermore, immunosuppression is generally known to lead to OM, for example, due to HIV infection (Ehlert et al., Dtsch Med Wochenschr, 138 (2013), 1601-1695) or treatment with immunosuppressants. Since S. salivarius has been shown to be safe and efficient in the treatment of CIOM, RIOM, and HSCT-induced OM in experiments conducted within the scope of the present invention, it is reasonable to consider that OM caused by the immunosuppressed state of a patient can also be treated with S. salivarius. Accordingly, the present invention also relates to Streptococcus salivarius for use in the treatment of OM in human subjects, wherein the human subjects are immunosuppressed. Immunosuppressants are generally known in the art and are outlined, for example, in Rathee et al., The Pharma Innovation Journal 1 (2013), 90-101. 【0056】 As mentioned above, S. salivarius is a probiotic microorganism, and generally, probiotics are defined as live microorganisms that provide health benefits to the host when administered in sufficient quantities. The beneficial effects of probiotics may be exerted through direct antagonistic effects against specific groups of undesirable organisms, by their effects on the metabolism of these microbial communities, or by their general stimulating effects on the immune system of an animal or human host. 【0057】 As shown in Example 4, thermally inactivated Streptococcus salivarius has beneficial effects on oral epithelial cells similar to those of live Streptococcus salivarius, which have not been previously shown. Therefore, the present invention also relates to inactivated Streptococcus salivarius for use in healing damaged oral epithelial cells induced by cancer treatment, particularly those damaged by chemotherapy. More specifically, the present invention also relates to inactivated Streptococcus salivarius for use in the treatment of cancer treatment-induced OM, preferably CIOM. Inactivation can be carried out by various methods, such as heat, pressure, or radiation. Preferably, inactivation is carried out by heat, so Streptococcus salivarius is preferably thermally inactivated (heat-killed) Streptococcus salivarius. Thermal inactivation is preferably carried out at a temperature in the range of 60°C to 120°C, more preferably at about 65°C for about 60 minutes, or at about 120°C for about 20 minutes. 【0058】 As mentioned above, OM is an ENT disease. ENT diseases, also known as ear, nose, and throat diseases, include problems involving the ears, nose and nasopharynx, sinuses, throat, mouth and oropharynx, larynx, trachea, and neck. The most common ENT diseases are outlined, for example, in Paradis and Messner, Pediatric Board Study Guide (2015), March 28, 469-89, the contents of which are incorporated herein by reference. ENT diseases are usually caused by viral or bacterial infections. 【0059】 Certain cancer treatments are known to temporarily weaken the immune system because they can reduce the number of white blood cells produced in the bone marrow. Therefore, cancer treatments typically make patients more susceptible to certain infections, such as the ENT diseases mentioned above, similar to general immunosuppressive therapies. Experiments conducted within the scope of this invention have shown that administration of S. salivarius is safe for patients who are currently being treated with or have been treated with anticancer drugs and are therefore immunosuppressed. 【0060】 Furthermore, clinical trials have already established the safety and efficacy of daily administration of Streptococcus salivarius in reducing respiratory infections and major respiratory pathogens. In particular, the K12 strain has been shown to be well-suited for use as an upper respiratory probiotic due to its natural tendency to inhabit the human oral cavity and to strongly compete with several potential pathogens. In addition, S. salivarius K12 has been shown to induce an anti-inflammatory response. For example, S. salivarius K12 antagonizes the growth of Streptococcus pyogenes, a major causative agent of human pharyngeal infections, which also lead to the development of acute otitis media. Prophylactic administration of S. salivarius K12 to children with a history of recurrent oral streptococcal disease resulted in a significant reduction in episodes of both streptococcal and viral infections. See Di Pierro et al., Drug Health Patient Saf. 6 (2014), 15-20. As outlined in detail in Example 2, pre-existing studies have also confirmed that Streptococcus salivarius can be used for the treatment and prevention of respiratory infections in patients undergoing cancer treatment, particularly chemotherapy and HSCT. 【0061】 Accordingly, the present invention also relates to Streptococcus salivarius for use not only in oral mucosal disorders but also in the treatment of further ENT diseases induced by cancer treatment in human subjects. Thus, patients treated with Streptococcus salivarius have ENT diseases and cancer, or have ENT diseases and autoimmune diseases and are undergoing or have completed cancer treatment. Furthermore, the present invention relates to Streptococcus salivarius for use in the treatment of ENT diseases in human subjects, wherein the subjects are immunosuppressed as described above. 【0062】 Since S. salivarius is particularly useful in the treatment of upper respiratory tract infections and otitis media as confirmed by Example 2, the ENT disease is preferably an upper respiratory tract infection or otitis media. The upper respiratory tract includes the throat, nose, pharynx, larynx, sinuses, and trachea, and bacterial-borne upper respiratory tract infections include, for example, pharyngitis, tonsillitis, sinusitis, and bronchitis. Severe respiratory infections can lead to unplanned hospitalization and may result in changes in treatment, such as interruption of radiotherapy or reduction of chemotherapy. As a result, treatment outcomes may be adversely affected. Furthermore, colonization of the upper respiratory tract by Streptococcus pneumoniae can lead to colonization of the lower respiratory tract, which can result in pneumonia and death of the patient, especially immunocompromised patients. Therefore, in preferred embodiments, the ENT disease is a respiratory infection. Accordingly, the present invention relates, in preferred embodiments, to Streptococcus salivarius for use in the prevention and treatment of respiratory infections during cancer treatment. 【0063】 In particular, in one embodiment, administration of Streptococcus salivarius during cancer treatment prevents the progression from upper respiratory tract infections to lower respiratory tract infections in subjects. In one embodiment, administration of Streptococcus salivarius during cancer treatment shortens the duration of respiratory infection symptoms in subjects compared to controls that did not receive Streptococcus salivarius. In one embodiment, administration of Streptococcus salivarius during cancer treatment reduces the mean duration of respiratory infection episodes in subjects compared to a control group. In one embodiment, administration of Streptococcus salivarius during cancer treatment reduces the need for antibiotic use in subjects compared to a control group. 【0064】 Therefore, experiments conducted within the scope of this invention have shown for the first time that Streptococcus salivarius can be used to treat respiratory infections in patients undergoing cancer treatment, and thus in immunocompromised patients. Prior to the conduct of this study, administering live bacteria to already immunocompromised patients would not have been recommended due to the high risk of undesirable side effects such as sepsis. Thus, it has been shown for the first time that Streptococcus salivarius can be used to treat oral mucosal disorders, particularly omya malformations (OM), and respiratory infections in patients undergoing cancer treatment and immunocompromised patients, respectively. 【0065】 In a preferred embodiment, cancer treatment includes HSCT, chemotherapy, and / or radiotherapy. As described in detail above, experiments conducted according to the present invention further demonstrated that severe omandibular malformation (OM) was prevented in patients who ingested oropharyngeal probiotics. Therefore, administration of S. salivarius is useful in preventing OM and maintaining healthy ear, nose, and throat (ENT) regions, preferably healthy oral mucosa, in human subjects undergoing cancer treatment. Accordingly, the present invention also relates to the use of Streptococcus salivarius in maintaining healthy ear, nose, and throat (ENT) regions, preferably healthy oral mucosa, in human subjects undergoing cancer treatment and in immunosuppressed patients, respectively. 【0066】 The present invention further relates to compositions for use in accordance with the present invention, comprising (an effective amount) of Streptococcus salivarius as an active ingredient and optionally one or more pharmaceutically acceptable excipients. The present invention also relates to compositions for use in the treatment of cancer treatment-induced oral mucosal disorders, particularly oral mucositis, comprising (an effective amount) of inactivated Streptococcus salivarius as an active ingredient and optionally one or more pharmaceutically acceptable excipients. 【0067】 The compositions according to the present invention are suitable for administering S. salivarius to patients requiring administration of S. salivarius, particularly patients susceptible to ENT diseases, more specifically to OM as defined above, and / or patients suffering from ENT diseases, particularly OM. Generally, the therapeutic compositions consist of S. salivarius and acceptable excipients. 【0068】 Excipients include bulking agents, diluents, carriers, binders, lubricants, disintegrants, anticaking agents, preservatives, colorants, or fragrances. The excipients are preferably pharmaceutically acceptable and / or suitable for human use. 【0069】 A carrier or extender is a vehicle for the delivery of probiotic microorganisms to an organism, and means a vehicle compatible with cell viability or the activity of probiotics, here in the inactivated form of S. salivarius. Acceptable carriers / extenders are well known to those skilled in the art. See, for example, Remington's Pharmaceutical Sciences, 18th edition, edited by Gennaro, 1990, Mack Publishing Co., Easton, Pa., incorporated herein by reference. Suitable carriers are generally inert and may be either solid or liquid. Acceptable carriers include, but are not limited to, water, buffered saline (e.g., phosphate-buffered saline), pharmaceutically acceptable culture media (e.g., BACa, TSBCaYE agar), or other solutions that maintain bacterial viability and, respectively, maintain the biologically active portion of the bacteria. Furthermore, such pharmaceutically acceptable carriers may be aqueous or non-aqueous solutions, suspensions, and emulsions. Various pharmaceutically acceptable carriers suitable for oral, topical, nasal, or respiratory administration of live or freeze-dried bacteria are well known in the art (see, for example, Remington). Suitable solid carriers known in the art include, but are not limited to, magnesium carbonate; magnesium stearate; celluloses; talc; sugars such as fructose, sucrose, mannitol, and lactose; starch; wheat flour; oligosaccharides and skim milk, and similar edible powders. Carriers for administering extracts are also well known. 【0070】 Acceptable carriers / extenders for use with probiotic microorganisms in compressed products such as lozenges are typically solid carriers known to those skilled in the art, but are not limited to, magnesium carbonate; magnesium stearate; celluloses; talc; sugars such as fructose, sucrose, mannitol, sorbitol, xylitol, and lactose; sugar substitutes such as isomalt; starch; maltodextrin; wheat flour; (fructose-)oligosaccharides and skim milk, as well as similar edible powders. 【0071】 Typical diluents include, for example, starch, lactose, mannitol, kaolin, calcium phosphate or calcium sulfate, inorganic salts such as sodium chloride, and powdered sugars and the above sugar substitutes or celluloses. 【0072】 Typical binders include starch, gelatin, sugars such as lactose, fructose, and glucose. Natural and synthetic rubbers are also suitable, such as acacia, alginate, locust bean gum, methylcellulose, e.g., hydroxypropyl methylcellulose (HMPC), polyvinylpyrrolidine (PVP) tragacanth, PVP K-30, PVP K-25, and xanthan gum. Polyethylene glycol (PEG 4000 or PEG 6000), ethylcellulose, and waxes can also function as binders and as Nu-BIND® and CompactCel® DIS. 【0073】 Examples of lubricants and anticaking agents for preventing adhesion and clump formation during formulation include talc, silica, magnesium stearate and calcium stearate, polyethylene glycol, stearic acid, hydrogenated vegetable oil, rice extract blends such as Nu-MAG®, CompactCel® LUB, potato starch, gum arabic, CompactCel® FLO, Nu-FLOW®, silicon dioxide, tricalcium phosphate, and solids with slippery properties such as rice husks, such as Nu-FLOW® or CompactCel® FLO. 【0074】 Disintegrants are substances that swell when wet, breaking down the composition and releasing S. salivarius or extracts. Examples of disintegrants include starch, clay, celluloses, alginates, and gums, more specifically, corn starch and potato starch, methylcellulose, agar, bentonite, wood cellulose, cation exchange resins, alginic acid, guar gum, citrus pulp, carboxymethylcellulose, powdered sponge, silica, and sodium lauryl sulfate. 【0075】 Fragrances are known to those skilled in the art and may be any kind that are good (or at least have a different taste) in the compressed product. Such fragrances include, but are not limited to, strawberry, mint, orange, banana, passion fruit, cocoa, menthol, yuzu, lemon and / or combinations thereof, preferably passion fruit, cocoa and menthol, yuzu and mint, orange and mint. 【0076】 S. salivarius can be formulated in any of a variety of compositions suitable for oral, nasal, topical, or respiratory tract administration. In a preferred embodiment, S. salivarius is formulated for oral administration. The composition may be in liquid, solid, or semi-solid form, but in a preferred embodiment, the composition is in solid form. In another preferred embodiment, the composition is an oil droplet composition, and S. salivarius is formulated with oil, preferably vegetable oil, preferably sunflower oil, in particular sunflower (Helianthus annuus L., alpha-tocopherol) seed oil. 【0077】 S. salivarius can be administered in the form of oral creams, oral films, wound dressings, lotions, gels, ointments, solutions, suspensions, emulsions, powders, granules, drops, especially oily drops, sachets, mouth washes, mouth rinses, toothpastes, dentifrices, sprays, gargles, capsules, troches, syrups, floss, films, chewing gums, or chewable tablets, but the form is not limited thereto. Administration can be carried out through the airway by a nebulizer, with or without a propellant. Preferably, the administration is carried out in a dosage form that remains in the oral cavity for a long time, such as in the form of troches, chewable tablets, chewing gums, oil suspensions, powder / sachets or sprays. In a preferred embodiment, the composition containing Streptococcus salivarius used according to the present invention is an oral solid dosage form selected from tablets, capsules, gels, troches, chewable tablets, and powders. Preferably, the oral composition is a troche or a powder, more preferably a troche. 【0078】 Generally, the amount of S. salivarius administered to a patient is an amount of the active ingredient that is high enough to provide the desired benefit and low enough to avoid serious side effects. 【0079】 In one embodiment, the daily dose of Streptococcus salivarius administered according to the present invention is 3×10 6 ~4×10 10 CFU, preferably 3×10 9 ~3×10 10 CFU or 4×10 9 ~4×10 10 CFU. The inactivated form of S. salivarius is preferably administered in an amount similar to that of viable S. salivarius, i.e., 3×10 6 ~4×10 10 cell equivalents, preferably 3×10 9 ~3×10 10 cell equivalents or 4×10 9 ~4×10 10 administered in an amount effective to provide cell equivalents. 【0080】 In one embodiment, the composition used according to the present invention, preferably in solid dosage form, more preferably in tablet form, most preferably in lozenge form, contains at least 1 mg of Streptococcus salivarius and may contain up to 1000 mg of Streptococcus salivarius. In a preferred embodiment, the composition used according to the present invention, preferably in solid dosage form, more preferably in tablet form, most preferably in lozenge form, contains 1 to 500 mg of Streptococcus salivarius, preferably 5 to 160 mg of Streptococcus salivarius, preferably 10 to 120 mg, more preferably 10 to 100 mg, or 30 to 120 mg, more preferably 40 mg, 50 mg, 60 mg, 100 mg, or 120 mg, most preferably 50 mg of Streptococcus salivarius. 【0081】 In one embodiment, the composition used according to the present invention, preferably in solid dosage form, more preferably in tablet form, most preferably in lozenge form, contains 50 mg each of Streptococcus salivarius and 10 6 ~10 10 CFU, preferably 1 × 10 9 ~10×10 9 CFU, fer6×10 9 ~7×10 9 The composition comprises CFU of Streptococcus salivarius, or a corresponding amount of the cell equivalent in an inactivated form. Preferably, the composition, preferably the solid dosage form, most preferably the lozenge, is administered to a human subject several times a day, preferably three or four times a day. In one embodiment, the composition is administered three times a day to patients who have undergone HSCT and / or been treated with chemotherapy, and four times a day to patients who have been treated with radiotherapy and / or whose cancer is NPC. 【0082】 In one embodiment, the composition and Streptococcus salivarius used according to the present invention are administered over a period of several days to several weeks, respectively, where the duration varies depending on the type of cancer treatment / autoimmune disease treatment and the type of cancer treatment administered. For example, as mentioned above, chemotherapy is usually administered for a short period, in which case damage to mucosal tissue tends to be acute. Chemotherapy-induced OM usually develops within 4 to 7 days after the start of treatment and peaks within 2 weeks. Therefore, to avoid the development of severe OM, it may be sufficient to administer the composition for about 2 weeks. In contrast, as also shown in Example 1, radiotherapy is usually applied over several weeks, and therefore, in one embodiment, the composition used according to the present invention is administered to human subjects treated with radiotherapy and / or whose cancer is NPC for a period of several weeks, preferably about 7 weeks. In a preferred embodiment, the composition is administered as a precautionary measure, even before the start of cancer treatment. For example, in the case of radiotherapy and CCRT treatment, the composition is administered 2 weeks before the start of treatment, respectively. 【0083】 In a preferred embodiment, the composition is administered four times a day. More preferably, administration is performed after breakfast, lunch, dinner, and before bedtime, after cleaning teeth (mouthwash or brushing). 【0084】 In another embodiment, the composition used according to the present invention and Streptococcus salivarius undergo HSCT, and after HSCT, the platelet count is 20 × 10 9 Individuals / L greater than 0.5 × 10 9 In human subjects in whom hematopoietic reconstitution, defined as exceeding 1 / L, has been confirmed, the drug is preferably administered three times a day for a period of approximately 100 days. More preferably, administration is performed after breakfast, after lunch, and before bedtime, following dental hygiene (mouthwash or brushing). 【0085】 S. salivarius can be administered before, during (concurrently with), and / or after cancer treatment, and can also be administered long-term to immunosuppressed patients. In preferred embodiments, S. salivarius is administered to patients with RIOM or CIOM, preferably at least during cancer treatment, and preferably after the completion of cancer treatment, as long as OM symptoms are still present and the patient remains immunosuppressed. With respect to HSCT-induced OM, S. salivarius is preferably administered after their hematopoietic system has been reconstituted, for example, after HSC when platelets >20 × 10 9 Cells / L and neutrophils > 0.5 × 10⁻¹⁰ 9 It is defined as a cell / L. 【0086】 In one embodiment, the composition used in accordance with the present invention comprises, in addition to Streptococcus salivarius, isomaltulose, and one or more excipients, preferably a lubricant and / or an anticaking agent, and / or a binder, and / or a fragrance, wherein the lubricant is preferably magnesium stearate or a natural substitute, the anticaking agent is silicon dioxide, tricalcium phosphate or a natural substitute, the binder is HMPC or a natural substitute, and / or the fragrance is a fragrance, preferably a strawberry fragrance, and optionally the product further comprises one or more active ingredients, preferably a vitamin, a mineral, and / or a fructooligosaccharide, wherein the vitamin is preferably vitamin D3. 【0087】 Natural lubricants are known in the art, for example, crude fat-containing soybean powder, rice extract blends, e.g., product Nu-MAG® or product CompactCel® LUB (CompactCel® F clear 290.02 LUB, Biogrund GmbH, Huenstetten, Germany), as described in International Publication No. 2013 / 165131, oat fiber blends, e.g., product CompactCel® LUB (CompactCel® F 200.28 LUB, Biogrund GmbH, Huenstetten, Germany), or further products such as potato starch or gum arabic. Natural anticaking agents are also known in the art, for example, powdered cellulose, e.g., JELUCEL®, natural potato starch, inulin, rice fiber, or rice husks, e.g., Nu-FLOW® or CompactCel® FLO. Natural binders include, for example, CompactCel® DIS, NuBind®, or pre-gelatinized corn starch. 【0088】 In one embodiment, a composition used in accordance with the present invention is (i) Compressed product, i.e., an oral solid dosage form, comprising, in addition to Streptococcus salivarius, isomaltulose and further a lubricant / anticaking agent, a fragrance, and optionally a natural sweetener and / or one or more active ingredients, preferably the lubricant / anticaking agent is magnesium stearate or a natural substitute, preferably the fragrance is a flavoring, preferably strawberry flavoring, and the natural sweetener and / or one or more active ingredients is preferably a vitamin, preferably vitamin D3, in an oral solid dosage form. (ii) Compressed product, i.e., an oral solid dosage form, comprising, in addition to Streptococcus salivarius, isomaltulose and further a lubricant / anticaking agent, binder, fragrance, and optionally a natural sweetener and / or one or more active ingredients, preferably the lubricant / anticaking agent is magnesium stearate or a natural substitute, preferably the binder is hydroxypropyl methylcellulose (HMPC) or a natural substitute, preferably the fragrance is a flavor, preferably strawberry flavor, and preferably the natural sweetener and / or one or more active ingredients is preferably a vitamin, preferably vitamin D3, in an oral solid dosage form. (iii) A powder product comprising, in addition to Streptococcus salivarius, isomaltulose and further an anticaking agent, a fragrance agent, and optionally a natural sweetener and / or one or more active ingredients, wherein the anticaking agent is preferably silicon dioxide, tricalcium phosphate or a natural substitute, the fragrance agent is preferably a flavoring, preferably strawberry flavoring, and the natural sweetener and / or one or more active ingredients is preferably a vitamin, preferably vitamin D3. 【0089】 Natural substitutes for magnesium stearate are preferably rice extract blends, preferably rice extract, microcrystalline cellulose, and refined sunflower oil, such as product CompactCel® F clear 290.02 LUB; preferably oat fiber blends, preferably oat fiber, microcrystalline cellulose, and refined sunflower oil, such as product CompactCel® F 200.28 LUB, Nu-MAG®, potato starch, gum arabic, Nu-FLOW®, or rice husks such as CompactCel® FLO, but most preferably rice extract blends. Natural substitutes for HMPC are preferably natural binders such as CompactCel® DIS, NuBind®, or pre-gelatinized corn starch, most preferably pre-gelatinized corn starch. 【0090】 Accordingly, in one embodiment, the composition used according to the present invention is a compressed product, i.e., an oral solid dosage form, and further comprises Streptococcus salivarius, in addition isomaltulose and a rice extract blend as a lubricant / anticaking agent, pre-gelatinized corn starch as a binder, a flavoring agent, preferably strawberry flavoring, a natural sweetener, preferably stevia, and further vitamin D3 as an additional active ingredient. The exemplary solid dosage forms contain the components shown in Tables 1-3. 【0091】 [Table 1] 【0092】 [Table 2] 【0093】 [Table 3] 【0094】 Magnesium stearate is used in rice extract blends, such as product Nu-MAG® or product CompactCel® LUB (CompactCel® F clear 290.02 LUB, Biogrund GmbH, Huenstetten, Germany), and oat fiber blends, such as product CompactCel® LUB (CompactCel® F 200.28 LUB, Biogrund HMPC can be substituted with natural lubricants / anticaking agents such as Huenstetten GmbH (Huenstetten, Germany), potato starch or gum arabic, powdered cellulose, e.g., JELUCEL®, natural potato starch, inulin, rice fiber or rice husk, e.g., Nu-FLOW® or CompactCel® FLO, but preferably with a rice extract blend, preferably with CompactCel® LUB, and / or HMPC can be substituted with natural binders such as CompactCel® DIS, NuBind®, or preferably with pre-gelatinized corn starch. 【0095】 The compressed product of the present invention may contain silicon dioxide, but silicon dioxide may be substituted with a natural anticaking agent such as rice husks, for example CompactCel® FLO or Nu-FLOW®. Preferably, the compressed product does not contain magnesium stearate and silicon dioxide, but instead contains a rice extract blend. The example powder product contains the ingredients shown in Table 4. 【0096】 [Table 4] 【0097】 Silicon dioxide can be substituted with tricalcium phosphate, or the aforementioned natural anticaking agents such as rice husks (Nu-Flow®), or CompactCel® FLO, and can also be omitted entirely. 【0098】 The composition is preferably made using the following steps: (i) A step of mixing S. salivarius, isomaltulose, one or more further excipients, and optionally one or more further active ingredients. (ii) A process of homogenizing the mixture, (ii) A step of compressing the mixture to make a compressed product, preferably a step of compressing with a pressure of 6 to 10 kN per compressed product, or a step of filling the mixture into individual packets, and optionally, (iii) A step of analyzing the CFU / g (cacheuscultivable count per gram) and / or water activity of the composition, and optionally a step of analyzing the disintegration time, average mass, crushing strength, appearance, abrasion, and / or water activity. It is manufactured by a method that includes [a specific process]. 【0099】 In a preferred embodiment, step (i) is the following step: (a) A step of mixing isomaltulose with one or more further excipients, preferably lubricants / anticaking agents, preferably magnesium stearate. (b) A step of spraying a binder solution onto a mixed excipient to form wet granules, wherein the binder solution preferably consists of water and a binder, and the binder preferably is HMPC. (c) A step of drying the granules at a temperature of preferably 40+2°C for about 30 minutes. (d) A process of crushing granules, (e) A step of adding S. salivarius to crushed granules, and optionally adding one or more additional active ingredients. It consists of. 【0100】 Details of the composition and the method for producing the same are described in European Patent Application EP 23 167 706.3, which is incorporated herein by reference. 【0101】 In another embodiment, the composition used according to the present invention, particularly the solid dosage form, comprises, in addition to Streptococcus salivarius, one or more excipients, preferably one or more fillers, lubricants, and optionally anticaking agents and / or fragrances. 【0102】 In one embodiment, the composition used according to the present invention, particularly the solid dosage form, comprises fructose and maltodextrin (bulking agents), magnesium stearate or a natural substitute (lubricant), a flavoring, preferably strawberry flavoring (fragrance), and optionally silicon dioxide or a natural substitute (anti-caking agent). The natural substitute is as described above. Exemplary compositions, particularly solid dosage forms, are shown in Tables 5 and 6. 【0103】 [Table 5] 【0104】 [Table 6] 【0105】 The compositions used in accordance with the present invention, particularly the solid dosage forms, are preferably manufactured by a "briquetting" method comprising the following steps. (i) A step of briquetting the excipient, preferably at a pressure of about 7.5 kN / compressed product, (ii) A process of classifying and crushing the briquettes, (iii) Adding S. salivarius to the crushed briquettes and mixing, (iv) Compressing the mixture into a compressed product, preferably at a pressure of about 4.8 kN / compressed product, and optionally, (vii) A step of analyzing the collapse time, average mass, resistance to crushing, appearance, abrasion, and / or water activity of the compressed product. 【0106】 In a preferred embodiment, the method further includes the following steps. (a) A step of drying, weighing, and sieving the excipient before step (i), wherein drying is preferably performed in a fluid granulator. (b) A step of mixing and homogenizing the excipients after step (a) and before step (i), (c) A step of weighing and sieving S. salivarius prior to step (iii), preferably by sieving through a 1 mm net, and / or (d) A step to homogenize the mixture after step (iii) and before step (iv). 【0107】 Details regarding the composition and the method of producing the same are referred to in international application PCT / EP2023 / 059635, which is incorporated herein by reference. 【0108】 In one embodiment, the composition used according to the present invention, particularly the solid dosage form, comprises isomalt (bulking agent), magnesium stearate or a natural substitute (lubricant), a fragrance, preferably strawberry fragrance (aromatic agent), and optionally silicon dioxide or a natural substitute (anti-caking agent). The natural substitute is as described above. Exemplary compositions, particularly solid dosage forms, are shown in Table 7. 【0109】 [Table 7] 【0110】 In all lozenges, the concentration of vitamin D3 (cholecalciferol) in the premix is ​​100,000 IU / g (1 IU = 0.025 mcg). 【0111】 As described above, the oral solid dosage form used in accordance with the present invention is, in a preferred embodiment, a lozenge. Therefore, the present invention further relates to lozenges containing Streptococcus salivarius for use in accordance with the present invention, and to the use of said lozenges in accordance with the present invention. In a preferred embodiment, the lozenge contains the components described above with respect to the oral solid dosage form. Most preferably, the lozenge contains the components listed in Table 6, namely Streptococcus salivarius, fructose, maltodextrin, magnesium stearate, and a flavoring, preferably a strawberry flavoring, and most preferably, the lozenge contains the components in the amounts listed in Table 6. 【0112】 In one embodiment, the lozenges used according to the present invention are manufactured by the “briquette” method described above. 【0113】 In one embodiment, the lozenges used according to the present invention are slowly sucked by the patient until they are completely dissolved and no longer chewable or directly swallowable, and preferably the patient should not eat, drink, or swallow for at least two hours after administration of the lozenges. In a preferred embodiment, the disintegration time of the lozenges is at least 4-5 minutes, which ensures that Streptococcus salivarius can be reliably fixed in the patient's oral cavity. The disintegration time can be measured according to the method described in Chapter 2.9.1 of the European Pharmacopoeia, 6th edition, published by the Council of Europe, 10 May 2008, ISBN-10:9287160546, ISBN-13:978-9287160546. 【0114】 The present invention further relates to S. salivarius and the corresponding compositions defined above, particularly lozenges for use according to the present invention, wherein S. salivarius or the corresponding compositions, particularly lozenges, are packaged in a kit including instructions for administering S. salivarius or the corresponding compositions, particularly lozenges. In a preferred embodiment, the lozenges are packaged in a blister pack. 【0115】 In another preferred embodiment, the composition is in the form of oil droplets, which is a preferred and convenient form of administration for patients who have already developed painful omandibular malformation (OM). An exemplary oil droplet composition consists of the probiotic S. salivarius K12, sunflower (Helianthus annuus L., alpha-tocopherol) seed oil, and cholecalciferol oil (vitamin D3, alpha-tocopherol, medium-chain triglyceride). 【0116】 The present invention further relates to a method for treating cancer treatment-induced ENT disease, preferably cancer treatment-induced OM, in patients as described above, comprising the administration of S. salivarius, a composition containing S. salivarius, or lozenges containing S. salivarius. The present invention further relates to a method for treating cancer treatment-induced ENT disease, preferably OM, in immunosuppressed patients as described above, comprising the administration of S. salivarius, a composition containing S. salivarius, or lozenges containing S. salivarius. 【0117】 In preferred embodiments, the Streptococcus salivarius species used in accordance with the present invention are Streptococcus salivarius K12 (American Type Culture Collection (ATCC), POBox 1549, Manassas, VA 20108, USA, accession number BAA-1024) and S. salivarius ENT-K12 (Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstraße 7B, 38124 Braunschweig, Germany, accession number DSM 34540). 【0118】 Throughout this specification, several documents are cited. The contents of all cited references (including, but not limited to, the background art section, references cited throughout this application, published patents, published patent applications, and manufacturer specifications, instructions, etc.) are expressly incorporated herein by reference. However, this does not constitute an admission that the cited documents are actually prior art relating to the present invention. 【0119】 A more complete understanding can be obtained by referring to the following specific examples, which are provided herein for illustrative purposes only and are not intended to limit the scope of the invention. [Examples] 【0120】 Example 1: Oral and pharyngeal probiotic ENT-K12 prevents severe radiation-induced oral mucositis in nasopharyngeal cancer patients undergoing radiotherapy. 【0121】 Concurrent chemoradiotherapy (CCRT) is the standard treatment strategy for nasopharyngeal cancer (NPC), and studies have shown that 59.4%–100% of patients undergoing CCRT for head and neck cancer experience oral mucositis (OM). Radiotherapy-induced oral mucositis (RIOM) is one of the most common complications in head and neck cancer (HNC) patients undergoing CCRT. The main symptom of RIOM is discomfort or pain in the mouth or throat, and RIOM can reduce patient tolerance to radiotherapy, lead to a reduction in the total dose of radiotherapy, and may even lead to radiotherapy discontinuation (Marta et al., Radiother Oncol 110 (2014), 9-15; Gobbo et al., Lasers Med Sci 31 (2016), 471-9). The development of RIOM increases the incidence and severity of anxiety and depression in NPC patients during radiotherapy. It also reduces quality of life (QoL) and adherence to treatment. In addition, RIOM can lead to numerous complications, including pain, sore throat, dysphagia, decreased oral intake, and systemic infections, often resulting in delays or interruptions in treatment. These interruptions can lead to reduced disease control and poor patient prognosis. This not only negatively impacts quality of life but also has a significant negative impact on tumor control and patient survival (Triarico et al., Pathogen 11 (2022), 448; Oronsky et al., Transl Oncol 11 (2018), 771-778). Therefore, the prevention and treatment of CCRT-induced RIOM are clinically very important. Currently, prophylactic measures are considered the most effective treatment for RIOM, and several studies and clinical trials on palliative care agents are being conducted with the aim of preventing and controlling RIOM. However, there is a lack of high-level evidence regarding single-target therapies in the treatment of RIOM. 【0122】 Radiotherapy-induced toxicity includes xerostomia and mucositis due to mucosal barrier disruption, leading to severe oropharyngeal dysbiosis throughout the entire duration of CCRT treatment, often accompanied by severe mucositis. While probiotic supplements that alter the gut microbiota have been studied to reduce the risk of cancer treatment-induced oral mucositis (Shu et al., Oral Oncology 102 (2020), 104559), recovery of the oropharyngeal microbiota during CCRT has not been reported. Previous studies have shown that oropharyngeal probiotics inhibit pathogen growth in periodontal biofilms and help maintain the physiological balance of the oral microbiota (Begic et al., Int J Mol Sci 24 (2023), 7249). This also improves the host's immune level, as evidenced by the reduction of anti-inflammatory and pro-inflammatory cytokines induced by this strain (Evivie et al., Front Microbiol 10 (2019), 782). Previous clinical trials have shown that oropharyngeal probiotics can effectively colonize the oropharynx / nasopharynx and reduce the risk of pharyngotonsillitis infection (Wilcox et al., Clin Microbiol Infect 25 (2019), 673-680). For these reasons, we designed a pilot trial to investigate the safety of oropharyngeal probiotics and whether they can reduce the incidence and progression of RIOM in NPC patients receiving CCRT. RIOM is thought to occur when radiation damages rapidly dividing basal cells in the submucosa, resulting in damage to epithelial cells. This condition is a common complication in nasopharyngeal carcinoma (NPC) patients undergoing concurrent chemoradiotherapy. 【0123】 Main purpose The objective of this trial is to evaluate the efficacy and safety of the oropharyngeal probiotic ENT-K12 for the incidence, severity, and disease progression of RIOM in NPC patients receiving CCRT. RIOM will be clinically scored five times per week by the same radiotherapist according to the Group of Radiation Therapy Oncology (RTOG) criteria (see Table 8) to track dynamic changes in RIOM throughout the entire treatment period. 【0124】 [Table 8] 【0125】 Secondary purpose • To evaluate the pathogenesis of RIOM (supragingival plaque, saliva, oral microbial dysbiosis, anaerobic bacterial overgrowth associated with increased expression of the nitrate reductase gene napA, and regulation of the TLR2 / TLR4 signaling pathway in the development of RIOM). • To evaluate the anti-inflammatory effect on oral mucosa or tissues (CD3+ T cells, CD4+ T cells, and CD8+ T cells). • Nutritional status (weekly monitoring of weight loss, BMI, albumin, prealbumin, and NRS2002 score) • Evaluate quality of life, such as a reduction in side effects caused by radiation therapy and improvement in quality of life (e.g., sore throat, loss of taste and appetite, dry mouth, peeling skin on the neck, difficulty swallowing, hair loss, ear discharge, nausea or vomiting, fatigue, diarrhea, etc.). • Confirm the safety of the subject as determined by adverse event reports. ·Recovery rate • To evaluate the efficacy of the oral-pharyngeal probiotic ENT-K12 against respiratory infection episodes during the study. • Number of days of antibiotic treatment • Number of days of antiviral drug treatment • Number of days of treatment with antipyretics • Number of days experienced oral mucositis and tongue ulcers • Number of days on which you have experienced gum bleeding or gum disease 【0126】 Exam Overview A randomized controlled clinical trial was conducted to evaluate the efficacy of the oropharyngeal probiotic ENT-K12 in preventing radiotherapy-induced oral mucositis (RIOM) in patients with stage II nasopharyngeal cancer (NPC). The study included 10 patients aged 18 to 65 years who were newly diagnosed with stage II or higher nasopharyngeal cancer (NPC) according to the International Union Against Cancer / American Joint Committee on Cancer (TNM) classification, 8th edition, and who had not previously received head and neck radiotherapy. All patients diagnosed with NPC received CCRT and basic oral hygiene instructions, and were randomly assigned (1:1) to either the oropharyngeal probiotic group (CCRT-P group) or the control group (CCRT group) without probiotic intervention. Patients in the oropharyngeal probiotic group received additional oropharyngeal probiotic interventions two weeks prior to and during CCRT. Furthermore, participants underwent a series of assessments including blood tests, abdominal ultrasound, chest computed tomography scans, head and neck magnetic resonance imaging scans, and whole-body bone scans. Exclusion criteria for patient enrollment included having an immunodeficiency, having OM or recurrent OM prior to CCRT, being unable to take OM, a history of tumors, and allergies to probiotics. This trial was conducted in accordance with the Declaration of Helsinki and approved by the Ethics Committee of Wuhan University of Science and Technology (registration number 2021101). Informed consent was obtained from patients to collect data. The principal investigators evaluated RIOM using the Group of Radiation Therapy Oncology (RTOG) guidelines (Table 8) and graded it from I to IV according to the severity of the OM. 【0127】 Test protocol All patients underwent CCRT in accordance with National Comprehensive Cancer Network guidelines. Prior to initiating radiation therapy, all patients received a full oral examination and oral hygiene instruction. Patients with caries, pulp disease, and gingivitis underwent professional cleaning, fillings, endodontic treatment, and surgical procedures including extraction of irreparable teeth. All participating patients received radiation therapy at 2 Gy per session, five times per week for seven weeks, for a total dose of 60–70 Gy, along with the concurrent chemotherapy listed in Table 9: 【0128】 [Table 9] 【0129】 Participants were randomly assigned to one of two groups: • Oral and pharyngeal probiotic ENT-K12 (CCRT-P) • Control group (no treatment) (CCRT) 【0130】 Participants in the CCRT-P group took one slow-dissolving lozenge four times a day: after brushing their teeth with breakfast, lunch, and dinner, and after applying mouthwash before bedtime, for seven weeks during radiotherapy and two weeks prior to CCRT. Participants were instructed to slowly inhale the lozenge until it was completely dissolved (approximately 4-5 minutes), and were cautioned not to chew or swallow it whole. They were also advised not to eat, drink, or swallow for two hours after administering the oropharyngeal probiotic. Uniformly trained physicians and nurses were designated as observers of clinical symptoms and recorded patients' RIOMs daily. The primary objectives were the safety of the oropharyngeal probiotic and the severity and time of RIOM occurrence during CCRT. Study physicians and nurses also recorded the type and severity of any oropharyngeal probiotic-related adverse events that occurred during the study, and survival, distant metastasis, and recurrent NPCs were followed up for 18 months after CCRT. 【0131】 Inclusion Criteria To be considered eligible to register for this examination, applicants must meet the following criteria: • Patients who were diagnosed with stage II nasopharyngeal cancer (NPC) for the first time at the time of registration. • Ages 18-65 • Never received radiation therapy • No active symptoms of respiratory infection. • No active symptoms of bacteremia. • No active symptoms of organ dysfunction • The ability to tolerate and complete this exam. • There is a signed written informed consent. 【0132】 Exclusion criteria Participants will be excluded from the test if they meet the following criteria. • Patients diagnosed with stage III or IV nasopharyngeal cancer (NPC) at the time of registration. • Patients who have previously received head and neck radiation therapy • Patients with poor oral hygiene or those suffering from severe periodontal disease. • Active symptoms of respiratory infection at the time of registration • Active symptoms of bacteremia • Active symptoms of organ dysfunction • Has a chronic respiratory condition that may affect observation. • Chronic oral mucositis and ulcer formation may affect the observation. 【0133】 Detailed test procedures outlined in the clinical trial protocol Screening visits and visits for the 1st to 35th radiation treatments During the initial screening visit (first visit), after confirming the diagnostic and radiotherapy schedule, participants will be given an overview of the study and the procedures to be followed. Participants will then be asked to read the participant information document and sign the informed consent form, and a signed copy will be provided. 【0134】 In detail, the following information is recorded and the procedure is executed. • Written informed consent • Target number and date of visit • Inclusion and exclusion criteria 【0135】 If the test subject meets the registration requirements, record the following information and follow the procedure. • Eligibility • Demographic data • Medication history ·Oral mucositis grading scale; WHO • Randomization 【0136】 Participants will be randomly assigned to one of two groups: a) oral pharyngeal probiotic ENT-K12; or b) control (no treatment). Participants in the probiotic group will be provided with a 7-week supply of the study product and instructed to take one slow-dissolving lozenge four times a day during the 7-week period of radiotherapy: after brushing their teeth with breakfast, lunch, and dinner, and after applying mouthwash before bedtime. Participants must slowly inhale the lozenge until it is completely dissolved (approximately 4-5 minutes) and ensure they do not chew or swallow the lozenge directly. Participants are advised not to eat, drink, or swallow for at least 2 hours after administering the probiotics. Participants will be instructed to return any unused study product on their final visit on the last day of their radiotherapy period. 【0137】 Participants in the study will be instructed to continue their usual medication, nursing care, diet, and exercise during the study period. 【0138】 Participants in the study will be instructed to contact their doctor at any time during the study if they experience symptoms of respiratory infection, such as sore throat, fever above 38°C, headache, muscle pain, shortness of breath, chest tightness, cough, nasal congestion or runny nose, swollen lymph nodes and / or the appearance of tonsil abscesses (pus) or white patches; or symptoms of oral mucositis, such as dry mouth, thickened saliva, increased mucus, shiny, swollen, erythematous gums, soft white patches or pus on the tongue, oral pain, oral bleeding, or mild burning while eating. If findings of respiratory infection or oral mucositis are observed, participants will be asked to visit the study site for confirmation of the diagnosis. 【0139】 Participants in the study will be asked to record the number of days they experienced oral mucositis, tongue ulcer formation, gingival bleeding, and gingival disease during the 15-week study period. • Self-assessment of overall health status Ask participants questions about their overall health status and rate the results on a 5-point scale. • Schedule appointments for radiation therapy visits 1 through 35. Participants in the study will be given appointments to visit the study site for 35 visits (five visits per week for seven weeks, coinciding with each radiotherapy procedure, throughout the entire duration of radiotherapy treatment). • Final appointment Participants in the study will be given an appointment to visit the study site for their final visit on the last day of their entire radiation therapy period. 【0140】 Visiting the clinic when symptoms are present (visiting for the diagnosis of respiratory infections and oral mucositis) ) Participants in the study will be instructed to contact their doctor at any time during the study if they experience any of the following symptoms: respiratory infection, oral mucositis, or tongue ulcer. ·sore throat, • Fever exceeding 38℃, ·headache, Muscle pain, ·shortness of breath, • Chest tightness, ·cough, • Nasal congestion or runny nose • Swollen lymph nodes and / or the appearance of tonsil abscesses (pus) or white patches. ·Dry mouth, • Increased viscosity of saliva • Increased mucus, • Shiny, swollen, erythematous gums, • Soft white patches or pus on the tongue • Pain in the mouth, • Bleeding in the mouth or mild burning sensation while eating 【0141】 If signs of respiratory infection are observed, the participant will be asked to visit the study site. A throat swab will be collected, and a rapid swab test will be performed to detect respiratory pathogens. The physician will advise on medication / medical treatment as needed. The participant will continue taking the test product throughout the study period as instructed. If antibiotic treatment becomes necessary, the participant will continue taking the test product during the antibiotic treatment period. 【0142】 Record the following information and perform the procedure during a "symptomatic visit." • Target number and date of visit • Diagnosis (respiratory infection, oral mucositis, tongue ulcer) (Please indicate the type of respiratory infection (e.g., streptococcal pharyngitis / tonsillitis) and whether the infection was confirmed by rapid pharyngeal swab test (RAD) or other microbiological tests.) • Prescribed medications (including dates and duration of treatment for antibiotics, antivirals, antipyretics / steroids, and antifungals) ·Oral mucositis grading scale; WHO • Number of days for oral mucositis and tongue ulcer • Number of days of gingivitis and gingival disease • Progression from upper respiratory tract infection to lower respiratory tract infection (e.g., ground-glass opacities on chest CT) • Severe symptoms and hypoxemia requiring hospitalization or ICU treatment. • Pulmonary complications (decreased FEV1, reduced oxygen diffusion capacity) • Adverse events (Confirm whether or not adverse events occurred in the test subjects. If adverse events occurred during the procedure, please describe their type and severity in the attached "Adverse Events" section.) • Self-assessment of overall health status (ask test subjects about their overall health status and have them indicate their results on a 5-point scale). 【0143】 Final visit (the date the entire period of radiation therapy ended) Record the following information and perform the procedure during your "last visit". • Target number and date of visit • Medical history over the past 7 weeks (upper respiratory tract infections, progression from URTi to LRTi, need for hospitalization, pulmonary complications, occurrence and etiology of diarrhea) (Please indicate the type and number of respiratory infections, oral mucositis, and tongue ulcers that were not diagnosed during the "symptomatic visit," and whether the infection was confirmed by rapid pharyngeal swab testing (RAD) or other microbiological tests) ·Oral mucositis grading scale; WHO • Number of days in the past 7 weeks during which oral mucositis and tongue ulcers were experienced. • Number of days in the past 7 weeks during which gingivitis and gingival disease have been experienced Adverse events • Compliance (return of unused test samples and evaluation of compliance) • Self-assessment of overall health status (ask participants about their overall health status and have them indicate their results on a 5-point scale). • Antibiotic treatment over the past 7 weeks • Treatment with antiviral drugs over the past 7 weeks • Treatment with antipyretics / steroids over the past 7 weeks • Treatment with antifungal medication over the past 7 weeks • Overall evaluation of tolerability and treatment efficacy • Treatment completed • Schedule a follow-up appointment (8 weeks after your last visit) 【0144】 Follow-up visit (8 weeks after the last visit) Record the following information and perform the procedure during your follow-up visit. • Target number and date of visit • Medical history over the past 8 weeks (upper respiratory tract infections, progression from URTi to LRTi requiring hospitalization, pulmonary complications, occurrence and etiology of diarrhea) (Please indicate the type and number of upper respiratory tract infections you have had in the past 8 weeks, and whether the infections were confirmed by rapid pharyngeal swab testing (RAD) or other microbiological tests) • Antibiotic treatment over the past 8 weeks • Treatment with antiviral drugs over the past 8 weeks • Treatment with antipyretics / steroids over the past 8 weeks • Treatment with antifungal medication over the past 8 weeks ·Oral mucositis grading scale; WHO • Number of days in the past 8 weeks during which oral mucositis and tongue ulcers were experienced. • Number of days in the past 8 weeks during which gingivitis and gingival disease have been experienced Adverse events • Self-assessment of overall health status (ask participants about their overall health status and have them indicate their results on a 5-point scale). • Exam completed 【0145】 Concomitant medications Participants in the study will be asked about their medication history. Details of the medications taken will be recorded in the case record and case report form. 【0146】 Test sample Participants in the probiotic group received oropharyngeal probiotic lozenges formulated in the form of oral lozenges containing the compositions described below. Each lozenge contained at least 1 billion colony-forming units (CFUs) of Streptococcus salivarius ENT-K12 (also known as Streptococcus salivarius subspecies thermophilus ENT-K12, DSM 34540) over its shelf life. 【0147】 [Table 10] 【0148】 [Table 11] 【0149】 Grading and evaluation of adverse events AE is assessed using a three-level scale and reported in detail as shown in the CRF. • Mild – Easily tolerable, with slight discomfort, but does not interfere with daily life. • Moderate - Discomfort that interferes with daily life. • Severe – results in incapacitation or inability to carry out normal daily activities. 【0150】 Relevance Assessment: The relevance of each adverse event (AE) to the treatment in the study shall be determined by the principal investigator based on the following explanation. • Not related - The event is clearly attributable to other factors such as the patient's clinical condition, therapeutic intervention, or concomitant medications, and is not consistent with known response patterns to the investigational drug. • Possible – The event has a reasonable temporal relationship to the timing of administration of the investigational drug and / or is consistent with known response patterns to the treatment in the study, but may also have been caused by other factors, such as the patient's clinical condition, therapeutic intervention, or concomitant medications. • Clear correlation exists - The event is reasonably time-related to the timing of administration of the investigational drug, consistent with known response patterns to the investigational drug, cannot be reasonably explained by other factors such as the patient's clinical condition or the therapeutic intervention administered to the patient, and meets one of the following conditions: the event occurs immediately after administration of the investigational drug, improves upon discontinuation of the investigational drug, recurs upon re-administration, or a positive reaction is observed at the administration site. 【0151】 statistical analysis For continuous numerical variables fitted to a normal distribution with variance based on a chi-squared distribution, t-tests were used to compare their means. For continuous numerical variables not fitted to a normal distribution, nonparametric tests were performed. Data for continuous variables were reported as mean ± standard deviation or median, as well as upper and lower quartiles. Fisher's exact test was used to compare proportions between two groups. Data were analyzed and plotted using SPSS 26.0 and GraphPad Prism 9 software. All tests were performed as two-tailed tests, and statistical significance was considered to be less than 0.05. 【0152】 result A total of 10 patients were selected to participate in this randomized controlled pilot trial. Five patients took four oropharyngeal probiotic lozenges daily for a total of nine weeks, including two weeks before the start of CCRT (concurrent chemoradiotherapy) and seven weeks during the CCRT period. The other five patients did not take oropharyngeal probiotics and were treated as a control group. Treatment compliance was 100% in this trial, and no patients dropped out. As shown in Table 11, there were no differences in basic characteristics between the two groups. The CCRT-P group consisted of four men and one woman, with clinical stages being newly diagnosed as stage II (two patients), stage III (two patients), and stage IV (one patient). The CCRT group consisted of three men and two women, all of whom were newly diagnosed as stage III. The median total target volume of radiation was 48.4 and 51.8 for the CCRT and CCRT-P groups, respectively. Baseline characteristics analysis showed no significant differences between the CCRT group and the CCRT-P group in terms of sex, age, total target volume, tumor stage / lymph node stage / metastasis stage (TNM staging classification), and clinical stage (P>0.05). There were no interruptions or delays in patient treatment due to RIOM in this trial. 【0153】 [Table 12] 【0154】 Initial evaluations demonstrated that oropharyngeal probiotic intervention was safe for NPC patients undergoing radiotherapy, while simultaneously reducing the incidence and severity of radiation-induced oral mucositis (RIOM), as shown in Tables 12 and 13. During 7 weeks of radiotherapy, the incidence of RIOM with RTOG ≥ 2 decreased. Furthermore, there was a 3-week delay in the onset of RIOM when over 60% of patients presented with RTOG ≥ 2 (week 3 in the control group, week 6 in the probiotic group). In contrast, severe RIOM classified as RTOG 3 and 4 was completely prevented in patients receiving oropharyngeal probiotics. 【0155】 [Table 13] 【0156】 [Table 14] 【0157】 In a more detailed assessment, 60% of patients experienced severe RIOM (RTOG ≥ 3), all of whom were in the CCRT (control) group. The incidence rates of RTOG grade 1, 2, 3, and 4 RIOM in the CCRT group and the CCRT-P group were 0%, 40%, 40%, and 20%, respectively, and 40%, 60%, 0%, and 0%, respectively. 【0158】 Figure 1 shows the mean RTOG scores experienced by patients during the 7-week CCRT period. RIOM (RTOG=1) onset began in the CCRT and CCRT-P groups at weeks 2 and 3, respectively. The CCRT-P group showed a 42.9% reduction in mean maximum RIOM severity at week 7 of CCRT, with mean RTOG scores of 2.8±0.84 and 1.6±0.55 for the CCRT and CCRT-P groups, respectively. The mean interval between CCRT initiation and maximum RIOM severity was 5 weeks and 4 weeks for the CCRT and CCRT-P groups, respectively. A significant difference in RIOM distribution between the two groups was observed at week 5 of the CCRT treatment period (p=0.0309), with mean RTOG scores of 2.6±0.55 and 1.0±0.71 for the CCRT and CCRT-P groups, respectively. Table 14 shows detailed mean RTOG scores for patients at different clinical stages during the 7-week CCRT treatment period. While there was no strong correlation between the mean change in RIOM severity and clinical stage, it was observed that RIOM severity gradually increased over time with the progression of CCRT treatment. Overall, RIOM severity was halved in patients in the CCRT-P group, and a tendency for RIOM onset to be delayed was observed. Furthermore, administration of oropharyngeal probiotics significantly protected patients from developing severe RIOM (RTOG ≥ 3) during the 7-week CCRT treatment period. Table 15 shows the mean time from CCRT initiation to onset for each RIOM severity during the 7-week CCRT treatment period. In the table, the mean time to RIOM initiation for RTOG I-IV was 17 days, 20 days, 27 days, and 41 days in the CCRT group, respectively, while the mean time to RIOM initiation for RTOG I-II was 26 days and 34 days in the CCRT-P group, respectively. No patients in the CCRT-P group experienced severe RIOM with RTOG ≥ 3. In the CCRT-P group, compared to the CCRT group, a median delay of 9 days in the onset of RTOG I RIOM and a median delay of 14 days in the onset of RTOG II RIOM were observed. In Figures 2-5, the Kaplan-Meier curves compare the time to the first onset of RTOG I-IV RIOM in each patient in both groups.In patients in the CCRT group, the time from the start of CCRT treatment to the first onset of RTOG level I RIOM was 14–23 days, with a median of 17 days. In contrast, in patients in the CCRT-P group who presented with mild pain, the time from the start of CCRT treatment to the first onset of RTOG level I RIOM was 19–37 days, with a median of 26 days (Figure 2). Similarly, in patients in the CCRT group, the time from the start of CCRT treatment to the first onset of RTOG level II RIOM was 18–25 days, with a median of 20 days. On the other hand, in 3 of the 5 patients in the CCRT-P group, the time from the start of CCRT treatment to the first onset of RTOG level II RIOM was 28–39 days, with a median of 34 days, and the other 2 patients in the CCRT-P group did not experience moderate pain (Figure 3). In three of the five patients in the CCRT group, the time from the start of CCRT treatment to the first onset of RTOG level III RIOM ranged from 24 to 32 days, with a median of 27 days. In contrast, none of the patients in the CCRT-P group experienced severe RIOM (Figure 4). Only one patient in the CCRT group experienced bleeding and necrosis, while none of the patients in the CCRT-P group experienced such pain (Figure 5). 【0159】 [Table 15] 【0160】 [Table 16] 【0161】 Throughout the study period, no adverse events related to oropharyngeal probiotics were reported. In this study, all 10 patients survived longer than 18 months after CCRT, and during the 18-month follow-up period, no distant metastases or recurrent NPCs were observed in any of the patients, and their disease activity remained stable. This demonstrates that the oropharyngeal probiotic DSM 34540 is safe for NPC patients without inducing caries during CCRT. 【0162】 RIOM negatively impacts patients' quality of life, physical function, mental function, and social role functioning, and also increases the risk of malnutrition, all of which affect treatment outcomes. Therefore, there is a need for advanced clinical practice that enables healthcare professionals to prioritize coping approaches to reduce the burden of cancer treatment and improve patients' clinical outcomes. However, managing RIOM is difficult, and while various preventive drugs and natural substances have been clinically validated, few are affordable or have consistent data. For example, drugs that promote epithelial cell proliferation are considered approaches that should be carefully considered because they may also promote cancer cell proliferation, which can result in significant additional costs for cancer patients, such as increased medical consultations and increased financial burden. In this study, supplementation with oropharyngeal probiotics two weeks prior to and during CCRT delayed the onset of RIOM and provided a significant benefit in substantially reducing the prevalence of severe RIOM in NPC patients undergoing CCRT. On the other hand, safety was confirmed by the fact that no adverse events related to oropharyngeal probiotics were reported during the 7-week study period and the subsequent 18-month follow-up period, and no distant metastases or recurrent NPCs were observed. 【0163】 Example 2: Oral and pharyngeal probiotic ENT-K12 for preventing oral mucositis in human subjects who have undergone hematopoietic stem cell transplantation. Autologous hematopoietic stem cell transplantation (autologous HSCT) is the standard treatment for multiple myeloma patients who are candidates for high-dose therapy, lymphoma patients receiving second-line treatment, and acute myeloid leukemia. Immune function impairment and chemotherapy prior to HSCT significantly increase the risk of respiratory infections, particularly early-onset respiratory infections that develop within 100 days post-transplant (Scarlata et al., Eur Respir J.49 (2017), 1601902). Therefore, respiratory viral infections are common in patients who have undergone HSCT, usually presenting as upper respiratory tract infections but rapidly progressing to lower respiratory tract infections, which can be associated with increased mortality. 【0164】 Approximately 10% of autologous HSCT patients experience signs or symptoms of a common respiratory viral infection after hospitalization. Common respiratory viral pathogens include influenza virus, parainfluenza virus, adenovirus, human rhinovirus, and human coronavirus. Approximately 45% of patients with respiratory viral infections develop lower respiratory tract infections, and 7% require intensive care hospitalization (Moret et al., Infect Dis (Lond). 53 (2021), 274-280; Marinelli et al., Biol Blood Marrow Transplant. 26 (2020), 782-788). The progression of respiratory viral infections and the associated high mortality rates in hematopoietic stem cell transplant (HSCT) recipients and patients with hematological malignancies are increasingly recognized as significant causes of morbidity and mortality. (Fontana and Strasfeld, Infect Dis Clin North Am.33(2019), 523-544. Prevention of respiratory infections in HSCT patients is crucial, and certain drugs have been reported to be effective as treatments based on molecular diagnostic tests and multiplex assays. However, nasopharyngeal swab examinations are rarely performed regularly after discharge following transplantation, and antibiotic prescriptions are generally based on the clinician's experience. Numerous studies have shown that administration of gut probiotics and prebiotics is a safe and well-tolerated treatment in adult and pediatric autologous and allogeneic HSCT patients regarding the reconstruction of gut microbiota dysbiosis and immunomodulatory effects after HSCT. These may contribute to a reduction in gastrointestinal toxicity and the incidence of graft-versus-host disease (GVHD).See, for example, Mizutani et al., Intern Med. 62 (2023), 2949-2958; Andermann et al., Transplant Cell Ther. 27 (2021), 932.e1-932.e11; Ladas et al., Bone Marrow Transplant. 51 (2016), 262-266; Gorshein et al., Clin Transplant. 31 (2017), 10.1111 / ctr.12947; Yazdandoust et al., Transpl Immunol. 78 (2023), 101836. In an attempt to discover other approaches to mitigate the risk of RTI and the progression from upper respiratory tract infections to lower respiratory tract infections in HSCT patients, this pilot randomized clinical trial was conducted to investigate whether oropharyngeal probiotics are associated with preventing or reducing the onset and severity of respiratory infections, and thus reducing antibiotic use after autologous HSCT. 【0165】 Furthermore, oral mucositis is a dose-limiting toxicity in HSCT that is generally associated with debilitation. The oral microbiome changed significantly after HSCT, returning to its original composition after 3 months, which is a risk factor for microbial-driven oral mucositis. In particular, the changes in microbial diversity and similarity were more pronounced and rapid in patients who developed ulcerative oral mucositis. It was reported that Streptococcus species were identified before HSCT but gradually decreased after transplantation and were replaced by coagulase-negative Staphylococcus, and that an increase in Candida species and the detection of Enterococcus species were significantly associated with ulcerative oral mucositis after HSCT. On the other hand, a more resilient microbial ecosystem was observed in patients who did not develop ulcerative oral mucositis. 【0166】 The test product contains the oral and pharyngeal probiotic strain Streptococcus salivarius subspecies thermophilus ENT-K12 (DSM 34540) – a probiotic strain for the oral and upper respiratory tract associated with ear, nose, and throat health. Previous clinical trials have established the safety and efficacy of daily administration of Streptococcus salivarius subspecies thermophilus ENT-K12 in reducing respiratory infections and major respiratory pathogens. The rationale for this study is to establish the safety and efficacy of oral and pharyngeal probiotic ENT-K12 administration in patients who have undergone hematopoietic stem cell transplantation (HSCT). The primary objective of this proposed study is to evaluate the efficacy of oral and pharyngeal probiotic ENT-K12 in reducing respiratory infections and oral mucositis in patients who have undergone hematopoietic stem cell transplantation. [Table 17] 【0167】 Main purpose A multicenter, open-label, randomized controlled pilot study was conducted to evaluate the efficacy of the oropharyngeal probiotic ENT-K12 in preventing respiratory infections and oral mucositis during the 100-day period following HSCT. 【0168】 Secondary purpose To evaluate the efficacy of the oropharyngeal probiotic ENT-K12 in reducing episodes of viral respiratory infections within 100 days after HSCT. • To evaluate the efficacy of the oropharyngeal probiotic ENT-K12 in reducing episodes of bacterial respiratory infections. • To evaluate the efficacy of the oropharyngeal probiotic ENT-K12 in reducing episodes of fungal respiratory infections. To evaluate the efficacy of the oral pharyngeal probiotic ENT-K12 in reducing the severity of oral mucositis and xerostomia (according to the Oral Mucositis Severity Scale; WHO). • To evaluate the effect of the oral pharyngeal probiotic ENT-K12 on the recovery of T lymphocyte subsets. • To evaluate the effects of the oral-pharyngeal probiotic ENT-K12 on immunomodulation mediated by salivary cytokines. • Number of days of antibiotic treatment • Number of days of treatment with antiviral drugs (ribavirin / parvizumab) • Number of days of treatment with antipyretics / steroids • Number of days of treatment with antifungal drugs • Number of days experienced oral mucositis • Number of days spent experiencing gingivitis • Progression from URTi to LRTi (ground-glass opacities on chest CT) • Severe symptoms and requiring hospitalization due to hypoxemia (ICU) • Pulmonary complications (decreased FEV1, reduced oxygen diffusion capacity) • Incidence and etiology of diarrhea • Incidence and grade of acute graft-versus-host disease (GvHD) ·mortality rate • Confirm the safety of the subject as determined by adverse event reports. 【0169】 Exam Overview A study to determine the efficacy of the oral pharyngeal probiotic ENT-K12 in preventing respiratory infections and oral mucositis in patients who have undergone hematopoietic stem cell transplantation (HSCT). Up to 60 patients aged 18 to 65 years were eligible for this trial if they were diagnosed as eligible for HSCT according to the 2016 WHO criteria, had an Eastern Cooperative Oncology Group (ECOG) performance status of 0-2, had adequately reconstructed hematopoietic systems after HSCT, and were free from active symptoms of lower respiratory tract infections, chronic respiratory disease, or organ failure. Ultimately, 16 patients who met these criteria participated in this trial. Participants were randomly assigned to one of the following two treatment groups: • Oral and pharyngeal probiotics ENT-K12 • Control (no treatment) 【0170】 The probiotic group included patients whose hematopoietic system showed a platelet count of 20 × 10 after HSCT. 9 Individuals / L greater than 0.5 × 10 9 After sufficient confirmation of hematopoietic reconstitution, defined as exceeding 1 / L, participants took one slow-dissolving lozenge three times a day for 100 days: after brushing their teeth at breakfast and lunch, and after applying mouthwash before bedtime. Participants were instructed to slowly inhale the lozenge until it was completely dissolved (approximately 4-5 minutes), and not to chew or swallow it whole. They were also advised to refrain from eating, drinking, or swallowing for two hours after administration of the oropharyngeal probiotics. 【0171】 Participant diagnoses and pre-treatment regimens were extracted from medical records from the start of pre-treatment to day 130, including the 100-day intervention period and the subsequent 30-day follow-up. Each episode of respiratory infection was recorded weekly. During the study period, participants were instructed to contact their study physician if they experienced symptoms of respiratory infection, such as sore throat, fever above 38°C, headache, muscle pain, shortness of breath, chest tightness, cough, nasal congestion or runny nose, lymphadenopathy, or abscesses or white patches on the tonsils. Adverse events were recorded during each consultation. Participants were required to return any unused probiotics at the end of the study, and compliance was assessed by calculating the number of remaining lozenges at the final consultation. Compliance was defined as the use of 90% or more of the distributed lozenges. 【0172】 Test group Patients diagnosed as eligible for HSCT according to the 2016 WHO criteria, aged 18 to 65 years, with a performance status of 0 to 2 according to the Eastern Cooperative Oncology Group (ECOG), who have had sufficient hematopoietic reconstitution after HSCT and have no active symptoms of lower respiratory tract infection, chronic respiratory disease, or organ dysfunction, are eligible for this trial. Therefore, a total of 16 patients who met the above criteria were enrolled in this trial and randomly assigned to the probiotic group or the control group. 【0173】 Inclusion Criteria To be considered eligible for enrollment in this trial, subjects must meet the following criteria: · Patients diagnosed as eligible for HSCT according to the 2016 WHO criteria · 18 - 65 years old · Performance status of 0 to 2 according to the Eastern Cooperative Oncology Group (ECOG) · No active symptoms of respiratory infection · No active symptoms of bacteremia · No active symptoms of organ dysfunction · Tolerate this trial and be able to complete it until the end · Have signed written informed consent Exclusion criteria Subjects will be excluded from the trial if they meet the following criteria. · Performance status of ≥3 according to the Eastern Cooperative Oncology Group (ECOG) · Incomplete hematopoietic reconstitution defined as platelet count < 20 × 10 9 cells / L and neutrophil count < 0.5 × 10 9 cells / L after HSCT · Active symptoms of respiratory infection at the time of enrollment · Active symptoms of bacteremia · Active symptoms of organ dysfunction · Presence of chronic respiratory disease that may affect observation 【0174】 Test Procedure At the time of the screening visit, the platelet count after HSCT was 20 x 10 9 cells / L or more, neutrophil count 0.5 × 10 9 After hematopoietic reconstitution is confirmed by a value of 1 / L or higher, the study subjects will be given an overview of the study and the planned procedures. Subsequently, the subjects will be asked to read the subject information document and sign the informed consent form, and a signed copy will be provided to them. 【0175】 In detail, the following information is recorded and the procedure is executed. • Written informed consent • Target number and date of visit • Inclusion and exclusion criteria 【0176】 If the test subject meets the registration requirements, record the following information and follow the procedure. • Eligibility • Demographic data • Medication history ·Oral mucositis grading scale; WHO • Randomization 【0177】 Participants will be randomly assigned to one of two treatment groups: a) oral pharyngeal probiotic ENT-K12; or b) control (no treatment). Participants will be provided with a 100-day supply of the test product and instructed to take one lozenge three times daily, after brushing their teeth and applying mouthwash: after breakfast, after lunch, and before bedtime. Participants must slowly inhale the lozenge until it is completely dissolved (approximately 4-5 minutes), ensuring they do not chew or swallow it directly. Participants are advised not to eat, drink, or swallow for at least two hours after administering the probiotics. Participants will be instructed to return any unused test product on their final visit. 【0178】 Participants will be instructed to continue their usual medication, nursing care, diet, and exercise routines throughout the study period. Participants will be instructed to contact their study physician at any time during the study if they experience symptoms of respiratory infection, such as sore throat, fever above 38°C, headache, muscle pain, shortness of breath, chest tightness, cough, nasal congestion or runny nose, swollen lymph nodes and / or the appearance of tonsil abscesses (pus) or white patches, or symptoms of oral mucositis, such as dry mouth, thickened saliva, increased mucus, shiny, swollen, erythematous gums, soft white patches or pus on the tongue, oral pain, oral bleeding, or mild burning while eating. If findings of respiratory infection or oral mucositis are observed, participants will be asked to visit the study site for confirmation of the diagnosis. • Blood sample collection for detection of T lymphocyte subsets (performed according to standard medical procedures) • Saliva collection for detection of immunomodulation (0, 1, 3, 7, 100, 130) • Swab collection of pharyngeal and oral mucus for the purpose of detecting major respiratory pathogens (including C. albicans) and reconstructing the oral microbiota. • Self-assessment of overall health status (ask participants about their overall health status and have them indicate their results on a 5-point scale). • Final appointment 【0179】 Participants will be given an appointment to visit the study site for their final visit (day 100). Participants will be asked to record the days they experienced symptoms of respiratory infection, oral mucositis, and gingivitis in the past 100 days (medical staff or medical students will contact participants weekly by phone to check on their health). 【0180】 Visiting the clinic when symptoms are present (visiting for the diagnosis of respiratory infections and oral mucositis) Participants in the study will be instructed to contact their study physician at any time during the study if they experience any of the following symptoms of respiratory infection or oral mucositis: ·Sore throat, • Fever exceeding 38℃, ·headache, Muscle pain, ·shortness of breath, · Chest compression · Cough · Nasal congestion or runny nose · Lymph node swelling and / or the appearance of tonsillar abscess or white coating. · Dry mouth · Thickening of saliva · Increase in mucus · Shiny, swollen, erythematous gums · Soft white patches or pus on the tongue · Pain in the mouth · Bleeding in the mouth or mild burning sensation during meals 【0181】 If findings of respiratory infection are recognized, the subject is required to visit the test facility. A throat swab is taken and a rapid swab test is conducted to detect respiratory pathogens. The physician advises on medication / medical treatment as necessary. The test subject continues to take the test article throughout the test period according to the given instructions. If treatment with antibiotics is required, the test subject continues to take the test article during the antibiotic treatment period. 【0182】 Record the following information and perform the procedure in "Visiting the hospital when symptomatic". · Subject number and date of hospital visit · Diagnosis (respiratory infection, oral mucositis, diarrhea, acute GvHD) (type of respiratory infection (e.g., streptococcal pharyngitis / tonsillitis, respiratory syncytial virus infection), and please indicate whether the infection was confirmed by rapid pharyngeal swab test (RAD) or other microbial tests) · Prescribed medications (describe antibiotics, antiviral drugs, antipyretic / steroids, antifungal drugs) · Oral mucositis grading scale; WHO · Number of days of oral mucositis · Number of days of gingivitis · Progression from upper respiratory infection to lower respiratory infection (e.g., ground-glass opacities on chest CT) · Requirement for hospitalization or ICU due to severe symptoms and hypoxemia · Pulmonary complications (decrease in FEV1, decrease in oxygen diffusion capacity) • Adverse events (Confirm whether or not adverse events occurred in the test subjects. If adverse events occurred during the procedure, please describe their type and severity in the attached "Adverse Events" section.) • Self-assessment of overall health status (ask test subjects about their overall health status and have them indicate their results on a 5-point scale). 【0183】 Follow-up visit (30 days after the last visit) Record the following information and perform the procedure during your follow-up visit. • Target number and date of visit • Medical history over the past 30 days (upper respiratory tract infection, progression from URTi to LRTi requiring hospitalization, pulmonary complications, occurrence and etiology of diarrhea, occurrence and grade of acute GvHD) ·Oral mucositis grading scale; WHO • Number of days in the past 30 days on which you have experienced oral mucositis • Number of days you experienced gingivitis in the past 30 days (Please indicate the type and number of respiratory infections and oral mucositis you have had in the past 30 days, and whether the infection was confirmed by a rapid pharyngeal swab test (RAD) or other microbiological test.) Adverse events • Self-assessment of overall health status (ask participants about their overall health status and have them indicate their results on a 5-point scale). • Antibiotic treatment in the past 30 days • Treatment with antiviral drugs (ribavirin / parvizumab) within the past 30 days • Treatment with antipyretics / steroids in the past 30 days • Treatment with antifungal medication in the past 30 days • Exam completed 【0184】 Exclusion from the test Participants will be excluded from the exam in the following cases: • If a test subject voluntarily wishes to withdraw from the test. • If the test subject develops a condition that contradicts the initial selection criteria • If the principal investigator determines, at their discretion, that a participant is unsuitable for continuing the study. 【0185】 Concomitant medications Participants in the study will be asked about their medication history. Details of the medications taken will be recorded in the case record and case report form. 【0186】 Test sample Participants in the probiotic group will take oropharyngeal probiotic lozenges formulated in the form of oral lozenges containing the following composition. Each lozenge will contain at least 1 billion colony-forming units (CFUs) of Streptococcus salivarius ENT-K12 (also known as Streptococcus salivarius subspecies thermophilus ENT-K12) over the shelf life. [Table 18] 【0187】 [Table 19] 【0188】 Grading and evaluation of adverse events AE is assessed using a three-level scale and reported in detail as shown in the CRF. • Mild – Easily tolerable, with slight discomfort, but does not interfere with daily life. • Moderate - Discomfort that interferes with daily life. • Severe – results in incapacitation or inability to carry out normal daily activities. 【0189】 Relevance Assessment: The relevance of each adverse event (AE) to the treatment in the study shall be determined by the principal investigator based on the following explanation. • Not related - The event is clearly attributable to other factors such as the patient's clinical condition, therapeutic intervention, or concomitant medications, and is not consistent with known response patterns to the investigational drug. • Possible – The event has a reasonable temporal relationship to the timing of administration of the investigational drug and / or is consistent with known response patterns to the treatment in the study, but may also have been caused by other factors, such as the patient's clinical condition, therapeutic intervention, or concomitant medications. • Clear correlation exists - The event is reasonably time-related to the timing of administration of the investigational drug, consistent with known response patterns to the investigational drug, cannot be reasonably explained by other factors such as the patient's clinical condition or the therapeutic intervention administered to the patient, and meets one of the following conditions: the event occurs immediately after administration of the investigational drug, improves upon discontinuation of the investigational drug, recurs upon re-administration, or a positive reaction is observed at the administration site. 【0190】 statistical analysis For continuous numerical variables that follow a normal distribution and satisfy the principle of equal variances, t-tests were used to compare their means. For continuous numerical variables that do not fit a normal distribution, nonparametric tests were performed. Data for continuous variables were reported as mean ± standard deviation or median, as well as upper and lower quartiles. The chi-squared test was used to compare proportions between two groups. Data were analyzed and plotted using SPSS 26.0 and GraphPad Prism 9 software. All tests were performed as two-tailed tests, and statistical significance was considered to be less than 0.05. 【0191】 result This study enrolled 16 patients (6 females, 10 males, mean age 39.8 years (28-53 years)) with hematopoietic malignancies who underwent autologous HSCT. Seven patients were randomly assigned to receive oropharyngeal probiotics, while the remaining nine were assigned to a control group that did not receive probiotics. Patient compliance was good in this trial, and no patients dropped out. The characteristics of the patients at the time of randomization are summarized in Table 18. 【0192】 [Table 20] 【0193】 The most common underlying disease was non-Hodgkin lymphoma, accounting for 100% in the probiotic group and 88.9% in the control group, with the remainder being Hodgkin lymphoma. Of the seven patients in the probiotic group, two, four, and one received BEAM, BuCyE, and BUCY pretreatment regimens prior to autologous HSCT, while five, one, and three patients in the control group received BEAM, BuCyE, and BUCY pretreatment regimens prior to autologous HSCT. Patient age, sex distribution, BMI, disease status at autologous HSCT, median number of CD34+ cells / kg infused, serum albumin levels, length of hospital stay, weight loss during hospitalization, duration of thrombocytopenia, duration of fever symptoms, and number of days of prophylactic antibiotic use prior to HSCT were similar between the two groups. The rates of febrile neutropenia, positive symptoms of sore throat, and dry mouth were similar between the two groups. Patients in the control group had higher mean c-reactive protein levels and longer duration of neutropenia at enrollment, but this was not statistically significant. Bloodstream infections (BSIs) were recorded in one patient in the probiotic group at enrollment, but there was no significant difference in BSI rates between the two groups. 【0194】 Results regarding the efficacy of the oral pharyngeal probiotic S. salivarius ENT-K12 for oral mucositis. Episodes of ulcerative oral mucositis were monitored three times monthly for 100 days after hematopoietic system reconstitution, followed by a 30-day follow-up. Initial evaluations showed that oropharyngeal probiotic intervention was safe (no adverse events were observed) in patients with sufficient hematopoietic system reconstitution after HSCT, and furthermore, a reduction in the incidence of ulcerative oral mucositis was observed throughout the entire 130-day study period. In particular, the incidence of oral mucositis with a WHO grade ≥2 was reduced in patients treated with oropharyngeal probiotics; that is, the incidence-free rate was higher in patients treated with oropharyngeal probiotics compared to the control group. No cases of ulcerative oral mucositis were observed in patients in the probiotic group during the 100-day oropharyngeal probiotic intervention. See Figure 6. 【0195】 Because HSCT patients are immunosuppressed, careful consideration and comprehensive safety assessment are necessary when administering probiotics. In this study, probiotic administration was initiated only after the hematopoietic system had been sufficiently reconstituted. Sepsis is one of the worst outcomes for patients with mucositis caused by cancer treatment, potentially leading to treatment interruption and even life-threatening complications. In this pilot study, no patients developed sepsis with probiotic administration, and no probiotic-related side effects were reported throughout the entire study period. Furthermore, patients taking oropharyngeal probiotics were completely protected from ulcerative oral mucositis. 【0196】 Results regarding the efficacy of the oral and pharyngeal probiotic S. salivarius ENT-K12 against respiratory infections. Following an evaluation of clinical trial results regarding oral mucositis, we analyzed the efficacy and safety of oropharyngeal probiotics for respiratory infection-like symptoms in lymphoma patients after autologous hematopoietic stem cell transplantation. 【0197】 In patients who received oropharyngeal probiotics for 100 days after hematopoietic stem cell transplantation (HSCT), the incidence of respiratory tract infections (RTi) tended to decrease by more than half (29% vs. 67%; p=0.131). The progression rate from upper respiratory tract infections (URTi) to lower respiratory tract infections (LRTi) was 0% in the probiotic group and 16.7% in the control group. Similarly, over 100 days after HSCT, the incidence of RTi tended to decrease by approximately two-thirds in the probiotic group (0.43% vs. 1.00%; p=0.094). Regarding RTi-like symptoms, the duration of cough, runny nose, and sore throat was shorter in the probiotic group than in the control group over 100 days after HSCT (3, 1, and 0 days vs. 17, 15, and 4 days, respectively; p=0.090, 0.059, and 0.102). Furthermore, over 100 days after HSCT, the probiotic group showed a more than tenfold reduction in the number of days of medication compared to the control group, with particularly significant statistical differences observed in the number of days of antibiotic use (0 days antiviral vs. 1 day antifungal, 0 days antibiotic vs. 1 day antiviral vs. 0 days antifungal, 15 days antibiotic; p=0.378, 0.257, and 0.024, respectively). On the other hand, in the 30-day follow-up, the average number of days of medication was similar in both groups (0 days of antibiotic use vs. 0.7 days of antibiotic use; p=0.170, respectively). Over 100 days after HSCT, the average duration of each RTi episode in the probiotic group was half that of the control group. However, as mentioned above, under conditions of reduced antibiotic use, the average duration of each RTi episode was almost the same in both groups (1.4 days vs. 0.7 days; p=0.816). As shown in Table 21, a statistically significant difference was observed in the average number of days of antibiotic use over the entire 130-day study period, with the average number of days of antibiotic use per person in the probiotic group being significantly lower than in the control group (0 days vs. 15 days; p=0.011). 【0198】 [Table 21] 【0199】 [Table 22] 【0200】 [Table 23] 【0201】 Patients who were diagnosed with bacteremia and had positive blood culture tests at the time of enrollment in the probiotic group did not develop sepsis throughout the study period. Sepsis with pneumonia after HSCT occurred in one of the nine patients in the control group. 【0202】 Kaplan-Meier analysis showed that patients in the probiotic group had a consistently higher probability of not developing an RTi episode compared to the control group (p=0.071; Figure 7), while the cumulative duration of RTi-like symptoms was consistently shorter in the probiotic group (p=0.131; Figure 8), and the cumulative number of days of antibiotic use in the probiotic group throughout the entire study period was significantly lower (p=0.011; Figure 9). In patients in the control group, the cumulative number of days of antibiotic use increased significantly with increasing cumulative duration of RTi-like symptoms. On the other hand, in patients in the probiotic group, RTi-like symptoms were mild, and antibiotics were not prescribed by the attending physician. Neither therapeutic nor prophylactic antibiotic use was required during the study period. No adverse events related to oropharyngeal probiotics were reported throughout the study period, all seven patients in the probiotic group survived, and no BSI, distant metastases, or lymphoma recurrences were observed during the 24-month follow-up period after the end of the study. These results indicate that oropharyngeal probiotics are safe in patients whose hematopoietic system has been reconstituted after HSCT. 【0203】 Prior to allogeneic or autologous HSCT, recipients must undergo chemotherapy or radiotherapy preconditioning to remove hematopoietic stem cells and eliminate as many residual leukemia cells as possible to facilitate transplant success during HSCT, although chemotherapy or radiotherapy can lead to immunosuppression, dysbiosis of the microbiome, bacterial migration, and pro-inflammatory cytokine responses. Common sources of inflammation among HSCT patients include bacterial, viral, and fungal infections, as well as chronic inflammatory / autoimmune / neurological disorders. Antibiotic prophylactic administration to protect HSCT recipients from infection has extended survival time, but its long-term effects have been debated for decades (Sepkowitz 29(., Bone Marrow Transplant. 2002), 367-371; Tabarraee et al., Iran J Pharm Res. 15(Suppl) (2016), 159-163; Cesar-Arce et al., Transplant Proc. 49 (2017), 1444-1448; Wang et al., J Microbiol Immunol Infect. 51 (2018), 123-131; Kimura et al., J Infect. 69 (2014), 13-25). Although antibiotic exposure is unrelated to infectious complications immediately after transplantation, it has also been reported to have long-term adverse effects on overall survival after HSCT (Gromowsky et al., Blood 140(Supplement). 1) (2022), 7716-7717; Mokhtar et al., Nutrients. 14 (2022), 112. Similarly, the wise use of appropriate antifungal treatments remains an important part of the treatment protocol. 【0204】 This study demonstrated that adjuvant treatment with oropharyngeal probiotics effectively reduced the prevalence of RTi in patients after HSCT, shortened the duration of respiratory symptoms, and reduced the number of days of antibiotic use. 【0205】 Example 3: Streptococcus salivarius ENT-K12 promotes healing of chemotherapy-induced oral mucositis in mice. After demonstrating the advantageous benefits of administering the probiotic S. salivarius ENT-K12—delaying the onset of RIOM (radiation-induced oral mucositis), significantly reducing the prevalence of severe RIOM (Example 1), and completely protecting patients who underwent HSCT from ulcerative OM (Example 2)—the effects of S. salivarius ENT-K12 on chemotherapy-induced oral mucositis, particularly the epithelial layer, were further analyzed in mouse studies. Furthermore, tests were conducted to demonstrate the effects of heat-killed S. salivarius ENT-K12 on oral epithelial cells damaged by chemotherapy. 【0206】 Initial trials were conducted using a mouse model, including chemotherapy with busulfan and cyclophosphamide as described below. Experimental grouping: (40 male C57BL / 6 mice, 6-8 weeks old) 1) Blank control group (n=10): No intervention, 2) Disease-injured group (n=10): Busulfan + cyclophosphamide combination chemotherapy, 3) Probiotic intervention group A (n=10): Busulfan + cyclophosphamide + heat-killed probiotic intervention. 4) Probiotic intervention group B (n=10): Busulfan + cyclophosphamide + live probiotic intervention. 【0207】 material: DMSO (to dissolve busulfan and cyclophosphamide), 4% paraformaldehyde, 0.1% heparin, PBS, pentobarbital sodium, busulfan, cyclophosphamide, probiotic tablets, heat-killed probiotics, 100 μl pipette, 1 ml syringe, etc. 【0208】 Reagent preparation: For use, weigh 100 mg of busulfan and dissolve it in 25 mL of DMSO to make a 4 mg / mL solution. For further use, dissolve 0.25 g of cyclophosphamide in 25 mL of DMSO to make a 10 mg / mL solution. Wrap all of the above solutions in tin foil and store in a cool, dark place. 【0209】 Mode of administration and dosage i) Busulfan + cyclophosphamide combination chemotherapy: Intraperitoneal injection: Once daily at a fixed time. Body weight was measured before administration and calculated after administration (dosage: Busulfan (BU) was administered at a dose of 80 mg / kg, and cyclophosphamide (CY) was administered at a dose of 200 mg / kg). ii) Intervention with live probiotics: Probiotic tablets (test samples described in Examples 1 and 2) were dissolved to prepare a probiotic solution. 100 μl of the probiotic solution was pipetteed and administered orally to mice from day 1 to day 6. After drug delivery, the mice were deprived of food and water for the next 30 minutes to allow the probiotics to remain in the oral cavity for as long as possible. A blank control group was administered 100 μl of saline as a placebo in the same manner. All mice were sacrificed on day 7. ii) Intervention with heat-killed probiotics: Administer orally as described above. 【0210】 Duration of chemotherapy administration (busulfan injection for the first 4 days, followed by cyclophosphamide injection for the next 2 days) 【0211】 [Table 24] 【0212】 Tissue collection At the assigned time (7 days after treatment of the mice), the mice were euthanized with pentobarbital sodium. After euthanasia, the animals were perfused with 0.1 M phosphate-buffered saline (PBS) mixed with 0.1% heparin, followed by 4% paraformaldehyde mixed with PBS, to immediately fix the tongue tissue. After dissection, the tongue was excised and fixed in 4% paraformaldehyde solution for 3 hours, after which it was divided into three parts. The tissue was then incubated with a series of sucrose concentrations (0.5 M, 1.0 M, 1.5 M) for cryoprotection, embedded in OCT compound, and stored at -80°C. The tissue sections were cut to a thickness of 12 μm and mounted directly onto slides. These slides were stored at -80°C until use in processing. 【0213】 Detection indicators HE staining We observed morphological changes in taste buds. 【0214】 Procedure: (1) Deparaffinize the paraffin sections and replace with water. Place the sections in xylene I for 10 minutes, xylene II for 10 minutes, anhydrous ethanol I for 5 minutes, anhydrous ethanol II for 5 minutes, 95% alcohol for 5 minutes, 90% alcohol for 5 minutes, 80% alcohol for 5 minutes, 70% alcohol for 5 minutes, and wash with distilled water. (2) Hematoxylin staining of the cell nuclei. Stain the sections with Harris hematoxylin for 3-8 minutes, wash with tap water, differentiate with 1% hydrochloric acid alcohol for a few seconds, rinse with tap water, return to blue with 0.6% ammonia water, and rinse with running water. (3) Eosin staining of the cytoplasm. Stain the sections with eosin staining solution for 1-3 minutes. (4) Dehydration and mounting. (5) Immerse the sections sequentially in 95% alcohol I for 5 minutes, 95% alcohol II for 5 minutes, anhydrous ethanol I for 5 minutes, anhydrous ethanol II for 5 minutes, xylene I for 5 minutes, and xylene II for 5 minutes. (5) Microscopic examination, image acquisition, and analysis. 【0215】 Toluidine blue staining The tongue was stained with 1% toluidine blue in 10% acetic acid for 1 minute, followed by repeated washing with acetic acid to reveal erosive or ulcerative lesions on the surface (Muanza et al., Clin Cancer Res, 11(2005), 5121). The percentage of toluidine blue-positive surface area (excluding resected trauma) was calculated using ImageJ software. The tissue was then fixed in a 4% (v / v) paraformaldehyde solution in PBS at 4°C for 2 hours and treated for paraffin or OCT embedding. The thickness of the tongue and esophageal epithelium, as well as the length of the jejunal villi, were measured in H&E-stained tissue using ImageJ. Analysis was performed in a blinded manner with 3 mice per group, 3 fields per sample, and 5 measurements per field. Images were obtained using a Zeiss Axio Imager M1 microscope and an EC-Plan-Neofluar 920-NA 0.5 air objective lens, with the AXIOVISION-SE64 Rel.4.9.1 program (Bertolini et al., Transl Oncol 10 (2017), 612-620). 【0216】 Immunohistochemical staining for Ki67 This study was conducted to compare the effects of probiotic intervention on cell proliferation. Ki67 is a protein expressed at all stages of the cell cycle except the G0 and early G1 phases. Procedure: For this procedure, slides were first washed with PBS, then immersed in 10 mM sodium citrate buffer (pH 6.0) at 95°C for 15 minutes for antigen retrieval, and then cooled to room temperature. After washing with PBS, the tissue was blocked at room temperature for 1 hour with a mixture of 5% NGS, 1% BSA, and 0.3% Triton-X 100. Rabbit anti-Ki67 primary antibody was added to the tissue, covered with a Hybridrip, and incubated overnight at 40°C. The following day, after washing the sections with PBS, they were incubated at room temperature for 2 hours with Alexa 546 goat anti-rabbit secondary antibody. Subsequently, the sections were washed with PB and immersed in Sytox green to double-label the nuclei. After further cleaning, the coverslip was attached using Fluoromount G (Delay et al., PLoS One, 14 (2019), e0214890; Mukherjee et al., PLoS One 12 (2017), e0185473; Mukherjee et al., PLoS One 8 (2013), e61607). 【0217】 E-cadherin detection Testing the integrity of adhesive bonds in the oral mucosa. Procedure: Paraffin-embedded (for E-cadherin) or frozen (for PMN) tissue sections were stained with an anti-E-cadherin polyclonal antibody, followed by a FITC-conjugated secondary antibody, or, in the case of PMN, with NIMP-R14, a monoclonal antibody highly specific to mouse Ly-6G and Ly-6C, followed by a secondary anti-rat antibody conjugated with Alexa 555. The nuclear stain Hoechst 33,258 was used to visualize all cells (Bertolini et al., Transl Oncol, 10(2017), 612-620). 【0218】 PLCβ2 staining Immunofluorescence labeling of PLCβ2 was used to identify type II cells in taste buds, and immunolabeling of type II cells was used to determine whether differentiated, mature taste bud cells were affected by probiotic intervention. 【0219】 Procedure: Slides were washed with PBS and then incubated with 5% NGS in blocking solution at room temperature for 1.5 hours. Tissue was then incubated overnight at 40°C with rabbit anti-PLCβ2 primary antibody at a 1:1000 dilution. Tissue was incubated in the dark for 2 hours using Alexa 546 goat anti-rabbit secondary antibody (1:1000). Sytox green was used as a nuclear marker (Delay et al., PLoS One, 14 (2019), e0214890; Mukherjee et al., PLoS One 12 (2017), e0185473; Mukherjee et al., PLoS One 8 (2013), e61607; Sarkar et al., Chem Senses (2021), 46). 【0220】 SNAP-25 staining Immunofluorescence labeling of SNAP-25 was used to identify type III cells in taste buds, and immunolabeling of type III cells was used to determine whether differentiated, mature taste bud cells were affected by probiotic intervention. 【0221】 Procedure: The SNAP-25 protocol was the same as the PLCβ2 protocol. Tissues were incubated overnight at 40°C in rabbit anti-SNAP-25 primary antibody. The concentration and processing time of Alexa 546 secondary antibody, as well as the Sytox labeling procedure, were all the same as described above (Delay et al., PLoS One 14 (2019), e0214890; Mukherjee et al., PLoS One 12 (2017), e0185473; Sarkar et al., Chem Senses (2021), 46). 【0222】 RNA extraction and reverse transcription quantitative polymerase chain reaction (RT-qPCR) IL-6, IL-8, IL-1β, TNF-α, NF-κB, and other pro-inflammatory cytokines were detected in the tongue. 【0223】 Procedure: Mouse tongues were homogenized using a POLYTRON homogenizer, and the supernatant was beaten with zirconia beads containing phenol:chloroform:isoamyl alcohol. RNA was purified using the QIAgen RNeasy® Mini Kit, and its concentration / quality was determined using a NanoDrop instrument. Complementary DNA was synthesized using the SuperScriptIII Cells Direct® cDNA Synthesis Kit. Reverse transcription quantitative polymerase chain reaction was performed using a Bio-Rad CFX96 cycler and iQ® SYBR Green Supermix (Bertolini et al., Transl Oncol 10 (2017), 612-620). 【0224】 statistical analysis Immunofluorescence images were captured using a color camera and spot image acquisition software mounted on a Nikon Eclipse E600 microscope. Cell counts were performed by observers unaware of the experimental conditions, using the criteria of Nguyen et al. To more clearly identify immunopositive cells, image brightness and RGB levels were adjusted using Adobe Photoshop CS6 (https: / / www.Adobe.com), and images were enhanced as needed, before quantitative analysis was performed using NIH ImageJ (https: / / imagej.nih.gov / ij). The Ki67 percentage score was calculated by dividing the number of immunopositive Ki67+ cells by the total number of basal epithelial cells or Sytox-positive taste cells. Only cells present in the basal layer and crypt walls of circumvallate papillae containing taste buds were counted. Data on PLCβ2 and SNAP-25 labeling were average values ​​for 5–12 taste buds per mouse. The number of immunopositive cells and the total number of Sytox green-labeled nuclei were counted within each taste bud. All experimental data was collected from 3 to 6 mice per group. 【0225】 Linear variance model analysis (ANOVA) was used to analyze data for Ki67, PLCβ2, and SNAP-25. For each cell label, separate ANOVAs were performed to assess the number of labeled cells, the total number of cells in the taste bud, and the proportion of labeled cells in the taste bud. All statistical tests were performed using SPSS version 26.0. Graphs were created using GraphPad Prism 8 (Delay et al., PLoS One 14 (2019), e0214890; Mukherjee et al., PLoS One 12 (2017), e0185473; Mukherjee et al., PLoS One 8 (2013), e61607). 【0226】 result Nine mice were administered a dose of busulfan at 80 mg / kg, and it was observed that three mice died after one day. Therefore, in subsequent experiments, busulfan was administered at a total dose of 120 mg / kg (30 mg / kg divided into 4 doses) and cyclophosphamide at a total dose of 200 mg / kg (100 mg / kg divided into 2 doses). The duration of chemotherapy administration (busulfan injection for the first 4 days, followed by cyclophosphamide injection for the next 2 days) is as follows. 【0227】 [Table 25] 【0228】 Initial results showed that Streptococcus thermophilus ENT-K12 promoted healing of chemotherapy-induced oral mucositis in mice. In particular, chemotherapy treatment in mice resulted in significant mucosal hypoplasia and ulceration of the tongue. Furthermore, basal layer cells were only loosely aligned, and nuclear condensation, which is irreversible condensation of chromatin in the nucleus of cells undergoing necrosis or apoptosis, was observed. In addition, the tongue tissue had fewer cells in the stratum spinosum and stratum granulosum compared to healthy tissue. See Figures 10A and 10B. Treatment with ENT-K12 during chemotherapy restored the integrity of the tongue mucosa and partially restored the basal layer, stratum spinosum, and stratum granulosum. See Figure 10C. It was further observed that chemotherapy treatment reduced mucosal thickness by almost 50%, and that when Streptococcus salivarius ENT-K12 was administered during chemotherapy, the reduction was less, i.e., about 25%. See Figures 11A-11C and 12. 【0229】 Before conducting further experiments, the mouse model was changed to one administered with 5-fluorouracil (5-Fu). This was because this chemotherapeutic agent is more commonly used in the chemotherapy of human patients. See Example 4. 【0230】 Example 4: Live and inactivated Streptococcus salivarius ENT-K12 promotes healing of chemotherapy-induced oral mucositis in mice. This experiment analyzed the effects of administering the chemotherapeutic agents 5-fluorouracil (5-Fu) and S. salivarius ENT-K12 to 5-Fu-damaged oral epithelial cells. 【0231】 material and method: Reagent preparation: 5 mg of 5-Fu was dissolved in 100 μl of DMSO solvent to prepare a 5 mg / 100 μl (50 μg / μl) stock solution of 5-Fu. Since DMSO is cytotoxic, 10 μl of the stock solution of 5-Fu was diluted in 10 ml of DMEM medium to prepare a 1000-fold dilution. That is, the solution concentration was 50 μg / ml. In the intervention group, in addition to preparing a 50 μg / ml 5-Fu solution, it was necessary to add heat-killed S. salivarius ENT-K12 solution and the supernatant of heat-killed S. salivarius ENT-K12. Preparation method for heat-inactivated S. salivarius ENT-K12 solution: Thermal inactivation was performed by incubating S. salivarius at 65°C for 60 minutes. 【0232】 Establishing animal models: Eighteen 8-week-old mice weighing approximately 20g were selected and assigned to a control group, a chemotherapy group, and a post-chemotherapy probiotic intervention group (n=6). Mice in the 5-FU group and the probiotic intervention group were injected with 5-FU at a dose of 50 mg / kg for 5 consecutive days. 1 × 10⁶ mice in the intervention group were collected by centrifuge. 9 Oral lavage was performed twice daily with the CFU probiotic S. salivarius ENT-K12, while mice in the 5-Fu group were orally lavaged twice daily with sterile pure water. Tissue collection and staining were performed as described in Example 3. 【0233】 Analysis of oral epithelial cell (HOK cell) proliferation and proliferative activity in the presence of 5-Fu In the first experimental setup, the proliferative activity of oral epithelial cells (HOK cells) in the presence of 5-Fu was analyzed. For this purpose, 5-Fu was added to 96-well plates at six different concentrations: 0, 0.08, 0.4, 2, 10, and 50 μg / mL. Each concentration was used to create six wells, and 5000 HOK cells were added to each well. After panel treatment, the HOK cells were cultured in a carbon dioxide incubator for 24 hours. Cell proliferation was detected by life cell analysis (LCA). The apparatus was set up with five pairs of wells per group, four fields of view per well, and a magnification of 10x, with imaging performed every two hours for two days. As shown in the results of the life cell imaging experiment in Figure 13, a decrease in the proliferative activity of HOK cells was confirmed 48 hours after 5-Fu treatment. 【0234】 Furthermore, the proliferation activity of HOK cells was analyzed. Again, 5-Fu was added to 96-well plates at six different concentrations: 0, 0.08, 0.4, 2, 10, and 50 μg / mL. Each concentration was used to create six wells, and 5000 HOK cells were added to each well. The cells in the 96-well plates were treated with 5-Fu for 48 hours, after which 10 μl of CCK8 reagent was added to each well. CCK8 refers to Cell Counting Kit-8, a kit for highly sensitive colorimetric quantification of cell viability in cell proliferation and cytotoxicity assays, and is available, for example, from MedChemExpress (product number: HY-K0301). 【0235】 After 4 hours of treatment, cells were detected using a standard microplate reader (e.g., Thermo Scientific's Multiskan® FC microplate photometer). Mitochondrial metabolic activity was measured as an indicator reflecting cell proliferation capacity, and treatment with 5-FU at concentrations greater than 10 μg / mL for 48 hours resulted in a significant decrease in the proliferation activity of HOK cells. See Figure 14. 【0236】 Effect of heat-killed S. salivarius ENT-K12 on the proliferation of oral epithelial cells (HOK cells) A solution of heat-inactivated S. salivarius ENT-K12 was prepared as described above, and the effect of heat-inactivated S. salivarius ENT-K12 on the growth of HOK cells in a 96-well plate (5000 HOK cells per well) was measured by adding it at different concentrations ranging from 0 mg / mL to 2 mg / mL. Incubation was performed for 48 hours, after which 10 μl of CCK8 reagent was added to each well. After 4 hours of treatment, cells were detected by a microplate reader. As shown in Figure 15, heat-inactivated S. salivarius ENT-K12 did not show growth toxicity to HOK cells at any concentration, and the cell viability of HOK cells was significantly increased by heat-inactivated DSM 34540 treatment at concentrations >1.5 mg / mL. 【0237】 Furthermore, the proliferative capacity of HOK cells was analyzed in the presence of 5-Fu to investigate the effect of S. salivarius ENT-K12 on HOK cells. For this purpose, a solution of heat-killed S. salivarius ENT-K12 was prepared as described above and added to HOK cells in a 96-well plate (5000 HOK cells per well) at different concentrations ranging from 0 mg / mL to 2 mg / mL. In addition, 10 μg / mL of 5-Fu was added to each well. Incubation was performed for 48 hours, after which 10 μl of CCK8 reagent was added to each well. After 4 hours of treatment, cells were detected by a microplate reader. As shown in Figure 16, the proliferation of HOK cells was significantly reduced after treatment with 10 μg / mL of 5-Fu. However, treatment of HOK cells with heat-killed S. salivarius ENT-K12 significantly and dose-dependently restored their proliferation. 【0238】 In vivo protection of the oral mucosal barrier with live and heat-killed S. salivarius ENT-K12 bacteria. As shown in Figures 17A and 17D, and Figures 18A and 18D, 5-Fu had significant adverse effects on the structural integrity and permeability of the oral mucosal barrier in mice. Clearly, 5-Fu treatment reduced the area of ​​the oral mucosal layer in mice, and treatment with both live and heat-killed S. salivarius ENT-K12 significantly increased it. 【0239】 Furthermore, Ki-67 expression was analyzed. Ki-67 is a protein located in the nucleus and expressed only during the stages of cell division associated with cell proliferation. As shown in Figures 19 and 20, Ki-67 expression in the 5-Fu group was lower than in the probiotic and control groups. Therefore, both live and heat-killed S. salivarius ENT-K12 significantly promoted the proliferation of oral mucosal cells. 【0240】 Heat-treated S. salivarius ENT-K12 improved HOK cell cycle damage induced by the chemotherapeutic agent 5-Fu. The cell cycle was analyzed by flow cytometry. For this purpose, a solution of heat-killed S. salivarius ENT-K12 was prepared, and cell proliferation was detected by CCK-8 analysis and LCA in the same manner as described above. 【0241】 The results are shown in Figure 21, where, compared to the control group, heat-killed S. salivarius ENT-K12 did not significantly alter the HOK cell cycle after 48 hours of treatment. On the other hand, 5-Fu treatment induced cell cycle arrest in the S phase and G2 / M phase, the period when cells undergo rapid growth and protein synthesis in preparation for mitosis and cytokinesis. Notably, heat-killed Streptococcus salivarius ENT-K12 restored the entire cell cycle after 5-Fu treatment. 【0242】 S. salivarius ENT-K12 reduced the level of DNA damage in HOK cells induced by chemotherapy 5-Fu. In further experiments, reactive oxygen species (ROS) levels in HOK cells were detected by flow cytometry as described above. As shown in Figure 22, ROS levels in HOK cells, which typically reflect DNA damage during chemotherapy, increased after 5-Fu treatment and significantly decreased after treatment with heat-killed S. salivarius ENT-K12. 【0243】 In summary, heat-killed S. salivarius ENT-K12 has been shown to have beneficial effects on oral epithelial cells damaged by chemotherapy, and therefore, in addition to live S. salivarius ENT-K12, it can be used for the prevention and treatment of oral mucositis.

Claims

[Claim 1] Streptococcus salivarius for use in the treatment or prevention of ear, nose, and throat (ENT) diseases induced by or associated with cancer treatment. [Claim 2] Streptococcus salivarius for use according to claim 1, wherein the ENT disease is cancer treatment-induced oral mucosal disorder. [Claim 3] Streptococcus salivarius for use according to claim 1, wherein the ENT disease is a cancer treatment-induced respiratory infection. [Claim 4] Streptococcus salivarius for use according to any one of claims 1 to 3, wherein the ENT disease is induced or associated with radiotherapy, chemotherapy, hematopoietic stem cell transplantation (HSCT), immune checkpoint inhibitor therapy, or any combination thereof. [Claim 5] Streptococcus salivarius for use according to any one of claims 1 to 4, wherein the ENT disease is induced by or associated with radiotherapy or concurrent chemoradiotherapy (CCRT). [Claim 6] Streptococcus salivarius for use according to any one of claims 1 to 4, wherein the ENT disease is induced by or associated with chemotherapy. [Claim 7] Streptococcus salivarius for use according to any one of claims 1 to 4, wherein the ENT disease is induced by or associated with hematopoietic stem cell therapy (HSCT). [Claim 8] Streptococcus salivarius for use according to any one of claims 1 to 4, wherein the ENT disease is induced or associated with HSCT preceded by chemotherapy. [Claim 9] Streptococcus salivarius for use according to any one of claims 1 to 4, wherein the ENT disease is induced by or associated with treatment of head and neck cancer, preferably nasopharyngeal cancer (NPC), by radiotherapy or CCRT. [Claim 10] Streptococcus salivarius for use according to any one of claims 1 to 4, wherein the ENT disease is induced or associated with HSCT, preferably chemotherapy and subsequent treatment of hematopoietic tumors with HSCT. [Claim 11] Streptococcus salivarius for use in the treatment or prevention of ear, nose, and throat (ENT) disorders induced by or associated with immunosuppression, preferably the ENT disorder being an oral mucosal disorder or a respiratory infection. [Claim 12] Streptococcus salivarius for use according to any one of claims 1 to 11, wherein the oral mucosal disorder is oral mucositis. [Claim 13] Streptococcus salivarius for use according to any one of claims 1, 2, and 4 to 12, wherein Streptococcus salivarius is provided in an inactive form, preferably Streptococcus salivarius is thermally inactivated. [Claim 14] The administration of Streptococcus salivarius, (i) Delay the onset of oral mucositis and reduce the risk of severe oral mucositis. (ii) dose-dependently restore the reduced proliferation and disruption of the cell cycle of oral epithelial cells after chemotherapy, and / or (iii) To prevent a decrease in the oral mucosal layer area after chemotherapy and to restore the integrity of the oral mucosa. Streptococcus salivarius for use according to claim 12 or 13. [Claim 15] The administration of Streptococcus salivarius, (i) To prevent the progression from upper respiratory tract infections to lower respiratory tract infections in the target population, (ii) Compared to controls that did not receive Streptococcus salivarius, the duration of respiratory infection symptoms in the subjects was reduced. (iii) Compared to the control group, reduce the mean duration of respiratory infection episodes in the subjects, and / or (iv) Reduce the need for the use of the target antibiotic compared to the control group. Streptococcus salivarius for use according to claim 3. [Claim 16] 3 x 10 ６ ~4 x 10 １０ CFU, preferably 3 × 10 ９ ~3 x 10 １０ CFU or 4x10 ９ ~4 x 10 １０ Streptococcus salivarius for use according to any one of claims 1 to 12 and 14 to 15, administered orally in a daily dose of CFU. [Claim 17] Streptococcus salivarius for use according to any one of claims 1 to 12 and 14 to 16, administered orally at a daily dose of 3 × 10 mg to 4 × 120 mg, preferably 3 × 50 mg or 4 × 50 mg. [Claim 18] Streptococcus salivarius for use according to any one of claims 1 to 17, administered orally to a subject four times a day for a period of about six to seven weeks during radiotherapy or chemoradiotherapy. [Claim 19] Streptococcus salivarius for use according to claim 18, administered orally four times a day for a period of about two weeks prior to the commencement of radiotherapy or chemoradiotherapy. [Claim 20] After undergoing HSCT, the platelet count was 20 x 10 ９ Individuals / L greater than 0.5 x 10⁻¹⁰ neutrophil count. ９ Streptococcus salivarius for use according to any one of claims 1 to 17, administered orally three times a day for a period of approximately 100 days to subjects in whom hematopoietic system reconstruction, defined as exceeding cells / L, has been confirmed. [Claim 21] A composition, preferably formulated as an oral composition, of Streptococcus salivarius for use according to any one of claims 1 to 20. [Claim 22] Streptococcus salivarius for use according to any one of claims 1 to 21, wherein the composition is an oral dosage form selected from tablets, capsules, gels, lozenges, chewable tablets, oil droplets, and powders, preferably an oil droplet, or a solid oral dosage form selected from tablets, capsules, chewable tablets, and lozenges, most preferably a lozenge. [Claim 23] The oral dosage form contains at least 10 ６ CFU of Streptococcus salivarius, preferably 10 ６ to 10 １０ CFU of Streptococcus salivarius, more preferably 10 ９ CFU or 10 １０ CFU of Streptococcus salivarius for use according to claim 22, comprising Streptococcus salivarius. [Claim 24] Streptococcus salivarius for use according to claim 22 or 23, wherein the oral dosage form comprises 1 to 500 mg, preferably 1 to 120 mg, preferably 10 to 120 mg of Streptococcus salivarius, most preferably 50 mg of Streptococcus salivarius. [Claim 25] Streptococcus salivarius for use according to any one of claims 21 to 24, wherein the oral dosage form is a lozenge. [Claim 26] The lozenge further comprises fructose, maltodextrin, magnesium stearate, and a flavoring, preferably a strawberry flavoring, for use according to claim 26. [Claim 27] The Streptococcus salivarius for use according to claim 25 or 36, wherein the lozenge is a slow-dissolving lozenge, preferably having a disintegration time of at least 4 to 5 minutes. [Claim 28] Streptococcus salivarius for use according to any one of claims 25 to 27, wherein the lozenge comprises about 50 mg or 5% Streptococcus salivarius, about 635 mg or 63.5% fructose, about 290 mg or 29% maltodextrin, about 15 mg or 1.5% magnesium stearate, and about 10 mg or 1% flavoring. [Claim 29] A composition comprising Streptococcus salivarius for use in the treatment or prevention of ear, nose, and throat (ENT) diseases induced by or associated with cancer treatment. [Claim 30] The composition for use according to claim 29, wherein the ENT disease is a cancer treatment-induced oral mucosal disorder. [Claim 31] The composition for use according to claim 29, wherein the ENT disease is a cancer treatment-induced respiratory infection. [Claim 32] A composition for use according to any one of claims 29 to 31, wherein the ENT disease is induced or associated with radiotherapy, chemotherapy, hematopoietic stem cell transplantation (HSCT), immune checkpoint inhibitor therapy, or any combination thereof. [Claim 33] The aforementioned ENT disease, (i) Head and neck cancer, preferably nasopharyngeal cancer (NPC), treated with radiotherapy or chemoradiotherapy, (ii) Hematopoietic tumors treated with HSCT, preferably chemotherapy followed by HSCT. A composition for use according to any one of claims 29 to 32, which is induced by or related to the treatment of the [Claim 34] A composition comprising Streptococcus salivarius for use in the treatment or prevention of an ear, nose, and throat (ENT) disorder induced by or associated with immunosuppression, wherein the ENT disorder is an oral mucosal disorder or a respiratory infection. [Claim 35] The composition for use according to any one of claims 29 to 34, wherein the oral mucosal disorder is oral mucositis. [Claim 36] A composition for use according to any one of claims 29, 30, and 32-35, comprising an inactivated, preferably thermally inactivated, Streptococcus salivarius. [Claim 37] An oral composition, the composition for use according to any one of claims 29 to 36. [Claim 38] The composition for use according to any one of claims 29 to 37, wherein the composition is an oral dosage form selected from tablets, capsules, gels, lozenges, chewable tablets, oil droplets, and powders, preferably an oil droplet, or a solid oral dosage form selected from tablets, capsules, chewable tablets, and lozenges, most preferably a lozenge. [Claim 39] at least 10 ６ CFU Streptococcus salivarius, preferably 10 ６ ~10 １０ CFU's Streptococcus salivarius, more preferably 10 ９ CFU or 10 １０ A composition for use according to any one of claims 29-35 and 37-38, comprising CFU Streptococcus salivarius. [Claim 40] A composition for use according to any one of claims 29 to 35 and 37 to 39, comprising 1 to 500 mg, preferably 1 to 120 mg, preferably 10 to 120 mg of Streptococcus salivarius, and most preferably 50 mg of Streptococcus salivarius. [Claim 41] A composition for use according to any one of claims 29-35 and 37-40, comprising 50 mg of Streptococcus salivarius, which is administered orally several times a day, preferably three or four times a day. [Claim 42] at least 10 ６ A composition for use according to any one of claims 29-35 and 37-41, comprising CFU of Streptococcus salivarius, which is administered orally several times a day, preferably three or four times a day. [Claim 43] A composition for use according to any one of claims 29 to 42, which is administered orally to a subject four times a day for a period of about six to seven weeks during radiotherapy or chemoradiotherapy. [Claim 44] The composition for use according to claim 43, which is administered orally four times a day for a period of about two weeks prior to the commencement of the radiotherapy or chemoradiotherapy. [Claim 45] After undergoing HSCT, the platelet count was 20 x 10 ９ Individuals / L greater than 0.5 x 10⁻¹⁰ neutrophil count. ９ A composition for use according to any one of claims 29 to 42, which is administered orally three times a day for a period of approximately 100 days to subjects in whom hematopoietic system reconstruction, defined as exceeding cells / L, has been confirmed. [Claim 46] A composition for use according to any one of claims 29 to 45, preferably a lozenge as defined in any one of claims 26 to 28. [Claim 47] Use of compositions containing Streptococcus salivarius in subjects undergoing treatment for cancer or autoimmune diseases, subjects undergoing radiotherapy, chemotherapy, hematopoietic stem cell transplantation (HSCT), immune checkpoint inhibitor therapy, or any combination thereof, or in immunosuppressed subjects. [Claim 48] The use according to claim 47, wherein the composition is the composition according to any one of claims 37 to 40 and 43 to 46. [Claim 49] A composition comprising Streptococcus salivarius or Streptococcus salivarius for use in the treatment or prevention of respiratory infections in human subjects, wherein the human subject is immunosuppressed. [Claim 50] The immunosuppression is caused by treatment for cancer or an autoimmune disease, and the Streptococcus salivarius or composition for use according to claim 49. [Claim 51] Streptococcus salivarius or composition for use according to claim 49 or 50, wherein the cancer treatment comprises HSCT and / or chemotherapy, preferably HSCT preceded by chemotherapy. [Claim 52] Streptococcus salivarius or composition for use according to any one of claims 49 to 51, wherein administration of Streptococcus salivarius prevents progression from upper respiratory tract infection to lower respiratory tract infection in the subject. [Claim 53] Streptococcus salivarius or composition for use according to any one of claims 49 to 52, wherein administration of Streptococcus salivarius shortens the duration of symptoms of respiratory infection in subjects compared to a control that was not administered Streptococcus salivarius or a composition containing Streptococcus salivarius. [Claim 54] Streptococcus salivarius or composition for use according to any one of claims 49 to 53, wherein administration of Streptococcus salivarius reduces the mean duration of respiratory infection episodes in the subject compared to a control group. [Claim 55] Streptococcus salivarius or composition for use according to any one of claims 49 to 54, wherein administration of Streptococcus salivarius reduces the need for the use of the antibiotic of the subject compared to a control group. [Claim 56] Streptococcus salivarius for use according to any one of claims 49 to 55, wherein Streptococcus salivarius is formulated as a composition, preferably an oral composition. [Claim 57] The composition for use according to any one of claims 49 to 55, wherein the composition is an oral composition. [Claim 58] Streptococcus salivarius or composition for use according to any one of claims 49 to 57, wherein the composition is an oral dosage form selected from tablets, capsules, gels, lozenges, chewable tablets, oil droplets, and powders, preferably the composition is a lozenge. [Claim 59] The composition contains at least 10 ６ CFU Streptococcus salivarius, preferably 10 ６ ~10 １０ CFU's Streptococcus salivarius, more preferably 10 ９ CFU or 10 １０ A composition of Streptococcus salivarius for use according to any one of claims 49 to 58, comprising CFU Streptococcus salivarius. [Claim 60] The composition comprises 1 to 500 mg, preferably 1 to 120 mg, preferably 10 to 120 mg of Streptococcus salivarius, most preferably 50 mg of Streptococcus salivarius, or Streptococcus salivarius for use according to any one of claims 49 to 59. [Claim 61] Streptococcus salivarius, 3 x 10 ６ ~4 x 10 １０ CFU, preferably 3 × 10 ９ ~3 x 10 １０ Streptococcus salivarius or composition for use according to any one of claims 49 to 60, administered orally to the subject in a daily dose of CFU. [Claim 62] Streptococcus salivarius or composition for use according to any one of claims 49 to 61, wherein Streptococcus salivarius is orally administered to the subject at a daily dose of 3 × 10 mg to 4 × 120 mg, preferably 3 × 50 mg. [Claim 63] The composition comprises 50 mg of Streptococcus salivarius, and the composition is administered orally to the subject several times a day, preferably three times a day, according to any one of claims 49 to 60. [Claim 64] The composition contains at least 10 ６ Streptococcus salivarius or composition for use according to any one of claims 49 to 60, comprising CFU of Streptococcus salivarius, wherein the composition is administered orally to the subject several times a day, preferably three times a day. [Claim 65] The composition undergoes HSCT, and after HSCT, the platelet count becomes 20 × 10 ９ Individuals / L greater than 0.5 x 10⁻¹⁰ neutrophil count. ９ Streptococcus salivarius or composition for use according to any one of claims 49 to 64, administered orally three times a day for a period of approximately 100 days to subjects in whom hematopoietic system reconstruction, defined as exceeding cells / L, has been confirmed. [Claim 66] Streptococcus salivarius or composition for use according to any one of claims 59 to 65, wherein the composition is an oral solid dosage form, preferably a lozenge, most preferably a lozenge as defined in any one of claims 26 to 28. [Claim 67] A method for treating or preventing ear, nose, and throat (ENT) disease in a subject requiring treatment or prevention of such ENT disease, comprising administering to the subject a composition containing Streptococcus salivarius or Streptococcus salivarius. [Claim 68] The method according to claim 67, wherein the ENT disease is cancer treatment-induced oral mucosal disorder or cancer treatment-related respiratory infection. [Claim 69] The method according to claim 67 or 68, wherein the ENT disease is an ENT disease as defined in any one of claims 2 to 12. [Claim 70] The method according to any one of claims 67 to 69, wherein Streptococcus salivarius is Streptococcus salivarius as defined in any one of the preceding claims. [Claim 71] The method according to any one of claims 67 to 69, wherein the composition is a composition defined in any one of the preceding claims. [Claim 72] The method according to any one of claims 67 to 71, wherein Streptococcus salivarius and the composition are administered as defined in any one of the preceding claims. [Claim 73] Use of Streptococcus salivarius or a composition comprising Streptococcus salivarius in the manufacture of a pharmaceutical product for treating or preventing ear, nose, and throat (ENT) diseases in subjects requiring treatment or prevention of such ENT diseases in patients requiring treatment or prevention of such ENT diseases. [Claim 74] The use according to claim 73, wherein the ENT disease is a cancer treatment-induced oral mucosal disorder or a cancer treatment-related respiratory infection, and preferably the ENT disease is an ENT disease as defined in any one of claims 2 to 12. [Claim 75] The use according to claim 73 or 74, wherein Streptococcus salivarius is Streptococcus salivarius as defined in any one of the preceding claims. [Claim 76] The use according to claim 73 or 74, wherein the composition is a composition defined in any one of the preceding claims. [Claim 77] The use according to any one of claims 73 to 76, wherein Streptococcus salivarius and the composition are administered as defined in any one of the preceding claims. [Claim 78] A method for treating or preventing a respiratory infection in a human subject, wherein the human subject is immunosuppressed, and the method comprises administering to the subject a Streptococcus salivarius or a composition containing Streptococcus salivarius. [Claim 79] The method according to claim 78, wherein the object is an object as defined in claim 50 or 51. [Claim 80] The method according to claim 78 or 79, wherein the Streptococcus salivarius is the Streptococcus salivarius defined in any one of claims 56 and 58-66. [Claim 81] The method according to claim 78 or 79, wherein the composition is a composition defined in any one of claims 57 to 66. [Claim 82] The method according to any one of claims 78 to 81, wherein Streptococcus salivarius and the composition are administered as defined in any one of claims 61 to 65, respectively. [Claim 83] A lozenge comprising Streptococcus salivarius for use in the treatment of ear, nose, and throat (ENT) disorders induced by or associated with cancer treatment, wherein the ENT disorder is a cancer treatment-induced oral mucosal disorder or a cancer treatment-associated respiratory infection, preferably an ENT disorder as defined in any one of claims 1 to 12. [Claim 84] A lozenge containing Streptococcus salivarius for use in the treatment or prevention of respiratory infections in human subjects, wherein the human subjects are immunosuppressed, and preferably as defined in claim 50 or 51. [Claim 85] A lozenge as defined in any one of claims 26 to 28, preferably administered as defined in any one of the preceding claims, according to claim 83 or 84. [Claim 86] Streptococcus salivarius is also known as Streptococcus salivarius K12 (American Type Culture Collection (ATCC), P.O. Box 1549, Manassas, VA 20108, USA, accession number BAA-1024) or Streptococcus salivarius ENT-K12 (Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkullturen GmbH, Inhofensträbe 7B, 38124 Braunschweig, Germany, accession number DSM Streptococcus salivarius for use according to any one of claims 1 to 28, 49 to 56 and 58 to 66, a composition for use according to any one of claims 29 to 46, 49 to 55 and 57 to 66, a use according to any one of claims 47 or 48 and 73 to 77, a method according to any one of claims 67 to 72 and 78 to 82, or a lozenge according to any one of claims 83 to 85.

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