Craig J. Cender, Dirk Haller, Charlotte Walters, Ryan B. Sartor, Chapel Hill, NC
Background & Aim: Clinical and experimental observations have demonstrated efficacy of the probiotic compound VSL#3 in human as well as animal models of inflammatory bowel diseases. Although the precise mechanism by which these bacteria exert their beneficial properties remains unknown, recent evidence supports modulation of the host immune response.
We investigated this hypothesis by evaluating the cytokine profile of an unfractionated spleen cell population upon stimulation with cecal bacterial lysate (CBL) preceded by incubation with VSL#3 lysate (VSL3-L).
Methods: A lysate of both the probiotic preparation VSL#3, containing 4 strains of lactobacilli, 3 strains of bifidobacteria, and Streptococcus thermophilus, and the cecal contents from a syngeneic Fisher rat were prepared per standard protocol. Isolated spleen cells (2 x 106) from wild type Fisher rats were incubated for 18h with VSL3-L (25?g/ml) in the presence or absence of anti-rat IL-10 antibody (10?g/ml), followed by a 6h stimulation period with CBL (100?g/ml). TNF and IL-10 concentrations of the supernatant were determined by ELISA. Cell viability was controlled by trypan blue exclusion.
Results: Rat splenocytes demonstrated a dose-dependent secretion of TNF and IL-10 when exposed to CBL and VSL3-L, respectively. Time-dependent analysis of cytokine response showed early upregulation (3-6h) of TNF that peaked by 12h while IL-10 increase was steadily progressive over 60h, irrespective of the stimuli utilized. Incubation of VSL3-L prior to stimulation with CBL reduced mean (±SEM, n=6) TNF levels from 1233.65±208.64 to 268.03±41.87 (p=.0005) and increased mean (±SEM, n=6) IL-10 levels from 138.82±26.91 to 252.11±55.71 (p=.05). The addition of neutralizing anti-rat IL-10 antibody in the incubation phase did not significantly alter the TNF reduction observed in the presence of VSL3-L alone, a 68% vs. 77% reduction respectively.
Conclusions: TNF reduction was independent of the IL-10 upregulation observed in stimulated rat splenocytes when pre-incubated with the non-viable lysate of VSL#3. Alteration of the proinflammatory and protective cytokines by VSL#3 supports an immunomodulatory mechanism for this probiotic preparation.
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