Immunohistochemical detection of cytokines in paraffin-embedded mouse tissues

Whiteland, Joanne L.; Shimeld, C.; Nicholls, Susan M.; Easty, D L; Williams, Neil; Hill, Terry

doi:10.1016/s0022-1759(97)00167-1

Cited by 22 publications

(20 citation statements)

References 13 publications

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“…Taken together, these prior studies suggest that the production of MCP proteins within the intestine, possibly by epithelial cells, contributes to a generally suppressive environment at this site. That this suppressive environment is mediated by IL-10 is supported by the findings of the current study and is consistent with the results of prior studies demonstrating the presence of high levels of IL-10 within the intestinal tract (43)(44)(45). In addition, it is consistent with the fact that IL-10 mice develop Th1-mediated enterocolitis (46,47).…”

Section: Discussionsupporting

confidence: 93%

Selective Suppression of IL-12 Production by Chemoattractants

Braun

Lahey

Kelsall

2000

The Journal of Immunology

151

100

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We investigated the ability of chemoattractants to affect IL-12 production by human monocytes and dendritic cells. We found that pretreatment of monocytes with macrophage chemoattractant proteins (MCP-1 to -4), or C5a, but not stromal-derived factor-1, macrophage inflammatory protein-1α, RANTES, or eotaxin, inhibited IL-12 p70 production in response to stimulation with Staphylococcus aureus, Cowan strain 1 (SAC), and IFN-γ. The production of TNF-α and IL-10, however, was minimally affected by any of the chemoattractants. The degree of inhibition of IL-12 p70 production by MCP-1 to -4 was donor dependent and was affected by the autocrine inhibitory effects of IL-10. In contrast, C5a profoundly suppressed IL-12 production in an IL-10-independent fashion. Neither TGF-β1 nor PGE2 was important for the suppression of IL-12 by any of the chemoattractants tested. The accumulation of mRNA for both IL-12 p35 and p40 genes was inhibited by chemokine pretreatment. Interestingly, MCP-1 to -4 and C5a did not suppress IL-12 production by monocyte-derived dendritic cells (DC) stimulated with CD40 ligand and IFN-γ or by SAC and IFN-γ, suggesting that these factors may act at the site of inflammation to suppress IL-12 and IFN-γ production rather than in the lymph node to affect T cell priming. Despite the inability of C5a to inhibit IL-12 production by DCs, the receptor for C5a (CD88) was expressed by these cells, and recombinant C5a induced a Ca2+ flux. Taken together, these results define a range of chemoattractant molecules with the ability to suppress IL-12 production by human monocytes and have broad implications for the regulation of immune responses in vivo.

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Section: Discussionsupporting

confidence: 93%

Selective Suppression of IL-12 Production by Chemoattractants

Braun

Lahey

Kelsall

2000

The Journal of Immunology

151

100

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“…During P. aeruginosa keratitis of the cornea, it was noted that the kinetics of TNF-␣ and IFN-␥ production paralleled the numbers of neutrophils found in the corneal tissue. It has been shown that neutrophils are a major source of TNF-␣ during P. aeruginosa keratitis (8) and have also been reported to produce IFN-␥ in the cornea during HSV keratitis (29,32). These findings suggest that the reduction in IFN-␥ and TNF-␣ may result from the reduced numbers of neutrophils present in the corneas of IL-10 Ϫ/Ϫ mice at 7 days postchallenge.…”

Section: Discussionmentioning

confidence: 88%

“…The sections were dewaxed and rehydrated through a graded series of ethanols. Sections were stained for IL-10 by a method modified from that of Whiteland et al (32). Controls for nonspecific binding were sections incubated with an irrelevant antibody of the same isotype and sections not incubated with the primary antibody.…”

Section: Reverse Transcription-pcr (Rt-pcr)mentioning

confidence: 99%

ExperimentalPseudomonas aeruginosaKeratitis in Interleukin-10 Gene Knockout Mice

et al. 2003

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Pseudomonas aeruginosa keratitis is one of the most destructive diseases of the cornea. The host response to this infection is critical to the outcome. The cytokine interleukin-10 (IL-10) is thought to play an important role in modulating excessive inflammation and antimicrobial defenses. We have found that in IL-10 ؊/؊ mice there is a significant decrease in bacterial load in corneas at 7 days postchallenge with P. aeruginosa. This decrease was accompanied by a reduction in neutrophil numbers in the cornea and changes in cytokine levels compared to those of wild-type mice. A characteristic increase in neovascularization in the cornea was found in the IL-10 ؊/؊ mice. This increased angiogenesis correlated with an increased expression of KC, whereas the kinetics of macrophage inflammatory peptide 2 expression correlated with neutrophil numbers. This finding suggests that KC may play a role in corneal angiogenesis. The source of IL-10 in mouse corneas was identified as a subpopulation of infiltrating cells and keratocytes. This study demonstrates that IL-10 plays an important role in regulating the balance of inflammatory mediators during P. aeruginosa infection of the cornea.

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“…2 and data not shown). The lower production of IL-12 by mesenteric LN DC might be due to the high levels of IL-10 in intestinal associated lymphoid organs, because IL-10 has been reported to have a negative priming effect on the IL-12-producing capacity of DC (34,35).…”

Section: Cd8mentioning

confidence: 99%

Differential Production of IL-12, IFN-α, and IFN-γ by Mouse Dendritic Cell Subsets

Hochrein

Shortman

Vremec

et al. 2001

The Journal of Immunology

443

410

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Dendritic cells (DC) not only stimulate T cells effectively but are also producers of cytokines that have important immune regulatory functions. In this study we have extended information on the functional differences between DC subpopulations to include differences in the production of the major immune-directing cytokines IL-12, IFN-α, and IFN-γ. Splenic CD4−8+ DC were identified as the major IL-12 producers in response to microbiological or T cell stimuli when compared with splenic CD4−8− or CD4+8− DC; however, all three subsets of DC showed similar IL-12 regulation and responded with increased IL-12 p70 production if IL-4 was present during stimulation. High level CD8 expression also correlated with extent of IL-12 production for DC isolated from thymus and lymph nodes. By using gene knockout mice we ruled out any role for CD8α itself, or of priming by T cells, on the superior IL-12-producing capacity of the CD8+ DC. Additionally, CD8+ DC were identified as the major producers of IFN-α compared with the two CD8− DC subsets, a finding that suggests similarity to the human plasmacytoid DC lineage. In contrast, the CD4−8− DC produced much more IFN-γ than the CD4−8+ or the CD4+8− DC under all conditions tested.

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Immunohistochemical detection of cytokines in paraffin-embedded mouse tissues

Cited by 22 publications

References 13 publications

Selective Suppression of IL-12 Production by Chemoattractants

Selective Suppression of IL-12 Production by Chemoattractants

ExperimentalPseudomonas aeruginosaKeratitis in Interleukin-10 Gene Knockout Mice

Differential Production of IL-12, IFN-α, and IFN-γ by Mouse Dendritic Cell Subsets

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