Differential cytokine response from dendritic cells to commensal and pathogenic bacteria in different lymphoid compartments in humans

2006 ◽  
Vol 290 (4) ◽  
pp. G839-G845 ◽  
Author(s):  
Liam O’Mahony ◽  
Louise O’Callaghan ◽  
Jane McCarthy ◽  
David Shilling ◽  
Paul Scully ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Immune System ◽  
Peripheral Blood ◽  
Pathogenic Bacteria ◽  
Mononuclear Cells ◽  
Cytokine Production ◽  
Transforming Growth Factor ◽  
Mesenteric Lymph Nodes ◽  
Tnf Α

Resident host microflora condition and prime the immune system. However, systemic and mucosal immune responses to bacteria may be divergent. Our aim was to compare, in vitro, cytokine production by human mononuclear and dendritic cells (DCs) from mesenteric lymph nodes (MLNs) and peripheral blood mononuclear cells (PBMCs) to defined microbial stimuli. Mononuclear cells and DCs isolated from the MLN ( n = 10) and peripheral blood ( n = 12) of patients with active colitis were incubated in vitro with the probiotic bacteria Lactobacillus salivarius UCC118 or Bifidobacterium infantis 35624 or the pathogenic organism Salmonella typhimurium UK1. Interleukin (IL)-12, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, and IL-10 cytokine levels were quantified by ELISA. PBMCs and PBMC-derived DCs secreted TNF-α in response to the Lactobacillus, Bifidobacteria, and Salmonella strains, whereas MLN cells and MLN-derived DCs secreted TNF-α only in response to Salmonella challenge. Cells from the systemic compartment secreted IL-12 after coincubation with Salmonella or Lactobacilli, whereas MLN-derived cells produced IL-12 only in response to Salmonella. PBMCs secreted IL-10 in response to the Bifidobacterium strain but not in response to the Lactobacillus or Salmonella strain. However, MLN cells secreted IL-10 in response to Bifidobacteria and Lactobacilli but not in response to Salmonella. In conclusion, commensal bacteria induced regulatory cytokine production by MLN cells, whereas pathogenic bacteria induce T cell helper 1-polarizing cytokines. Commensal-pathogen divergence in cytokine responses is more marked in cells isolated from the mucosal immune system compared with PBMCs.

Polymyxin-B Stimulates Tumor Necrosis Factor-α Production by Human Peripheral Blood Mononuclear Cells

1998 ◽  
Vol 21 (5) ◽  
pp. 269-273 ◽  
Author(s):  
B.L. Jaber ◽  
S. Sundaram ◽  
M. Cendoroglo Neto ◽  
A.J. King ◽  
B.J.G. Pereira
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Tumor Necrosis ◽  
Mononuclear Cells ◽  
Cytokine Production ◽  
Polymyxin B ◽  
Peripheral Blood Mononuclear ◽  
Gram Negative ◽  
Blood Mononuclear Cells

Gram-negative bacterial lipopolysaccharide (LPS) is a well known stimulus for cytokine production, particularly interleukin-1 (IL-1) and tumor necrosis tactor alpha (TNFα). Polymyxin B (PMX-B) is a cationic polypeptide that binds to LPS, neutralizing its biological effects. PMX-B also disrupts gram-negative bacterial cell membrane phospholipids but is highly toxic to mammalian cells, therefore is of limited use. PMX-B is used as additive to media, as a way to handle LPS contamination. To derive benefit from the ability of PMX-B to neutralize lipid A in vivo while avoiding its systemic toxicity, PMX-B was covalently bound to polystyrene-derivative fibers, creating a hemoperfusion column (PMX-F) for the selective removal of circulating ET In vitro PMX-F hemoperfusion studies have demonstrated effective ET removal, using either the Limulus amebocyte lysate assay or TNFα production by peripheral blood mononuclear cells (PBMC) as an index of ET removal. However, the question whether PMX-B itself could stimulate human PBMC to produce cytokines has not been adequately addressed. We examined the effect of increasing concentrations of PMX-B on cytokine production by PBMC in vitro. PBMC harvested from healthy volunteers were incubated for 24 hours at 37°C with control (tissue culture media RPMI), or 5 µg/ml, 10 µg/ml, 20 µg/ml or 100 µg/ml PMX-B. At the end of 24 hours, PBMC were subjected to three freeze-thaw cycles, and total TNFα production (pg/2.5x106 PBMC) was measured by radioimmunoassay. Total TNFα production by PBMC was 163 ± 3 pg, 171 ± 9 pg, 164 ± 4 pg, 323 ± 63 pg and 331 ± 58 pg, in the control, PMX-B 5 µg/ml, 10 µg/ml, 20 µg/ml and 100 µg/ml conditions, respectively. Compared to controls (RPMI), the percentage increase in TNFα production by PBMC was 5 ± 6% (P=0.23), 1 ± 3% (P=0.45), 99 ± 40% (P=0.03) and 103 ± 36% (P=0.02) in the presence of 5 µg/ml, 10 µg/ml, 20 µg/ml and 100 µg/ml of PMX-B, respectively. Furthermore, total TNFα production correlated significantly with increasing concentrations of PMX-B (R=0.53, P=0.007). We conclude that the use of PMX-B in in vitro studies as an LPS-neutralizing agent, or in the experimental treatment of endotoxic or septic shock can lead to erroneous interpretations of cytokine production by PBMC, and should be used cautiously in in vitro systems at high concentrations.

Ginseng Total Saponin Enhances the Phagocytic Capacity of Canine Peripheral Blood Phagocytes in Vitro

2008 ◽  
Vol 36 (02) ◽  
pp. 329-341 ◽  
Author(s):  
K. A. Kang ◽  
J. H. Kang ◽  
M. P. Yang
Keyword(s):  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Culture Supernatant ◽  
Polymorphonuclear Cells ◽  
Phagocytic Capacity ◽  
Total Saponin ◽  
Tnf Α ◽  
Vitro Effect ◽  
Ginseng Total Saponin

The clinical and pharmacological activities of ginseng are known to modulate immune function, metabolic processes and neuro-endocrine system activities. Ginseng saponins are the principle active ingredients in the formation of immune stimulating complexes. The objective of this study was to evaluate the in vitro effect of ginseng total saponin (GTS) on the phagocytic capacity of canine peripheral blood phagocytes. GTS itself did not cause any direct effect on the phagocytic capacity of peripheral blood mononuclear cells (PBMC) and polymorphonuclear cells (PMN) but not peripheral blood monocytes. However, the phagocytic capacity of PMN and monocytes, but not PBMC, was enhanced by the culture supernatant from PBMC treated with GTS. The phagocytic capacity of PMN and monocytes was also increased by treatment with recombinant canine (rc) tumor necrosis factor (TNF)-α. The ability of the culture supernatant from GTS-treated PBMC to stimulate the phagocytic capacity of phagocytes was inhibited by addition of anti-rc TNF-α polyclonal antibody (pAb) prior to the culture. The amount of TNF-α in the culture supernatant from PBMC was shown to increase upon treatment of GTS as compared with that of vehicle-treated PBMC culture supernatant. These results suggest that GTS has an immunoenhancing effect on the phagocytic capacity of canine peripheral blood phagocytes, which is mainly mediated by TNF-α released from GTS-stimulated PBMC.

scholarly journals Interaction of Rotavirus with Human Myeloid Dendritic Cells

2005 ◽  
Vol 79 (23) ◽  
pp. 14526-14535 ◽  
Author(s):  
Carlos F. Narváez ◽  
Juana Angel ◽  
Manuel A. Franco
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Transforming Growth Factor Beta ◽  
Mononuclear Cells ◽  
Transforming Growth Factor ◽  
Myeloid Dendritic Cells ◽  
Peripheral Blood Mononuclear ◽  
Necrosis Factor Alpha ◽  
Stimulation Of

ABSTRACT We have previously shown that very few rotavirus (RV)-specific T cells that secrete gamma interferon circulate in recently infected and seropositive adults and children. Here, we have studied the interaction of RV with myeloid immature (IDC) and mature dendritic cells (MDC) in vitro. RV did not induce cell death of IDC or MDC and induced maturation of between 12 and 48% of IDC. Nonetheless, RV did not inhibit the maturation of IDC or change the expression of maturation markers on MDC. After treatment with RV, few IDC expressed the nonstructural viral protein NSP4. In contrast, a discrete productive viral infection was shown in MDC of a subset of volunteers, and between 3 and 46% of these cells expressed NSP4. RV-treated IDC secreted interleukin 6 (IL-6) (but not IL-1β, IL-8, IL-10, IL-12, tumor necrosis factor alpha, or transforming growth factor beta), and MDC released IL-6 and small amounts of IL-10 and IL-12p70. The patterns of cytokines secreted by T cells stimulated by staphylococcal enterotoxin B presented by MDC infected with RV or uninfected were comparable. The frequencies and patterns of cytokines secreted by memory RV-specific T cells evidenced after stimulation of peripheral blood mononuclear cells (PBMC) with RV were similar to those evidenced after stimulation of PBMC with RV-infected MDC. Finally, IDC treated with RV strongly stimulated naive allogeneic CD4+ T cells to secrete Th1 cytokines. Thus, although RV does not seem to be a strong maturing stimulus for DC, it promotes their capacity to prime Th1 cells.

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