Kombucha is a healthy and becoming popular beverage worldwide. It is produced from tea leaves using symbiotic cultures of bacteria and yeast. Herein, we explored the use of pineapple peels and cores (by-products), as alternative ingredients for kombucha production. We examined crude pineapple kombucha (PK) for its microbial diversity, metabolites, and its impact on the cellular immune response in PBMCs older individual ex vivo. We observed a substantial shift in bacterial diversity, with higher percentages of Blautia and Lactobacillus compared with the original kombucha. This microbial variation may be attributed to the PK's bioactive compounds. The expression of CD4+CD28null NKG2D+ senescent CD4+ T cells and pathogenic CD4+ T cells decreased, particularly in the high natural killer group 2 member D (NKG2D) expression group. This indicates the presence of bioactive substances in PK that can reduce the number of senescent CD4+ T cells. PK may be a health-promoting alternative beverage, especially for older individuals. All rights reserved, Elsevier.