Effects of burn with and without Escherichia coli infection in rats on intestinal vs. splenic T-cell responses.
OBJECTIVE
To evaluate the effect of burn injury with and without an Escherichia coliseptic complication on T-cell proliferation, interleukin-2 production, and Ca(2+) signaling responses in intestinal Peyer's patch and splenic T cells.


METHODS
Prospective, randomized, sham-controlled animal study.


METHODS
University medical center research laboratory.


METHODS
Adult male Sprague-Dawley rats.


METHODS
Rats were subjected to a 30% total body surface area, full skin thickness burn. Infection in rats was induced via intraperitoneal inoculation of E. coli, 10(9) colony forming units/kg, with or without a prior burn.


RESULTS
Rat Peyer's patch and splenic T lymphocytes were isolated by using a nylon wool cell purification protocol. T-cell proliferation, interleukin-2 production, and Ca(2+) signaling responses were measured after stimulation of cells with the mitogen, concanavalin A. T-cell proliferation was determined by measuring incorporation of (3)H-thymidine into T-cell cultures. Interleukin-2 production by T-cell cultures was measured by using enzyme-linked immunosorbent assay. Intracellular T-cell Ca2(+ )concentration, [Ca(2+)](i), was measured by the use of Ca(2+)-specific fluorescent label, fura-2, and its fluorometric quantification. [Ca(2+)](i) was also evaluated by the use of digital video imaging of fura-2 loaded individual T cells. T-cell proliferation and interleukin-2 production were suppressed substantially in both Peyer's patch and splenic T cells 3 days after either the initial burn alone or burn followed by the E. coli inoculation at 24 hrs after the initial burn. There seemed to be no demonstrable additive effects of E. coli infection on the effects produced by burn injury alone. The T-cell proliferation and interleukin-2 production suppressions with burn or burn-plus-infection insults were correlated with attenuated Ca(2+) signaling. E. coli infection alone suppressed T-cell proliferation in Peyer's patch but not in splenic T cells at 2 days postbacterial inoculation; E. coli infection had no effect on Peyer's patch or splenic T cells at 1 day postinjury. On the other hand, burn injury alone caused a substantial T-cell proliferative suppression at 2 days postburn in both Peyer's patch and splenic cells and a significant suppression in T-cell proliferation on day 1 postburn in Peyer's patch but not in the spleen.


CONCLUSIONS
An initial burn injury suppressed T-cell proliferation at a level that it would not be further affected by a subsequent infection even if the infection by itself has the potential of suppressing T-cell proliferation. An earlier onset of T-cell suppression in Peyer's patch cells than in the spleen with burn could be attributable to an initial hypoperfusion-related intestinal mucosal tissue injury. Overall, our study supports the concept that burn injury per se can significantly suppress T-cell mediated immunity and that the intestine is an early tissue site of such suppression.
Ravindranath
T
T
Department of Pediatrics, Ronald McDonald Children's' Hospital, Loyola University Medical Center, Maywood, IL 60153, USA.
Al-Ghoul
W
W
Namak
S
S
Fazal
N
N
Durazo-Arvizu
R
R
Choudhry
M
M
Sayeed
M M
MM
D000704:Analysis of Variance; D000818:Animals; D002056:Burns / Q000276:immunology* / Q000382:microbiology; D020013:Calcium Signaling; D003208:Concanavalin A; D004927:Escherichia coli Infections / Q000209:etiology / Q000276:immunology*; D007108:Immune Tolerance; D007376:Interleukin-2 / Q000378:metabolism; D008297:Male; D010581:Peyer's Patches / Q000276:immunology*; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D018805:Sepsis / Q000209:etiology / Q000276:immunology*; D013154:Spleen / Q000276:immunology*; D018709:Statistics, Nonparametric; D013601:T-Lymphocytes / Q000378:metabolism*
D007376:Interleukin-2; D003208:Concanavalin A