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Half of all postmenopausal women report symptoms of vulvar, vaginal or urinary discomfort with significant impact on sexual function and quality of life; underlying mechanisms leading to symptoms are poorly understood. To examine the possibility that the vaginal microbiota and/or mucosal immune response contribute to the severity of bothersome vaginal symptoms we conducted a substudy of samples from a randomized trial of vaginal treatment for genitourinary syndrome of menopause (GSM) to compare these features between women whose symptoms improved vs. those who did not.