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. 2003 Aug;71(8):4271-7.
doi: 10.1128/IAI.71.8.4271-4277.2003.

Coordinate cytokine gene expression in vivo following induction of tuberculous pleurisy in guinea pigs

Shannon Sedberry Allen  1 , David N McMurray
Affiliations

Coordinate cytokine gene expression in vivo following induction of tuberculous pleurisy in guinea pigs

Shannon Sedberry Allen et al. Infect Immun. 2003 Aug.

Abstract

Tuberculous pleurisy is a severe inflammatory response induced by Mycobacterium tuberculosis organisms that have escaped from lung granulomata into the pleural space during pulmonary infection. We have used the guinea pig model of tuberculous pleurisy to examine several aspects of the immune response to this antigen-specific inflammatory event. Pleurisy was induced by injection of heat-killed M. tuberculosis H37Rv directly into the pleural space of guinea pigs previously vaccinated with M. bovis BCG. Four animals were euthanized each day over a period of 9 days. Fluid in the pleural cavity was analyzed for transforming growth factor beta 1 (TGF-beta 1) and total interferon (IFN) protein levels. In addition, RNA was obtained from pleural cells and examined for TGF-beta 1, tumor necrosis factor alpha (TNF-alpha), IFN-gamma, and interleukin-8 (IL-8) expression by real-time PCR. Finally, pleural cells were examined for the ability to proliferate in response to concanavalin A and purified protein derivative (PPD) in vitro. In the pleural fluid, TGF-beta 1 protein concentrations increased over the course of the inflammatory response while IFN protein levels were not significantly altered. Expression of TGF-beta 1 mRNA peaked on days 3 and 4, and IFN-gamma mRNA expression peaked on day 3 and then returned to background levels. TNF-alpha mRNA expression was highest on days 2 to 4, and IL-8 mRNA levels remained elevated between days 2 and 5, peaking on day 3 before returning to background levels. PPD-induced proliferative responses were evident by day 3 and remained present throughout the study. Analysis of cytokine expression during tuberculous pleurisy may lead to a better understanding of the self-healing nature of this manifestation of tuberculosis.

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Figures

FIG. 1.
FIG. 1.
Cell and fluid accumulation in the pleural space of guinea pigs with tuberculous pleurisy. Cells (A) and pleural fluid (B) in the pleural space of pleuritic guinea pigs were quantified between days 1 and 9 of the inflammatory response. Cells were stained with trypan blue, and four squares of a hemocytometer were counted and averaged to obtain cell numbers, which are expressed in millions. Total milliliters of pleural fluid were assessed after the fluid was centrifuged and removed from cells. Data and error bars represent average cell number or fluid accumulation for three or four animals ± the standard error of the mean. An asterisk indicates a significant difference in the cell number compared with that on any other day.
FIG. 2.
FIG. 2.
Cytokine protein concentrations in pleural fluid. TGF-β1 (A) and total bioactive IFN (B) protein levels in pleural fluid of guinea pigs during 9 days of tuberculous pleurisy were measured. TGF-β1 protein levels were analyzed by ELISA and are expressed in nanograms per milliliter. Total IFN protein levels were analyzed by bioassay as described in Materials and Methods and are expressed in units per milliliter. Data and error bars represent the mean ± the standard error of the mean for three or four animals per day. An asterisk indicates a significant increase in TGF-β1 levels above that on all days except day 5.
FIG. 3.
FIG. 3.
Expression of TGF-β1 and IFN-γ mRNAs during tuberculous pleurisy. Expression of TGF-β1 (A) and IFN-γ (B) mRNAs was quantified in pleural fluid cells obtained from guinea pigs with tuberculous pleurisy between days 1 and 9. Fold induction was determined from the threshold cycle values normalized for 18S expression and then normalized to the values derived from healthy, pleuritis-free animals. Data and error bars represent the mean of three or four animals ± the standard error of the mean. An asterisk indicates a significant increase in mRNA expression compared to that on all other days.
FIG. 4.
FIG. 4.
Expression of IL-8 and TNF-α mRNAs during tuberculous pleurisy. Expression of IL-8 (A) and TNF-α (B) mRNAs was quantified in pleural fluid cells obtained from guinea pigs with tuberculous pleurisy euthanized on days 1 to 9. Fold induction was calculated as described in the legend to Fig. 3. Data and error bars represent the mean of three or four animals ± the standard error of the mean. An asterisk indicates a significant increase in mRNA expression compared with that on days 6, 7, and 9.
FIG. 5.
FIG. 5.
Proliferation of pleural exudate cells in tuberculous pleurisy. Pleuritic guinea pigs were euthanized on days 1 to 9, and cells were removed from the pleural cavity. Cells were cultured for 96 h in the presence of ConA (10 μg/ml) or PPD (12.5 or 25 μg/ml). Proliferative responses are reported as stimulation indices (counts per minute of stimulated cells divided by counts per minute of unstimulated cells). Data and error bars represent the mean of three or four animals ± the standard error of the mean. An asterisk indicates a significant increase in proliferation compared to similarly stimulated cells on day 1.
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References

    1. Allen, J. C., and M. A. Apicella. 1968. Experimental pleural effusion as a manifestation of delayed hypersensitivity to tuberculin PPD. J. Immunol. 101:481-487. - PubMed
    1. Antony, V. B., J. W. Hott, S. L. Kunkel, S. W. Godbey, M. D. Burdick, and R. M. Strieter. 1995. Pleural mesothelial cell expression of C-C (monocyte chemotactic peptide) and C-X-C (interleukin 8) chemokines. Am. J. Respir. Cell Mol. Biol. 12:581-588. - PubMed
    1. Antony, V. B., S. A. Sahn, A. C. Antony, and J. E. Repine. 1985. Bacillus Calmette-Guérin-stimulated neutrophils release chemotaxins for monocytes in rabbit pleural spaces and in vitro. J. Clin. Investig. 76:1514-1521. - PMC - PubMed
    1. Assoian, R. K., B. E. Fleurdelys, H. C. Stevenson, P. J. Miller, D. K. Madtes, E. W. Raines, R. Ross, and M. B. Sporn. 1987. Expression and secretion of type β transforming growth factor by activated human macrophages. Proc. Natl. Acad. Sci. USA 84:6020-6024. - PMC - PubMed
    1. Baggiolini, M., A. Walz, and S. L. Kunkel. 1989. Neutrophil-activating peptide-1/interleukin 8, a novel cytokine that activates neutrophils. J. Clin. Investig. 84:1045-1049. - PMC - PubMed

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