Use of non-steroidal anti-inflammatory drugs in infectious disease by M9fCD9

VIEWS: 9 PAGES: 2

									                                Colorado State University
                                 Animal Care Program
 Guidelines on Use of Non-Steroidal Anti-Inflammatory Drugs in Immunological Research

The Institutional Animal Care and Use Committee requires, and consideration of animal welfare
dictates, that investigators minimize pain and suffering in all experimental procedures. Measures
to minimize suffering include use of local or systemic analgesics, provision of symptomatic care,
and reducing the amount of time an animal is ill with an induced disease.

Non-steroidal anti-inflammatory medications are a class of drugs commonly used to treat the
pain and discomfort associated with inflammation. “Inflammation” refers to the early stages of
an immune response before the adaptive phase (B and T cell responses) come into play. As
such, any manipulation of the inflammatory response will affect the downstream immune
response. In particular, non-steroidal anti-inflammatory drugs inhibit, in various ways, the
actions of cyclooxygenases, which generate prostaglandins. Prostaglandins mediate numerous
aspects of the inflammatory response, as well as the subsequent immune response. Some recent
examples from top tier immunology journals are given below (search phrases were “T cell” and
“prostaglandin”).

The IACUC recognizes that non-steroidal anti-inflammatory medications may affect experiments
in which immune responses are directly measured, and therefore may not be appropriate
analgesia for these experiments. The IACUC therefore requires that investigators minimize the
amount of time that animals are ill with infectious, autoimmune and neoplastic diseases, and
encourages investigators to explore the use of alternatives to non-steroidals (opiates) if their
experimental system involves significant or prolonged disease burden. The need for analgesia as
a measure to alleviate pain and distress during experiments that induce potentially painful
procedures will continue to be evaluated on a case-by-case basis by the IACUC.


European Journal of Immunology. 38(2):459-69, 2008
We evaluated the role of regulatory T cells (CD4(+) CD25(+) Foxp3(+) cells, Tregs) in human Mycobacterium
tuberculosis infection. Tregs were expanded in response to M. tuberculosis in healthy tuberculin reactors, but not in
tuberculin-negative individuals. The M. tuberculosis mannose-capped lipoarabinomannan (ManLAM) resulted in
regulatory T cell expansion, whereas the M. tuberculosis 19-kDa protein and heat shock protein 65 had no effect.
Anti-IL-10 and anti-TGF-beta alone or in combination, did not reduce expansion of Tregs. In contrast, the
cyclooxygenase enzyme-2 inhibitor NS398 significantly inhibited expansion of Tregs, indicating that prostaglandin
E2 (PGE2) contributes to Treg expansion. Monocytes produced PGE2 upon culturing with heat-killed M.
tuberculosis or ManLAM, and T cells from healthy tuberculin reactors enhanced PGE2 production by monocytes.
Expanded Tregs produced significant amounts of TGF-beta and IL-10 and depletion of Tregs from PBMC of these
individuals increased the frequency of M. tuberculosis-responsive CD4(+) IFN-gamma cells. Culturing M.
tuberculosis-expanded Tregs with autologous CD8(+) cells decreased the frequency of IFN-gamma(+)cells. Freshly
isolated PBMC from tuberculosis patients had increased percentages of Tregs, compared to healthy tuberculin
reactors. These findings demonstrate that Tregs expand in response to M. tuberculosis through mechanisms that
depend on ManLAM and PGE2.

Journal of Immunology. 180(3):1390-7, 2008
PGE(2) inhibits mature T cell proliferation and protects T cells from activation-induced cell death (AICD). We have
previously demonstrated that human follicular dendritic cells (FDC) strongly express PGI synthase. In this study, the
hypothesis that FDC have regulatory roles on germinal center T cells by controlling production of PGE(2) and
PGI(2) was tested. Confocal microscopic analyses of human tonsil tissues revealed that FDC indeed expressed PGE
synthase in addition to PGIS. To confirm these results, we studied the regulation mechanism of PG production in
FDC, using an established human FDC-like cell line, HK. Specifically in response to TNF-alpha, TGF-beta, and
LPS, protein expression of cyclooxygenase (COX)-2 and downstream PGE synthase was up-regulated with
coordinate kinetics, whereas COX-1 and PGIS were constitutively expressed. The increase of these enzymes was
reflected in actual production of PGE(2) and PGI(2). Interestingly, IL-4 almost completely abrogated the stimulatory
activity of TNF-alpha, TGF-beta, and LPS in PG production. Furthermore, the up-regulation of PGE(2) and PGI(2)
production was markedly down-regulated by indomethacin and a selective COX-2 inhibitor. PGI(2) analog and
PGE(2) inhibited proliferation and AICD of T cells in dose- and time-dependent manners. Finally, coculture
experiments revealed that HK cells indeed inhibit proliferation and AICD of T cells. Put together, these results show
an unrecognized pathway of FDC and T cell interactions and differential mechanisms for PGE(2) and PGI(2)
production, suggesting an important implication for development and use of anti-inflammatory drugs.

Immunology. 123(2):290-303, 2008
Inflammatory responses mediated by antigen-presenting dendritic cells (DCs), can be modulated by the presence of
prostaglandins (PG), including prostaglandin E2 (PGE2). PGE2 modifies the production of an immune response
by altering DC function through PGE2 receptors. PGE2 is produced by epithelial cells lining the murine female
reproductive tract during Chlamydia muridarum infection and likely manipulates the antichlamydial immune
response during antigen uptake in the genital mucosa. Our data demonstrate that the PGE2 present locally in the
genital tract upon chlamydial genital infection enhanced the recruitment of CD11b+ conventional DCs, but not
CD45R+ plasmacytoid DCs, to infected genital tract tissue and draining lymph nodes in vivo. Furthermore,
exposure to PGE2 in vitro during infection of murine bone-marrow-derived conventional DCs (cDCs) boosted
interleukin-10 mRNA and protein while not influencing interleukin-12p40 production. Infection of cDCs markedly
increased mRNA production of the costimulatory molecules CD86, CD40 and a member of the C-type lectin family,
DEC-205, but addition of PGE2 increased other costimulatory molecules and C-type lectins. Also, exposure of
PGE2 to infected cDCs increased FcgammaRIII and FcgammaRIIb, suggesting that PGE2 enhances the uptake and
presentation of C. muridarum and augments production of the antichlamydial adaptive immune response. Taken
together, the data suggest that exposure of infected cDCs to PGE2 drives production of a diverse adaptive immune
response with implications for regulating tissue inflammation.

Journal of Experimental Medicine. 204(12):2865-74, 2007
Prostaglandin E2 (PGE2) exerts its actions via four subtypes of the PGE receptor, EP1-4. We show that mice
deficient in EP1 exhibited significantly attenuated Th1 response in contact hypersensitivity induced by
dinitrofluorobenzene (DNFB). This phenotype was recapitulated in wild-type mice by administration of an EP1-
selective antagonist during the sensitization phase, and by adoptive transfer of T cells from sensitized EP1-/- mice.
Conversely, an EP1-selective agonist facilitated Th1 differentiation of naive T cells in vitro. Finally, CD11c+ cells
containing the inducible form of PGE synthase increased in number in the draining lymph nodes after DNFB
application. These results suggest that PGE2 produced by dendritic cells in the lymph nodes acts on EP1 in naive T
cells to promote Th1 differentiation.




Last Modified: May 23, 2008

								
To top