Effect of LGG Probiotic on Human Whole Blood Transcriptome

Open label study to evaluate the safety of Lactobacillus rhamnosus GG ATCC 53103 (LGG) in elderly subjects. Fifteen healthy elderly volunteers, ages 65-80 were enrolled in a study in which they received LGG capsules containing 1 x 1010 CFU, twice daily for 28 days and were followed through day 56. The study subjects completed a daily diary, a telephone call on study days 3, 7 and 14 and study visits at the Massachusetts General Hospital Clinical Research Center at baseline, day 28 and day 56. During each visit, the subject diary, interim history, potential adverse effects and concomitant medications were reviewed and vital signs and a physical examination were performed. Routine blood tests were obtained to monitor for safety during visits and nasopharyngeal and stool samples were collected for microbiome analysis. Volunteers interested in participating in the effect of LGG probiotic on the Human whole Blood Transcriptome substudy also had blood drawn for DNA and RNA extraction, after signing the substudy consent form. The main objective of the study was to assess the safety and tolerability of 2 x 1010 CFU LGG administered orally to elderly subjects for 28 days. Secondary objectives were to evaluate the richness and microbial diversity in nasopharyngeal and stool specimens using pyrosequencing, and to compare cytokine production in response to bacterial stimulation by following the kinetics of mRNA expression of pro and anti-inflammatory genes and different signaling pathways, in relation to changes in stool Bifidobacterium and Lactobacillus spp. The study was reviewed and approved by the Partners Human Research Committee (IRB # 2010P001695) and was registered at ClinicalTrials.gov (NCT01274598). The main results regarding the clinical signs and safety of intervention is published in PLoS One. 2014 Dec 1;9(12):e113456. doi: 10.1371/journal.pone.0113456. eCollection 2014. PMID: 25438151. RNA sequencing for transcriptome analysis was only done in 11 of the 15 subjects as RNA integrity and quantity had to be optimal for the three times where blood was collected in order for a subject to be included in the study. The information regarding the microbiome data associated with this study is presented in a separate paper, which can be found at dbGaP study accession, phs000896, and PMID: 25873374.]]>
Partners HealthCare System Research Consent FormPartners HealthCare System Research Consent Form - SubStudyInclusion Criteria Subjects must meet all of the following inclusion criteria to be eligible to participate in the study: Age 65-80 years Willing to complete the informed consent process Able and willing to participate for the planned duration of the study, including availability for follow-up telephone contact Is community-dwelling for the past two years Has received routine physical in the past two years Has no new chronic conditions in the past two years Identifies a primary care clinician Has received recommended preventive services (Task Force for Clinical Preventive Services) for vaccination and cancer prevention/detection, e.g.; Pneumococcal vaccination Mammography Screening colonoscopy for colon cancer Willing to comply with protocol and report on compliance and side effects during the study period Informed consent obtained and signed prior to screening. Exclusion Criteria Subjects meeting any of the exclusion criteria at baseline will be excluded from the study: Vaccination with any vaccine within the one month period prior to study enrollment or intent to receive any other vaccine during the study period, other than TIV in fall 2012 History of hypersensitivity to any influenza vaccine components including eggs, egg proteins, gentamicin, gelatin or arginine History of avoidance of egg and / or egg based products for any reason, unless they have previously received TIV or LAIV without hypersensitivity History of Guillain-Barre syndrome Acute febrile illness within the week prior to immunization - immunization deferred until illness resolved Consumption of supplements or food products containing LGG or probiotics for 28 days prior to the start of the study or consumption of yogurt that has the "live and active cultures" seal Known or suspected allergies to probiotics, Lactobacillus, microcrystalline cellulose, gelatin, or antibiotics that may be used to treat LGG bacteremia or infection (i.e. subject able to tolerate at least 2 of the following regimens - Ampicillin or other beta lactam antibiotic, and Clindamycin, and Moxifloxacin) Received oral or parenteral antibiotics within 4 weeks of enrollment or prescribed antibiotics on the day of enrollment Drug or alcohol abuse, defined as the continued use of alcohol despite the development of social, legal, or health problems, within the previous 12 months Hospitalization, major surgery or endoscopy within the last 3 months Scheduled hospital admission or surgery within 3 months of enrollment Resident of a nursing home or rehabilitation center Close and/or household contact with severely immunocompromised persons (e.g. transplant recipients, cancer patients) Receipt of any of the following medications within the specified time frame Antiviral agents for influenza A and B in the prior 2 weeks (anti-influenza medications are unlikely to be needed after LAIV immunization since LAIV will be administered at the end of the influenza season, but should be prescribed if clinically indicated) Immune Globulin the prior 6 months Immune modulating drugs such as Belimumab, azathioprine, mercaptopurine, methotrexate, hydroxychloroquine or leflunomide at any time or any oral or parenteral corticosteroid in the prior 12 months Fingolimod at any time Salicylates at doses higher than 163 mg/day for primary prevention of myocardial infarction. (Reye's Syndrome in adults is extremely rare - to our knowledge the oldest reported patient was aged 61 and as of 1989 when Reye Syndrome was more common, it had only been reported in 25 adults. In addition, increasing numbers of adults are being placed on low dose aspirin for primary and secondary prevention of cardiovascular disease, some of whom may also have influenza, but there has been no increase in reports of Reye's Syndrome in adults.) Presence of any of the following: Grade 2 or higher abnormal vital signs or abnormalities on physical exam during screening (Appendix A) or presence of any wheezing on physical examination at baseline Indwelling catheter or implanted hardware/prosthetic device or feeding tube Current or within the last 2 years, any episode of bowel leak, acute abdomen, diverticulitis, colitis, bloody bowel movements or peptic ulcer disease, including any surgical procedure or current prescription medications for any of these conditions Current or within the last four weeks, active bowel disease such as an episode of infectious or non-infectious diarrhea, constipation, or vomiting lasting more than 12 hours or current prescription medication for any of these conditions Any history of gastric or intestinal dysmotility, slowed transit time, variable small intestinal permeability, pancreatitis, history of gastrointestinal tract cancer or metastasis, or inflammatory bowel disease or current prescription medications for any of these conditions Any history of Hepatitis B or Hepatitis C infections, cirrhosis, or chronic liver disease Underlying structural heart disease such as abnormal native heart valve or congenital abnormality, previous history of endocarditis or valve replacement, Stage IV congestive heart failure or chronic cardiovascular conditions (except hypertension) History of peripheral vascular disease or stroke Immunosuppression including HIV positive, solid organ or stem cell transplant recipient, neutrophil count <500/mm3, or an anticipated drop in the neutrophil count to <500/mm3 during the study period, or active or planned chemotherapy or radiotherapy History of collagen vascular or autoimmune disease History of renal disease History of chronic obstructive pulmonary disease or asthma History of neurologic or neuromuscular diseases History of hematologic diseases Diabetes or thyroid disease or metabolic disorder Active tuberculosis (TB), defined as undergoing work up for suspected active TB infection or currently on treatment for active TB or scheduled for tuberculin test in the next 4 weeks. Fever or acute illness or nasal congestion during the baseline visit (LAIV administration) Positive drug or alcohol testing at screening or positive breathalyzer at baseline or an unwillingness to undergo drug and alcohol testing Abnormal laboratory tests defined as any of the following: White blood cell (WBC) < 3.3 or > 12.0 K/µL Neutrophil < 500/mm3 Platelets < 125 K/µL Hemoglobin Males: < 12.0 g/dL; Females: < 11.0 g/dL Creatinine > 1.8 mg/dL Blood urea nitrogen (BUN) > 27 mg/dL Aspartate aminotransferase (AST) > 1.25 ULN Alanine aminotransferase (ALT) > 1.25 ULN Alkaline phosphatase > 2.0 ULN Bilirubin (total) > 1.5 ULN Glucose (non-fasting ) > 126 mg/dL Positive HIV, Hepatitis B surface antigen or Hepatitis C antibody Any other condition that in the opinion of the investigator would jeopardize the safety or rights of the subject participating in the study or would make it unlikely the subject could complete the study. ]]>
Telephone screening for study subjects started on December 1, 2010 and the last volunteer exited the study on August 5, 2011.]]>