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The Journal of Immunology, 1998, 161: 3237-3241.
Copyright © 1998 by The American Association of Immunologists

Urocanic Acid Enhances IL-10 Production in Activated CD4+ T Cells1

Vladimír Holán2,*, Lucia Kuffová*,{dagger}, Alena Zajícová*, Magdaléna Krulová*, Martin Filipec{dagger}, Petr Holler{ddagger} and Alexander Jancárek§

* Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, {dagger} Second Department of Ophthalmology, Charles University, {ddagger} Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, and § Faculty of Nuclear Sciences and Physical Engineering, Prague, Czech Republic


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
The immunosuppressive effects of UV radiation have been well documented. This suppression has been attributed to the action of the cis form of urocanic acid (UCA), a photoproduct of trans-UCA, a natural constituent of the skin. Here, we show that mouse spleen cells preincubated with cis-UCA have a diminished proliferative response to allogeneic cells in MLC and to stimulation with anti-CD3 mAb. Cells preincubated with cis-UCA also had a decreased ability to serve as APC and to stimulate the proliferation of allogeneic lymphocytes in MLC. Simultaneously, the production of IL-2 and IFN-{gamma} by cells preincubated with cis-UCA was decreased. However, IL-10 gene expression and IL-10 protein secretion by spleen cells stimulated in the presence of cis-UCA were significantly enhanced. The principal cell population displaying the cis-UCA-induced elevated production of IL-10 was CD4+ T cells, which were shown to be a direct target of cis-UCA action. This was also supported by the observation that production of IL-10 by stimulated splenic non-T cells or by macrophages was not altered by cis-UCA. The enhanced production of IL-10 by activated CD4+ T cells may represent a novel pathway of UVB radiation-induced, cis-UCA-mediated immunosuppression. We suggest that the elevated production of IL-10 by activated CD4+ T cells may account for the suppressor T cell phenomena described in UV-irradiated recipients.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
The immunosuppressive effects of UV radiation have been well documented in many systems (1, 2). After UV irradiation, contact hypersensitivity (CH)3 is impaired (3), delayed-type hypersensitivity (DTH) and allotransplantation reactions are more weakly expressed (4, 5), and the activity of NK cells is decreased (6).

The mechanism of the suppressive action of UV radiation on the immune system has, for a long time, remained unexplained. De Fabo and Noonan (7) proposed that the immunosuppression induced by UV radiation is mediated by cis-urocanic acid (UCA). UCA is naturally present in the skin in trans configuration and undergoes UVB irradiation-induced isomerization from the trans to the cis isomer. It has been shown that systemic administration of cis-UCA inhibits murine DTH reactions (8, 9), CH (10), and allograft rejection (11, 12, 13).

However, it has remained unclear whether cis-UCA is a final effector molecule with an inhibitory effect on the immune system or whether cis-UCA induces another molecule responsible for the immunosuppression. Some data have suggested that cis-UCA could alter APC function (14, 15) and thus inhibit the immune system functions. Kurimoto and Streilein (10) proposed that cis-UCA can cause the local release of TNF-{alpha}, which thwarts sensitization by blocking epidermal Langerhans cells (LC) function. On the other hand, Rattis et al. (16), who studied the proliferative response to alloantigens, did not find any direct effect of cis-UCA on human LC Ag-presenting function. Similarly, Beissert et al. (15) did not observe changes in the expression of MHC class II, CD80, or CD86 molecules on an LC-like dendritic cell line treated with cis-UCA in vitro. Therefore, cis-UCA does not appear to be a direct effector molecule causing immune suppression.

We present another explanation herein for cis-UCA-induced immunosuppression. We found that cis-UCA significantly enhances the production of IL-10, a cytokine with immunosuppressive actions, by direct action on activated, IL-10-producing CD4+ T cells. Consequently, IL-10 can inhibit the Ag-presenting function of LC (17) or induce Ag-specific anergy in T cell clones (18), thus suppressing the immune response.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Animals

BALB/c and C57BL/10Sn (B10) mice of both sexes, obtained from the breeding colony of the Institute of Molecular Genetics (Prague, Czech Republic) were used at the age of 8 to 12 wk.

Urocanic acid

A commercially available trans-UCA (Sigma Chemical, St. Louis, MO) was dissolved at a concentration of 2 mg/ml in NaCl solution. Aliquots of this solution were irradiated by an XeCl excimer laser beam pulsed with a wavelength of 308 nm, frequency of 10 Hz, spot diameter of 3.5 cm, and fluence of 1.8 mJ/cm2 for 14 min. While unirradiated samples contained 100% trans isomer, irradiated samples contained 50 to 56% cis-UCA and 44 to 50% trans-UCA as determined by HPLC. All samples were stored at 4°C in dark bottles to ensure their stability. Samples of irradiated UCA are referred to as cis-UCA.

Cell separation

Enriched T cell populations were prepared by two cycles of spleen cell separation on a nylon wool column according to the technique of Julius et al. (19). Any remaining B cells were removed by a "panning" technique using dishes coated with swine anti-mouse Ig (20). Non-T cell populations were prepared by passing spleen cells through a nylon wool column, then eliminating any remaining T cells from nylon wool-adherent cells with anti-Thy-1.2 mAb (21) and C. CD4-, CD8-, and Thy-1.2- cell negative cell populations were prepared by two cycles of treating spleen cells with cytotoxic anti-CD4 (clone GK1.5 (22)), anti-CD8 (clone TIB 150 (23)), or anti-Thy-1.2 (clone F7D5 (21)) mAb, respectively, and C.

Preincubation of cells with UCA

Spleen cells at a concentration of 3 x 106/ml were preincubated in a volume of 3 ml of RPMI 1640 medium (Sigma Chemical) containing 10% heat-inactivated FCS (Sigma), antibiotics (100 U/ml of penicillin, 100 µg/ml of streptomycin), 10 mM HEPES buffer, and 5 x 10-5 M 2-ME (hereafter referred to as complete RPMI 1640 medium) in six-well tissue culture plates (Sterilin, Feltham, U.K.) with 200 µg/ml of trans- or cis-UCA. After a 2- or 24-h preincubation, cells were washed twice and resuspended at an appropriate concentration.

Proliferative assay

Freshly isolated or preincubated spleen cells at a concentration of 0.5 x 106/ml were incubated in 200 µl of complete RPMI 1640 medium in flat-bottom, 96-well plates (Nunclon, Roskilde, Denmark) with 2 µg/ml of anti-CD3 mAb (24) in the presence or absence of 200 µg/ml of cis- or trans-UCA. Cell proliferation was determined by adding 0.5 µCi of [3H]thymidine per well for the last 6 h of the 72-h incubation period.

MLC reaction

Reactive spleen cells (1.5 x 105 per well) were incubated in 96-well tissue culture plates (Nunclon) with stimulatory spleen cells (2 x 105 per well) in a volume of 200 µl of complete RPMI 1640 medium, as described elsewhere (25). Stimulatory cells were preincubated for 24 h alone or with 200 µg/ml of cis- or trans-UCA, then were irradiated with 3000 rad. Neutralizing rat anti-mouse IL-10 mAb (IgG1) (Genzyme, Cambridge, MA) or normal rat IgG1 (Sevac, Prague, Czech Republic) at a concentration of 10 µg/ml were added to cultures containing cells preincubated with cis-UCA. Cell proliferation was determined by adding 0.5 µCi of [3H]thymidine for the last 6 h of the 96-h incubation period.

Cytokine production and determination

Unseparated or purified spleen cells (1 x 106) were incubated with 2 µg/ml of anti-CD3 mAb in a volume of 1 ml of complete RPMI 1640 medium, with or without 200 µg/ml of cis- or trans-UCA. Cell-free supernatants were harvested after 24, 48, or 72 h. After stimulation with alloantigens (1.5 x 106 reactive cells and 2 x 106 irradiated stimulatory cells in 1 ml of culture medium), the supernatants were harvested after 48, 72, and 96 h.

The presence of IL-2, IL-4, IL-10, and IFN-{gamma} in the supernatants was measured by ELISA (26) using sets of cytokine-specific capture and detection mAb purchased from PharMingen (San Diego, CA) and precisely following the manufacturer’s instructions. For quantitation of cytokine levels, standards for IL-2, IL-4, IL-10, and IFN-{gamma} (all purchased from Genzyme) were included in all ELISA determinations.

RNA preparation and RT-PCR

Using the guanidine isothiocyanate method (27), total RNA was extracted from BALB/c spleen cells that were cultivated for 20 h while unstimulated or stimulated with 2 µg/ml of anti-CD3 mAb. The cultures were set with or without 200 µg/ml of cis- or trans-UCA. Two micrograms of total RNA were reverse transcribed into cDNA in a 50-µl reaction mixture containing 0.5 µl (10 mg/ml) of BSA, 5 µl (10 mM) of dNTPs, 2 µl (0.5 µg/ml) of p(dT)15, 1 µl (27250 U/ml) of RNAsin, 1 µl (15600 U/ml) of M-MuLV RT, and 10 µl of 5x RT buffer. Five microliters of the cDNA preparation was amplified during 29 cycles (MJ Research, Watertown, MA) in a reaction mixture containing 5 µl of 10 x PCR buffer, 1 µl (10 mM) of dNTPs, 2 µl (25 µM) of 5' and 3' primers (Strategene, La Jolla, CA), 0.5 µl (5000 U/ml) of Taq polymerase, and distilled water to a final volume of 50 µl. Cycling conditions were 94°C for 30 s, 60°C for 45 s, 72°C for 1 min, and a final extension of 72°C for 10 min. After PCR, the products were electrophoresed on an ethidium bromide-stained agarose gel.

Statistics

All results are expressed as means ± SEM. The statistical significance of differences between the means of individual experimental groups was calculated using Student’s t test.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Proliferative response of cells preincubated with UCA

Spleen cells preincubated for 24 h in the presence of 200 µg/ml of cis-UCA had a significantly decreased proliferative response after stimulation with anti-CD3 mAb or irradiated allogeneic cells compared with the response of cells preincubated with trans-UCA or medium alone (Fig. 1Go). To obtain a significant suppression of the proliferative responses, a 24-h preincubation of cells with cis-UCA was required; preincubation for 12 h or for a shorter period of time did not result in a significant decrease in the proliferative response.



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  		FIGURE 1. Proliferative response of cells preincubated with UCA. Spleen cells from BALB/c mice were preincubated for 24 h either alone ({square}) or with 200 µg/ml of cis-UCA () or trans-UCA ({blacksquare}) and then stimulated with anti-CD3 mAb (2 µg/ml) or with irradiated (3000 rad) spleen cells from allogeneic B10 donors. Cell proliferation was determined by adding 0.5 µCi/well of [3H]thymidine for the last 6 h of the 72-h (anti-CD3) or 96-h (allogeneic cells) incubation period. Each bar represents mean ± SEM for one of five similar experiments. Values marked with asterisks are significantly (p < 0.001) different from those of control cultures without UCA.

 
Ag-presenting function of cells preincubated with UCA

Spleen cells preincubated for 24 h with cis-UCA had a significantly decreased ability to stimulate proliferation of allogeneic cells in an MLC reaction in comparison with cells preincubated with trans-UCA or medium only (Fig. 2Go). This immunosuppression was significantly blocked by adding neutralizing anti-IL-10 mAb, but not control rat IgG1, to the MLC reaction (Fig. 2Go).



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  		FIGURE 2. Ability of spleen cells preincubated with UCA to stimulate proliferation of untreated allogeneic cells. Spleen cells from BALB/c mice were preincubated for 24 h either with or without ({square}) 200 µg/ml of cis-UCA () or trans-UCA ({blacksquare}), washed, and irradiated (3000 rad). One hundred microliters of untreated spleen cells (1.5 x 106 cells/ml) from B10 mice were cultivated alone () or with preincubated cells (2 x 105 cells in 100 µl). Neutralized anti-IL-10 mAb () or normal rat IgG1 (NRIg) () at a concentration of 10 µg/ml were added to the cultures containing cells preincubated with cis-UCA. Cell proliferation was determined by adding 0.5 µCi/well of [3H]thymidine for the last 6 h of the 96-h incubation period. One of five similar experiments is shown. Values marked with an asterisk (*) are significantly (p < 0.001) different from those of control cultures without UCA. Value with two asterisks (**) is significantly different (p < 0.001) from those of cultures preincubated with cis-UCA.

 
Production of IL-10 by cells stimulated in the presence of UCA

Preliminary experiments showed that spleen cells preincubated for 24 h with cis- but not trans-UCA and stimulated with anti-CD3 mAb or allogeneic cells had an enhanced production of IL-10, but the production of IL-2 and IFN-{gamma} was somewhat decreased (data not shown). To test the possibility that cis-UCA enhances the production of IL-10, we measured production of IL-10 in cell cultures stimulated with anti-CD3 mAb or with irradiated allogeneic cells in the presence of cis- or trans-UCA. Figure 3GoA shows that the production of IL-10 was significantly increased in the cultures containing cis- but not trans-UCA. This increase in IL-10 production was apparent at all time points tested (i.e., after 48, 72, and 96 h of incubation; data not shown) and was significant in a range of cis-UCA concentrations from 50 to 400 µg/ml (Fig. 3GoB). Neither cis- nor trans-UCA itself induced a significant IL-10 production (i.e., >5 pg/ml, which is the sensitivity limit of the assay) in spleen cells (Fig. 3GoA).



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  		FIGURE 3. Production of IL-10 by spleen cells stimulated with anti-CD3 or allogeneic cells in the presence of UCA (A) and UCA dose response on IL-10 production (B). Spleen cells from BALB/c mice were cultured alone, with anti-CD3 mAb, or with irradiated (3000 rad) allogeneic B10 spleen cells either in medium alone ({square}) or in the presence of 200 µg/ml of cis-UCA () or trans-UCA ({blacksquare}). The supernatants were harvested after a 72-h (anti-CD3 stimulation) or 96-h (with allogeneic cells) incubation, and the level of IL-10 in the supernatants was determined by ELISA. Each value represents the mean ± SEM of four similar experiments. Values marked with asterisks are significantly (* p < 0.05, ** p < 0.01,*** p < 0.001) different from those of control cultures without UCA.

 
Effects of UCA on IL-10 production by cell subpopulations

The elimination of CD4+ cells or total T cells from the spleen cell population completely abolished the ability of the remaining cells to produce IL-10 after stimulation with anti-CD3 (Fig. 4Go). However, spleen cells treated with anti-CD8 mAb and C produced even more IL-10 than untreated cells, and this production was considerably increased in the presence of cis-UCA (Fig. 4Go). In addition, both spleen cells depleted of T cells and stimulated with the B cell mitogen LPS as well as purified peritoneal macrophages stimulated with LPS produced IL-10; but the production of IL-10 by these cells was not altered in the presence of cis-UCA (data not shown).



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  		FIGURE 4. Production of IL-10 by spleen cell subpopulations stimulated in the presence of UCA. Spleen cells from BALB/c mice were either untreated or treated by two cycles of incubation with cytotoxic anti-CD4, anti-CD8, or anti-Thy-1.2 mAb and C. Then, the cells were stimulated with 2 µg/ml of anti-CD3 mAb with or without ({square}) 200 µg/ml of cis-UCA () or trans-UCA ({blacksquare}). The level of IL-10 in supernatants was determined after 72 h. One of 3 similar experiments is shown. Values marked with asterisks are significantly (p < 0.001) different from those of control cultures without UCA.

 
To identify a cellular target of cis-UCA action, spleen cells were separated into T cell and non-T cell populations. Neither of these populations alone produced detectable IL-10 after stimulation with anti-CD3 (Fig. 5Go). Separated T cell and non-T cell populations were preincubated for 2 h either alone or in the presence of 200 µg/ml of cis- or trans-UCA. Then, untreated T cells were mixed with non-T cells that were preincubated either alone, with cis-UCA, or with trans-UCA; vice versa, untreated non-T cells were mixed with T cells preincubated either alone, with cis-UCA or trans-UCA. As shown in Figure 5Go, enhanced IL-10 production was observed only in cultures containing untreated non-T cells and T cells preincubated with cis-UCA.



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  		FIGURE 5. UCA acts directly on T cells. Spleen cells from BALB/c mice, either unseparated or separated into T and non-T cell subpopulations, were stimulated for 72 h with 2 µg/ml of anti-CD3 mAb with or without ({square}) 200 µg/ml of cis-UCA () or trans-UCA ({blacksquare}). In addition, each subpopulation was preincubated for 2 h either alone or with 200 µg/ml of cis- or trans-UCA. After washing, each of these populations was mixed at a ratio of 1:1 with fresh, untreated cells of the opposite population (i.e., treated T cells with untreated non-T cells and vice versa) and incubated with 2 µg/ml of anti-CD3 mAb. The level of IL-10 in the supernatants was determined after a 72-h incubation. One of three similar experiments is shown. Values marked with asterisks are significantly (p < 0.001) different from those of control cultures without UCA.

 
Cis-UCA enhances expression of IL-10 gene

To test whether cis-UCA acts on the level of IL-10 gene expression, spleen cells were cultivated unstimulated for 20 h or were stimulated with anti-CD3 mAb and the cultures were set in the presence or absence of cis- or trans-UCA. The level of IL-10 mRNA was measured by RT-PCR. As demonstrated in Figure 6Go, neither cis-UCA nor trans-UCA increased production of IL-10 mRNA in unstimulated cells, but the level of IL-10 mRNA was considerably increased in cultures stimulated with anti-CD3 mAb in the presence of cis-UCA.



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  		FIGURE 6. Expression of IL-10 mRNA by spleen cells stimulated in the presence of UCA. Spleen cells from BALB/c mice were cultivated unstimulated for 20 h alone (lane 1) or in the presence of 200 µg/ml of cis-UCA (lane 2) or trans-UCA (lane 3); or were stimulated with 2 µg/ml of anti-CD3 mAb alone (lane 4) or in the presence of 200 µg/ml of cis-UCA (lane 5) or trans-UCA (lane 6). Total RNA was isolated, reverse transcribed, and amplified by PCR with 5' and 3' primers for IL-10 and actin. PCR products were electrophoresed; the ethidium bromide-stained agarose gel is shown.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
The immunosuppressive effects of UV radiation have been well documented in various systems, but the molecular mechanism of the suppression has remained unclear. It has been proposed that the immunosuppressive effects of UVB radiation are mediated via cis-UCA, a photoproduct of naturally occurring trans-UCA (7). However, we demonstrate here that cis-UCA is not the effector molecule that inhibits immune system function. Rather, cis-UCA stimulates the production of IL-10 by activated CD4+ T cells through an increase in the expression of the IL-10 gene. Consequently, IL-10 can inhibit the activity of APC (17) or anergize T cells (18) and thus suppress the immune response.

We have shown that spleen cells preincubated with cis-UCA, but not trans-UCA, have a diminished ability to respond by proliferation to stimulation with allogeneic cells in a MLC reaction and to stimulation with anti-CD3 mAb. Simultaneously, the ability of cells preincubated with cis-UCA to stimulate the proliferation of allogeneic cells in MLC was decreased. Previous studies have shown the harmful effects of UV radiation on the activity of APC (28, 29). Since direct effects of cis-UCA on APC function (16) or on the expression of costimulatory molecules (15) have not been shown, another molecule mediating the effects of UV radiation and UCA on APC function has been hypothesized. Since IL-10 can inhibit Ag presentation by epidermal APC (30) and possesses an ability to inhibit the production of Th1 cytokines through modulating APC function (31), IL-10 has appeared as a candidate molecule for cis-UCA-mediated immunosuppression. Indeed, we also observed a decreased ability of spleen cells preincubated with cis-UCA to produce IL-2 and IFN-{gamma}.

Several lines of evidence support our conclusion that cis-UCA enhances IL-10 production by activated CD4+ T cells. First, IL-10 gene expression was increased in cultures of spleen cells stimulated with anti-CD3 mAb in the presence of cis-UCA. Second, the production of IL-10 protein by spleen cells was significantly increased in the presence of cis-UCA. The enhanced production of IL-10 was completely inhibited by treating spleen cells with anti-Thy-1.2 or anti-CD4 mAb, but was not impaired by the elimination of CD8+ cells. In addition, the production of IL-10 by stimulated splenic non-T cells or by LPS-stimulated macrophages was not altered by cis-UCA (data not shown). Thus, activated CD4+ T cells appear to be the principal cells responding to cis-UCA by enhancing IL-10 production. These cells might be responsible for the Ts cell phenomena described after UV irradiation (32, 33). This conclusion is also supported by the findings of Rivas and Ullrich (34), who showed that the activity of UV radiation-induced Ts cells can be blocked by anti-IL-10 mAb.

The observation that cis-UCA increases IL-10 production in activated T cells and the finding of decreased IL-2 and IFN-{gamma} production in cultures pretreated with cis-UCA suggest a shift to a Th2-type response in cells preincubated with cis-UCA. This observation is consisted with the report of Brown et al. (35), who showed that UV irradiation in vivo down-regulates Th1 immune responses while leaving Th2 responses intact.

To exclude the possibility that cis-UCA enhances IL-10 production in T cells through an effect on APC, we divided spleen cells into T and non-T subpopulations and preincubated them with cis-UCA or trans-UCA. Then, the cells of each subpopulation were mixed with untreated cells of the opposite population, and the mixtures were stimulated with anti-CD3 mAb. This experiment showed that T cells were a direct target of cis-UCA.

We are aware that the described pathway of cis-UCA-mediated immunosuppression is not the only one operating after UV irradiation. UV radiation can have a direct deleterious effect on DNA (36). It has been shown that the extent of DNA damage after UV irradiation correlates with UV-induced systemic suppression of CH and DTH (36, 37). Cis-UCA arising after UVB irradiation of naturally occurring trans-UCA may induce a local increase in the synthesis of TNF-{alpha}, which suppresses Ag presentation by LC (10). This observation is supported by findings that UVB radiation-induced suppression of contact sensitivity in mice can be blocked by anti-TNF-{alpha} Abs (10, 33). However, TNF-{alpha} appears as a molecule involved in the UV-induced suppression of CH, but only to a much lesser extent in the suppression of DTH (34). Ullrich and coworkers reported that UVB radiation may act directly on keratinocytes by inducing IL-10 production in these cells (38). These authors also showed that immunosuppression induced by UVB irradiation can be mimicked by the administration of IL-10 or reversed by anti-IL-10 Abs (38, 39, 40). However, it seems improbable that cis-UCA induces a significant production of IL-10 in keratinocytes. Moodycliffe et al. (41) showed that injecting anti-cis-UCA mAb into UV-irradiated mice reversed UV-induced suppression of DTH but did not block IL-10 production in the epidermis. Similarly, we found in our system that anti-IL-10 mAb blocked the immunosuppressive effect of cis-UCA observed in MLC. In accordance with the other authors who have shown that IL-10 inhibits allogeneic proliferative responses in vitro (42, 43), we observed that IL-10 added to the cultures can mimic the immunosuppressive effects of cis-UCA (data not shown).

IL-10 secreted by CD4+ T cells activated with Ag in the presence of cis-UCA appears to be the main effector molecule responsible for UVB radiation-induced immunosuppression. It has been shown that IL-10 can decrease APC function (44) and, together with Ag, can induce anergy in responding T cell clones (18). We suggest that IL-10 secreted by CD4+ T cells in the presence of cis-UCA might be responsible for the suppressor T cell phenomena observed after UV irradiation.


   Footnotes
 
1 This work was supported by Grant 3964-3 from the Grant Agency of the Ministry of Health of the Czech Republic, Grant 310/97/1261 from the Grant Agency of the Czech Republic, and Project VS 97099 from the Ministry of Education of the Czech Republic. Back

2 Address correspondence and reprint requests to Dr. Vladimír Holán, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 37 Prague 6, Czech Republic. E-mail address: Back

3 Abbreviations used in this paper: CH, contact hypersensitivity; DTH, delayed-type hypersensitivity; UCA, urocanic acid; LC, Langerhans cells; Ts, Ag-specific suppressor T cells. Back

Received for publication February 17, 1998. Accepted for publication May 26, 1998.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Elmets, C. A., J. Krutmann. 1988. Recent studies on mechanisms in photoimmunology. Photochem. Photobiol. 48:787.[Medline]
  2. Noonan, F. P., E. C. De Fabo. 1992. Immunosupression by ultraviolet B radiation: initiation by urocanic acid. Immunol. Today 13:250.[Medline]
  3. Streilein, J. W., P. R. Bergstresser. 1988. Genetic basis of ultraviolet-B effects on contact hypersensitivity. Immunogenetics 27:252.[Medline]
  4. Mottram, P. L., A. Mirisklavos, G. J. A. Clunie, F. P. Noonan. 1988. Single dose of UV radiation suppresses delayed type hypersensitivity responses to alloantigens and prolongs heart allograft survival in mice. Immunol. Cell Biol. 66:377.
  5. Ullrich, S. E.. 1995. Modulation of immunity by ultraviolet radiation: key effect on antigen presentation. J. Invest. Dermatol. 105:30S.[Medline]
  6. Schacter, B., M. M. Lederman, M. J. Le Vine, J. J. Ellner. 1983. Ultraviolet radiation inhibits human natural killer activity and lymphocyte proliferation. J. Immunol. 130:2484.[Abstract]
  7. De Fabo, E. C., F. P. Noonan. 1983. Mechanism of immune suppression by ultraviolet irradiation in vivo. I. Evidence for the existence of a unique photoreceptor in skin and its role in photoimmunobiology. J. Exp. Med. 157:84.
  8. Ross, J. A., S. E. Howie, M. Norval, J. Maingay, T. Simpson. 1986. Ultraviolet-irradiated urocanic acid suppresses delayed type hypersensitivity to herpes simplex virus in mice. J. Invest. Dermatol. 87:630.[Medline]
  9. Norval, M., T. J. Simpson, E. Bardshiri, S. E. M. Howie. 1989. Urocanic acid analogues and the suppression of the delayed type hypersensitivity response to herpes simplex virus. Photochem. Photobiol. 49:633.[Medline]
  10. Kurimoto, I., J. W. Streilein. 1992. cis-urocanic acid suppression of contact hypersensitivity induction is mediated via tumor necrosis factor-{alpha}. J. Immunol. 148:3072.[Abstract]
  11. Gruner, S., W. Diezel, H. Stoppe, H. Oesterwitz, W. Henke. 1992. Inhibition of skin allograft rejection and acute graft-versus-host disease by cis-urocanic acid. J. Invest. Dermatol. 98:459.[Medline]
  12. Guymer, R. H., T. E. Mandel. 1993. Urocanic acid as an immunosuppressant in allotransplantation in mice. Transplantation 55:35.
  13. Filipec, M., E. Letko, Z. Hasöková, D. Jencíèková, P. Holler, A. Jancèárek, V. Holán. 1998. The effect of urocanic acid on graft rejection in an experimental model of orthotopic corneal transplantation in rabbits. Graefe’s Arch. Clin. Exp. Ophthalmol. 236:65.[Medline]
  14. Noonan, F. P., E. C. DeFabo, H. Morrison. 1988. Cis-urocanic acid, a product formed by ultraviolet B irradiation of the skin, initiates an antigen presentation defect in splenic dendritic cells in vivo. J. Invest. Dermatol. 90:92.[Medline]
  15. Beissert, S., T. Mohammed, H. Torri, A. Lonati, Z. Yan, H. Morrison, R. D. Granstein. 1997. Regulation of tumor antigen presentation by urocanic acid. J. Immunol. 159:92.[Abstract]
  16. Rattis, F. M., J. Peguet-Navarro, P. Courtellemont, G. Schmitt. 1995. Cis-urocanic acid failed to affect in vitro human Langerhans cell allostimulatory function. Photochem. Photobiol. 62:914.
  17. Enk, A. H., V. Angeloni, M. C. Udey, S. I. Katz. 1993. Inhibition of Langerhans cell antigen-presenting function by IL-10: a role for IL-10 in induction of tolerance. J. Immunol. 151:2390.[Abstract]
  18. Groux, H., M. Bigler, J. E. De Vries, M.-G. Roncarolo. 1996. Interleukin-10 induces a long-term antigen-specific anergic state in human CD4+ T cells. J. Exp. Med. 184:19.[Abstract/Free Full Text]
  19. Julius, M. H., E. Simpson, L. A. Herzenberg. 1973. A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur. J. Immunol. 3:645.[Medline]
  20. Wysocki, L. J., V. L. Sato. 1978. "Panning" for lymphocytes: a method for cell selection. Proc. Natl. Acad. Sci. USA 75:2844.[Abstract/Free Full Text]
  21. Lake, P., E. A. Clark, M. Khorshidi, G. H. Sunshine. 1979. Production and characterization of cytotoxic Thy-1 antibody secreting hybrid cell lines. Detection of T-cell subsets. Eur. J. Immunol. 9:875.[Medline]
  22. Dialynas, D. P., D. B. Wilde, P. Marrack, A. Pierres, K. A. Wall, W. Havran, G. Otten, M. R. Loken, M. Pierres, J. Kappler, W. W. Fitch. 1983. Characterization of the murine antigenic determinant designated L3T4a, recognized by monoclonal antibody GK1.5: expression of L3T4a by functional T-cell clones appears to correlate primarily with class II MHC antigen reactivity. Immunol. Rev. 74:29.[Medline]
  23. Gottlieb, P. D., A. Marshak-Rothstein, K. Auditore-Hargreaves, D. B. Berhoben, D. A. August, R. M. Posche, J. D. Benetto. 1980. Construction and properties of new Lyt-congenic strains and anti-Lyt-2.2 and anti-Lyt-3.1 monoclonal antibodies. Immunogenetics 10:545.
  24. Tomonari, K.. 1988. A rat antibody against a structure functionally related to the mouse T-cell receptor/T3 complex. Immunogenetics 28:455.[Medline]
  25. Holán, V., P. Demant, M. Lipoldová. 1996. Identical genetic control of MLC reactivity to different MHC incompatibilities, independent of production of and response to IL-2. Immunogenetics 44:27.[Medline]
  26. Holán, V., K. Sedlácková, M. Ruzicková. 1996. Production of high levels of Th1 and Th2 cytokines in mice with acquired transplantation tolerance. Cell. Immunol. 174:7.[Medline]
  27. Chirgwin, J. M., A. E. Przybyla, R. J. MacDonald, W. J. Rutter. 1979. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294.[Medline]
  28. Greene, M. I., M. S. Sy, M. L. Kripke, B. Benacerraf. 1979. Impairment of antigen-presenting cell function by ultraviolet radiation. Proc. Natl. Acad. Sci. USA 76:6591.[Abstract/Free Full Text]
  29. Stingl, L. A., D. N. Sauder, M. Iijima, K. Wolff, K. Pehamberger, G. Stingl. 1983. Mechanism of UV-B induced impairment of antigen-presenting capacity of murine epidermal cells. J. Immunol. 130:1586.[Medline]
  30. Beissert, S., J. Hosoi, S. Grabbe, A. Asahina, R. D. Granstein. 1995. IL-10 inhibits tumor antigen presentation by epidermal antigen-presenting cells. J. Immunol. 154:1280.[Abstract]
  31. Fiorentino, D. F., M. W. Bond, T. R. Mosmann. 1989. Two types of mouse of mouse T helper cell clone. IV. Th2 clones secrete a factor that inhibits cytokine production by Th1 clones. J. Exp. Med. 170:2081.[Abstract/Free Full Text]
  32. Fisher, M. S., M. L. Kripke. 1982. Suppressor T lymphocytes control the development of primary skin cancers in UV-irradiated mice. Science 216:1133.[Abstract/Free Full Text]
  33. Elmets, C. A., P. R. Bergstresser, R. E. Tigelaar, P. J. Wood, J. W. Streilein. 1983. Analysis of the mechanism of unresponsiveness produced by haptens painted on skin exposed to low dose ultraviolet radiation. J. Exp. Med. 158:781.[Abstract/Free Full Text]
  34. Rivas, J. M., S. E. Ullrich. 1994. The role of IL-4, IL-10, and TNF-{alpha} in the immune suppression induced by ultraviolet radiation. J. Leukocyte Biol. 56:769.[Abstract]
  35. Brown, E. L., J. M. Rivas, S. E. Ullrich, C. R. Young, S. J. Norris, M. L. Kripke. 1995. Modulation of immunity to Borrelia burgdorferi by ultraviolet irradiation: differential effect on Th1 and Th2 immune responses. Eur. J. Immunol. 25:3017.[Medline]
  36. Kripke, M. L., P. A. Cox, L. G. Alas, D. B. Yarosh. 1992. Pyrimidine dimers in DNA initiate systemic immunosuppression in UV-irradiated mice. Proc. Natl. Acad. Sci. USA 89:7516.[Abstract/Free Full Text]
  37. Applegate, A., R. D. Ley, J. Alcalay, M. L. Kripke. 1989. Identification of the molecular target for the suppression of contact hypersensitivity by ultraviolet radiation. J. Exp. Med. 170:1117.[Abstract/Free Full Text]
  38. Rivas, J. M., S. E. Ullrich. 1992. Systemic suppression of delayed type hypersensitivity by supernatants from UV-irradiated keratinocytes: an essential role for keratinocyte-derived IL-10. J. Immunol. 149:3865.[Abstract]
  39. Jeevan, A., S. E. Ullrich, M. De-Gracia, R. Shah, Y. Sun. 1996. Mechanism of UVB-induced suppression of the immune response to Mycobacterium bovis bacillus Calmette-Guerin: role of cytokines on macrophage function. Photochem. Photobiol. 64:259.[Medline]
  40. Beissert, S., S. E. Ullrich, J. Hosoi, R. D. Granstein. 1995. Supernatants from UVB radiation-exposed keratinocytes inhibit Langerhans cell presentation of tumor-associated antigens via IL-10 content. J. Leukocyte Biol. 58:234.[Abstract]
  41. Moodycliffe, A. M., C. D. Bucana, M. L. Kripke, M. Norval, S. E. Ullrich. 1996. Differential effect of a monoclonal antibody to cis-urocanic acid on the suppression of delayed and contact hypersensitivity following ultraviolet irradiation. J. Immunol. 157:2891.[Abstract]
  42. Bejerano, M. T., R. De Waal Malefyt, J. S. Abrams, M. Bigler, R. Bacchetta, J. E. de Vries, M. G. Roncarolo. 1992. Interleukin 10 inhibits allogeneic proliferative and cytotoxic T cell responses generated in primary mixed lymphocyte cultures. Int. Immunol. 4:1389.[Abstract/Free Full Text]
  43. Caux, C., C. Massacrier, B. Vandervliet, C. Barthelemy, Y. J. Liu, J. Banchereau. 1994. Interleukin 10 inhibits T cell alloreactions induced by human dendritic cells. Int. Immunol. 6:1177.[Abstract/Free Full Text]
  44. Ullrich, S. E.. 1994. Mechanism involved in the systemic suppression of antigen-presenting function by UV irradiation: keratinocyte-derived IL-10 modulates antigen-presenting cell function of splenic adherent cells. J. Immunol. 152:3410.[Abstract]



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